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From the Editor’s Desk

Dear Readers,

On behalf of, the Editorial Board and the Editorial Team of our journal, I would like to
wish all authors, patrons and readers a wonderful and prosperous year ahead.

With a thought that has been enduring in mind becomes real, it is truly an interesting and
exciting experience. Our journal continues to enjoy a pan India reach and also excellent
international presence. In the year that has gone by, our publications continued to maintain
a subscriber base that is unparalleled by any other journal.

I shall assure all our readers that our consistent efforts will be aimed toward increasing the
visibility, impact, editorial cycle time, citations and the overall quality of our journals. We
very much look forward to strengthening the reputation of our publications, and we want
to attract more higher-quality submissions. I hope our readers and patrons share a similar
vision, and we look forward to a productive, challenging and successful year ahead. In the
spirit of continuous improvement, any constructive input on streamlining our processes
is very welcome.

Dr. Bhagwant Singh


ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Council Members APDF/APRO
Executive Officers (2018-2019)
President Dr Fernando Fernandez (Philippines)
President Elect Dr Cheol- Sooo Kim (Korea)
Secretary General Dr Oliver Hennedige (Singapore)
Chairman College Dr Jeffrey Y S Tsang, MBE (Hong Kong)
Treasurer Dr Yang Chun-Chieh (Chinese Taipei)

Council Officers (2018-2019)

Vice Presidents Dr Asif Arain (Pakistan)
Vice Presidents Prof Chia Tze Kao (Chinese Taipei)
Vice Presidents Dr Seung-Mok Na (Korea)
Vice Presidents Dr Carlos Buendia (Philippines)
Vice Presidents Dr Kenny Lau (Hong Kong)

Imm Past President Dr Cristina Antonio (Macau)

Editor Dr Bhagwant Singh (India)
Chairman, Dental Education Commission Dr Mark Villalobos (Philippines)
Chairman, Oral Diseases Commission Dr Mahmood Shah (Pakistan)
Chairman, Dental Public Health Dr Lin-Yang Chi (Chinese Taipei)
Chairman, General Dental Practice Dr Sudin Shakya (Nepal)
Chairman, Defence Forces Dentistry Air Vice Marshall (Retired) Dr A M B Amunugama (Sri Lanka)

ICCDE Board Members

President Dr Jeffrey Y S Tsang
Executive Director Dr Oliver Hennedige
Vice-President Dr Cristina Antonio
Prof Dr S M Balaji
Prof Dr Amish Mehta
Dr Sudin Shakya
Dr Kuan Chee Keong
Dr Century Tsang
Finance Director Dr Yang Chun Chieh
Editor Dr Bhagwant Singh
Board of Directors Dr Asif Niaz Arain
Dr Hermogenes P Villareal
Dr Mahmood Shah
Dr Fernando “Andy” Fernandez
Dr S P Aggarwal
Dr Floren Ly
Dr Roberto M Tajonera
Dr Mui Sau Fung Michael
Dr Chan Chi Chun Andrew
Dr Ritika Arora
Dr Anusak Sintapnont
Dr Saurabh Arora
Dr Vikash J. Singh
Dr Kelvin Chuan Hee Chye
Dr Hilary W.M. Cooray
Regent For South Asia Dr Bhagwant Singh
Regent For East Asia Prof. Allen Ming-Lun Hsu
Regent For Middle East Dr Aisha Sultan Alsuwaidi
Regent For South East Asia Dr Mirza Zamzami Djasri M A
Regent for Australiasia Dr Patrick J Colgan

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Dr. Bhagwant Singh
A-6, Gurudwara Shaheedan Road, Model Town Ludhiana (Pb.), India – 141002
M. +91-98142-45608
Email : dr.bs_ldh@rediffmail.com

Associate Editors
Dr. Saurabh Arora Dr. Amish Mehta Dr. Vikas Jindal

Assistant Editors
Dr. Ravneet Arora Dr. Pallvi Goomer Dr. Arpit Sikri

Advisory Editorial Board

Dr. Aisha Sultan (U.A.E.) Dr. Oliver Hennedige (Singapore) Lt. Gen. Vimal Arora (India)
Jeffre Y.S. Tasang (Hong Kong) Dr. Anil Kohli (India) Dr. Keki Mistry (India)
Dr. Boy Vallareal (Philippines) Dr. James Lee (Chinese Taipei) Dr. Arturo De Leon (Philippines)

Community Dentistry Esthetic Dentistry Oral Medicine

Dr. R.K. Bali Dr. Sandesh Mayekar Dr. S.Y. Rajan
Dr. Ajith Krishnan Dr. Rumpa Wig Dr. Soheyl Sheikh
Dr. Ankur Singh (Australia) Dr. Sushant Umre Dr. Ankur Aggarwal
Dr. Sanjeet S. Risam
Pedodontics Periodontics
Dr. S.G. Damle Dr. Arunachalam Orthodontics
Dr. I.K.Pandit Dr. Ritika Arora Dr. D.N. Kapoor
Dr. Neru Singh Dr. Pradeep Shukla Dr. Krishna Nayak
Dr. Nikhil Sivastava Dr. Mayur Kaushik Dr. O.P. Kharbanda
Dr. Vivek Gaurav Dr. Chandresh Shukla
Gen. Dentistry Dr. Diki Tsering Lasquite (Philippines)
Conservative Dentistry Dr. Asif Niaz Arian Dr. Mauricio Gonzalez Balut (Mexico)
Dr. Vimal Sikri Dr. Christina Antonio Dr. Anand Marya (Philippines)
Dr. Rajiv Bali Dr. Anwar Saeed
Dr. Parvin Kumar Dr. Sudin Shakya Oral Pathology
Dr. Jaidev Singh Dhillon Dr. Amar Singh Dr. R.M. Mathur
Dr. Vijita Mehta Dr. K.S. Ghai Dr. Ish Paul Singh
Dr. Nikhil Bahuguna Dr. Vivek Vij (New York)
Oral Surgery
Dr. R. Vemareddy Dr. Shikha Kanotra (Boston)
Dr. S.P.S. Sodhi
Dr. Sachin Dev Mehta Dr. Anureet Dhillon
Dr. Vimal Kalia
Dr. Sukhpash Sandhu
Dr. Rahul Thakkur
Dr. Puneet Girdhar
Implants Dr. Mahesh Verma
Dr. Amreen Kaur
Dr. Sanjay Kalra Dr. Padmanabhan
Dr. S.P. Aggarwal Dr. Himanshu Aeran Allied Medical Sciences
Dr. Minas Leventis (Greece) Dr. Ramanpreet Ranauda Dr. L.S. Chawla (Medicine)
Dr. Rohan Sikka Dr. Rajesh Bhanot Dr. Robert Patricia (Dermatologist, Sweden)
Dr. Andrea Mastrorosa (New York) Dr. Manu Rathee Dr. Rohan Arora (Neurologist, USA)
Dr. Sunil Arora Dr. Carl Brown (Cardiologist, Canada)
Dr. R.S. Bhatia (Pulmonologist)

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018


Two Phase Treatment for the correction of Skeletal Class II Malocclusion – A Case Report 1
Prof. (Dr.) U.S. Krishna Nayak, Dr. Rajshekhar Banerjee, Dr. Megha Parikh, Dr. Adarsh N.K.,
Dr. Harshit Atul Kumar, Dr Upasak Mukherjee 1

Changing face of history-virtual anthropology 4

Dr Saurabh Arora, Dr Ravneet Arora

Shear bond strength assessment of composite – tooth interface following caries excavation by different 7
chemomechanical caries removal agents
Dr. Jyotika Grover

Ill effects of technology: dehiscence encountered following laser depigmentation: A case report 11
Dr Anamika Sharma

Recent trends in regenerative dentistry: A review 14

Dr. Jyotsana Sikri and Dr. Arpit Sikri

Biofilms in endodontics 24
Dr. Nikenlemla, Dr. Naman Vaidya, Dr. Rucha Shinde, Dr. Swati Bali

Mandibular canine index 35

Dr Ravneet Arora, Dr Saurabh Arora

A Paradigm shift from micro to nano in dentistry 37

Dr. Atulana Roy and Dr. Arpit Sikri

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Two Phase Treatment for the correction of Skeletal
Class II Malocclusion – A Case Report
Prof. (Dr.) U.S. Krishna Nayak, Dr. Rajshekhar Banerjee, Dr. Megha Parikh,
Dr. Adarsh N.K., Dr. Harshit Atul Kumar, Dr Upasak Mukherjee

Case Report: Diagnosis:

A 12 year old male patient presented with a chief complaint of From clinical and cephalometric analysis, the case was
forwardly placed upper front teeth. diagnosed to be a case of

Extraoral Examination 1. Skeletal and Dental Class II with a retrognathic

mandible and a normal maxilla
Patient had a Brachycephalic skull type and a Mesoprosopic
facial type with a convex facial profile, posterior facial 2. Horizontal growth pattern
divergence, Class II relation of apical bases and a flat 3. Proclined Maxillary and
mandibular plane angle. [Figures 1A-1E] Mandibular Incisors
4. Generalised upper and lower
anterior spacing
5. Procumbent lips
Visual Treatment Objective
The Visual Treatment Objective
(VTO) was positive. Hence a
treatment plan involving mandibular
advancement using a functional
appliance was considered. (Figure 3) Figure 3: Visual
Treatment Objective
Treatment Objectives:
1. To achieve Class I skeletal relation.
2. To achieve Class I molar and canine relation
3. Closure of spaces
Figure 1: Extraoral Photographs 4. Uprighting of the anteriors
Intraoral Findings 5. To achieve harmonious soft tissue profile.
Patient had a Class II molar and canine relation with Treatment Plan:
generalised spacing. An overjet of 10 mm and overbite of A Two=Phase treatment was considered to achieve the
5mm was present. Distolingual rotation with respect to 14 and treatment objectives
24 was also present. (Figures 2A-2D)
1. Phase I: Forward repositioning of the mandible by Twin
Block with incisor capping
2. Phase II: Fixed orthodontic therapy with 0.022 Slot
MBT prescription
Closure of Anterior spacing
Finishing and Detailing.
Phase I Therapy
The Twin Block functional appliance consists two bite
blocks, upper and lower (twin-block) with inclined planes
interlocking at 70 degrees. The interlocking of the two blocks
positions the mandible in a protrusive manner.1 It is designed
Figure 2: Intraoral Photographs

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 1

to be worn full time. The appliance works by transmitting
occlusal forces to the occlusal inclined planes covering the
posterior teeth.2 It induces supplementary lengthening of the
mandible by stimulating increased growth at the condylar
cartilage.3 For children with skeletal Class II discrepancy, a
two phase therapy can be beneficial as skeletal growth can
be modified and subsequent fixed appliance therapy can be
simplified when compared to a one phase treatment.4 Not Figure 6: Fixed Orthodontic Therapy
addressing the skeletal discrepancy in childhood may result
in the need arising for surgical treatment later in life.
The twin block appliance was cemented in place and kept for
8 months. The mandibular incisors were capped by extending
the acrylic of the twin block over their incisal edges onto
the buccal surface, thereby preventing them from further
proclination. (Figure 4) Figure 7: Anterior Space closure and Class II elastics

Figure 8: Intrusion Utility Arch

Following this, an Intrusion Utility Arch was attached from

the auxiliary tubes of the maxillary molar tubes and was used
Figure 4: Twin Block with Incisor Capping to intrude 12-22. Class II elastics were continued in this stage
as well. (Figure 8)
Occlusion at the end of Functional Therapy (Twin Block
Appliance) Subsequently 0.019x0.025” SS wires were engaged in both
upper and lower arches. A Reverse Curve of Spee was
Post Twin Block therapy, the mandibular arch was visibly incorporated in the mandibular archwire.
placed forward. A Lateral Open Bite was present. (Figure 5)
Anterior crown build-ups were done utilizing the spacings
remaining at the end of Fixed Orthodontic therapy.

Phase II Therapy: Fixed Orthodontic Therapy

MBT 0.022” slot pre-adjusted edgewise appliance was used
for the fixed orthodontic therapy. Initial levelling and aligning
was done using NiTi wires. (Figure 6A and 6B) The wires
Figure 9: Post treatment
were progressively stepped up to 0.017x0.025” Stainless
Steel. Class II elastics were engaged. The anterior space
closures were done using E-Chains from 13 to 23 and 33 to
43. (Figure 7A and B)
2 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018
the fact that full time wear of the appliance was essential to
elicit its maximum therapeutic effect.7

Rondeau B. Twin Block appliance. Part II. The Functional
orthodontist. 1996;13(2):4-10.
Clark W. Twin block functional therapy. JP Medical Ltd; 2014 Sep
Baccetti T, Franchi L, Toth LR, McNamara Jr JA. Treatment timing
for Twin-block therapy. American Journal of Orthodontics and
Figure 10: Intraoral - Pre and Post treatment Dentofacial Orthopedics. 2000 Aug 1;118(2):159-70.
Tulloch JC, Phillips C, Proffit WR. Benefit of early Class II
treatment: progress report of a two-phase randomized clinical
trial. American Journal of Orthodontics and Dentofacial
Orthopedics. 1998 Jan 1;113(1):62-74.
Gianelly AA. One-phase versus two-phase treatment. American
Journal of Orthodontics and Dentofacial Orthopedics. 1995
Nov 1;108(5):556-9.
McNamara Jr JA. Neuromuscular and skeletal adaptations to altered
function in the orofacial region. Am J Orthod 1973;64:578-606.
Aggarwal P, Kharbanda OP, Mathur R, Duggal R, Parkash H. Muscle
response to the twin-block appliance: an electromyographic
study of the masseter and anterior temporal muscles. American
journal of orthodontics and dentofacial orthopedics. 1999 Oct

Prof. (Dr.) U.S. Krishna Nayak

Principal and Professor
Dept. of Orthodontics and Dentofacial Orthopaedics, A.B.
Comparison of Pre and Post treatment - Intraoral Shetty Memorial Institute of Dental sciences, Mangalore

Comparison of Pre and Post treatment - Extraoral Dr. Rajshekhar Banerjee

Post Graduate Student
Summary Dept. of Orthodontics and Dentofacial Orthopaedics, A.B.
The current case presented at a stage when the patient was Shetty Memorial Institute of Dental sciences, Mangalore
still in the growing phase and the remaining growth could
Dr. Megha Parikh
be harnessed. Skeletal discrepancies such as this with a
Ex-Post Graduate Student
mandibular retrognathism are well indicated for a two-phase
Dept. of Orthodontics and Dentofacial Orthopaedics, A.B.
therapy.5 The first phase of the therapy not only substantially
Shetty Memorial Institute of Dental sciences, Mangalore
minimizes the skeletal problem, but also reduces the
complexity of the phase of the fixed orthodontic treatment. Dr. Adarsh N.K.
Further, failure to correct the skeletal discrepancy during the Post Graduate Student
time when growth is still remaining will warrant a surgical Dept. of Orthodontics and Dentofacial Orthopaedics, A.B.
correction of the problem. The option of orthognathic surgery Shetty Memorial Institute of Dental sciences, Mangalore
was presented more often in the cases of children who did not
undergo early treatment.4 Dr. Harshit Atul Kumar
Post Graduate Student
The mode of action of functional therapy is linked to Dept. of Orthodontics and Dentofacial Orthopaedics, A.B.
neuromuscular and skeletal adaptations to altered function Shetty Memorial Institute of Dental sciences, Mangalore
in the orofacial region.6 Aggarwal, in a study in 1999 studied
the muscle response to twin block therapy and concluded Dr Upasak Mukherjee
that the main force for Twin-block treatment was provided Post graduate student
through increased active tension in the stretched muscles and Dept of Orthodontics,AB Shetty Memorial Institute of
initiation of myotatic reflex activity and not through passive Dental Sciences
tension (viscoelastic properties) of jaw muscles. It reaffirmed

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 3

Changing face of history-virtual anthropology
Dr Saurabh Arora, Dr Ravneet Arora

Cadavers speak their own language. Moving back in history in human skulls, detection of influencial landmarks7and
human identification was one of the most challenging subjects confidence intervals8.
that man had confronted. The concept of identity, with few
Laser facial scanning: Technologies for generating a
significant variations, is the same as the assertion of Alves1
3-dimensional model of a human face with registered texture
that identity is a set of physical characteristics, functional or
include laser scanning and visible light techniques. The laser
psychic, normal or pathological, that define an individual.
scanners produce a detailed model Researchers have explored
The determination of race, sex, age, and stature of the bone
the use of laser surface scanning for assessment of facial
gives valuable information in establishing the identity of a
asymmetry9. The authors divided the face in different regions
and then classified the pre-surgical and post-surgical areas
Another skill that is receiving increasing attention is the according to different surface type primitives: valley, ridge,
virtual anthropology saddle surface, etc. The quantitative changes per region are
expressed in terms of area size changes and their movement
Early methods used were
on the face9.
 Anthropometry; ‘measurement of mankind’
Stereo-photogrammetry: In 1960 dot stereograms and the
 Osteometrics; measurements of skeleton idea that stereoscopic vision is a cooperative process were
 Craniometrics; measurements of skull introduced10. A algorithm for stereo reconstruction followed.
 Anthroposcopy: visual differences Stereo-photogrammetry has also been used to find the optimal
Now Virtual anthropology is becoming a fundamental tool plane of reference for assessment of craniofacial anomalies
for anthropological analysis, it allows researchers to deal Commercial visible-light imaging systems: Fixed viewpoint
with problems that could not be resolved using traditional depth maps are created from stereo-photogrammetry systems
anthropological approaches without compromising the (Geometrix’s Face 200 and 800) and structured-light camera
integrity of the physical remains (i.e., analysis of mummies, systems (such as Eyetronics, 3DMD and Inspeck requiring a
reconstruction of deformed fossils, etc.). Models of the slide projector and one or more cameras). To produce a full
physical object allow for virtual manipulation, simulation, and face model (from ear to ear) with these systems, two or three
bone sectioning, etc., in a virtual space, therefore preserving depth maps are obtained for a subject from varying view
the original object from invasive procedures. points (e.g., left-side, right-side, and frontal) and stitched
Facial imaging technologies for virtual antropology together with manual assistance. The only commercially
available system that acquires an ear-to-ear model is Face
Imaging modalities for clinical evaluation of the face, such 1200 from Geometrix with 12 cameras. Given the current
as photographs and two-dimensional radiographic films state, a fast, efficient, reliable, non-invasive solution to 3-D
(used since 1931) were developed decades ago and are still facial image acquisition would be a very significant step
in mainstream clinical use. However, the information they forward. A system such as this would eliminate many of the
provide is limited in perspective, accuracy, and contains barriers for clinicians to obtain and use such as system. Today,
information voids. For these reasons, in the last decades, no 3-D systems are in common clinical use, while traditional
threedimensional techniques such as 3D CT, laser surface photography is the standard.
scanning, photogrammetry (conversion of photographs taken
from different views into 3D models), Moire’ stripes18, and Cranial form analysis
Computer Assisted Design (CAD) manipulation of these Landmarks have been used for over a century by
models have been explored.5 anthropometrists interested in quantifying cranial variation.
CT Scans: Computed tomography (CT scan) has much of A new field, morphometrics, has grown around the statistical
its history in general medicine while its use in craniofacial analysis of shape and size for comparison of biological shapes11
assessment is more recent. In this area, the bulk of the research A great body of work in craniofacial anthropometry is that of
work is focused upon bony cranial landmarks. Previous Farkas12 who established a database of anthropometric norms
work reported the use of 3D CT for craniofacial surgical by measuring and comparing more than 100 dimensions
planning and comprehensive assessment. Richtsmeier and her (linear, angular and surface contour’s) and proportions
research group6 developed mathematical tools such as EDMA in hundreds of people over a period of many years. These
(Euclidean Distance Matrix Analysis) to assess asymmetry measurements include 47 landmark points to describe the
face (Figure 1 shows some of the Farkas’ landmarks)
4 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018
Figure 1: Subset of Farkas’s anthropometric landmarks (frontal and side picture of the mannequin).

Farkas’s inventory of facial measurements has been used in 10. The final step is to cover the clay muscles with a layer
computer graphics to automatically create new “plausible” of clay skin ,which is smoothened over such that it
computer graphic faces12. resembles the real skin.
11. Color :
Steps for reconstruction
12. - Hair, skin & eye color are added by borrowing the
1. Click photograph and digitize it using digital camera
physical features of a living person of similar age, racial
2. Place digitized markers on anthropometric landmarks qualities & built by a process called “3-D mapping.”
• Head: g – glabella, tr – trichion, ft – frontotemporale. F.A.C.E.S & C.A.R.E Softwares.
• Face: zy – zygion, go – gonion, sl – sublabiale, pg– Conclusion
pogonion, gn – gnathion (or menton, not visible), cdl –
Forensic arts has to deal with many ambiguous variables (such
condylion laterale.
as the shape of the eyes, the lips and the nose), cranio-facial
• Orbits: en – endocanthion, ex – exocanthion. reconstruction cannot claim to provide with absolute certainty
• Nose: n – nasion, prn – pronasale, sn – subnasale, sbal – the look of the personage. On the other hand, we can assume
sub-alare (sbal’), ac – alar curvature (ac’). that facial reconstruction based on forensic procedures is the
• Lips and mouth: cph – crista philtri (cph’), ch – most scientific approach to obtain an approximated aspect of
cheilion (ch’), sto – stomion, ls – labiale superius, li– the face, at least regarding the overall shape.
• Ears: obi – otobasion inferius, obs – otobasion superius,
Alves ES – Medicina Legal E Deontologia. Curitiba, Ed. Do Autor,
sa – superaurale, sba – subaurale, pa – postaurale,pra –
preaurale. 12
Aulsebrook AW, Iscan MY, Slabbert JH and Becker P. Superimposition
3. Depth of skin that overlays the skull is estimated. and reconstruction in forensic facial identification - A survey:
4. Small pegs are used as facial depth indicators & are fixed Forensic Sci Int. 1995; 75: 101-120.
into the skull. Phillips VM, Rosendorff S, Scholtz HJ. Identification of a suicide
5. The mimic muscles are made of plastilin or clay, eyes are victim by facial reconstruction: Forensic Odontostomatol.
of marble & the nose is formed from paraffin or wax. 1996; 14: 34-38.
Vanezis M, Vanezis P. Cranio-facial reconstruction in forensic
6. Started with 20-35 tissue layers usually, scattered all over
identification - Historical development and a review of current
the face. Main heavily concentrated depths are situated
practice. Med Sci Law. 2000; 40 : 97- 205.
around the mouth & in between the eyes
T. Kawai, N. Natsume, H. Shibata, and T. Yamamoto, “Three-
7. Work on the eyes, mouth, ears, nose, chin, jaws & cheeks dimensional analysis of facial morphology using moire
is now started. stripes.Part I. Methods,” Int J Oral Maxillofac Surg, 19, 356-358,
8. Next a mould from clay head is made using plaster of 1990.
paris S. Lele and J. T. Richtsmeier, “Euclidean Distance matrix analysis:
9. Muscles are approximated by noting the shape & size a coordinate-free approach for comparing biological shapes
of certain facial bones. Shaping & fixing of each muscle using landmark data,” Am J Phys Anthropol, 86, 415-427, 1991.
onto the skull in it’s place is important criteria. S. Lele and J. T. Richtsmeier, “On comparing biological shapes-
detection of influential landmarks.,” Am J Phys Anthropol,

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 5

S. Lele and J. T. Richtsmeier, “Euclidean Distance Matrix Analysis:
confidence intervals for form and growth comparison.Amer. J. Dr Saurabh Arora
Phys. Anthropol. 98: (1) 73-86, 1995 Reader
J. P. Moss, A. Linney, S. Grindrod, A. SR, and C. JS, “Three- Department of Conservative and Endodontics
dimensional visualization of the face and skull using Vananchal Dèntal College and Hospital
computerized tomography and laser scanning techniques,” Garwa, Jharkhand
European Journal of Orthodontics, 9, 247-53, 1989.
Dr Ravneet Arora
D. Marr and T. A. Poggio. Cooperative Computation of Stereo
Disparity. Science, 194[4262], 283-287, 197 Reader
Department of Oral Medicine and Radiology
F. Rohlf, “Morphometrics,” Annual Rev Ecol Syst, 21, 299-316,
Vananchal Dèntal College and Hospital
Garwa, Jharkhand
L.Farkas,Anthropometry of the Head and Face, Raven Press, New
York, 1994.

6 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Shear bond strength assessment of composite – tooth
interface following caries excavation by different
chemomechanical caries removal agents
Dr. Jyotika Grover

Background: Chemomechanical agents are routinely used in the excavation of caries in primary teeth. However, to
achieve clinical success of the final restoration, the bond strength of the affected dentin to the composite is essential.
Aim: The purpose of this study was to assess the shear bond strength between composite and caries affected dentin
after caries excavation by different chemomechanical caries removal agents. Settings and Design: Thirty human primary
molars with moderate caries were selected and randomly divided into 3 groups according to the CMCR agent used for
caries excavation; Group I Carisolv, Group II Cariecare and Group III Papacarie. The teeth were then restored with light
cured microhybrid composite and sectioned at tooth restoration interface for the assessment of shear bond strength using
universal testing machine. Results: The observations were put to descriptive statistics to find out the mean and standard
deviation of all 3 Groups. Further, independent t-test was used for intergroup comparisons. Conclusion: Maximum shear
bond strength was found in Group II (Cariecare - 15.21 + 4.45 N/mm2) while minimum shear bond strength was observed
in Group I (Carisolv). There was statistically significant difference in shear bond strength between Group I and Group III.
Keywords: Cariecare, Papacarie, Carisolv, Shear bond strength.

Introduction Several restorative materials have been compared (nanohybrid,

The peculiarity of pediatric dentistry is that it focuses on microhybrid, and microfilled), out of which microhybrid resin
eradicating anxiety, pain, and fear from the child towards based composite demonstrated highest shear bond strength.7
dental treatment and inspiring the child towards it by behavior However the literature is scarce regarding the studies on the
modification and by providing comfort and relief environment bonding of resin composite (microhybrid) to affected dentin
in dental office. following caries removal through various chemomechanical
Traditional methods which were based on the ideology of caries removal agents. Hence, the present study was aimed
extension for prevention have been shifted to minimally to evaluate and compare shear bond strength of new hybrid
invasive technique due to which less tooth cutting is achieved.1 composite to affected dentin following caries excavation with
Carisolv, Cariecare and Papacarie.
Several new technologies came into light which supported
minimal intervention technique such as ozone therapy, Methodology
ultrasonic instrumentation, lasers, air abrasions and A total of 90 extracted human primary teeth (samples) with
chemomechanical caries removal agents. Out of all, moderate caries were included in the study and stored in
chemomechanical caries removal agent fitted best for caries deionized water with 0.1% thymol. Roots were removed
removal without any excessive tooth cutting and preventing perpendicular to the long axis of the tooth at CEJ with a
from causing any anxiety or fear for dental treatments in 0.3mm thick diamond blade. The crowns were washed in
patient.2 distilled water and dried with a triplex syringe.
Carisolv, Papacarie and Cariecare are some of the The collected samples were equally and randomly divided
chemomechanical caries removal agents used to excavate into 3 groups according to CMCR agents were to be used for
unmineralized infected dentin which is rich in bacteria. These caries excavation.
also help in removal of smear layer of varying degree, leaving
behind affected dentin which is the remineralizable.3-5 a. GROUP 1 : 30 samples, caries excavation using Carisolv
b. GROUP 2 : 30 samples, caries excavation using Cariecare
The presence of smear layer has been known to act as a
barrier between restorative material and prepared cavity c. GROUP 3 : 30 samples, caries excavation using Papacarie
compromising the bond strength as it prevents the formation In group I (Carisolv), caries was excavated using instruments
of resin tags. The type of restorative material used is another provided by manufacturer. Required amount of gel was
factor which contributes to bond strength with the prepared dispensed into dappen dish and was applied to carious lesion
cavity walls.6 using multistar instrument and left it over for 30 seconds
for chemical process to soften the caries, which was then

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 7

excavated with the multi star instrument and the process was to remove with an 11 no. scalpel blade, leaving a 2 mm high
repeated until the solution remained clear without turning resin composite rod (diameter= 1 mm) bonded on the restored
cloudy. After completion of excavation, the surface was composite. All segments were then stored in tap water at 37°C
wiped out with a moist cotton pellet. for 12 hours prior to testing.
In Group II (Cariecare), gel was applied directly into cavity The bonded interface was subjected to shear bond strength
using disposable applicator tip; soon gel changed the color in testing in a universal testing machine. A metal intender was
the affected area. After 1 minute using sharp spoon excavator used to deliver a force parallel to the bonded surface at a
the gel along with dissolved caries was removed. The crosshead speed of 4.0 mm/minute (Fig 2).The microshear
procedure was repeated until caries was completely removed. bond strength value in megapascals (MPa) at fracture site was
calculated for each segment using the formula:
In Group III (Papacarie), the gel was dispensed in dappen
dish and was dispersed over the carious lesion using spoon Microshear bond strength (MPa) = Shear force (N)/Cross-
excavator and was left in place for 30 seconds undisturbed. sectional area (mm^)
When gel became cloudy, without applying pressure it was
removed gently by scraping with the spoon excavator. The Results
removal of carious dentin was continued until the gel was no The 30 segments of each Group (I, II and III) were evaluated
longer cloudy. The gel was then removed and the cavity was for shear bond strength. The shear bond strength was found to
wiped with a moistened cotton pellet and rinsed. be highest in Group II (15.21 + 4.45) and was lowest in Group
I (12.88 + 3.20) but difference in shear bond strength amongst
After excavation of caries from each sample, the crowns
groups was statistically not significant according to one way
were sectioned parallel to the occlusal plane to provide a flat
ANOVA test.
coronal dentinal surface and each sample was then segmented
vertically in buccolingual and mesiodistal directions to get 4 When intergroup comparison was made using independent
segments of each sample, using diamond disc (Fig 1). These t test (Table 1) between Groups I and II (Fig: 3), the shear
segments were mounted on dental stone for convenient bond strength was found to be more in group II (Cariecare)
handling. compared to group I (Carisolv) and the difference was
statistically significant (p= 0.023). When shearbond strength
All the segments were stored in 0.1% Thymol in deionized
between Group I and III were compared, the difference was
water until preparation for bonding. After dentinal etching
not statistically significant (p= 0.098) though the shear bond
with 37% phosphoric acid for 10 seconds (Scotchbond; 3M
strength of Group III (Papacarie) was more than Group I
ESPE), washing and minimal drying, an adhesive dentinal
(Carisolv) (Fig: 4). In intercomparison between Group III
bonding agent, 3M ESPE Single Bond (SB; 3M ESPE, St
and Group II (Fig: 5), the shear bond strength of Group II
Paul, Minn) was applied to all prepared areas of segments
(Cariecare) was more compared to Group III (Papacarie),
according to the manufacturer’s directions and restored with
though here also the difference was statistically non significant
resin composite (GC solare x microfine hybrid resin, A2
(p= 0.574).
A PVC tube (internal diameter= 1 mm, 2 mm high) was Discussion
placed on the restored composite surface and light-cured Frank carious lesions are restored with various esthetic and
together with the adhesive. The tube was filled with resin non esthetic materials with varying degree of success and
composite (GC solare x microfine hybrid resin, A2 shade) failures but with the advent of adhesive dentistry, composites
and light-cured for 40 seconds. After that the tube was cut have become material of choice owing to their esthetics,

Fig. 3 Fig. 4 Fig. 5

Fig. 3, 4 and 5: Comparison of Shear bond strength (N/mm2) using independent t-test.

8 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Table 1: Mean shear bond strength of microfilled hybrid composite to affected dentin following excavation using CMCR
Measure Distribution of primary tooth dentin segments
Group 1: Group 2: Group 3:
Carisolv Cariecare Papacarie
No. of specimen tested 30 30 30
Mean microshear bond strength 12.88 + 3.20 15.21 + 4.45 14.56 + 4.46
(MPa) ± (SD)
P values:
Group I vs Group II 0.023*
Group I vs Group III 0.098**
Group II vs Group III 0.574**
* Statistically significant
** Statistically non significant

strength and ease of restoring the teeth. Restorative material which acts only on damaged tissues due to the absence of
which was used in this study was microfilled hybrid composite an antiplasmatic protease, alpha-1-antitrypsin, which hinders
material, due to its good strength, wear resistance and for its proteolytic action on the normal tissues. The absence
permitting excellent esthetics. of alpha-1-antitrypsin in infected tissues allows Papain
to break down partially degraded molecules. It contains
Complete excavation of infected dentin, moisture free
Chloramine which acts as a disinfectant and softens the
environment, proper isolation and restorative material having
carious dentin due to chlorination of the partially degraded
good adhesion properties are some of the factors which
collagen because of which the smear layer is completely
determine the success of restoration.8 Another important factor
removed. Jawa et al pointed out that due to thorough removal
that can affect bond strength is the presence of smear layer.
of smear layer, highly irregular surface or high roughness is
Whenever dentin is cut using hand or rotary instruments, the
achieved, providing a suitable surface for strong bonding with
mineralized tissues are not shredded or cleaved but shattered to
produce considerable quantities of debris. Much of this, made restorative materials.14
up of very small particles of mineralized collagen matrix, Various other studies have shown that dentin surfaces
spread over the surface to form what is called the smear layer. undergoing excavation by bur and Co2 laser result in a
Smear layer consists of organic and hydroxyapatite particles surface characterized by crazing, undercuts, irregularities
that cover dentin which interferes with adhesion between and presence of smear layer which influence the strength
bonding resin and underlying dentin.9 required for bonding of resin adhesion to prepared cavity.
Chemomechanical caries removal agents, used in the study
In our study, Cariecare showed highest shear bond strength
has been found effective in removing smear layer partially or
out of the 3 chemomechanical caries removal agents used.
Similar results were found in the study by Shehab FAHE
and El-Tekeya et al who found that shear bond strength after Conclusions
excavating with Carisolv agents was less when compared with
Based on the results of present study, the following conclusions
other chemomechanical caries removal agent like Papacarie
were drawn:
and traditional techniques.10,11 The reason attributed was
that both Caricare and Papacarie contain an active ingredient 1. All the 3 chemomechanical caries removal agents used in
Papain which dissolves the infected dentin along with the study; Carisolv, Cariecare and Papacarie were found
complete removal of smear layer resulting in higher shear to be effective in removing infected carious dentin.
bond strength. 2. Shear bond strength was found to be maximum between
When the shear bond strength of Carisolv and Papacarie were composite restoration and tooth dentin when caries was
compared, Papacarie showed better results. Similar results excavated using Cariecare (Group II).
were also seen in the studies done by Hamama HH et al12 and 3. Minimum shear bond strength was observed between
Viral PM et al.13 The better shear bond strength of Papacarie composite restoration and tooth dentin when caries was
group could be explained by the fact that Papacarie has the removed with Carisolv (Group I).
ability to remove complete smear layer with less marked 4. Caries excavation using Cariecare and Papacarie did not
destruction to dentinal tubules resulting in better shear bond result in any significant change in the shear bond strength
strength.14 Papacarie also has an active ingredient, papain of composite restoration with tooth dentin.

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 9

However, further studies should be conducted using a larger Yazici AR, Atílla P, Ozgünaltay G, Müftüoglu S. In vitro comparison
sample size to evaluate the shear bond strength of composite of the efficacy of Carisolv and conventional rotary instrument
with caries affected dentin following excavation by various in caries removal. J Oral Rehabil. 2003 Dec;30(12):1177-82.
chemomechanical caries removal agents and to authenticate Moshonov J, Stabholz A, Bar-Hilel R and Peretz, B: Chemical
the results of the present study. analysis and surface morphology of enamel and dentin
following 9.6 micron CO2 laser irradiation versus high speed
References drilling. J Dent. 2005;33(5):427-32.
Bijle MNA, Patil S, Mumkekar SS, Arora N, Bhalla M, Murali
KV. Awareness of Dental Surgeons in Pune and Mumbai,
India, regarding Chemomechanical Caries Removal System. J
Contemp Dent Pract 2013;14(1):96-9.
Rajakumar S, Mungara J, Joseph E, Philip J, Guptha V, Mangalan
Pally SP. J Clin Pediatr Dent. 2013 Nov;38(1):23-6.
Shehab FAHE. Morphological and structural changes of dentin after
caries removal by different caries removal techniques and their
effect on the Shear bond strength to poly acid modified resin
composite. Cairo Dental Journal.2008 Jan;24 (I):99-110.
Corrêa FN, Filho LE, Rodrigues CR. Evaluation of residual dentin
after conventional and chemomechanical caries removal using
SEM. J Clin Pediatr Dent. 2008 Winter; 32(2):115-20.
Ramamoorthi S, Nivedhitha MS, Vanajassun PP. Effect of two
different chemomechanical caries removal agents on dentin
microhardness: An in vitro study. J Conserv Dent. 2013
Violich DR, Chandler NP. The smear layer in endodontics - a review.
Int Endod J. 2010 Jan;43(1):2-15.
Moncada G, Angel P, Fernandez E, Alonso P, Martin J, Gordan VV.
Bond strength evaluation of nanohybrid resin-based composite
repair. Gen Dent. 2012 May-Jun;60(3):230-4.
Shono Y, Ogawa T, Terashita M, Carvalho RM, Pashley EL, Pashley
DH. Regional measurement of resin-dentin bonding as an array.
J Dent Res. 1999 Feb;78(2):699-705.
Yamada Y,  Kimura Y,  Hossain M,  Kinoshita JI,  Shimizu
Y, Matsumoto K. Caries removal with Carisolv system: criteria
evaluation and microleakage test. J Clin Pediatr Dent. 2005
Winter; 30(2):121-6.
Shehab FAHE. Morphological and structural changes of dentin after
caries removal by different caries removal techniques and their
effect on the Shear bond strength to poly acid modified resin
composite. Cairo Dental Journal.2008 Jan;24 (I):99-110.
El-Tekeya M, El-Habashy L, Mokhles N, El-Kimary E. Effectiveness
of 2 chemomechanical caries removal methods on residual
bacteria in dentin of primary teeth. Pediatr Dent. 2012 Jul-
Hamama HH, Yiu CK, Burrow MF, King NM. Chemical,
morphological and microhardness changes of dentine
after chemomechanical caries removal. Aust Dent J. 2013
Viral PM, Nagarathna C, Shakuntala BS. Chemomechanical Caries
Removal in Primary Molars : Evaluation of Marginal Leakage
and Shear Bond Strength in Bonded Restorations” - An  in
Vitro Study. J Clin Pediatr Dent. 2013 July;37(3):269-74.
Jawa D, Singh S, Somani R, Jaidka S, Sirkar K, Jaidka R. Comparative
evaluation of the efficacy of chemomechanical caries removal
agent (papacarie) and conventional method of caries removal:
An in vitro study. J Indian Soc Pedod Prev Dent 2010;28:73-7.

10 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Dr Anamika Sharma

Deformities of the alveolar process discovered during mucogingival procedures can present surgical dilemmas which
may seriously affect the outcome of treatment Fenestration and dehiscence are localized defects of the cortical bone which
covers the teeth. Etiologic factors include developmental anomalies, frenum attachments, orthodontic tooth movement,
periodontal and endodontic pathosis, trauma from occlusion, tooth size, and tooth position. The present report describes a
case of alveolar dehiscence in relation to maxillary canine encountered three months following diode laser depigmentation

INTRODUCTION: of pain (Fig 5).

An alveolar dehiscence denotes a lack of the facial or lingual
alveolar cortical plate resulting in a denuded root surface,
while an alveolar fenestration is a circumscribed defect Alveolar bone architecture may vary from patient to patient
of the cortical plate which exposes the underlying root in point of thickness, contour and configuration, and all
surface, not involving the marginal bone1. Deformities of the these variations may be both normal and healthy1. The cause
alveolar process can present surgical dilemmas which are of these differences is the unique dependence between the
technically demanding and may seriously affect the outcome morphology of the alveolar process and the teeth. The bone
of treatment.2 contour normally conforms to the prominence of the roots
with intervening vertical depressions that taper toward the
The overall prevalence ranges from 0.99 to 13.4% for margin4.
dehiscences and 0.23% to 16.9% for fenestrations. Elliott and
Bowers3 reported that alveolar dehiscences and fenestrations Previous studies have reported the prevalence of alveolar
were associated with 9.19% and 10.93%, respectively. dehiscences and fenestrations in human skulls and autopsy
specimens from a variety of populations including Japanese4,
CASE REPORT: Bedouins5, Italians & Australians6, Mexican Indians7,
A 25 year old male patient reported with the complaint of Egyptians1, Britons8 and South African Blacks9.
dark coloured gums. Gingival depigmentation was performed Clinical predictors of dehiscences and fenestrations have not
with diode laser in both maxillary and mandibular arches been well defined. A number of possible etiologic factors have
in the intercanine region. Three months postoperatively, been examined, including developmental anomalies, frenum
the patient reported with pain in the attached gingiva of the attachments, orthodontic tooth movement, periodontal and
maxillary right lateral incisor and canine. On examination, a endodontic pathosis, trauma from occlusion, tooth size,
bony sequestrum surrounded by erythema was observed in and tooth position2. Davies et al9found dehiscences and
the gingiva overlying the maxillary right lateral incisor and fenestrations in 5.36% and 8.45% in teeth of 398 nineteenth
canine. (Fig 1) century British skulls. Dehiscences were most commonly
A submarginal incision was given with a 15 no.B.P.blade in associated with mandibular canines and first premolars and
the attached gingiva extending from the maxillary right lateral maxillary canines and first molars. Fenestrations were most
incisor to the maxillary right first premolar. A full thickness frequently associated with maxillary first molars. Edel6
flap was reflected to perform alveoloplasty for removal of evaluated 87 dry Bedouin jaws and found the prevalence
the bony sequestrum and reshaping of the underlying bone. of dehiscences and fenestrations to be 4.40% and 9.70%,
On removal of the sequestrum, an alveolar dehiscence was respectively, in 990 teeth examined. Our patient presented a
found in relation to the maxillary right canine, measuring dehiscence in the maxillary canine. Possible etiologic factor
upto 6 mm from the gingival margin (Fig 2). Alveoloplasty for the existence of this defect may be the prominent root
was performed; the flap was repositioned and sutured with of canine in combination with a thin alveolar bone plate.
5-0 black silk and postoperative instructions given (Fig 3). This observation is consistent with previous reports by
Antibiotic and analgesic was prescribed for 5 days. Erythema Abdelmaleket al1 and Edel6.
was absent one week postoperatively but slight pain was Presence of the alveolar defect observed in our patient may
reported (Fig 4). On one month follow up, the gingiva was also be attributed to the use of laser beam deep into the
pink, firm and resilient with stippled appearance and absence connective tissue in an attempt to remove the pigmented

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 11

Figure 1. Preoperative Figure 2. Alveolar Dehiscence Seen on Figure 3. Sutures Placed
Flap Reflection

Figure 4. 1 Week Postoperative Figure 5. 1 Month Postoperative

tissues completely. Also the soft tissues overlying the canine CONCLUSION:
region are thin, therefore laser ablation may have been Alveolar dehiscence is a defect which may be encountered
inadvertently carried out in close proximity of bone, leading over the maxillary canine as an intraoperative discovery.
to necrosis and eventually sequestration of bone. Similar Possible etiologic factors include malpositioned tooth
findings were reported by Atsawasuwan et al11 in 4 cases of with thin alveolar bone which may undergo necrosis and
gingival pigmentation treated with Nd:YAG laser. eventually sequestration of bone by laser ablation carried
Lost12 reported that in most cases the bone crest was found deep into connective tissue.
to be about 3 mm from the position of the gingival margin;
however, in 14% of instances, the bone crest was observed
located more than 4 mm from the position of the pre-surgical AbdelmalekRG, Bissada NF. Incidence and distribution of alveolar
soft tissue margin, most often associated with mandibular bone dehiscence and fenestration in dry human Egyptian jaws.
J Periodontol 1973;44:586-588.
canines. Similarly, in our patient, the distance of crestal bone
from gingival margin was 6 mm, though no recession was Rupprecht RD, Horning GM, Nicoll BK and Cohen ME.
Prevalence of Dehiscences and Fenestrations in Modern
present over the canine.
American Skulls. J Periodontol 2001;72:722-729.
Clinically, the intraoperative discovery of an alveolar Elliott JR, Bowers GM. Alveolar dehiscence and fenestration.
dehiscence or fenestration requires special consideration. If a Periodontics 1963;1:245-248.
defect is unintentionally exposed, root instrumentation which Nimigean VR, Nimigean V, Bencze MA, Dimcevici-Poesina
may remove radicular connective tissue fibers should be N, Cergan R, Moraru S. Alveolar bone dehiscences and
avoided, the root surface should not be permitted to become fenestrations: an anatomical study and review. Romanian
desiccated, and soft tissue coverage of the defect must be Journal of Morphology and Embryology2009, 50(3):391–397
ensured. The soft tissue used to cover these defects, whether Ezawa T, Sano H, Kaneko K, Huruma S, Fufikawa K, Murai
obtained by use of a free autogenous graft or mobilized soft S. The correlation between the presence of dehiscence or
tissue flap, must have an adequate blood supply to overcome fenestration and the severity of tooth attrition in contemporary
the lack of perfusion from the avascular root surface. The dry Japanese adult skulls. Part I. Nihon UnivSch Dent
absence of crestal bone reduces support for the soft tissue flap 1987;29:27-34.
and may increase the likelihood of undesirable post-operative Edel A. Alveolar bone fenestrations and dehiscences in dry Bedouin
gingival recession. Flaps must be carefully positioned and jaws. J ClinPeriodontol 1981; 8: 491-499.
sutured to optimally support the flap margin in the desired Larato DC. Alveolar plate fenestrations and dehiscences of human
position2. skull. Oral Surg Oral Med Oral Pathol1970;29: 816-819.

12 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Davies RM, Downer MC, Hull PS, Lennon MA. Alveolar defects
in human skulls. J Clin Periodontol 1974; 1: 107-111.
Dr Anamika Sharma
Professor and HOD, Department of Periodontology,
Tal H. Alveolar dehiscence and fenestrae in dried South African
Subharti Dental College and Hospital, Meerut, U.P., India
Negro mandibles. Am J Phys Anthropol1983;61:173-179.
Email: prof_anamika@hotmail.com
Atsawasuwan P, Greethong K, Nimmanon V. Treatment of
gingival hyperpigmentation for esthetic purposes by Nd:YAG
laser: Report of 4 cases. J Periodontol 2000; 71: 315-321.
Lost C. Depth of alveolar bone dehiscences in relation to gingival
recessions.J Clin Periodontol 1984; 11: 583-589.

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 13

Dr. Jyotsana Sikri and Dr. Arpit Sikri

Although humanmouth benefits fromremarkable mechanical properties, it is very susceptible to traumatic damages,
exposure to microbial attacks, and congenital maladies. Since the human dentition plays a crucial role in mastication,
phonation and esthetics, finding promising and more efficient strategies to reestablish its functionality in the event of
disruption has been important. Dating back to antiquity, conventional dentistry has been offering evacuation, restoration,
and replacement of the diseased dental tissue. However, due to the limited ability and short lifespan of traditional restorative
solutions, scientists have taken advantage of current advancements in medicine to create better solutions for the oral health
field and have coined it “regenerative dentistry.” This new field takes advantage of the recent innovations in stem cell
research, cellular and molecular biology, tissue engineering, and materials science etc. In this review, the recently known
resources and approaches used for regeneration of dental and oral tissues were evaluated using the databases of Scopus
and Web of Science. Scientists have used a wide range of biomaterials and scaffolds (artificial and natural), genes (with
viral and non-viral vectors), stem cells (isolated from deciduous teeth, dental pulp, periodontal ligament, adipose tissue,
salivary glands, and dental follicle) and growth factors (used for stimulating cell differentiation) in order to apply tissue
engineering approaches to dentistry. Although they have been successful in preclinical and clinical partial regeneration of
dental tissues, whole-tooth engineering still seems to be far-fetched, unless certain shortcomings are addressed.

1. Introduction from congenital maladies to chemical, physical, and microbial

After the first successful kidney transplant between two attacks [11]. While the oral cavity plays an essential role in
nongenetically identical patients was performed by Murray, daily life, it is severely exposed to microbial infections—
the Nobel prize winner and scientist in the early 1960s [1], therefore, any defect, induced by infections, decay or
transplantation has been the treatment for most of organ trauma and all other oral diseases including autoimmune and
injuries and failures. However, transplantation has major malignancies in the dental tissue should be addressed quickly
drawbacks such as severe shortage in organ donors, gradual [12]. More specifically, any large size defect that is close to
crescendo in the number of organ failure cases, indeterminate pulp exposure, including moderate to advance decay, needs
immune responses, and unreliable organ acceptability [1]. to be treated urgently. Often, trauma induced by mastication,
accidents or even pathogens can disrupt the oral epithelium
Therefore, scientists with backgrounds in cellular and protective barrier.
molecular biology, materials science, and stem cell
engineering came together and developed a newfield called The reports outline that 41% of the children aged 2–11 years
Tissue Engineering and RegenerativeMedicine (TERM). (in their primary teeth), 42% of children and adolescents
As a rapidly growing field of research, TERM offers novel aged 6–19 years, and approximately 90% of human adults (in
treatments for patients suffering fromslight injuries to end- their permanent teeth) suffer from at least one of the dental
stage organ failure for nearly every type of human body tissue diseases, such as caries [13], which makes it important to
and organ. The clinically available treatments include but are find approaches that can restore oral tissue to normal function
not limited to strategies for urethral tissue [2], bladder wall and form. Although the techniques used in conventional
tissue [3], genital tissues and organs [4], female reproductive dentistry—such as restoration with filling materials, whole
tissue [5], blood vessel [6], heart valves [7], liver [8] tooth replacement with synthetic restorative materials, and
and tracheal tissue [9]. In all of these cases, there are still teeth removal—date back to antiquity, they havemajor
substantial problems which need to be resolved; however, drawbacks that necessitate exploration of more effective
the recent advancements and their potential benefits seem to approaches and novel technologies in modern dentistry
be revolutionary. In dentistry, scientists have always placed [14]. The current efforts are focused on the investigation of
significant emphasis on the study of novel strategies that the possibility of engineering the whole tooth, as well as all
apply TERM to the dental practice [10]. Human teeth and of the individual dental structures separately. Both of these
orofacial tissues are responsible for phonation, mastication, routes require utilization and development of stem cells,
esthetics, respiration, and emotional and facial expressions. biomaterials, scaffolds, and growth factors. However, before
Although teeth have high abrasion resistance and lifelong outlining the details, grasping a better understanding of the
architectural durability, oral tissue, as one of the excessively human tooth structure and development is necessary.
used parts of the body, is prone to several common diseases

14 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

2. Current approaches in regenerative dentistry which can be harvested through relatively less-invasive
The recent scientific advancements in reprogramming procedures, have shown the capacity for multi-lineage
and guideddifferentiation of human embryonic and adult differentiation [35]. One-hundredth of white adipose cells are
stemcells, producing biocompatible materials, and scaffolding ADSCs with mesenchymal properties [36]. Tobita et al. [37]
systems that support cell growth have convinced scientists to has shown the possibility of periodontal tissue regeneration
apply these technologies to modern dentistry. using ADSCs [37]. Before the discovery of iPS by Takahashi
et al. [38], embryonic stemcells were the only available
Here, the major approaches recently used are discussed. pluripotent cells used in dentistry. Research on human and
mouse embryonic stem cells led to great accomplishments
2.1. Dental stem cells and growth factors
in differentiation of stemcells into oral tissues and organs
In the past few decades, a lot of progress has been made in [39,40]. Dental iPS cells have shown to be readily accessible
understanding, extracting and utilizing human embryonic from various dental stem cells [41,42] and fibroblasts [43,44].
and adult stem cells [17]. Self-renewal, programmability, The aim in using stem cells is to explore the possibility of
and the potential to produce various cell types are the main craniofacial, tooth, pulp, periodontal ligament, enamel, and
factors which make these cell types attractive for any field of dentin regeneration[45]. So far extensive studies on cementum
medicine [18]. In RD, scientists have used both pluripotent matrix by Handa et al. [46], periodontal ligament by Lin et al.
and adult stem cells derived from embryo, bone marrow, [47], soft dental pulp regeneration by Cordeiro et al. [48] and
dental tissues, oral tissues, and glands. They have also used Huang et al. [48,49], and enamel regeneration by Honda et al.
induced pluripotent stem (iPS) cells [19]. After discovery [50] have shown great promise in the future of stem cells in
of the potential of extracting stem cells from dental pulp by RD. Recently, Iglesias-Linares et al. [51] have investigated the
Gronthos et al. [20], dental pulp stem cells (DPSCs) and stem revascularization and apexogenesis induced by stem cells and
cells from human exfoliated deciduous teeth (SHED) were demonstrated the latest advancements in apical regeneration
the first cell lines derived from human dental pulp [20,21]. [51]. Although the scientists have established the preclinical
They are both favored because of their non-invasive harvest safety, efficacy and feasibility of pulp regeneration derived
and potential for multi-lineage differentiation [22]. Shi et al. fromdental stemcells [52], clinical trials cannot be launched,
[23] compared human dental pulp stem cells and bone marrow until certain challenges, such as the difficulty to handle
mesenchymal stromal stem cells (BMSCs) and showed critically-sized defects, are addressed [53].
distinct gene expression patterns for DPSCs [23]. In 2008
Growth factors can help tissue regrowth by regulating the
stem cells derived from apical papilla (SCAP), which were signaling between the cells, their environment, and their
harvested from wisdom teeth, were shown to have potential in neighbors [54]. Through making an information-conducive
dentin regeneration [24,25]. Although the abundant presence and extra-cellular matrix, growth factors play a crucial
of progenitor cells in the periodontal ligament was proven a role in the regeneration of dental tissues [55]. Tayalia et al.
long time ago [26], Seo et al. [27] investigated the stem cells demonstrated howscientists can take advantage of growth
harvested from the periodontal ligament (PDLSCs) of the factors to improve specifically guided differentiation of cells
third molar and found out that these stem cells are capable [56]. It is extremely important to knowwhich growth factors
of developing a tissue similar to their extraction site [27]. are suitable for specific types of cells and have the ability to
Morsczeck et al. [28] isolated precursor cells from the dental orchestrate the cell type’s proliferation and differentiation
follicle (DFSCs) of wisdom teeth and developed them into a into the anticipated cell lineage [57]. A wide spectrum of
mature periodontium [28]. Honda et al. [29] studied DFSCs growth factors is required in order to control each step of
and showed their osteogenic potential [29]. The progenitor tissue regeneration and the fate of the stemcell [58]. The
cells extracted from the tooth germ of the third molar during growth factors entrapped in the dentin matrix, which are
the bell stage (TGPCs), by Ikeda et al. [30] demonstrated the actively protected in the dentinmatrix, are responsible
ability to differentiate into osteoblasts, hepatocytes, and neural for the stimulation of processes—such as odontoblast
cells [30]. Scientists have also reported the extraction of stem differentiation—that lead to dentin formation [59]. Dental
cells from human dental epithelium tissue. Oral epithelial growth factors are also in charge of differentiation of adult pulp
stem cells [19], gingiva-derived mesenchymal stromal cells stem cells [60] and dentin bridging [61]. Although important
(GMSCs) [31], and periosteum-derived stem cells (PSCs) prerequisite steps for utilizing growth factors—delivery,
[32] have shown the potential to differentiate into lineages of immobilization, and release—are currently undergoing active
all three germ layers. On the other hand, stem cells derived investigations, preclinical and stage I/II clinical trials have
from human salivary glands have not shown the potential to demonstrated howgrowth factors can accelerate and improve
proliferate into all forms of epithelial cells [33]. Fig. 2 [19] periodontal and bone regeneration [62,63].
depicts the various oral and dental sources of adult stem cells.
2.2. Biomaterials and scaffolds
Adipose is loose, connective tissue which controls body
Scientists have used three major categories of materials in
energy resources in order to keep the body warm against cold
TERM: namely, naturally derived materials (such as chitosan,
environments[34]. Adipose-derived stromal cells (ADSCs),
elastin, and collagen), acellular tissue matrices, and synthetic

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 15

materials [1]. Since application of natural materials is limited, metallic implants have beenwidely used for medical and
FDA-approved synthetic polymers, such as polylactic acid dental applications and have been tailored for specific
(PLA), polyglycolic acid (PGA), and poly(lactic-co-glycolic reconstruction of small or large hard-tissue defects [102,103].
acid) (PLGA) have wide applications in many TERM However, scaffolds are suggested to be made of biodegradable
fields including RD [64,65]. Pre-clinical studies on animal materials with a degradation rate close to the tissue generation
models using all of the aforementioned categories have rate [11,104–106]. Therefore, the utilization of metallic
shown promising results in dental tissue regeneration [66]. scaffolds can be limited as they are mostly non-degradable,
Besides the conventional mechanical and chemical routes for and thus may require second surgery to be removed from the
synthesizing materials with biomedical applications [67–72], body. Scaffolds can be used in RD as structural templates for
scientists have always tried to explore innovative biomaterial stemcell differentiation and proliferation. Recently, Song et
synthesis techniques, such as green in situ synthesis of al. demonstrated the successful production of hard
silver particle encapsulated gelatin-based scaffolds, in situ
dental tissues on the periphery of macro-porous biphasic
encapsulation of iron nanoparticles in hydroxyapatite/chitosan
calcium phosphate scaffolds. Synthetic polymeric scaffolds
matrix, and particulate sol–gel and cellulose templating of
have shown great potential in promoting dental pulp tissue
nanostructured zirconium titanate fibers [73–77]. Recently,
regeneration [107]. Recent experiments on platelet-rich
novel biomaterials with more sophisticated designs that
plasma scaffolds have demonstrated that they can effectively
can be reinforced by bioactive elements have appealed to
improve the healing induction and tissue regeneration
scientists [78–81]. Some examples include coating of bone
in regenerative endodontic treatments [108,109]. Useful
scaffolds with fluoridated hydroxyapatite [80], adding various
implications of these scaffolds have been shown on a group
ion substitutes to bioactive glasses [79], and incorporation
of pediatric patients clinically or radiographically, although
of bonemorphogenetic protein into various bio-matrices
not significantly better than conventional blood clot scaffolds
to enhance osteogenesis[82]. Moreover, biodegradable
[110]. These studies can justify the investigations that are
hydrogels that profit from their tissuelike properties and cross-
currently being carried out for the design and improvement
linking potential can also be used for efficient incorporation
of dental scaffolds. Marine sponge skeletons [111], diatom
of biological agents [83,84]. In general, biomaterials that
skeletons [112], and Foraminifera micro-skeletons [113] have
are used in RD are artificial and must be able to promote the
been used as scaffolds, bioactive molecule delivery devices,
epithelial and mesenchymal interactions [85]. Trombelli and
and bone substitutes. These biomimetic structures can be
Farina [86] demonstrated how using calcium phosphate bone
interesting options for dental bone regeneration. Several
substitutes and collagen derivatives can encourage alveolar
pre-clinically successful collagenbase periodontal tissue
bone tissue rebuilding [86]. Marine biomaterials have also
regeneration strategies and also clinically available scaffold
started to attract a lot of attention in TERM and RD. A broad
materials have shown promising results to be used in RD
spectrum of biomaterials with high bio-availability can be
extracted from marine products. In 2011, Addad et al. isolated
collagen from jellyfish [87]. Two years later, Wysokowski 3.3. Other approaches
et al. extracted chitin from marine sponges [88]. Marine
There are over 700 genetic syndromes that cause approximately
biomimetics can be put into action in RDthrough either
75% of the congenital defects occurring in the United States.
deploymentwithout cellular content or in vitro culturing of
Beside the significant impact of these genetic disorders on
mature tissues inside their matrices [89]. Another approach
the quality of life, the estimated yearly treatment cost for
in delivering bioactive factors is called small molecule
these patients is more than $750 million [117]. For a long
technique, which involves utilizing carbon-based compounds
time, transferring manipulated genes for clinical applications
comprising only a small sequence of natural protein ligands
has been a dream, but nowadays with the recent advances in
[90]. In the past fifteen years, several small molecules have
biotechnology, gene therapy has shown promising preclinical
been designed and investigated for their osteoblast-promoting
results in curing non-hereditary and hereditary diseases [118].
and osteoclast-inhibiting properties [91–93].
In gene therapy, by either using a viral or non-viral vector
Due to their relatively smaller molecular size (b1000 Da),
as a carrier molecule, functional genes replace the abnormal
these molecules neither induce unwanted immune responses,
and malfunctioning mutant alleles after the insertion into
nor necessitate structural integrity for bioactivity [94].
the patient’s cells [119]. Unlike somatic gene therapy, in
Researchers have performed a number of preclinical animal
which functional genes are inserted into the patient’s somatic
studies on bone defects in order to reduce the nonspecific
cells, germline gene therapy targets genetic modification
adverse effect of small molecules [95–98]. Scaffolds provide
of spermand egg and would be heritable to the offspring
3-D support for cells, biological agents, and biomaterials
[120,121]. When using viral vectors, namely retroviruses,
in order to accomplish different missions—such as cell
adenoviruses, adenoassociated viruses, or herpes simplex
adhesion, stem cell differentiation, guided tissue regeneration,
virus—even though the forms of the genetic materials are
and permanent mechanical support [99–101]. For decades,
different—the transportation takes place after the virus

16 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

infects the host cell. For non-viral gene transfection, scientists scaffolds for cartilage regeneration, Schek et al. [142] used
have tried direct transfer of naked DNA [122], inactivation composite bio-printed scaffolds seeded with fibroblasts and
of diseased genes using oligonucleotides [123], liposomal reported a remarkable growth of cartilaginous tissue in the
delivery of plasmid DNA [124], application of cationic craniofacial region [142]. Kimet al. [143] made a 3-D printed
dendrimers and endocytosis [125], and the combination of tooth replica to performin vitro and in vivo experiments on
two or more techniques [126]. Over the past two decades, thewhole-tooth regeneration process [143]. Other groups
scientists have passionately worked on applying gene therapy have also tried the same route of research and have narrowed
to dentistry and as a result they have made tremendous down the fundamentals of whole-tooth regeneration via 3-D
progress in periodontal bone regeneration [127]. The salivary bio-printing [144, 145]. All of these technological advances
gland is another target of gene therapy and this area has shown showpromise for a hopeful future in 3-D bio-printing of the
promising results in both curing salivary gland diseases and whole tooth and other oral tissues for future generations.
even serious systemic pathologies [128]. Recently, genetically
In the last fifteen years, scientists have started to apply
modified cell therapy, by combining the benefits of direct
their knowledge of micro-electronics and achievements in
gene delivery and cell therapy [129], has been explored for
semiconducting materials to cellular differentiation and its
periodontal ligament in rabbits [130,131]. Showing good
microenvironment [146]. These technologies can potentially
patient specific adaptability, this novel therapy has potential
solve some of the challenges that other TERM approaches
for a bright future in this field.
face—for example, they can shed light on the reconstruction
Dental tissues have complex architecture, anisotropic of ectodermal and mesodermal interactions. They can provide
mechanical properties, and heterogeneous cell distribution; nano-resolution for building patterns to develop various cell
hence, it is hard to mimic their complex 3-D structure using types; hence, making these technologies useful for producing
the conventional techniques. To overcome this challenge, scaffolds carrying several stem cells. Moreover, microscale
recently 3-D bio-printing of dental and craniofacial tissues technologies provide the possibility of isolating, seeding, and
has been proposed [132,133]. 3-D bio-printed scaffolds combining various cell types, which makes them suitable
can be designed for each individual patient and have for in vitro assessments of cell behaviors inwell-controlled
shown remarkable controllability over cell and biomaterial environments [147]. This can enable rapid evaluation of
positioning, while maintaining great accuracy in internal the effects of biomaterials, drugs, and biological agents as
and external details [134]. In general, printers use computed a result of performing patterned single or multi-culturing
designs and follow the basic concept of layer-by-layer in vitro experiments. Flaim et al. [148] investigated the
deposition of materials to produce 3-D volumetric structures. potential synergistic effects of the simultaneous utilization
Based on the type of their ink dispenser, bio-printers of growth factors and extracellular matrix proteins on stem
can be grouped into three categories: Inkjet 3-D printers cell activity [148]. These novel technologies have improved
(capable of applying low-viscosity bio-inks using thermal growth factor delivery by offering precise cell control and
or piezoelectrical controlling system), laser-assisted printers regulation. For example, Ennett et al. [149] performed long-
(capable of using cell and biomaterial sources with various lasting growth factor release using PLGAmicro-spheres in
viscosities for pulse laser deposition of 3-D structures), and vitro and in vivo [149]. The so called “microscale technology
extrusion printers (capable of extruding high-viscosity and approach” can be carried out through either soft lithography
stiff polymeric sources at relatively high temperatures) [133]. or photolithography [15]. Kane et al. [150] and Rozkiewicz et
Due to their excellent biocompatibility and outstanding al. [151] used soft lithography tomold templates and pattern
tenability, polymeric hydrogels have been the best nominees selective cells [150,151]. Zhang et al. [152] and Kimet al. [153]
to be used as materials for 3-D bio-printing [135]. As the used photolithography to fabricate 3-D micro-vascularized
bio-printing process usually involves high-temperature steps, scaffolds and structures [152,153]. These techniques’ ability
cells and growth factors (temperature susceptiblematerials) to form 3-D microchannels can help in supporting the cell
are not initially amalgamated in the polymeric mixture [136]. metabolism [154]. This advantage can play a crucial role
Both ceramic (such as hydroxyapatite) [137] and composite in achieving a reliable technique for tooth regeneration.
materials (such as polymer composite hydrogels) [138] are Hydrogels, with approximately 99% water content, are
considered as alternative bio-ink materials. the best materials to use in the microscale approach [155].
These materials can provide controllability in the structural
Applying this novel technique to TERM, Reichert et al. [139]
formation with great detail. Microscale technologies can
used 3-D bio-printed scaffolds to study the bone formation
fabricate micro-structures, provide open channels, support
in a sheep model in which the sheep was suffering froma
vascularization, enhance diffusion, help regulate the cell
critically-sized bone defect and eventually showed significant
activity, and facilitate high-throughput approaches; hence,
bone formation improvement [139]. In RD, scientists have
they have a huge potential for both the in vitro and in vivo
made several attempts to mimic the intricate architecture of
constructions of tooth-like structures [156].
the periodontium in order to improve the regeneration of
the periodontal complex [140,141]. In an investigation on

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 17

Even after reaching the advanced technology of building cavernosa replacement using tissue engineering techniques, J.
patientspecific tooth substructures, the major challenges Urol. 168 (4) (2002) 1754–1758.
of the application of TE in dentistry range from the cost- [5] R.E. De Philippo, C.E. Bishop, L. Freitas Filho, J.J. Yoo, A.
efficiency of these approaches to their availability to public Atala, Tissue engineering a complete vaginal replacement
(in terms of well-equipped health centers and institutes). froma small biopsy of autologous tissue, Transplantation 86 (2)
Moreover, RD inherits the controversial ethical challenge of (2008) 208–214.
choosing which cell source (patient’s own or donors’) and cell [6] T. Shin’oka, Y. Imai, Y. Ikada, Transplantation of a tissue-
type (adult or fetal) for TE. However, the ongoing research on engineered pulmonary artery, N. Engl. J. Med. 344 (7) (2001)
TE and RD opens the venue to future investigations toward 532–533.
the development of whole-tooth structure during the next [7] P. Amrollahi, L. Tayebi, Bioreactors for heart valve tissue
decades; which furtherly can shed light on the regeneration engineering: a review, J. Chem. Technol. Biotechnol. (2015).
of other organs. [8] R.N. Bhandari, L.A. Riccalton, A.L. Lewis, J.R. Fry, A.H.
Hammond, S.J. Tendler, et al., Liver tissue engineering: a role
4. Conclusion for co-culture systems in modifying hepatocyte function and
Although a lot of advancements in RD have revolutionized viability, Tissue Eng. 7 (3) (2001) 345–357.
modern dentistry, there are still several steps left to take [9] S. Baiguera, M.A. Birchall, P. Macchiarini, Tissue-engineered
before declaring RD as a reliable alternative to conventional tracheal transplantation, Transplantation 89 (5) (2010) 485–
dentistry. RD owes plenty to stem cell science and growth 491.
factor engineering. However, a good source of totipotent [10] M.K. Marei, Regenerative dentistry, Synth. Lect. Tissue Eng. 2
stem cells is not yet readily accessible and extracting human (1) (2010) 1–178.
embryonic stem cells is a problematic and controversial issue. [11] K.M. Galler, R.N. D’Souza, Tissue engineering approaches for
Moreover, it is not easy to control the stemcell differentiation. regenerative dentistry, Regen. Med. 6 (1) (2011) 111–124.
Delivery of active growth factor to the desired site is [12] K. Singh, N. Mishra, L. Kumar, K.K. Agarwal, B. Agarwal, Role
challenging and might provoke side effects. Biomaterials and of stem cells in tooth bioengineering, J. Oral Biol. Craniofacial
scaffolds have played fundamental roles in facilitating partial Res. 2 (1) (2012) 41–45.
dental tissue regeneration, but until today, none of thematerials [13] E.D. Beltrán-Aguilar, L. Barker, M. Canto, B. Dye, B. Gooch,
have met all the mechanical and biological standards required S. Griffin, et al., Centers for Disease Control and Prevention
for RD. Gene therapy has opened up new directions to curing (CDC). Surveillance for dental caries, dental sealants,
dental congenital diseases in individuals and their offspring; tooth retention, edentulism, and enamel fluorosis—United
States,1988–1994 and 1999–2002, MMWR Surveill. Summ.
however, viral vectors used in this technique might trigger
54 (3) (2005) 1–43.
immune responses and side effectswith irreversible damage.
These genes live for a short period of time, which makes them [14] S.K. Majumdar, History of dentistry: an overview, Bull. Indian
Inst. Hist. Med. (Hyderabad) 32 (1) (2001) 31–42.
ineffective in some cases. Furthermore, 3-D bio-printing and
microscale technologies are pushing the boundaries of RD, [15] S. Hacking, A. Khademhosseini, Applications of microscale
technologies for regenerative dentistry, J. Dent. Res. 88 (5)
but both are costly and are still in their early developmental
(2009) 409–421.
stages. Though there is much work left, these are areas with
great promise for the future of RD. The future of dentistry [16] S.E. Duailibi, M.T. Duailibi, J.P. Vacanti, P.C. Yelick, Prospects
for tooth regeneration, Periodontol. 41 (1) (2006) 177–187.
is in the hands of cellular biologists, geneticists, biomedical
engineers, and materials scientists that strive to find and [17] C.-G. Fan, Z. Q-j, Z. J-r, Therapeutic potentials of mesenchymal
perfect novel approaches and techniques to address the stem cells derived from human umbilical cord, Stem Cell Rev.
Rep. 7 (1) (2011) 195–207.
aforementioned issues. Although the partial regeneration of
human dental tissues and structures seems to be attainable near, [18] Y. Sakaguchi, I. Sekiya, K. Yagishita, T.Muneta, Comparison of
considering the obstacles ahead,whole-tooth regeneration human stem cells derived from various mesenchymal tissues:
superiority of synovium as a cell source, Arthritis Rheum. 52
may be achievable in the farther future.
(8) (2005) 2521–2529.
References [19] H. Egusa, W. Sonoyama, M. Nishimura, I. Atsuta, K. Akiyama,
Stem cells in dentistry–part I: stem cell sources, J. Prosthodont.
[1] A. Atala, Regenerative medicine strategies, J. Pediatr. Surg. 47
Res. 56 (3) (2012) 151–165.
(1) (2012) 17–28.
[20] S. Gronthos, M.Mankani, J. Brahim, P.G. Robey, S. Shi,
[2] A. Raya-Rivera, D.R. Esquiliano, J.J. Yoo, E. Lopez-Bayghen,
Postnatal human dental pulp stem cells (DPSCs) in vitro and in
S. Soker, A. Atala, Tissueengineered autologous urethras for
vivo, Proc. Natl. Acad. Sci. 97 (25) (2000) 13625–13630.
patients who need reconstruction: an observational study,
Lancet 377 (9772) (2011) 1175–1182. [21] M. Miura, S. Gronthos, M. Zhao, B. Lu, L.W. Fisher, P.G. Robey,
et al., SHED: stem cells from human exfoliated deciduous teeth,
[3] F. Oberpenning, J. Meng, J.J. Yoo, A. Atala, De novo
Proc. Natl. Acad. Sci. 100 (10) (2003) 5807–5812.
reconstitution of a functional mammalian urinary bladder by
tissue engineering, Nat. Biotechnol. 17 (2) (1999) 149–155. [22] S. Gronthos, J. Brahim,W. Li, L. Fisher, N. Cherman, A. Boyde,
et al., Stem cell properties of human dental pulp stem cells, J.
[4] T.G. Kwon, J.J. Yoo, A. Atala, Autologous penile corpora
18 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018
Dent. Res. 81 (8) (2002) 531–535. factors, Cell 126 (4) (2006) 663–676.
[23] S. Shi, P. Robey, S. Gronthos, Comparison of human dental [39] Y. Shamis, K.J. Hewitt, M.W. Carlson, M. Margvelashvilli,
pulp and bone marrow stromal stem cells by cDNA microarray S. Dong, C.K. Kuo, et al., Fibroblasts derived from human
analysis, Bone 29 (6) (2001) 532–539. embryonic stem cells direct development and repair of 3D
[24] W. Sonoyama, Y. Liu, T. Yamaza, R.S. Tuan, S.Wang, S. Shi, et human skin equivalents, Stem Cell Res. Ther. 2 (10) (2011).
al., Characterization of the apical papilla and its residing stem [40] F. Ning, Y. Guo, J. Tang, J. Zhou, H. Zhang,W. Lu, et al.,
cells from human immature permanent teeth: a pilot study, J. Differentiation of mouse embryonic stem cells into dental
Endod. 34 (2) (2008) 166–171. epithelial-like cells induced by ameloblasts serumfree
[25] G.T.-J. Huang, W. Sonoyama, Y. Liu, H. Liu, S. Wang, S. Shi, conditioned medium, Biochem. Biophys. Res. Commun. 394
The hidden treasure in apical papilla: the potential role in pulp/ (2) (2010) 342–347.
dentin regeneration and bioroot engineering, J. Endod. 34 (6) [41] X. Yan, H. Qin, C. Qu, R.S. Tuan, S. Shi, G.T.-J. Huang, iPS
(2008) 645–651. cells reprogrammed from human mesenchymal-like stem/
[26] C.McCulloch, Progenitor cell populations in the periodontal progenitor cells of dental tissue origin, Stem Cells Dev. 19 (4)
ligament ofmice, Anat. Rec. 211 (3) (1985) 258–262. (2010) 469–480.
[27] B.-M. Seo, M. Miura, S. Gronthos, P.M. Bartold, S. Batouli, J. [42] Y. Oda, Y. Yoshimura, H. Ohnishi, M. Tadokoro, Y. Katsube, M.
Brahim, et al., Investigation of multipotent postnatal stem cells Sasao, et al., Induction of pluripotent stem cells from human
from human periodontal ligament, Lancet 364 (9429) (2004) third molar mesenchymal stromal cells, J. Biol. Chem. 285 (38)
149–155. (2010) 29270–29278.
[28] C. Morsczeck,W. Götz, J. Schierholz, F. Zeilhofer, U. Kühn, [43] K. Miyoshi, D. Tsuji, K. Kudoh, K. Satomura, T. Muto, K. Itoh,
C. Möhl, et al., Isolation of precursor cells (PCs) from human et al., Generation of human induced pluripotent stem cells from
dental follicle of wisdom teeth, Matrix Biol. 24 (2) (2005) oral mucosa, J. Biosci. Bioeng. 110 (3) (2010) 345–350.
155–165. [44] N. Wada, B.Wang, N.H. Lin, A.L. Laslett, S. Gronthos, P.M.
[29] M.J. Honda, M. Imaizumi, H. Suzuki, S. Ohshima, S. Tsuchiya, Bartold, Induced pluripotent stem cell lines derived from human
K. Satomura, Stem cells isolated from human dental follicles gingival fibroblasts and periodontal ligament fibroblasts, J.
have osteogenic potential, Oral Surg. Oral Med. Oral Pathol. Periodontal Res. 46 (4) (2011) 438–447.
Oral Radiol. Endod. 111 (6) (2011) 700–708. [45] M. Sethi, A. Dua, V. Dodwad, Stem cells: a window to
[30] E. Ikeda, K. Yagi, M. Kojima, T. Yagyuu, A. Ohshima, S. regenerative dentistry, Int. J. Pharm. Biomed. Res. 3 (2012)
Sobajima, et al., Multipotent cells from the human thirdmolar: 175–180.
feasibility of cell-based therapy for liver disease, Differentiation [46] K. Handa, M. Saito, A. Tsunoda, M. Yamauchi, S. Hattori, S.
76 (5) (2008) 495–505. Sato, et al., Progenitor cells fromdental follicle are able to form
[31] Q. Zhang, S. Shi, Y. Liu, J. Uyanne, Y. Shi, S. Shi, et al., cementummatrix in vivo, Connect. Tissue Res. 43 (2–3) (2002)
Mesenchymal stem cells derived from human gingiva are 406–408.
capable of immunomodulatory functions and ameliorate [47] N.H. Lin, S. Gronthos, P. Bartold, Stem cells and periodontal
inflammation-related tissue destruction in experimental colitis, regeneration, Aust. Dent. J. 53 (2) (2008) 108–121.
J. Immunol. 183 (12) (2009) 7787–7798. [48] M.M. Cordeiro, Z. Dong, T. Kaneko, Z. Zhang, M. Miyazawa,
[32] C. De Bari, F. Dell’Accio, J. Vanlauwe, J. Eyckmans, I.M. S. Shi, et al., Dental pulp
Khan, C.W. Archer, et al., Mesenchymal multipotency of adult tissue engineering with stem cells fromexfoliated deciduous teeth, J.
human periosteal cells demonstrated by single-cell lineage Endod. 34 (8) (2008) 962–969.
analysis, Arthritis Rheum. 54 (4) (2006) 1209–1221. [49] G.T.-J. Huang, T. Yamaza, L.D. Shea, F. Djouad, N.Z. Kuhn,
[33] A. Sato, K. Okumura, S. Matsumoto, K. Hattori, S. Hattori, R.S. Tuan, et al., Stem/ progenitor cell-mediated de novo
M. Shinohara, et al., Isolation, tissue localization, and cellular regeneration of dental pulp with newly deposited continuous
characterization of progenitors derived from adult human layer of dentin in an in vivo model, Tissue Eng. A 16 (2) (2009)
salivary glands, Cloning Stem Cells 9 (2) (2007) 191–205. 605–615.
[34] L.E. Kokai, K. Marra, J.P. Rubin, Adipose stem cells: biology [50] M.J. Honda, Y. Shinmura, Y. Shinohara, Enamel tissue
and clinical applications for tissue repair and regeneration, engineering using subcultured enamel organ epithelial cells in
Transl. Res. 163 (4) (2014) 399–408. combination with dental pulp cells, Cells Tissues Organs 189
[35] N. Arceo, J.J. Sauk, J. Moehring, R.A. Foster, M.J. Somerman, (1–4) (2009) 261–267.
Human periodontal cells initiate mineral-like nodules in vitro, [51] A. Iglesias-Linares, R.-M. Yáñez-Vico, E. Sánchez-Borrego,
J. Periodontol. 62 (8) (1991) 499–503. A.M. Moreno-Fernández, E. Solano-Reina, A. Mendoza-
[36] W.K. Ong, S. Sugii, Adipose-derived stem cells: fatty potentials Mendoza, Stem cells in current paediatric dentistry practice,
for therapy, Int. J. Biochem. Cell Biol. 45 (6) (2013) 1083– Arch. Oral Biol. 58 (3) (2013) 227–238.
1086. [52] M. Nakashima, K. Iohara, Mobilized dental pulp stem cells for
[37] M. Tobita, A.C. Uysal, R. Ogawa, H. Hyakusoku, H. Mizuno, pulp regeneration: initiation of clinical trial, J. Endod. 40 (4)
Periodontal tissue regeneration with adipose-derived stem (2014) S26–S32.
cells, Tissue Eng. A 14 (6) (2008) 945–953. [53] G.T.J. Huang, M. Al-Habib, P. Gauthier, Challenges of stem
[38] K. Takahashi, S. Yamanaka, Induction of pluripotent stem cells cell-based pulp and dentin regeneration: a clinical perspective,
from mouse embryonic and adult fibroblast cultures by defined
ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 19
Endod. Top. 28 (1) (2013) 51–60. [69] P. Amrollahi, A. Ataie, A. Nozari, S. Sheibani, Synthesis and
[54] D.E. Discher, D.J. Mooney, P.W. Zandstra, Growth factors, characterization of CuNi magnetic nanoparticles by mechano-
matrices, and forces combine and control stem cells, Science thermal route, J. Supercond. Nov. Magn. 27 (2) (2014) 481–
324 (5935) (2009) 1673–1677. 485.
[55] F.-M. Chen, Y. Jin, Periodontal tissue engineering and [70] E. Salahinejad, M. Hadianfard, D. Macdonald, I. Karimi, D.
regeneration: current approaches and expanding opportunities, Vashaee, L. Tayebi, Aqueous sol–gel synthesis of zirconium
Tissue Eng. B Rev. 16 (2) (2010) 219–255. titanate (ZrTiO4) nanoparticles using chloride precursors,
Ceram. Int. 38 (8) (2012) 6145–6149.
[56] P. Tayalia, D.J. Mooney, Controlled growth factor delivery for
tissue engineering, Adv. Mater. 21 (32−33) (2009) 3269–3285. [71] E. Salahinejad, M. Hadianfard, D. Macdonald, M. Mozafari,
D. Vashaee, L. Tayebi, Zirconium titanate thin film prepared
[57] F.-M. Chen, Y. An, R. Zhang, M. Zhang, New insights into
by an aqueous particulate sol–gel spin coating process using
and novel applications of release technology for periodontal
carboxymethyl cellulose as dispersant, Mater. Lett. 88 (2012)
reconstructive therapies, J. Control. Release 149 (2) (2011)
[72] E. Salahinejad, M. Hadianfard, D. Macdonald, M. Mozafari,
[58] F.J. Hughes, W. Turner, G. Belibasakis, G. Martuscelli, Effects
D. Vashaee, L. Tayebi, Multilayer zirconium titanate thin films
of growth factors and cytokines on osteoblast differentiation,
prepared by a sol–gel deposition method, Ceram. Int. 39 (2)
Periodontol. 41 (1) (2006) 48–72.
(2013) 1271–1276.
[59] A. Smith, P. Murray, A. Sloan, J. Matthews, S. Zhao, Trans-
[73] M. Yazdimamaghani, D. Vashaee, S. Assefa, M.
dentinal stimulation of tertiary dentinogenesis, Adv. Dent. Res.
Shabrangharehdasht, A.T. Rad, M.A. Eastman, et al., Green
15 (1) (2001) 51–54.
synthesis of a new gelatin-based antimicrobial scaffold for
[60] K. Iohara, L. Zheng, M. Ito, A. Tomokiyo, K.Matsushita, tissue engineering, Mater. Sci. Eng. C 39 (2014) 235–244.
M. Nakashima, Side population cells isolated from porcine
[74] M. Yazdimamaghani, T. Pourvala, E. Motamedi, B. Fathi,
dental pulp tissue with self-renewal and multipotency
D. Vashaee, L. Tayebi, Synthesis and characterization of
for dentinogenesis, chondrogenesis, adipogenesis, and
encapsulated nanosilica particles with an acrylic copolymer
neurogenesis, Stem Cells 24 (11) (2006) 2493–2503.
by in situ emulsion polymerization using thermoresponsive
[61] H. Lovschall, O. Fejerskov, A. Flyvbjerg, Pulp-capping with nonionic surfactant, Materials 6 (9) (2013) 3727–3741.
recombinant human insulin-like growth factor I (rhIGF-I) in rat
[75] F. Heidari, M.E. Bahrololoom, D. Vashaee, L. Tayebi,
molars, Adv. Dent. Res. 15 (1) (2001) 108–112.
In situ preparation of iron oxide nanoparticles in natural
[62] R. Rutherford, C. Niekrash, J. Kennedy, M. Charette, Platelet- hydroxyapatite/chitosan matrix for bone tissue engineering
derived and insulinlike growth factors stimulate regeneration of application, Ceram. Int. 41 (2) (2015) 3094–3100.
periodontal attachment in monkeys, J. Periodontal Res. 27 (4)
[76] E. Salahinejad, M. Hadianfard, D. Macdonald, M. Mozafari,
(1992) 285–290.
D. Vashaee, L. Tayebi, A new double-layer sol–gel coating to
[63] T.H. Howell, J.P. Fiorellini, D.W. Paquette, S. Offenbacher, improve the corrosion resistance of a medicalgrade stainless
W.V. Giannobile, S.E. Lynch, A phase I/II clinical trial to steel in a simulated body fluid, Mater. Lett. 97 (2013) 162–165.
evaluate a combination of recombinant human platelet-derived
[77] P. Rouhani, E. Salahinejad, R. Kaul, D. Vashaee, L. Tayebi,
growth factor-BB and recombinant human insulin-like growth
Nanostructured zirconium titanate fibers prepared by particulate
factor-I in patients with periodontal disease, J. Periodontol. 68
sol–gel and cellulose templating techniques, J. Alloys Compd.
(12) (1997) 1186–1193.
568 (2013) 102–105.
[64] R. Langer, D.A. Tirrell, Designing materials for biology and
[78] N. Huebsch, D.J. Mooney, Inspiration and application in the
medicine, Nature 428 (6982) (2004) 487–492.
evolution of biomaterials, Nature 462 (7272) (2009) 426–432.
[65] M. YazdiMamaghani, S.M. Davachi, P. Amrollahi, D. Vashaee,
[79] S.M. Rabiee, N. Nazparvar, M. Azizian, D. Vashaee, L. Tayebi,
L. Tayebi, Conducting polymers: developments, Encyclopedia
Effect of ion substitution on properties of bioactive glasses: a
of Biomedical Polymers and Polymeric Biomaterials, Taylor &
review, Ceram. Int. 41 (6) (2015) 7241–7251.
Francis 2016, pp. 1997–2010.
[80] M. Razavi, M. Fathi, O. Savabi, D. Vashaee, L. Tayebi,
[66] F.-M. Chen, H.-H. Sun, H. Lu, Q. Yu, Stem cell-delivery
Biodegradable magnesium alloy coated by fluoridated
therapeutics for periodontal tissue regeneration, Biomaterials
hydroxyapatite using MAO/EPD technique, Surf. Eng. 30 (8)
33 (27) (2012) 6320–6344.
(2014) 545–551.
[67] P. Amrollahi, A. Ataie, A. Nozari, E. Seyedjafari, A. Shafiee,
[81] M. Yazdimamaghani,M. Razavi, D. Vashaee, V.R. Pothineni, J.
Cytotoxicity evaluation and magnetic characteristics of
Rajadas, L. Tayebi, Significant degradability enhancement in
mechano-thermally synthesized CuNi nanoparticles for
multilayer coating of polycaprolactonebioactive glass/gelatin-
hyperthermia, J. Mater. Eng. Perform. 24 (3) (2015) 1220–1225.
bioactive glass on magnesium scaffold for tissue engineering
[68] P. Amrollahi, J.S. Krasinski, R. Vaidyanathan, L. Tayebi, D. applications, Appl. Surf. Sci. 338 (2015) 137–145.
Vashaee, Electrophoretic deposition (EPD): fundamentals and
[82] B.D. Ratner, S.J. Bryant, Biomaterials: wherewe have been and
applications from nano-to microscale structures, Handbook of
wherewe are going, Annu. Rev. Biomed. Eng. 6 (2004) 41–75.
Nanoelectrochemistry: Electrochemical Synthesis Methods,
Properties, and Characterization Techniques 2016, pp. 561– [83] G.D. Nicodemus, S.J. Bryant, Cell encapsulation in
591. biodegradable hydrogels for tissue engineering applications,
Tissue Eng. B Rev. 14 (2) (2008) 149–165.

20 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

[84] V. Shabafrooz, M. Mozafari, G.A. Köhler, S. Assefa, D. Vashaee, Open Access Publisher, 2012.
L. Tayebi, The effect of hyaluronic acid on biofunctionality [100] M. Yazdimamaghani, M. Razavi, D. Vashaee, L. Tayebi,
of gelatin–collagen intestine tissue engineering scaffolds, J. Surface modification of biodegradable porous Mg bone scaffold
Biomed. Mater. Res. Part A 102 (9) (2014) 3130–3139. using polycaprolactone/bioactive glass composite, Mater. Sci.
[85] T. Ohara, T. Itaya, K. Usami, Y. Ando, H. Sakurai,M.J. Honda, et Eng. C 49 (2015) 436–444.
al., Evaluation of scaffold materials for tooth tissue engineering, [101] M. Yazdimamaghani, M. Razavi, D. Vashaee, L. Tayebi,
J. Biomed. Mater. Res. Part A 94 (3) (2010) 800–805. Microstructural and mechanical study of PCL coated Mg
[86] L. Trombelli, R. Farina, Clinical outcomes with bioactive scaffolds, Surf. Eng. 30 (12) (2014) 920–926.
agents alone or in combination with grafting or guided tissue [102] K. Alvarez, H. Nakajima, Metallic scaffolds for bone
regeneration, J. Clin. Periodontol. 35 (s8) (2008) 117–135. regeneration, Materials 2 (3) (2009) 790–832.
[87] S. Addad, J.-Y. Exposito, C. Faye, S. Ricard-Blum, C. Lethias, [103] W.-E. Yang, M.-L. Hsu, M.-C. Lin, Z.-H. Chen, L.-K. Chen,
Isolation, characterization and biological evaluation of jellyfish H.-H. Huang, Nano/submicron-scale TiO2 network on titanium
collagen for use in biomedical applications, Mar. Drugs 9 (6) surface for dental implant application, J. Alloys Compd. 479 (1)
(2011) 967–983. (2009) 642–647.
[88] M. Wysokowski, M. Motylenko, V.V. Bazhenov, D. Stawski, [104] M. Razavi, M.H. Fathi, O. Savabi, D. Vashaee, L. Tayebi,
I. Petrenko, A. Ehrlich, et al., Poriferan chitin as a template Biodegradation, bioactivity and in vivo biocompatibility
for hydrothermal zirconia deposition, Front. Mater. Sci. 7 (3) analysis of plasma electrolytic oxidized (PEO) biodegradable
(2013) 248–260. Mg implants, Phys. Sci. Int. J. 4 (5) (2014) 708.
[89] D.W. Green, W.-F. Lai, H.-S. Jung, Evolving marine biomimetics [105] M. Razavi, M. Fathi, O. Savabi, S.M. Razavi, F. Heidari,
for regenerative dentistry, Mar. Drugs 12 (5) (2014) 2877–2912. M. Manshaei, et al., In vivo study of nanostructured diopside
[90] H. Egusa, Y. Kaneda, Y. Akashi, Y. Hamada, T. Matsumoto, (CaMgSi2O6) coating on magnesiumalloy as biodegradable
M. Saeki, et al., Enhanced bone regeneration via multimodal orthopedic implants, Appl. Surf. Sci. 313 (2014) 60–66.
actions of synthetic peptide SVVYGLR on osteoprogenitors [106] A. Tahmasbi Rad, N. Ali, H.S.R. Kotturi, M. Yazdimamaghani,
and osteoclasts, Biomaterials 30 (27) (2009) 4676–4686. J. Smay, D. Vashaee, et al., Conducting scaffolds for liver tissue
[91] H. Egusa, M. Saeki, M. Doi, S. Fukuyasu, T. Matsumoto, Y. engineering, J. Biomed. Mater. Res. A 102 (11) (2014) 4169–
Kamisaki, et al., A smallmolecule approach to bone regenerative 4181.
medicine in dentistry, J. Oral Biosci. 52 (2) (2010) 107–118. [107] L. Casagrande, M.M. Cordeiro, S.A. Nör, J.E. Nör, Dental
[92] C.T. Laurencin, K.M. Ashe, N. Henry, H.M. Kan, K.W.-H. Lo, pulp stem cells in regenerative dentistry, Odontology 99 (1)
Delivery of small molecules for bone regenerative engineering: (2011) 1–7.
preclinical studies and potential clinical applications, Drug [108] M. Torabinejad, M. Turman, Revitalization of tooth with
Discov. Today 19 (6) (2014) 794–800. necrotic pulp and open apex by using platelet-rich plasma: a
[93] C.-Y.E. Han, Y. Wang, L. Yu, D. Powers, X. Xiong, V. Yu, et case report, J. Endod. 37 (2) (2011) 265–268.
al., Small molecules with potent osteogenic-inducing activity [109] T. Bezgin, A. Yılmaz, B. Celik, H. Sönmez, Concentrated
in osteoblast cells, Bioorg. Med. Chem. Lett. 19 (5) (2009) platelet-rich plasma used in root canal revascularization: 2 case
1442–1445. reports, Int. Endod. J. 47 (1) (2014) 41–49.
[94] K.W. Lo, K.M. Ashe, H.M. Kan, C.T. Laurencin, The role of [110] T. Bezgin, A.D. Yilmaz, B.N. Celik, M.E. Kolsuz, H. Sonmez,
small molecules in musculoskeletal regeneration, Regen. Med. Efficacy of platelet-rich plasma as a scaffold in regenerative
7 (4) (2012) 535–549. endodontic treatment, J. Endod. 41 (1) (2015) 36–44.
[95] T. Ito, M. Takemasa, K. Makino, M. Otsuka, Preparation [111] S. Heinemann, H. Ehrlich, C. Knieb, T. Hanke, Biomimetically
of calcium phosphate nanocapsules including simvastatin/ inspired hybrid materials based on silicified collagen, Int. J.
deoxycholic acid assembly, and their therapeutic effect in Mater. Res. 98 (7) (2007) 603–608.
osteoporosis model mice, J. Pharm. Pharmacol. 65 (4) (2013) [112] M.S. Aw, S. Simovic, J. Addai-Mensah, D. Losic,
494–502. Silicamicrocapsules fromdiatoms as new carrier for delivery of
[96] K. Gellynck, R. Shah, M. Parkar, A. Young, P. Buxton, P. Brett, therapeutics, Nanomedicine 6 (7) (2011) 1159–1173.
Small molecule stimulation enhances bone regeneration but not [113] J. Chou, J. Hao, H. Hatoyama, B. Ben-Nissan, B. Milthorpe,M.
titanium implant osseointegration, Bone 57 (2) (2013) 405–412. Otsuka, The therapeutic effect on bone mineral formation from
[97] Y. Qi, T. Zhao, W. Yan, K. Xu, Z. Shi, J. Wang, Mesenchymal biomimetic zinc containing tricalcium phosphate (ZnTCP) in
stem cell sheet transplantation combined with locally released zinc-deficient osteoporotic mice, PLoS One 8 (8) (2013).
simvastatin enhances bone formation in a rat tibia osteotomy [114] T. Nakahara, T. Nakamura, E. Kobayashi, K.-I. Kuremoto,
model, Cytotherapy 15 (1) (2013) 44–56. T. Matsuno, Y. Tabata, et al., In situ tissue engineering of
[98] B. Bostan, T. Güneş, M. Aşçı, C. Sen,M. Keleştemur,M. Erdem, periodontal tissues by seeding with periodontal ligament-
et al., Simvastatin improves spinal fusion in rats, Acta Orthop. derived cells, Tissue Eng. 10 (3–4) (2004) 537–544.
Traumatol. Turc. 45 (4) (2010) 270–275. [115] Y. Liu, Y. Zheng, G. Ding, D. Fang, C. Zhang, P.M. Bartold,
[99] M.Mozafari, D. Vashaee, L. Tayebi, M. Mehraien, et al., Periodontal ligament stem cell-mediated treatment for
Electroconductive Nanocomposite Scaffolds: A New Strategy periodontitis in miniature swine, Stem Cells 26 (4) (2008)
into Tissue Engineering and Regenerative Medicine, INTECH 1065–1073.

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 21

[116] M. Taba, Q. Jin, J. Sugai, W. Giannobile, Current concepts [134] N.E. Fedorovich, J. Alblas, W.E. Hennink, F.C. Öner, W.J.
in periodontal bioengineering, Orthod. Craniofacial Res. 8 (4) Dhert, Organ printing: the future of bone regeneration? Trends
(2005) 292–302. Biotechnol. 29 (12) (2011) 601–606.
[117] R.J. Gorlin, M.M. Cohen, R.C. Hennekam, Syndromes of the [135] N. Annabi, A. Tamayol, J.A. Uquillas, M. Akbari, L.E.
Head and Neck, Oxford University Press, New York, 1990. Bertassoni, C. Cha, et al., 25th anniversary article: rational
[118] K. Gupta, S. Singh, K.N. Garg, Gene therapy in dentistry: design and applications of hydrogels in regenerative medicine,
tool of genetic engineering. Revisited, Arch. Oral Biol. 60 (3) Adv. Mater. 26 (1) (2014) 85–124.
(2015) 439–446. [136] D.W. Hutmacher, M. Sittinger, M.V. Risbud, Scaffold-based
[119] T. Friedmann, R. Roblin, Gene therapy for human genetic tissue engineering: rationale for computer-aided design and
disease? Science 175(4025) (1972) 949–955. solid free-form fabrication systems, Trends Biotechnol. 22 (7)
(2004) 354–362.
[120] A. Bank, Human somatic cell gene therapy, BioEssays 18 (12)
(1996) 999–1007. [137] S. Michna,W. Wu, J.A. Lewis, Concentrated hydroxyapatite
inks for direct-write assembly of 3-D periodic scaffolds,
[121] Q.L. Matthews, D.T. Curiel, Gene therapy: human germline
Biomaterials 26 (28) (2005) 5632–5639.
genetic modifications–assessing the scientific, socioethical, and
religious issues, Gene Ther. 100 (1)(2007). [138] S.E. Bakarich, R. Gorkin III,M. in het Panhuis, G.M.
Spinks, Three-dimensional printing fiber reinforced hydrogel
[122] J.A.Wolff, R.W. Malone, P.Williams,W. Chong, G. Acsadi,
composites, ACS Appl.Mater. Interfaces 6 (18) (2014) 15998–
A. Jani, et al., Direct gene transfer into mouse muscle in vivo,
Science 247 (4949) (1990) 1465–1468.
[139] J.C. Reichert, A. Cipitria, D.R. Epari, S. Saifzadeh, P.
[123] E. Pierce, Q. Liu, O. Igoucheva, R. Omarrudin, H. Ma, S.
Krishnakanth, A. Berner, et al., A tissue engineering solution
Diamond, et al., Oligonucleotide-directed single-base DNA
for segmental defect regeneration in load-bearing long bones,
alterations in mouse embryonic stem cells, Gene Ther. 10 (1)
Sci. Transl. Med. 4 (141) (2012) 141ra93.
(2003) 24–33.
[140] P.F. Costa, C. Vaquette, Q. Zhang, R.L. Reis, S. Ivanovski,
[124] N.J. Caplen, E.W. Alton, P.G. Mddleton, J.R. Dorin, B.J.
D.W. Hutmacher, Advanced tissue engineering scaffold design
Stevenson, X. Gao, et al., Liposome-mediated CFTR gene
for regeneration of the complex hierarchical periodontal
transfer to the nasal epithelium of patients with cystic fibrosis,
structure, J. Clin. Periodontol. 41 (3) (2014) 283–294.
Nat. Med. 1 (1) (1995) 39–46.
[141] B.T. Goh, L.Y. Teh, D.B.P. Tan, Z. Zhang, S.H. Teoh, Novel
[125] S.P. Chaplot, I.D. Rupenthal, Dendrimers for gene delivery–a
3D polycaprolactone scaffold for ridge preservation–a pilot
potential approach for ocular therapy? J. Pharm. Pharmacol. 66
randomised controlled clinical trial, Clin. Oral Implants Res.
(4) (2014) 542–556.
26 (3) (2015) 271–277.
[126] S. Huang, M. Kamihira, Development of hybrid viral vectors
[142] R. Schek, J. Taboas, S.J. Hollister, P. Krebsbach, Tissue
for gene therapy, Biotechnol. Adv. 31 (2) (2013) 208–223.
engineering osteochondral implants for temporomandibular
[127] O. Anusaksathien, W.V. Giannobile, Growth factor delivery to joint repair, Orthod. Craniofacial Res. 8 (4) (2005) 313–319.
re-engineer periodontal tissues, Curr. Pharm. Biotechnol. 3 (2)
[143] K. Kim, C. Lee, B. Kim, J. Mao, Anatomically shaped tooth
(2002) 129–139.
and periodontal regeneration by cell homing, J. Dent. Res. 89
[128] A. Voutetakis, I. Bossis, M.R. Kok, W. Zhang, J. Wang, A.P. (8) (2010) 842–847.
Cotrim, et al., Salivary glands as a potential gene transfer target
[144] E. Ikeda, R. Morita, K. Nakao, K. Ishida, T. Nakamura, T.
for gene therapeutics of some monogenetic endocrine disorders,
Takano-Yamamoto, et al., Fully functional bioengineered tooth
J. Endocrinol. 185 (3) (2005) 363–372.
replacement as an organ replacement therapy, Proc. Natl. Acad.
[129] D. Sheyn, O. Mizrahi, S. Benjamin, Z. Gazit, G. Pelled, D. Sci. 106 (32) (2009) 13475–13480.
Gazit, Genetically modified cells in regenerative medicine
[145] W. Zhang, I.P. Ahluwalia, P.C. Yelick, Three dimensional
and tissue engineering, Adv. Drug Deliv. Rev. 62 (7) (2010)
dental epithelialmesenchymal constructs of predetermined size
and shape for tooth regeneration, Biomaterials 31 (31) (2010)
[130] Y. Chen, P.K. Chen, L. Jeng, C. Huang, L. Yang, H. Chung, 7995–8003.
et al., Periodontal regeneration using ex vivo autologous stem
[146] G.M.Whitesides, E. Ostuni, S. Takayama, X. Jiang, D.E.
cells engineered to express the BMP-2 gene: an alternative to
Ingber, Soft lithography in biology and biochemistry, Annu.
alveolaplasty, Gene Ther. 15 (22) (2008) 1469–1477.
Rev. Biomed. Eng. 3 (1) (2001) 335–373.
[131] T. Yokoi, M. Saito, T. Kiyono, S. Iseki, K. Kosaka, E. Nishida,
[147] A. Rosenthal, A. Macdonald, J. Voldman, Cell patterning chip
et al., Establishment of immortalized dental follicle cells for
for controlling the stem cell microenvironment, Biomaterials
generating periodontal ligament in vivo, Cell Tissue Res. 327
28 (21) (2007) 3208–3216.
(2) (2007) 301–311.
[148] C.J. Flaim, D. Teng, S. Chien, S.N. Bhatia, Combinatorial
[132] S.V. Murphy, A. Atala, 3D bioprinting of tissues and organs,
signaling microenvironments for studying stem cell fate, Stem
Nat. Biotechnol. 32 (8) (2014) 773–785.
Cells Dev. 17 (1) (2008) 29–40.
[133] F. Obregon, C. Vaquette, S. Ivanovski, D. Hutmacher, L.
[149] A.B. Ennett, D. Kaigler, D.J. Mooney, Temporally regulated
Bertassoni, Threedimensional bioprinting for regenerative
delivery of VEGF in vitro and in vivo, J. Biomed. Mater. Res.
dentistry and craniofacial tissue engineering, J. Dent. Res. 94
A 79 (1) (2006) 176–184.
(9) (2015) 143S–152S.

22 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

[150] R.S. Kane, S. Takayama, E. Ostuni, D.E. Ingber,
G.M.Whitesides, Patterning proteins and cells using soft Dr. Jyotsana Sikri
lithography, Biomaterials 20 (23) (1999) 2363–2376. M.D.S, Senior lecturer,
[151] D.I. Rozkiewicz, Y. Kraan, M.W. Werten, F.A. de Wolf, Department of Conservative Dentistry & Endodontics,
V. Subramaniam, B.J. Ravoo, et al., Covalent microcontact Sudha Rustagi College of Dental Sciences and Research,
printing of proteins for cell patterning, Chem. Eur. J. 12 (24) Faridabad, Haryana.
(2006) 6290–6297.
Dr. Arpit Sikri
[152] J.-Y. Zhang, B.A. Doll, E.J. Beckman, J.O. Hollinger, Three- M.D.S, Senior Resident,
dimensional biocompatible ascorbic acid-containing scaffold
Department of Prosthodontics, Maulana Azad Institute of
for bone tissue engineering, Tissue Eng. 9 (6) (2003) 1143–
Dental Sciences, New Delhi.
[153] P. Kim, H.E. Jeong, A. Khademhosseini, K.Y. Suh, Fabrication
of non-biofouling polyethylene glycol micro-and nanochannels
by ultraviolet-assisted irreversible sealing, Lab Chip 6 (11)
(2006) 1432–1437.
[154] Y. Ling, J. Rubin, Y. Deng, C. Huang, U. Demirci, J.M. Karp,
et al., A cell-laden microfluidic hydrogel, Lab Chip 7 (6) (2007)
[155] N.W. Choi, M. Cabodi, B. Held, J.P. Gleghorn, L.J. Bonassar,
A.D. Stroock, Microfluidic scaffolds for tissue engineering,
Nat. Mater. 6 (11) (2007) 908–915.
[156] A. Khademhosseini, R. Langer, J. Borenstein, J.P. Vacanti,
Microscale technologies for tissue engineering and biology,
Proc. Natl. Acad. Sci. U. S. A. 103 (8) (2006) 2480–2487.

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Dr. Nikenlemla, Dr. Naman Vaidya, Dr. Rucha Shinde, Dr. Swati Bali

“Biofilm can be defined as a sessile multicellular microbial structural unit microcolonies or cell clusters formed be
community characterized by cells that are firmly attached surface adherent bacterial cells. Microcolonies comprises a
to a surface and enmeshed in a self-produced matrix discrete unit of densely packed bacterial cells aggregates.
of extracellular polymeric substance (EPS), usually a
Three factors essential for biofilm are:
1. Microorganisms
The term biofilm was introduced to designate the thin layered 2. Solid substrate
condensations of microbes (e.g. bacteria, fungi, protozoa) 3. Fluid channels.
that may occur on various surface structures in nature. Free- Composition of biofilm
floating bacteria existing in an aqueous environment, so-called
planktonic microorganisms, are a prerequisite for biofilm Biofilm consists of matrix material 85% volume and 15%
formation. In dental contexts, a well-known and extensively cells. A fresh biofilm is made up of biopolymers such
studied biofilm structure is established during the attachment as polysaccharides, proteins, nucleic acids, and salts. A
of bacteria to teeth to form dental plaque. Here, bacteria free glycocalyx matrix is made up of EPS, which surrounds the
in saliva (planktonic organisms) serve as the primary source microcolonies and anchors the bacterial cells to the substrate.
for the organization of this specific biofilm.[2] Tower or mushroom shaped structure is typically characteristic
feature of a viable fully hydrated biofilm.
Development of biofilm is influenced by physiochemical
property of components involved in the biofilm. pH, There is an important aspect when we see the composition
temperature, surface energy of substrate, nutrient availability, of biofilm these are water-filled channels which act as
and length of the time the bacteria is in contact with the a primitive circulatory system in a biofilm, intersect the
surface and bacterial cell surface charge may play a key role structure of biofilm to establish connections between the
in biofilms. Studies have shown that biofilm is made up of microbial colonies. Biofilm community comprises efficient
single cells and microcolonies, all embedded in a highly exchange between bacterial cells and fluid. Detachment has
hydrated, predominantly anionic exopolymer matrix. [3] been understood to play an important role in shaping the
morphological characteristics. It is also an “active dispersal
Biofilm formation is a step-wise procedure its formation mechanism” or “seeding dispersal” where a detached cell
occurs in the presence of microorganisms, fluid and solid forms resistance traits which is the source of persistent
surface. Biofilm is considered as community as it possesses infections.[3]
following criteria: Autopoiesis; Haemostasis; Synergy;
Communality[4] Oral diseases as consequences of ecological changes in
Stages in the Development of Biofilm While seemingly a contradictory quality, dental biofilms are
1. First step is the interaction and adsorption of inorganic essential for maintenance of both oral health and oral disease
and organic molecules to solid surface creating the conditions. Indeed, caries, gingivitis and chronic periodontitis
conditioning layer are caused by commensal microorganisms and not by
2. Once the conditioning layer is formed; finally, it classical microbial pathogens. Currently their development
undergoes adhesion of microbial cells to this layer. is considered to be a consequence of ecologically driven
In addition, the microbial adherence to substrate is also imbalances in dental microbial biofilms . In the case of caries
mediated by bacterial surface structure such as fimbriae, pili, for example, a low pH environment caused by microbial
flagella, and extra polymeric substances (EPS). The bacterial fermentation of carbohydrates selects populations of acid-
cell surface structures form bridges between the bacteria and tolerant and acid-producing strains that in turn increase acid
conditioning layer. Initially, the bond between the bacteria formation and may result in demineralization of the tooth
and the substrate may not be strong. However with time, structure. As far as endodontic infections are concerned the
these bonds gains in strength, making the bacteria substrate flare-up lesion could have a similar mechanism. Hence, acute
attachment irreversible.[3] exacerbations of endodontic lesions may be explained by a
shift in the flow of nutrients to the root canal space, giving rise
Ultrastructure of Biofilm to ecological changes, which promote growth of proteolytic
A fully developed biofilm is described as a heterogeneous bacteria.
arrangement of microbial cells on a solid surface. Basic Example, following the initial instrumentation of a primary

24 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

infected root canal, injury of the periapical tissue by over- the latter case, the inflammatory lesion front may recede
instrumentation may release entry of inflammatory exudates successively towards the apex, possibly in bursts, and provide
into the root canal and cause growth of proteolytic bacteria the fluid vehicle by which invading planktonic organisms can
that may have survived the endodontic treatment procedure. multiply and start attaching to the root canal walls.
The necrotic pulp tissue becomes a favorable environment Quorum sensing, a bacterial cell-to-cell communication
for microbial proliferation due to the presence of organic mechanism for controlling cellular functions, is of particular
residue or nutrients, which act as substrate or culture medium. interest because of the presence of dense aggregates of bacteria
Gram-negative bacteria are more frequent than Grampositive in biofilms. The signaling is mediated by diffusible molecules
bacteria. Facultative or strict anaerobic microorganisms are which, when present in sufficient concentrations, serve to
more frequent than aerobic microorganisms, and the presence modify gene expression in neighboring microorganisms.
of bacilli and filaments is equivalent to that of cocci.[2] Quorumsensingsignaling is known to be involved in the
regulation of several microbial properties, including virulence
BIOFILMS IN ENDODONTIC INFECTIONS and the ability to form biofilms, incorporate extracellular
Biofilm formation in root canals is probably initiated at some DNA and cope with environmental stress. The known peptide-
time after the first invasion of the pulp chamber by planktonic signal molecules produced by oral streptococci are primarily
oral organisms after some tissue breakdown.[4] used for intra-species communication. However, other
signals of unknown nature and actual function released by
An ischemic injury by trauma, leading to pulpal necrosis,
oral microorganisms serve as inter-species communication.
is likely to provide totally different prerequisites for the
Because many of the oral bacteria found in root canals, e.g.
colonization phase than in a caries exposure of the pulp. In

Table 1: Bacterial species/phylotypes commonly detected in endodontic infections

Species Gram staining Phylum Type of infection
Bacteroidetes oral clone X083 Gram negative Bacteroidetes Primary
Campylobacter rectus Gram negative Proteobacteria Primary
Dialister invisus Gram negative Firmicutes Primary; persistent/secondary
Fusobacterium Nucleatum Gram negative Fusobacteria Primary; persistent/secondary; extraradicular
Porphyromonas Endodontalis Gram negative Bacteroidetes Primary; extraradicular
Porphyromonas Gingivalis Gram negative Bacteroidetes Primary; extraradicular
Prevotella intermedia Gram negative Bacteroidetes Primary; extraradicular
Prevotella nigrescens Gram negative Bacteroidetes Primary
Prevotella baroniae Gram negative Bacteroidetes Primary
Pyramidobacter Piscolens Gram negative Synergistetes Primary; persistent/secondary
Tannerella forsythia Gram negative Bacteroidetes Primary
Treponema denticola Gram negative Spirochaetes Primary
Treponema socranskii Gram negative Spirochaetes Primary
Treponema maltophilum Gram negative Spirochaetes Primary
Veillonella parvula Gram negative Firmicutes Primary
Actinomyces israelii Gram positive Actinobacteria Primary; persistent/secondary; extraradicular
Enterococcus faecalis Gram positive Firmicutes Persistent/secondary
Filifactor alocis Gram positive Firmicutes Primary; persistent/secondary
Olsenella uli Gram positive Actinobacteria Primary; persistent/secondary
Parvimonas micra Gram positive Firmicutes Primary; persistent/secondary
Propionibacterium acnes Gram positive Actinobacteria Primary; persistent/secondary
Propionibacterium Propionicum Gram positive Actinobacteria Primary; persistent/secondary; extraradicular
Pseudoramibacter alactolyticus Gram positive Firmicutes Primary; persistent/secondary
Streptococcus anginosus Gram positive Firmicutes Primary; persistent/secondary
Streptococcus constellatus Gram positive Firmicutes Primary; persistent/secondary
Streptococcus intermedius Gram positive Firmicutes Primary; persistent/secondary
Streptococcus mitis Gram positive Firmicutes Primary; persistent/secondary

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 25

Fig 1: Stages of biofilm Formation

Fig 3: Variable thickness of biofilm adherent to the root canal Fig 2: Scanning electron microscope showing biofilm (a) on
predentine with moribound pulp tissue towards the canal the root canal wall (b) within dentinal tubules

Fig 4: Microorganisms observed in biofilms on root canals walls of teeth with primary endodontic infections. Cocci (a), spirochetes (b),
filaments (c), and bacilli (d) could be observed, although a direct correlation between bacterial morphological differences and clinical/radio-
graphic findings was not established by means of SEM evaluation

26 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Fig 5:. Biofilm within the root canal. (a) Radiograph showing maxillary left first premolar. Crown delivered 3 years previously.
Patient presented with intermittent pain, particularly upon chewing, and absence of deep probing. Diagnosis: necrotic pulp and
symptomatic apical periodontitis. Patient elected extraction. (b) SEM image of middle section of the root canal wall showing
necrotic pulp tissue and debris covering dentinal tubules; (c) Close-up of circled area in (b); (d) Close-up of circled area in (c)
showing dentinal collagen fibers and bacteria inside the dentinal tubule; (e) Close-up of circled area in (d) showing bacteria
inside dentinal tubule.

Fig 6:. Biofilm within the root canal. (a) Radiograph showing mandibular left second molar. Patient presented with no symptoms
and absence of deep probing. Diagnosis: necrotic pulp and asymptomatic apical periodontitis. Patient elected extraction. (b)
SEM image of middle section of the root canal wall showing dentinal wall as a niche for biofilms; (c and d) Close-up images of
left circled area labeled in (b) showing dentinal collagen fibers and bacteria inside dentinal tubules; (e and f) Close-up images
of right circled area in (b) showing multi-species biofilms with cocci, rods, and long filaments

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 27

Fig 7:External surface of the root apex of tooth with pulp necrosis and chronic periapical lesion. (a) Morphological changes in
the radicular cementum close to the apical foramen showing areas of intact cementum (arrowhead) between areas of resorption
(arrow) (_400). (b) Higher magnification of cementum resorption in (a) with areas of microorganisms (arrow) (_1650); (c)
Higher magnification of (b) (arrow), showing the presence of cocci forming apical biofilm (_6000)

Fig. 8 : Bacteria forming dense biofilms on the root canal walls of a lower canine with extensive caries and a periapical lesion.
Biofilm, mainly comprised of cocci and rods (*), can be observed at cervical (a), middle (b), and apical (c, d) thirds of the canal.
In some areas, bacteria could also be detected into dentinal tubules (~). (DC refers to defense cells.)

28 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Fig 9 : Biofilm on extruded gutta-percha cone recovered from previously treated tooth with refractory periapical periodontitis.
(a) Radiograph of tooth. Filling material (gutta-percha point) is observed outside the palatal root canal; (b) SEM image of gutta-
percha specimen after removal. The apically extruded material (arrows) is approximately 2 mm in length (original magnification
_50; bar¼500 um); (c) SEM image of extruded gutta-percha (original magnification _350; bar¼50 um); (d) High magnification
of arrowhead area in (c). Glycocalyx structure is present in the upper right area but not in lower left area. Filamentous or
spirochete-shaped bacteria are observed in lower left

Fig 10 : Schematic diagram showing different methods by which bactaria in a biofilm gain resistance against antimicrobials

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 29

Fig 11 : Comparison of ESEM (a) and SEM (b) images demonstrating differences in resolution. (a) Fusobacterium nucleatum
species (arrows) in mixed culture with Enterococcus faecalis; (b) Colonies of F. nucleatum and E. faecalis. Both species could
clearly be distinguished

Fig 12 : Schematic diagram showing different antibiofilm stratergies

30 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

S. gordonii, Streptococcus mitis, Porphyromonas gingivalis, III] Periapical microbial biofilms
Fusobacterium nucleatum and Prevotella intermedia possess These are isolated biofilms found in the periapical region of an
the ability to communicate through quorum sensing, it is likely endodontically infected tooth. The microbiota in the majority
that signal molecules operate as additional environmental of teeth associated with apical periodontitis is restricted to
factors that alter gene expression to optimize phenotypic root canal as most of the microbial species that infect the root
properties of biofilm bacteria in root canals.[2] canal are opportunistic pathogens that do not have the ability
to survive the host defense mechanisms in periapical tissues.
Types of Endodontic Infections
Rarely, microbial species or even strains within species may
Endodontic infections can be classified as intraradicular and possess strategies to survive and thus infect periapical tissues.
extraradicular according to the anatomic location of infection
in relation to the root canal. IV] Biomaterial centered infection (BCI)
It is caused when microorganisms adhere to an artificial
I] Intraradicular Infection biomaterial surface and form biofilms structures.
Primary intraradicular infections: Biomaterials play an important role, and hence presence
Microorganisms that initially invade and colonize the necrotic of biomaterials in close range to host immune systems can
pulp tissue cause primary intraradicular infection. Primary increase the susceptibility to biofilms. BCI is one of the major
infections are characterized by a mixed consortium composed complications associated with prosthesis and or implant
of 10 to 30 bacterial species and 10 to 108 bacterial cells per related infections. [3]
canal. The involved microbiota is conspicuously dominated
Mechanisms of antimicrobial resistance
by anaerobic bacteria, but some facultative or microaerophilic
species can also be commonly found in primary intraradicular There are several mechanisms for biofilm/s to resist
infections. antimicrobial agents. The polysaccharide matrix of biofilms
can retard diffusion of the antibiotics. In addition, extracellular
Secondary Intraradicular Infections: enzymes such as β‑lactamase may become trapped and
Microorganisms that were not present in the primary concentrated in the matrix, thereby inactivating β‑lactam
infection but that were introduced into the root canal system antibiotics. Furthermore, quorum sensing (communication
at some time after professional intervention cause secondary with one another) can influence the structure of the biofilm by
intraradicular infections. The entry can be during treatment, encouraging the growth of species beneficial to the biofilm.
between appointments, or even after root canal filling. Species It has been shown that subpopulations of bacteria in biofilm
involved can be oral or nonoral microorganisms, depending form a phenotypic state (altered gene expression) where
on the cause of infection. they are highly protected. Bacterial cells protect themselves
by being located within the interior part of a biofilm; hence,
Persistent Intraradicular Infection: medicaments will only act on the microorganisms in the
Microorganisms that can resist intracanal antimicrobial peripheral portion of the biofilm. Additionally, bacterial cells
procedures and endure periods of nutrient deprivation in a residing within a biofilm grow more slowly than planktonic
prepared canal cause persistent intraradicular infections. This cells, and as a result, antimicrobial agents act more slowly.
is also termed recurrent infection. Involved microorganisms Depletion of nutrients or accumulation of waste products can
are remnants of a primary or secondary infection. E faecalis result in bacteria entering a non‑growing state which protects
are predominant and is a persistent organism. It is commonly the bacteria from the antibiotics. Pajkos et al. revealed that
found in a high percentage of root canal as a single organism biofilm bacteria exist in a low metabolic state, with a slower
or as a major component of the flora. Persistent and growth rate and production of exopolysaccharides. Chemical
secondary infections are clinically indistinguishable and are changes to the environment in the biofilm are lack of oxygen
responsible for persistent exudation, persistent symptoms inhibits some antibiotics and accumulated acidic waste leads
interappointment exacerbation and failure of endodontic to a difference in pH which has an antagonizing effect on the
treatment characterized by persistent apical periodontitis. antibiotic.[5]


Microbial invasion of the inflamed periradicular tissue is Enterococcus faecalis are Gram positive cocci, facultative
invariably a sequel of interradicular infection. Acute alveolar anaerobes. They produce various infections in human and
abscess is an example of extraradicular extension or a sequel most commonly infect urinary tract, blood stream, biliary
to interradicular infection. Sometimes extraradicular infection tract, endocardium , burn wounds and also associated with
can be independent of intraradicular infections. For example, foreign body induced infections. They are associated with
apical actinomycosis caused by Actinomyces species and P. infections in root canal and also they are seen in chronic
propionicum is a pathological disease which can be treated periapical pathology, failed root canal cases.
only by periapical surgery.[7]

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 31

They have many survival and virulence factors: they physically removing the biofilm.
are capable of causing mono-infection, opportunistic
Chavez de Paz et al. found that 1% NaOCl affected the
microorganisms which can withstand nutritional deprivation,
membrane integrity of E. faecalis, Lactobacillus paracasei,
they utilize serum as nutritional source, binds to dentinal
Streptococcus anginosus, and Streptococcus gordonii, and
tubules, produces collagen –binding protein and serine
removed most biofilm cells. Exposure to EDTA (50 mmol/l)
protease alter host responses, suppresses the action of
affected the membrane integrity in all organisms, but failed to
lymphocytes, possesses lytic enzymes, cytolysin, aggregation
remove more than a few cells in the biofilms of E. faecalis,
substance ,phenomones and lipoteichoic acid,maintains pH
L. paracasei, and Str. anginosus. Chlorhexidine (2.5%) had
homeostasis due to proton pump mechanism and they resist
a mild effect on the membrane integrity of E. faecalis and
the activity of antimicrobial agents due to biofilm formation.
removed only 50% of its biofilm cells. The effects were
Among all the survival and virulence factors Enterococcus substratum‑dependent, and most organisms displayed
faecalis the unique property of this organisms is biofilm increased resistance to the antimicrobials on collagen‑coated
formation and the physiochemical properties of organisms surfaces. Furthermore, Triphala, green tea polyphenols (GTP)
help them to modify according to the prevailing environmental and MTAD showed statistically significant antibacterial
and nutrient conditions. Biofilms formed by Enterococcus activity.
faecalis are able to resist destruction by enabling the bacteria to Table 2: Relative efficacy of root canal irrigant agents in their
become 1000 times more resistant to phagocytosis, antibodies different properties
and antimicrobials than nonbiofilm producing bacteria.
Action/ Sodium Iodine Chlorhex- EDTA
Enterococcus faecalis forms biofilm in various stages which agent hypochlo- idine
includes adherence of the organisms and formation of rite
microcolonies followed by bacterial mediated dissolution of
Bacterial +++++ +++++ +++ +
the mineral fraction from the dentin which results in release
of calcium and phosphate ions leading to initial calcification.
Dissolution +++++ + – –
At later stages E. Faecalis biofilm shows carbonated –apatite
Biofilm +++++ ++++ ++ ++
structure as compared to natural dentine which contains
carbonated flor-apatite structure which are more resistant
and difficult to eradicate.Enterococcus faecalis can inactivate Biofilm +++++ – – ++++
antimicrobial agents such as metronidazole and also can break-up
protects organisms like B. Fragilis found in multispecies +relative degree of potency, – no potency
biofilm producing synergistic effect. Enterococcus faecalis ERADICATION OF BIOFILM:
coaggregates with F.nucleatum which results in further The structural dense organization of the biofilm within the
aggravate endodontic infection. polymeric matrix restricts the penetration of any agent
A recent study conducted showed that Enterococcus faecalis, into them. Biofilm are found to be resistant to amoxicillin,
S. sanguinis,S. intermedius, S.pyogenes and S. aureus were doxycycline and metronidazole. Sodium hypochlorite is an
capable of producing biofilm on gutta percha. Enterococcus effective irrigant to destroy all forms of Enterococcus faecalis
faecalis, S. sanguinis biofilm were thicker than other biofilms including its biofilm form. Chlorhexidine 2% gel or liquid
produced by other organisms[6] form is effective to eliminate Enterococcus faecalis from the
superficial layers of dentinal tubules up to 100micrometer.
Effects of endodontic irrigants and medicaments on
The new techniques include use of ultrasonic irrigation,
ozone, plasma dental probe, photoactivated disinfection with
Spratt et al. evaluated the effectiveness of NaOCl (2.25%), low-energy laser for biofilm removal.
0.2% chlorhexidine gluconate (CHX), 10% povidone
iodine, 5 ppm colloidal silver, and phosphate‑buffered saline Ultrasonic Irrigation
(PBS) solution (as control) against monoculture biofilms 1-minute use of ultrasonically activated irrigation, followed
of five root canal isolates including Prevotella intermedia, by root canal cleaning and shaping has been shown to
Peptostreptococcus micros, Streptococcus intermedius, improve canal and isthmus cleanliness in terms of necrotic
Fusobacterium nucleatum, and E. faecalis. Results showed debris/biofilm removal.
that NaOCl was the most effective antimicrobial, followed by
the iodine solution. Clegg et al. evaluated the effectiveness Ozone
of three concentrations of NaOCl (6%, 3%, and 1%), 2% High concentrated gaseous and aqueous ozone is strain,
CHX, and BioPure MTAD on apical dentin biofilms in dose and time dependently effective against the tested
vitro. Their findings indicated that 6% NaOCl was the only microorganisms in suspension and biofilm test model.
irrigant capable of both rendering the bacteria nonviable and However, NaOCl was the only method that eliminated biofilm.

32 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Plasma Dental Probe properties. Previous authors attributed the antibacterial
Plasma dental probe is effective for tooth disinfection. mechanism of BAG to its high pH, osmotic effects and Ca/P
Scanning electron microscopy shows complete destruction precipitation. Another author demonstrated that compared
of endodontic biofilms for a depth of 1 mm inside a root CH, BAG showed significantly less antibacterial effects as an
canal after plasma treatment for 5 min. Plasma emission intracanal medicament. In addition, another author showed
spectroscopy identifies atomic oxygen as one of the likely that BAG did not effectively prevent recontamination of
active agents for the bactericidal effect. instrumented root canals.[4]

The Er:YAG laser have produced excellent results due to its Current understanding emphasizes that endodontic disease
capacity for ablating hard tissue with very less thermal effects. is a biofilm-mediated infection and that elimination of
They are considered to be effective tool for the removal of bacterial biofilm from the root canal system and the exterior
apical biofilm. root surface may be necessary to maximize the prospects
for a favorable treatment outcome in the management of
Photoactivated disinfection this disease. Unfortunately, the root canal environment is a
Photodynamic therapy/ Light Activated Therapy is the latest challenging locale to accomplish this goal. It has been for
method used to destruct endodontic biofilm. It involves the this reason that different protocols ranging from antimicrobial
killing of microorganisms when a photo sensitizer selectively root canal irrigation to advanced methods that incorporate
accumulated in the target is activated by a visible light of lasers, photoactivated disinfection, and nanoparticles have
appropriate wavelength. been tried. While several of the advanced antimicrobial
protocols have shown a significant inhibitory effect against
PAD is a unique combination of a photosensitizer solution several types of microbial biofilms under in vitro conditions,
and low-power laser light. The photosensitizer, which is more in vivo studies are required to evaluate whether they
mostly colored, adheres to or gets absorbed by microbial are clinically practical and effective.. In bringing this new
cells. The low-power laser will destruct the target area and technology to the marketplace, it is important to combine
inactivate the microbial invaders. The photosensitizer then potent antibiofilm strategies with an effective and easy-to-use
binds to microbial cell walls or even enters the cells. Further, delivery system, if it is to enjoy broad acceptance.
the Laser light activates the photosensitizer and creates a
cascade of energy transfer and variable chemical reactions in References:
which singlet oxygen and free radicals play an important role. Cohen’s Pathways of the Pulp 11th edition.
The other advantage is the time taken. Usually sodium Gunnel Svensa¨Ter & Gunnar Bergenholtz. Biofilms in endodontic
hypochlorite, antibiotics and other methods against microbial infections.Endodontic Topics 2004, 9, 27– 36
threats need a lot of time to inactivate the microbes. Sindhu Haldal, K P Muhammed Yazar Arafath, K Subair, Kiran
PAD needs a maximum of 150 seconds. PAD is effective Joseph, Rajesh. Biofilms in Endodontics. Journal of
against Enterococcus faecalis, Streptococcus intermedius, International Oral Health 2016; 8(7):827-829
Fusobacterium nucleotum, Peptostreptococcus micros, Shruti Sharma, Varun Rajkumar, Sipra Sarin, Sahil Sarin, Charanpreet
Prevotella intermedia)[6] Singh Chugh, Harmeet Kaur. Root canal biofilms: review. Hecs
int j com health and med res 2017;3(3):93-95
Nanoparticles Magar S, Palekar A , Magar S. , Mosby S. Endodontic Biofilm: A
Nanoparticles are microscopic particles with one or more Review. NJDSR Number 2, Volume 1, January 2014
particle dimensions in the range of 1–100 nm. Nanoparticles Usha H.L, Anjali Kaiwar, Deepak Mehta. Biofilm In Endodontics:
are recognized to have properties that are very unique compared New Understanding To An Old Problem. IJCD • December,
to their bulk or powder counterparts. In root canal therapy, 2010 • 1(3)
nanoparticles may be applied as slurry or in combination Luis E. Chavez de Paz ,Christine M. Sedgley ,Anil Kishen. The Root
with sealers. They have the ability to diffuse antimicrobial Canal Biofilm.
components deep in dentin tissue. The successful application Gary B. Carr, DDS, Richard S. Schwartz, DDS, Christoph Schaudinn,
of nanoparticles in endodontics will depend on both the PhD,Amita Gorur, MSc,and J. William Costerton, PhD.
effectiveness of antimicrobial nanoparticles and the delivery Ultrastructural Examination of Failed Molar Retreatment with
method used to disperse these particles into the anatomical Secondary Apical Periodontitis: An Examination of Endodontic
complexities of the root canal system. Biofilms in an Endodontic Retreatment Failure. American
Association of Endodontists. doi:10.1016/j.joen.2009.05.035
Bioactive glass Francisco Correa Toral1, Leylin Delgado Hernández1, Carolina
Echavarría González1, Fátima Serna Varona1, Adriana
Bioactive glass (BAG) consists of SiO2, Na2O, CaO2, and
Rodríguez Ciodaro1, Hugo Díez Ortega2, .Ex vivo model for
P2O5 at different concentrations. It has received considerable
studying polymicrobial biofilm formation in root canals. Univ.
interest in root canal disinfection due to antibacterial

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 33

Sci. 22 (1): 31-43, 2017.
L Lakshmi Narayanan, Vaishnavi C. Endodontic microbiology. Dr. Nikenlemla
Journal of Conservative Dentistry.Oct-Dec 2010. Vol 13.Issue 4 (Asst. Professor)
Pankaj Yadav 1, Sarika Chaudhary 2, Rajendra K. Saxena 3, Sangeeta Department of Conservative Dentistry and Endodontics,
Talwar 4, Sudha Yadav 5.Evaluation of antimicrobial and Darshan Dental College, Loyara, Udaipur, Rajasthan
antifungal efficacy of chitosan as endodontic irrigant against
Dr. Naman Vaidya
enterococcus faecalis and candida albicans biofilm formed on
tooth substrate J Clin Exp Dent. 9(3):e361-7. 2017.
(III yr. Post Graduate Student)
Department of Conservative Dentistry and Endodontics,
Kim Lewis. Riddle of Biofilm Resistance. Antimicrobial agents and
Darshan Dental College, Loyara, Udaipur, Rajasthan
chemotherapy. 45( 4) . 2001,
Vytaute Peciuliene, Rasmute Maneliene, Estera Balcikonyte, Saulius Dr. Rucha Shinde
Drukteinis, Vygandas Rutkunas. Microorganisms in root canal (II yr. Post Graduate Student)
infections: a review. Baltic Dental and Maxillofacial Journal, Department of Conservative Dentistry and Endodontics,
10:4-9, 2008 Darshan Dental College, Loyara, Udaipur, Rajasthan
Zahed Mohammadi, Flavio Palazzi, Luciano Giardino, Sousan
Shalavi. Microbial Biofilms in Endodontic Infections: An Dr. Swati Bali
Update Review. Biomed J 2013;36:59-70) (II yr. Post Graduate Student)
Santos AL, Siqueira JF Jr, Roˆ c¸as IN, Jesus EC, Rosado AS, et al. Department of Conservative Dentistry and Endodontics,
Comparing the Bacterial Diversity of Acute and Chronic Dental Darshan Dental College, Loyara, Udaipur, Rajasthan
Root Canal Infections. PLoS ONE 6(11): (2011)
Kapil Jhajharia,  Abhishek Parolia,  K Vikram Shetty,  and  Lata
Kiran Mehta. Biofilm in endodontics: A review. J Int Soc Prev
Community Dent. 2015 Jan-Feb; 5(1): 1–12
Goran Sundqvist & David Figdor. Life as an endodontic pathogen
Ecological differences between the untreated and root-filled
root canals. Endodontic Topics 2003, 6, 3–28
H. J. Rolph, A. Lennon, M. P. Riggio, W. P. Saunders, D.
Mackenzie, L. Coldero, And J. Bagg.molecular identification
of microorganisms from endodontic infections. Journal Of
Clinical Microbiology, Sept. 2001, Vol. 39, No. 9
Shaikha Al-Samahi, Mohammad A. Al-Omari, Detection of bacteria
in endodontic samples and its association with defined clinical
signs and symptoms of endodontic infection. S J Oral Sci Vol
1 No 2, July 2014

34 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

Dr Ravneet Arora, Dr Saurabh Arora

The mandibular canines are not only exposed to less plaque, TABLE IX – MEASUREMENTS OF MANDIBULAR
calculus, abrasion from brushing, or heavy occlusal loading CANINE
than other teeth, they are also less severely affected by
Cervicoincisal length of crow 11mm
periodontal disease and so, usually are the last teeth to be
extracted with respect to age. These findings indicate that Length of root 16mm
mandibular canines can be considered as the key teeth for Mesiodistal diameter of crown 7mm
personal identification.115 Mesiodistal diameter of crown at cervix 5.5mm
Gran et al in 1967, Dekock in 1972 have found that male Labiolingual diameter of crown 7.5mm
canines were larger than the females and the inter canine arch Labiolingual diameter of crown at cervix 7mm
width was larger in males when compared to females and
Curvature of cervical line mesially 2.5mm
that those dimensions could be utilized for the purpose of sex
determination.116 Sherfuddin, Abdullaah and Khan in 1996 Curvature of cervical line distally 1mm
and Harris and Mohammed Q Al Rifaiy in 1997, Yadav et al in The dimensions of canine teeth have been studied by several
2002, Kaushal et al in 2003,Vandana M Reddy in 2008,Aliaa methods, including Fourier analysis, Moire topography and
Omar et al , Karen Boaz and Chavi Gupta, Ashith B. Acharya the measurement of linear dimensions such as mesiodistal
et al in 2009, Maneesha Sharma and R. K Gorea , Irfan width, buccolingual width, and inciso cervical height. The
Ahmed Mughal, Bindu Aggarwal et al in 2010, KS Nagesh et uses of Fourier analysis and Moire topography are limited to
al, Dhara Parekh et al, Rishabh Kapila et al, in 2011, Carlos small sample where as measurements of linear dimensions of
Sassi et al in 2012 conducted their studies and have found that canine teeth are used in large population.105
male canines were larger than the females and the inter canine
arch width was larger in males when compared to females and These methods can be applied directly on the Mandibular
that those dimensions could be utilized for the purpose of sex canine within the oral cavity, and also they can be best applied
determination on the cast models. Dimorphism in right and left mandibular
canines can be calculated using formula given by Garn &
Mandibular Canines Lens (1967) as follows.
It is the third tooth in the mandibular dental arch from the Sexual Dimorphism = Xm/Xf x 100
midline. It bears a close resemblance the maxillary canine.
Where, Xm = Mean value of males
The mesiodistal diameter of the crown is smaller than that of
its maxillary counter. Xf = Mean value for females

LABIAL ASPECT Further the mandibular canine index was calculated based on
the formula used by Rao et al
When the tooth is viewed from the labial aspect, the crown of
the Mandibular teeth resembles maxillary canines except for Mandibular Canine Index (MCI) = Mesio - distal crown width
the crown appearing longer and narrower mesio-distally. The of mandibular canine / Mandibular canine arch width or inter
mesial outline is more or less straight and is in line with the - canine distance
mesial outline of the root. The distal contact point is placed
Standard Mandibular Canine Index
more incisally than the maxillary canine.101
The standard Mandibular Canine Index of the population
PALATAL ASPECT studied was obtained from the measurements taken in the
The lingual surface of the mandibular canine crown is sample by applying the following formula Std. MCI = (Mean
relatively flat when compared to the maxillary canine. This male MCI SD) + (Mean female MCI +SD) / 2
is a result of the poorly developed cingulum and marginal According to Rao et al if the calculated Mandibular Canine
ridges. The lingual aspect of the root is relatively more is Index for the individual was higher than the Standard
relatively narrower than the maxillary canine.101 Mandibular Canine Index the individual was considered to
be male. If it was the other way round the subject was taken
as females.

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 35

Vandana M Reddy, Susmita Saxena, Puja Bansal. Mandibular canine Dr Ravneet Arora
index as a sex determinant: A study on the population of western Reader
Uttar Pradesh. Journal of Oral and Maxillo Facial Pathology Department of Oral Medicine and Radiology
Vananchal Dèntal college and Hospital
Shishir Yadav, Nagabhushana D, B. Balaji Rao, G. P. Mamatha.
Mandibular canine index in establishing sex identity. IJDR Garwa, Jharkhand
2002; 13(3&4):143-146. Dr Saurabh Arora
H. Sherfudhin, M.A. Abdullahm and N. Khan. A cross- sectional
study of canine dimorphism in establishing sex identity:
comparison of two statistical methods. Journal of Oral Department of Conservative and Endodontics
Rehabilitation 1996; 23:627-631. Vananchal Dèntal college and Hospital
Mohammed Q. Al-Rifaiy, M. Aleem Abdullah, Igbal Ashraf, Nazeer Garwa, Jharkhand
Khan. Dimorphism of mandibular and maxillary canine teeth
in establishing sex identity. The Saudi Dental Journal 1997; 9
Kaushal, S., Patnaik, V.V.G., Agnihotri, G. Mandibular Canines in
Sex Determination. J Anat. Soc. India 2003; 52(2):119-124.
S. Kaushal, V.V.G Patnaik, V. Sood, and G Agnihotri. Sex
determination in north Indians using Mandibular canine index.
JIAFM, 2004; 26(2):45-49.
Aliaa Omar and Sonia Azab. Applicability of Determination of
Gender from Odontometric Measurements of Canine Teeth in
a Sample of Adult Egyptian Population. Cairo Dental Journal
2009; 25(2):167:180.
Karen Boaz and Chhavi Gupta. Dimorphism in human maxillary
and madibular canines in establishment of gender. Journal of
Forensic Dental Sciences 2009; 1(1):42-44.
Ashith B. Acharya, and Sneedha Mainali. Limitations of the
mandibular canine index in sex assessment. Journal of Forensic
and Legal Medicine 2009; 16:67–69.
Maneesha Sharma and R. K. Gorea. Importance of Mandibular and
Maxillary Canines in Sex Determination. Journal of Punjab
Academy of Forensic Medicine & Toxicology 2010; 10:27-30.
P. C. Srivastava. Correlation of Odontometric Measures in Sex
Determination. J Indian Acad Forensic Med 2010 32(1):56-61.
Irfan Ahmed Mughal, Anwar Saood Saqib, Farida Manzur.
Mandibular Canine Index; its role in determining gender.
Professional Med J 2010; 17(3): 459-463.
Bindu Aggaewal, Subhash Kaushak, Kamlesh Vasudeva, Usha
Chhabre, Sanjay Singla. Significance of mandibular canine
index in sexual dimorphism. Journal Indo-Pacific Academy of
Forensic Odontology 2010; 1:1-4.
KS Nagesh, Asha R Iyengar, Rishabh Kapila, Sushma Mehkri.
Sexual Dimorphism in Human Mandibular Canine Teeth: A
Radiomorphometric Study. JIOMR 2011; 23(1):33-35.
Dhara Parekh, Ankur Zalawadia, Srushti Ruparelia, Shailesh Patel,
S. P. Rathod, S. V. Patel. Study of Mandibular Canine Teeth
Dimorphism in Establishing Sex Identity in Gujarat Region.
NJIRM 2011; Vol. 2(2):6-9.
Ashith B. Acharya, Punnya V. Angadi, Sudeendra Prabhu, Shweta
Nagnur. Validity of the mandibular canine index (MCI) in sex
prediction: Reassessment in an Indian sample. Forensic Science
International 2011; 204:207.e1–207.e4.

36 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

A Paradigm shift from micro to nano in dentistry
Dr. Atulana Roy and Dr. Arpit Sikri

The concept of “nanomaterials” formed in the early 1980’s, has captured a considerable amount of attention because of its
unique structures and properties. At present, the revolutionary development of nanotechnology has become the most highly
energized discipline in science and technology. The word ‘nano’ originates from the Greek word “dwarf”. The concept of
nanotechnology was first elaborated in 1959 by Richard Feynman . Nanometer is defined as ‘unit of length used to measure
the wavelength of light. It is equivalent to 1ˣ 10-9 m or 1 angstroms.’ As the size of the system decreases there is increase
in the ratio of surface area to volume and a number of physical phenomena becomes noticeably pronounced which tends
to alter the macroscopic properties of the material. Nanomaterials produced by using nanotechnology have revolutionized
dentistry by providing us with ‘nanofillers’.

HISTORICAL BACKGROUND OF cobalt-chromium alloy or cobalt-chromium-molybdenum

NANOTECHNOLOGY alloy and titanium alloy. The initial cobalt-based alloy is
Even long before the start of “nanoera”, people were coming cobalt –chromium binary alloy which was then developed
across various nanosized objects and the related nanolevel into cobalt chromium- tungsten alloy and later developed
processes and used them in practice. Thousands of years BC into cobalt chromium-molybdenum alloy. Its mechanical
people knew and used natural fabrics: flax, cotton, wool, properties and corrosion resistance are better than stainless
silk. They were able to cultivate them and process them into steel or gold alloy.
products. What makes these fabrics special is the fact that Another metal that is often used in dentistry are titanium
they have a developed network of pores with the size of 1-20 alloys because of its outstanding properties which are close
nanometers, i.e., they are typical nanoporous materials. Due to natural human bones, such as high specific strength, good
to their nanoporous structure, natural fabrics possess high biological security, high corrosion resistance, and elastic
utilities: they absorb sweat well, quickly swell and dry. modulus.
The word “nanotechnology” was introduced for the first time Modification of titanium implant surfaces into nanostructures
into a scientific world by N. Taniguchi at the international has been found to be able to improve their biological
conference on industrial production in Tokyo in 1974 so as to integration with surrounding soft tissues.
describe the super thin processing of materials with nanometer
accuracy and the creation of nano-sized mechanisms. Yao et al. created nanometer surface features on titanium
and Ti6Al4V implants by anodization, which was a quick
Ideas of nanotechnological strategy, which were put forward and relatively inexpensive electrochemical method.It
by Feynman, were developed by E. Drexler in his book demonstrated that anodization of Ti-based metals might create
“Vehicles of creation: the arrival of the nanotechnology era” nanometer surface features that could promote osteoblast
published in 1986. adhesion.
In 1991, the first nanotechnological program of National
2. Nanocomposites
Scientific Fund started to Operate in USA.
Addition of nanoparticles in composite resin can increase
In 2001, the National Nanotechnological Initiative (NNI) of strength and toughness of the composite resin. Due to
the USA was approved. In this way, according to the definition small particle size, composite resins with nanoparticles
of the National Nanotechnology Initiative, nanotechnology is significantly reduce the effect of polymerization shrinkage
the manipulation of different materials for different purposes and dramatically improve physical properties. In addition
at the nanoscale. The ability to arrange atoms as we desire composites containing nanofillers resulted in smooth surfaces
and subsequently to achieve effective, complete control of the with their ease of polish ability, increased abrasion resistance
structure of matter is possible through nanotechnology. and surface hardness.
NANOMATERIALS WITH THEIR CHARACTERISTIC Nanofilled composite resins provide a better finish on the
PROPERTIES surface. This characteristic could improve surface texture and
material biodegrading over time in this way, may be reduced.
1. Nanometals :
These composite resins have also achieved good mechanical
Currently, most metal stents of partial denture are applying properties, indicating possible use in the anterior and posterior

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 37

teeth. shows very high hardness,improved toughness over
conventional microcrystalline diamond, low friction and
3. Nanorobotic dentrifices good adhesion to titanium alloys.
Effective prevention has reduced caries in children, and a II. Nanostructured processing of HA coatings: This
caries vaccine may soon be available but a subocclusal- coating material is used to provide the desired mechanical
dwelling nanorobotic dentifrice delivered by mouthwash or characteristics and enhanced surface reactivity and has
toothpaste could patrol all supragingival and subgingival been shown to increase osteoblast adhesion, proliferation,
surfaces at least once a day, metabolizing trapped organic and mineralization.
matter into harmless and odorless vapors and performing
III. Nanostructured metalloceramic coatings: These
continuous calculus debridement.
coatings provide continuous variation from a
These invisibly small (1-10 μ) dentifrobots(FIGURE 1), nanocrystalline metallic bond at the interface to the hard
perhaps, numbering 103-105 nanodevices per oral cavity and ceramic bond on the surface.
crawling at 1-10 μ/s would be inexpensive purely mechanical Nanostructured ceramics, carbon fibers, polymers, metals,
devices that would safely deactivate themselves if swallowed and composites enhance osteoblast adhesion and calcium/
and would be programmed with strict occlusal avoidance phosphate mineral deposition. Studies have suggested that
protocols. nanophase ZnO and TiO2 may reduce Staphylococcus
epidermidis adhesion and increase the osteoblast functions
that are necessary to promote the efficacy of orthopedic

Although at present the impact of nanotechnology on dentistry
is somewhat limited due to the necessity to use materials
that are currently available, such as composite resin-based
materials, ongoing and future investigations will ensure that
developments that seem unbelievable today are possible
during the following years. The utilization of nanotechnology
in the future could facilitate improvements for the oral
health. Modified restorative materials, new diagnostic and
FIGURE1: Nanorobotic dentrifice acting on oral microflora therapeutic techniques, and pharmacologic approaches will
4. Nanotoothpaste improve overall dental care.
Metals have been used for centuries as antimicrobial agents Nanotechnology and its restless struggle for developing new
such as Ag, gold (Au) and metal oxides, such as zinc oxide better nanomaterials with applications in the biomedical
(ZnO) and titanium dioxide and amorphous calcium phosphate field has had an impact upon dentistry in terms of improving
nanoparticles which have gained significant interest over the the quality of materials and also in the conceptualizing
years due to their remarkable antimicrobial properties. of upgraded shaping technologies used for implant
manufacturing. Sequentially, oral health will probably be
Nano whitening toothpaste is a toothpaste that contains
maintained through an advanced level of biotechnology and
synthesized hydroxyapatite, a key component of tooth enamel,
nanorobot-assisted life-long processes. Nonetheless, despite
as nanosized crystals. It has been proven to freshen breathe
the advantages of nanotechnology, it does not come without
as well as whiten teeth. Ag nanoparticles are incorporated
risks when employed.
in these toothpastes which has a broad spectrum of activity
against bacteria, fungi, viruses, and even some protozoa. It remains yet to be established whether the nano patterning
Its antimicrobial action becomes even greater when in the or micron-scale patterning is the most convenient choice,
form of nanoparticles. Ag nanoparticles have been used to taking into account that nanopillars have been shown to
inactivate enzymes and prevent the replication of bacterial increase the hydrophobicity to an extent that might not favor
DNA and its action occurs because the nanoparticles adhere their use in implantology. Hence, further studies should be
to the outer membrane and so, promote changes in bacterial performed for establishing adequate coating composition
cell structures, modifying its permeability. and a correlation between the composite features and the
specifics of implantation area, in terms of thickness, surface
5. Nanoparticles on dental implant interface topography and cell response.
Recently three different nanostructured implant coatings have Straightforward directions should be established when the
been developed: scientific community reaches a high level of understanding of
I. Nanostructured diamond: This kind of coating the processes that happen and can be strictly controlled at the

38 ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018

nanoscale, when there is undoubted proof of its safe use for 11. Elias, KL, Price, RL & Webster, TJ 2002. ‘Enhanced functions
the benefits of human health. of osteoblasts on carbon nanofiber compacts.’ Biomaterials, 23,
Nanotechnology can be applied in almost all fields of human
12. Feynman RP. There’s plenty of room at the bottom. Eng Sci
activity. As Feynman, a Nobel-Prize-winning physicist Feb. 1960;23:22-36.Available at: “www.zyvex.com/nanotech/
explained briefly the important point of view with the feynman.html.
following words: “Concept of nanotechnology is an inevitable 13. Freitas Jr, RA 2000. ‘Nanodentistry.’ The Journal of the
development in the progress of science.” American Dental Association, 131(11), pp.1559-1565.
Particularly, nanotechnology has made progress in the fields 14. Gambhir, RS, Sogi, GM, Nirola, A, Brar, R, Sekhon, T. & Kakar,
of medicine and dentistry. The field of nanotechnology has H 2013. ‘Nanotechnology in dentistry: Current achievements
tremendous potential, which, if harnessed efficiently, can and prospects.’ Journal of Orofacial Sciences, 5(1), p.9.
bring out significant benefits such as improved health and 15. He, G, Dahl, T, Veis, A & George, A 2003. ‘Nucleation of
better use of natural resources. apatite crystals in vitro by self-assembled dentin matrix protein
1.’ Nature materials, 2(8), p.552.
Actually, it may be concluded that among these, the most
16. Jhaveri, HM & Balaji, PR, 2005. ‘Nanotechnology: The future
substantial contribution of nanotechnology to dentistry is the of dentistry.’ Journal of Indian Prosthodontic Society, 5(1).
enhancement in tooth restoration with nanocomposites.
17. Ji, B & Gao, H, 2004. ‘Mechanical properties of nanostructure
As final words, the combination of philosophies of science of biological materials.’ Journal of the Mechanics and Physics
and art will come together in this scene. In nanotechnology, of Solids, 52(9), pp.1963-1990.
someone will imagine it and then someone will do it. As an 18. Karimi, M, Ghasemi, A, Zangabad, PS, Rahighi, R, Basri,
example, Jules Gabriel Verne imagined the moon trip and SMM, Mirshekari, H, Amiri, M, Pishabad, ZS, Aslani, A,
Neil Armstrong and other scientists did it. Bozorgomid, M & Ghosh, D 2016. ‘Smart micro/nanoparticles
in stimulus-responsive drug/gene delivery systems.’ Chemical
REFERENCES Society Reviews, 45(5), pp.1457-1501.
1. Arora, R & Kapoor, H 2014. ‘Nanotechnology in dentistry-
Hope or hype.’ Oral Health Dent Manag, 13, pp.928-33.
2. Bhardwaj, A, Bhardwaj, A, Misuriya, A, Maroli, S, Manjula, S
& Singh, AK 2014. ‘Nanotechnology in dentistry: Present and
future.’ Journal of international oral health: JIOH, 6(1), p.121.
3. Bunker, BC, Rieke, PC, Tarasevich, BJ, Campbell, AA, Fryxell,
GE, Graff, GL, Song, L, Liu, J, Virden, JW & McVay, GL
1994.’Ceramic thin-film formation on functionalized interfaces
through biomimetic processing.’ Science, 264(5155), pp.48-55.
4. Campoccia, D, Montanaro, L & Arciola, CR 2013. ‘A review of
the biomaterials technologies for infection-resistant surfaces.’
Biomaterials, 34(34), pp.8533-8554.
5. De Jong, WH & Borm, PJ 2008. ‘Drug delivery and
nanoparticles: applications and hazards.’ International journal
of nanomedicine, 3(2), p.133.
6. Gao, H, Ji, B, Jäger, IL, Arzt, E, Fratzl, P 2003. ‘Materials
become insensitive to flaws at nanoscale: lessons from nature.’
Proceedings of the national Academy of Sciences, 100(10),
7. Gittens, RA, Scheideler, L, Rupp, F, Hyzy, SL, Geis-Gerstorfer,
J, Schwartz, Z & Boyan, BD 2014. ‘A review on the wettability
of dental implant surfaces II: biological and clinical aspects.’ Dr. Atulana Roy
Acta biomaterialia, 10(7), pp.2907-2918. PG Student
8. Gopinadh, A, Prakash, M, Lohitha, K, Kishore, KK, Chowdary, Department of Prosthodontics, Dasmesh Institute of
AS & Dev, JRR 2015. ‘The changing phase of prosthodontics: Research & Dental Sciences and Research, Faridkot,
Nanotechnology.’ Journal of Dental and Allied Sciences, 4(2), Punjab.
Dr. Arpit Sikri,
9. Grumezescu, AM 2016. ‘Nanobiomaterials in Drug Delivery:
Applications of Nanobiomaterials’. William Andrew.
M.D.S, Senior Resident,
Department of Prosthodontics, Maulana Azad Institute of
10. Eric, DK, 1986. ‘Engines of creation: the coming era of
Dental Sciences, New Delhi.
nanotechnology.’ Anchor Book.

ASIA PACIFIC DENTAL JOURNAL, Vol. 5, issue 3, October to December 2018 39

About the Journal 2. The title of the article, in full, (Concise and Informative);
It is the official Publication of Asia Pacific 3. Running title or short title not more than 50 characters;
Dental Federation (heretofore referred to as 4. The name by which each contributor is known with
APDF): Asia Pacific Regional Organization his or her highest academic degree(s) and institutional
of the FDI World Dental Federation. In each affiliation;
Issue, its readers have access to original Peer-
5. The name of the department(s) and institution(s) to which
reviewed Articles that examine all phases
the work should be attributed;
of Dental Health and Treatment, well Illustrated, including
Tables, Photos and statistical data. Coverage also includes 6. The name, address, phone numbers, facsimile numbers
successful diagnostic procedures, Imaging Techniques, and e-mail address of the contributor responsible for
Dental materials, Endodontics, Periodontics, Conservative correspondence about the manuscript;
and Operative dentistry, Paedodontics and Child Dental Care, 7. The total number of pages, total number of photographs
Oral Medicine and Radiology, Orthodontics, Prosthodontics and word counts separately for abstract and for the text
and Crown and bridge, Extraction and Impaction concerns, (excluding the references and abstract);
Maxillo-Facial Surgery, TMJ disorders, and Geriatric 8. Source(s) of support in the form grants, equipment,
Dentistry, and any other related Dental Subjects. drugs, or all of these;
Objectives 9. Acknowledgment, if any; and
Collect, Compile and edit, Articles for Publication, of 10. If the manuscript was presented as part at a meeting, the
Dental and related fields in the form of Original Articles, organization place and exact date on which it was read.
Case Reports, Literature Review, Viewpoints, Essays and
any other established Form in Print as well Online (Soft ABSTRACT PAGE:
Copy) format for the Education of the Medical and Dental The second page should carry the full title of the manuscript
personnel and Specialists including general Medical and and an abstract (of no more than 150 words for case reports,
Dental Practitioners and Paramedical & technical Staff of brief reports and 250 words for original articles). The abstract
hospital, clinic, or a diagnostic lab as well as for generating should be structured and state the Context, Aims, Methods and
and spreading Awareness of Dental Field in the Asia Pacific Materials, Statistical Analysis used, results and Conclusions.
Region and Worldwide. Below the abstract should provide 3 to10 keywords.
Dr. Bhagwant Singh
Gurudwara Shaheedan Road, Model Town, Ludhiana, INTRODUCTION:
Punjab-141002 Brief write the purpose of the article, summarize the rationale,
Phone: +91 981 424 5608, +91 161 501 5588 the study or observation.
Email: drbhagwantsingheditor@gmail.com ;
apdfdentaljournal@yahoo.com METHODS:
The methods section should include only information that
GUIDELINES FOR AUTHORS was available at the time the plan or protocol for the study
Manuscripts must be prepared in accordance with “Uniform was written; all information obtained during the conduct
requirements for Manuscripts submitted to Biomedical of the study belongs in the Result section. When reporting
Journal” developed by International Committee of Medical experiments on human subjects, state whether the procedures
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abstract and up to 10 references. Clearance has been obtained or not.
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Uniformly American or U K English (But not mixture Tables, and Illustrations, with the main and important findings
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TITLE PAGE: Summarize the key findings (Primary Outcome Measures,
1. Type of manuscript (e.g. Original article, Research, Secondary Outcome Measures, results as they relate to a prior
Systemized Review, Case report) hypothesis); Discuss the Strengths and limitations (design,
data Assimilation, analytic methods and the analysis);

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Interpretation and implications in the context of the totality We hereby transfer(s), assign(s), or otherwise convey(s) all
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Write References ONLY in Vancouver Style. References are electronic format.
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