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Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-

polycaprolactam) Blends: Potential Biodegradable Scaffold for Bone
Tissue Regeneration

Article  in  Journal of Biomaterials and Tissue Engineering · June 2011

DOI: 10.1166/jbt.2011.1004

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 Copyright © 2011 American Scientific Publishers Journal of
All rights reserved Biomaterials and Tissue Engineering
Printed in the United States of America Vol. 1, 30–39, 2011

Electrospun Polycaprolactone/Poly(1,4-butylene
adipate-co-polycaprolactam) Blends: Potential
Biodegradable Scaffold for Bone Tissue Regeneration
Rajkamal Balu1 , T. S. Sampath Kumar1
∗ , Murugan Ramalingam2 , and Seeram Ramakrishna3
1
Medical Materials Laboratory, Department of Metallurgical and Materials Engineering,
Indian Institute of Technology Madras, Chennai 600036, India
2
Faculty of Medicine, University of Strasbourg, Strasbourg 67085, France
3
Nanoscience and Nanotechnology Initiative, Division of Bioengineering,
National University of Singapore, Singapore 117576

Electrospun nanofibers have attracted much attention in recent years as scaffolds for
improved osteo-regeneration. In the present study, polycaprolactone/poly(1,4-butylene adipate-co-
polycaprolactam) blends were electrospun as potential biodegradable scaffolds for bone tissue engi-
neering. The morphology of the scaffold was observed to be bead-free nanofibers with average
diameter of about 400 nm by scanning electron microscopy. The semi crystalline nature, molec-
ular interactions among the polymers, lowering of thermal degradation temperature and improved
wetability of the blends compared to polycaprolactone were confirmed by wide angle X-ray diffrac-
tion analysis, Fourier transform infrared spectroscopy, simultaneous thermal analysis and contact
angle measurements. Enzymatic in vitro degradation study using lipase enzyme showed the electro-
spun blend fiber mat to be biodegradable with degradation rate higher than that of polycaprolactone.
Direct contact in vitro cytotoxicity test and MTT reduction calorimetric assay using mouse fibroblast
cells indicated the non-cytotoxic reactivity and cell viability of the scaffold. Human osteoblast cell
culture studies of the polymer blend fibers showed improved cellular response with good adhesion
and proliferation, demonstrating the viability of the electrospun blend mat for bone tissue engineer-
ing applications.
Keywords: Electrospinning, Bone Scaffold, Degradation, Biocompatibility, Osteoblast Cell
Culture.

1. INTRODUCTION the form of fibers that are collected over a grounded

plate. The large surface areas to volume ratio of elec-
Bone tissue engineering seeks to address the needs of trospun fibers, interconnected porous structure and good
viable substitutes that restore and maintain the function of mechanical properties have made them a potential can-
RESEARCH ARTICLE

human bones by applying the principles of biology and
engineering. Selecting biocompatible, functional, porous
scaffolds that mimic the biological environment plays a
key role in bone tissue engineering. Several techniques
including freeze drying, gas foaming, rapid prototyping,
electrospinning, ultra violet, laser irradiation etc. have been
used to fabricate porous scaffolds.1 Electrospinning (ES) is
a bottom-up technique widely used for producing ultra fine
fibers in order of few nanometers to micrometers.2 This
technique involves applying high voltage to the droplet
of polymeric solution, which causes the liquid to become
charged leading to electrostatic repulsion and ejection in
∗
Author to whom correspondence should be addressed.
didate for tissue engineering applications. The beneficial
features of a nanofibrous structure for bone tissue engi-
neering were first realized with polymers, which stimulate
cells into osteogenic pathway assisted via well-controlled
differentiation.3 The various electrospun nanofiber sys-
tems including polymers of biological origin and synthetic
polymers for bone tissue engineering are summarised in
Table I.4–15
Synthetic biodegradable polymers are of great interest
in the biomedical area particularly in the field of tissue
engineering and drug delivery. The greatest advantage of
these degradable polymers is that they are broken down
into biologically acceptable molecules that are metabo-
lized and removed from the body via normal metabolic

30 J. Biomater. Tissue Eng. 2011, Vol. 1, No. 1 2157-9083/2011/1/030/010 doi:10.1166/jbt.2011.1004

Balu et al. Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends

Table I. List of polymer nanofiber systems electrospun for bone tissue engineering.

Functional assay
Polymers Solvents Fiber diameter cell lines Remarks
4
Silk fibroin Formic acid 150 to 300 nm Mouse MC3T3-E1 cells showed
preosteoblast stellate shape and
(MC3T3-E1) broad cytoplasmic
extensions on the
membrane.
Chitosan5 Acetic acid 200 nm Human MG63 cells proliferated
osteosarcoma for 28 days on the
(MG63) chitosan nanofiber
membranes and
expressed ALP,
collagen, OCN, and
GAPDH at 2 weeks.
Type I Collagen6 HFIP 50 to 1000 nm Human bone MSCs showed osteogenic
marrow-derived gene expression of
mesenchymal osteocalcin,
stem cells osteonectin, and
(MSCs) ostepontin.
Polycaprolactone (PCL)7 Chloroform 20 nm to 5
m Human mes- Abundant extracellular
enchymal stem matrix was observed
cells (MSC) after 1 week.
Mineralization and
type I collagen was
observed at 4 weeks.
Poly(lactic-co-glycolic acid) Mixture of tetrahydrofuran 760 ± 210 nm Human hMSCs cells
(PLGA)8 (THF) and dimethyl mesenchymal differentiated into
formamide (DMF) stem derived chondrogenic cells and
osteoblast osteogenic cells after
(hMSC-Ob) 2 weeks.
Poly(3-hydroxybutyrate) Chloroform 3.7
m Human Increased alkaline
(PHB)9 osteoblasts phosphatase (ALP)
(SaOS-2) activity.
Poly(3-hydroxybutyrate-co- Chloroform 2.3
m SaOS-2 SaOS-2 cells maintained
3-hydroxyvalerate) phenotype.
(PHBV)9
PolyL-lactic acid) (PLLA)10 Trifluoroethanol (TFE) 450 ± 50 nm Osteoblast-like MG63 exhibited
MG63 cells shuttle-like shapes on
the parallel scaffolds
Silk/Poly (ethylene oxide) Water 700 ± 50 nm Human Extensive hMSC cell
(PEO)11 bone marrow- proliferation and
derived mes- matrix coverage after
enchymal stem 14 days.
cells (hMSCs)
PCL/gelatine12 TFE tens of nm to approx. Bone-marrow BMSC infiltration into
1
m stromal cell the scaffold up to
(BMSC) 114
m.
4.0
m
RESEARCH ARTICLE
PHB/PHBV9 Chloroform SaOS-2 The 1:1 blend showed
the highest ALP
activity.
PLA/gelatine13 TFE 190 to 390 nm MC3T3-E1 Increased
MC3T3-E1 cell growth
with introduction of
gelatin to the pure
PLA substrate
PCL/collagen14 Chloroform tens of nm–1000 nm pig bone marrow Matrix mineralization
mesenchymal was observed on
cells osteogenically induced
(pBMMCs). constructs.
Chitosan/PEO15 Acetic acid 30 to 74 nm MG-63 MG-63 cells maintained
cell morphology and
phenotype.

J. Biomater. Tissue Eng. 1, 30–39, 2011 31

 Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends Balu et al.

pathways. Examples of US Food and Drug Administration
(FDA) approved biodegradable polymers are polylactic
acid (PLA), polyglycolic acid (PGA), poly(lactic-co-
glycolic acid) (PLGA) etc. thereof listed in Table II. These
polymers degrade in the body hydrolytically to produce
lactic acid and glycolic acid, respectively. However, these
polymers are too weak to be used in load-bearing appli-
cations and exhibit bulk degradation which might lead to
an inflammatory foreign body response as the body is
unable to cope with the vast amounts of implant degra-
dation products.16–18 In addition, bulk degradation causes
both a loss in mechanical properties and lowering of
the local solution pH which accelerates further degra-
dation in an autocatalytic manner. Other biodegradable
polymers currently being studied for tissue engineering
applications include polycaprolactone (PCL), polyanhy-
drides, polyphosphazenes (PPF) etc.19 The PCL (chemi-
cal structure shown in Fig. 1) is hydrolytic biodegradable
polyester approved by FDA as a compatible polymer with
both soft and hard tissues. Unlike other FDA approved
biopolymers like PLA, PGA, etc., PCL undergo surface
degradation by hydrolysis of its ester linkages in phys-
iological conditions and hence received a wide range
of applications such as biodegradable packaging materi-
als, implantable biomaterials, carriers for drug delivery
etc. Additionally, PCL releases non-toxic by-products like
caproic acid upon degradation and the degradation prod-
ucts are easily resorbed through metabolic pathways and
do not produce local acidic environments as opposed to
polylactides and glycolides.20 Based on the literature data
obtained from ISI Web of Knowledge, PCL is the most
extensively used electrospun polymer scaffolds for bone
tissue engineering contributing nearly to one fifth of the
total scaffolds. However, PCL having lower degradation
rate, lower hydrophilicity and the lack of surface cell
recognition sites causes a reduction in cell adhesion, prolif-
eration, and differentiation compared to other biocompati-
ble polymers.12 The tendency of PCL to form compatible
blends with a wide range of other polymers provides a
promising polymer platform to tailor its cell affinity to suit
a specific anatomical site.
In this study, a biodegradable semi-crystalline
thermoplastic polyester poly(1,4-butylene adipate-co-
polycaprolactam) (PBAPCL) (chemical structure shown in
Fig. 1) is blended with PCL and electrospun to fabricate
a biodegradable scaffold with a higher degradation rate
than PCL. The PBAPCL being an ester, it is expected to
degrade by hydrolysis of ester bonds. The authors have no
knowledge of any previous study on PBAPCL in bone tis-
sue engineering applications. The electrospun blends were
characterized for its physico-chemical properties, enzy-
matic degradation and biocompatibility. Human osteoblast
cell culture studies were performed to determine the cell
attachment and proliferation for the blend to be used in
bone tissue engineering applications.
2. MATERIALS AND METHODS
2.1. Electrospinning
The polymeric solutions were prepared separately by dis-
solving 20 Wt% of PBAPCL (Sigma Aldrich, USA) in
1,1,1,3,3,3-Hexafluoroisopropanol (HFIP) (Sigma Aldrich,
USA) and 15 Wt% of PCL (Mn ∼ 80,000, Sigma Aldrich,
USA) in chloroform respectively. The solutions were then
mixed in volumetric ratios of 3:1, 1:1 and 1:3 for 48 h
to get a homogenous solution. Electrospinning was car-
ried out at room temperature using a custom made electro-
spinning set-up fabricated with a syringe pump (NE1000,
New Era Pump Systems Inc., USA) and a high-voltage
DC generator (Model 2-A, Zeonics Systech, India). The
anode of the power supply was connected with the spin-
neret (typically a hypodermic syringe needle) and the col-
lector plate was grounded. After several trials, the mixed
solutions were fed at 200
L h−1 by the syringe pump into
a metallic capillary of 22 gauges with tip-target distance

Table II. Properties of various FDA approved biodegradable polymers.

Approximate Approximate degradation

RESEARCH ARTICLE

Polymer strength (GPa) time (months) Degradation products Applications

Poly(glycolic acid) 7.0 6 to 12 Glycolic acid Therapeutic device

Poly(l-lactic acid) 2.7 >24 l-lactic acid Sutures, stents, dialysis
media and drug delivery
devices
Poly(d,l-lactic acid) 1.9 12 to 16 d,l-lactic acid Orthopedic
Poly(d,l-lactic-co-glycolic 2.0 5 to 6 d,l-lactic acid and glycolic Therapeutic device, drug
acid)(85/15) acid delivery
Poly(d,l-lactic-co-glycolic 2.0 4 to 5 d,l-lactic acid and glycolic Therapeutic device, drug
acid)(75/25) acid delivery
Poly(d,l-lactic-co-glycolic 2.0 3 to 4 d,l-lactic acid and glycolic Therapeutic device, drug
acid)(65/35) acid delivery
Poly(d,l-lactic-co-glycolic 2.0 1 to 2 d,l-lactic acid and glycolic Therapeutic device, drug
acid)(50/50) acid delivery
Polycaprolactone 0.4 >24 Caproic acid Drug delivery device, suture,
orthopedic

32 J. Biomater. Tissue Eng. 1, 30–39, 2011

Balu et al. Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends
Fig. 1. Chemical structure of PCL and PBAPCL polymers.
fixed at 20 cm to form fibers. The as-spun fibers were col-
lected over aluminium foil on the ground, dried overnight
at 37  C and stored in a desiccator.
2.2. Scanning Electron Microscopy (SEM)
The electrospun fiber mat was sputter coated with gold
using a precession coating system (Gatan, USA) and
the morphology of electrospun PCL/PBAPCL mats were
observed using a scanning electron microscope (FEI-
Quanta 200, Netherlands).
2.3. Wide Angle X-ray Diffraction (WAXD) Analysis
The crystalline nature of PCL, PBAPCL and their blend
was studied by WAXD analysis. The studies were carried
out using a X-ray diffractometer (D8 DISCOVER, Bruker,
USA) with Cu K radiation ( = 154 Å) at scanning rate
of 1 step per second and 0.15  /step.
2.4. Fourier Transform Infrared (FT-IR) Spectroscopy
The molecular interactions of the blend electrospun fibers
were assayed using a Fourier transform infrared spectrom-
eter (Spectrum One FT-IR spectrometer, Perkin-Elmer,
USA). Spectra were collected in the spectral range of
4000–400 cm−1 and the functional groups were character-
ized accordingly.
2.5. Thermal Analysis (TG-DTA)
The thermo gravimetric analysis (TGA) and differential
thermal analysis (DTA) of PCL, PBAPCL and the blend
was carried out using a high resolution simultaneous ther-
mal analyser (Q500, TA Instruments, USA). Approxi-
mately 3 mg of the samples were heated from ambient to
800  C at a heating rate of 10  C/min with nitrogen flow
rate of 60 ml/min.
2.6. Contact Angle Analysis
The static contact angle of the polymer mats were mea-
sured by sessile drop method using a contact angle
analyser (GBX, France) with distilled water as the contact-
ing solvent at room temperature. The measurements were
made after allowing the droplet on thoroughly cleaned and
dried sample surface for 30 s. Averages of 5 measurements
were taken from 3 replicates for each sample.
J. Biomater. Tissue Eng. 1, 30–39, 2011
2.7. Enzymatic Degradation
A preliminary study about the enzymatic degradation of
the PCL/PBAPCL blend was carried along with the native
polymers. Each experiment was carried out twice, where
the dry electrospun mat of PCL/PBAPCL blend along with
PCL and PBAPCL were weighed and then placed into a
capped bottle containing 10 ml of phosphate buffer solu-
tion (PBS) (0.05 mol/L, pH = 74). Lipase enzyme from
Candida cylindracea (Sigma Aldrich, USA) of concentra-
tion 0.1 mg/ml and 0.02% w/v sodium azide were added
as degradation and bacteriostatic agents. The bottles were
incubated at 37  C and at predetermined time intervals
samples were withdrawn from the degradation medium,
gently rinsed with distilled water and then dried to constant
weight in vacuum desiccators at 37  C. Weight loss was
determined gravimetrically by comparing the dry weight
remaining at a specific time with the initial weight using a
high precision weighing balance. The buffer solution was
changed every 24 h to maintain the initial activity of the
enzyme.
2.8. Direct Contact In Vitro Cytotoxicity Test
An in vitro cytotoxicity test using direct contact method
was performed for electrospun PCL/PBAPCL scaffold as
per ISO 10993-5. Briefly, the samples were sterilized using
70% alcohol and rinsed with minimum essential medium
supplemented with foetal bovine serum. Test samples, neg-
ative and positive controls in triplicates were placed on
subconfluent monolayer of L-929 mouse fibroblast cells
(received from National Centre for Cell Sciences, India).
After incubation of cells with test samples at 37 ± 1  C for
24 to 26 h, cell culture was examined microscopically for
cellular response around and under the test samples. The
cytotoxic reactivity were graded as 1, 2, 3 and 4 based
on zone of lysis, vacuolization, detachment and membrane
disintegration as given in Table III.
2.9. MTT Cell Viability Assay
The cell viability of electrospun blend scaffold was
quantitatively determined by methylthiazolyldiphenyl- RESEARCH ARTICLE
tetrazolium bromide (MTT) colorimetric assay. The assay
was carried out in triplicate using L-929 mouse fibrob-
last cells. Scaffolds were placed into a 24-well culture
Table III. Grades of direct contact cytotoxic reactivity.
Grade Reactivity Description of reactivity zone
0 None No detectable zone around or under specimen
1 Slight Some malformed or degenerated cells
under specimen
2 Mild Zone limited to area under specimen
3 Moderate Zone extending specimen size upto 0.33 cm
4 Severe Zone extending farther than 0.33 cm
beyond specimen
33
 Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends
plate and 100 ml of cell suspension with a cell density of
5 × 105 cells/ml was seeded onto the scaffolds. The cell-
seeded scaffolds were incubated at 37  C in a humidified
atmosphere containing 5% CO2 for 24 h and then 1.5 ml
of culture medium was added to each well. At predeter-
mined intervals 15 ml of MTT solution (5 mg/ml in PBS)
was added to each well followed by incubation at 37  C
for 4 h to MTT formazan formation. After incubation, the
medium was removed from the well and the solution was
then aspirated. The dimethylsulfoxide (DMSO) containing
125 ml per well of glycine buffer (pH = 10) was added
to dissolve any insoluble formazan crystals to a coloured
solution. After 10 min of rotary agitation, the absorbance
of the DMSO solution at 570 nm was measured using
a UV/Visible spectrophotometer (Cary 5E, Varian, USA)
and cell viability was calculated as the percentage relative
to the untreated control cells using the equation
% Cell viability = Mean OD/Control OD × 100
2.10. Human Osteoblast Cell Culture Studies
Cell culture studies were carried out for electrospun
PCL/PBAPCL blend fibrous mat with the human pri-
mary osteoblast (hOB) cells at Sree Chitra Tirunal Insti-
tute for Medical Sciences and Technology (SCTIMST,
Trivandrum, India). The hOB cells of concentration
5000 cells/cm2 surface area cultured in osteogenic medium
for 1 week were seeded on the surface of the control and
the samples in triplicates, sterilized with 70% ethanol. The
RESEARCH ARTICLE
Fig. 2. SEM micrograph of electrospun PCL/PBAPCL fibers (A) 3:1 blend, (B) 1:1 blend and (C) 1:3 blend and respective fiber size distribution.
34
Balu et al.
culture media was added gently along the sides of the wall
so that cell suspension will be on the top of the material.
The culture plate was then incubated at 37  C in a fully
humidified atmosphere at 5% CO2 for 2 and 4 days. After
2nd and 4th day, the medium was removed and the samples
were rinsed with phosphate buffer solution. The cells were
then fixed on samples by 2.5% glutaraldehyde followed by
rinsing with phosphate buffer solution. After rinsing, the
samples were dehydrated in different grades of methanol
and the samples were then immersed in iso-amylacetate
for 2 min and dried.21 The dried samples were gold sputter
coated and observed in SEM for hOB cell attachment and
proliferation.
3. RESULTS AND DISCUSSION
3.1. Morphology of Scaffold
(1) The effect of concentration of PBAPCL on the morphol-
ogy and diameter of the as-spun fibers was investigated
by mixing 15 Wt% PCL with 20 Wt% PBAPCL solutions
at varied volumetric ratios (i.e., 3:1, 1:1 and 1:3 v/v %)
followed by electrospinning of blends. An addition of
PBAPCL solution to PCL solution in volumetric ratio of
1:1 corresponding to PCL:PBAPCL of 1.4:2 w/w% pro-
duced narrow distributed PCL/PBAPCL blend fibers. Fur-
ther, increasing or decreasing the ratio of blend to 3:1 or
1:3 v/v % showed widely distributed fibers with beads.
The morphology and distributions of fiber diameters by
the scanning electron micrograph for respective condi-
tions are shown in Figure 2. The bead-free, narrow size
J. Biomater. Tissue Eng. 1, 30–39, 2011
Balu et al. Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends
distributed 1:1 v/v% PCL/PBAPCL blend fiber mat was
considered for further characterization. There after the
1:1 v/v% PCL/PBAPCL composite fibers can be referred
as 1:1 blend. The detailed structure of the 1:1 blend fibrous
mat by SEM micrographs indicated that the electrospun
fibers were homologous and uniform over a large area
with interconnected pores throughout the scaffold. How-
ever, fibers were observed to be attached to each other with
an average fiber diameter of 400 nm in the scaffold. The
3D structure of the scaffold provides a large surface area to
volume ratio for cell attachment, proliferation with space
for nutrient transportation and bone in-growth.21 Attempts
were also made to electrospin 20 Wt% PBAPCL solution
for comparison, however the solution was found to be elec-
trospraying leading to formation of polymer droplets.
3.2. Crystalline Nature
The WAXD pattern of PCL, PBAPCL polymers and the
1:1 blend nanofiber mat is shown in Figure 3. It was
known that the PCL homopolymer crystallizes easily and
the diffraction pattern of the crystalline PCL mat reveals
the presence of significant crystallinity with peaks at 2 =
219 and 24.2 corresponding to the (110) and (200)
planes of the orthorhombic crystal structure.22 On the other
hand, PBAPCL showed broad diffraction peaks at 2 =
212 and 24 which is characteristic of semi-crystalline or
mostly an amorphous polymer. The diffractogram of the
PCL/PBAPCL 1:1 blend fiber mat showed a broad peak at
Fig. 3. XRD pattern of PCL, PBAPCL polymers and blend (1:1)
nanofiber mat.
J. Biomater. Tissue Eng. 1, 30–39, 2011
2 = 212 corresponding to diffraction peak of PBAPCL.
Peaks were also observed at 2 = 219 corresponding to
diffraction peak of PCL and 2 = 24 due to the merge of
the peaks at 24.2 and 24 of PCL and PBAPCL respec-
tively. The XRD results thus suggest that the blend fiber
mat to be semi crystalline in nature.
3.3. Molecular Interactions
Figure 4 shows the FT-IR spectra of the polymers and
their corresponding 1:1 blend. The blend spectrum showed
CH3 , CH2 , C O, C–O–C, C–H vibrational peaks cor-
responding to PCL and PBAPCL at 2923, 2853, 1618
1028 and 836 cm−1 respectively.23 The peak shifts from
1723 to 1727, 1123 to 1135 and 617 to 575 corresponds to
stretching of C O, C–O–C and C–H bending vibrations
of blend mat in comparison to the native polymers. The
ester peaks in the range of 1500 to 1200 were observed to
be similar for both the polymers and the blend. In addition,
a new peak at 3060 cm−1 which corresponds to stretching
of N–H group bonded with carbonyl (N–H · · · O C) was
also observed in the blend.24 The shifting of the peaks and
formation of a new peak suggest the molecular interaction
of polymers in the blend.
3.4. Thermal Degradation
The thermal analysis of the PCL/PBAPCL 1:1 blend was
carried out to study the effect of PBAPCL on the thermal
stability of PCL matrix and for further characterization
of the polymer interactions. Figure 5(A) shows the differ-
ential thermal analysis of PCL, PBAPCL and 1:1 blend.
The DTA thermogram of PCL showed exothermic peak at
443  C whereas the blend showed two exothermic peaks
between 300 and 425  C. The first peak at 340  C was due
to PBAPCL and the second peak at 360  C was due to
RESEARCH ARTICLE
Fig. 4. FTIR spectra of (a) PCL (b) blend (1:1) nanofiber mat and (C)
PBAPCL.
35
 Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends Balu et al.

Fig. 6. Contact angle micrograph of (A) PCL, (B) electrospun 1:1 blend
fiber mat and (C) PBAPCL.

Fig. 7. Degradation profile of PCL, PBAPCL and their blend (1:1) fiber
mat by lipase enzyme.

3.5. Wetability
Fig. 5. (A) DTA thermogram of PCL, PBAPCL and PCL/PBAPCL
blend (1:1) fiber mat; (B) TGA thermogram of PCL, PBAPCL and The contact angle values of PCL (90 ± 01) with dis-
PCL/PBAPCL blend (1:1) fiber mat. tilled water showed improved wetability upon blending
with PBAPCL as shown in Figure 6. The 74 ± 01 water
contact angle value of the 1:1 blend is comparatively
PCL. The PCL peak was slightly shifted due to the graft- closer to that of PBAPCL (68 ± 01) in spite of having
ing reaction. These results suggest that PBAPCL and PCL equal volumetric fraction of PCL respectively. This also
chains were mixed well at a molecular level. Decomposi- indicates the possibility of PBAPCL blending with the
tion curves of PCL, PBAPCL and the polymer blend are
shown in Figure 5(B).
The polymers displayed a simple decomposition pro-
file corresponding to single step degradation. The thermal
RESEARCH ARTICLE

onset degradation temperature at 10 % Wt loss (T10%  and

50% Wt loss (T50%  are given in Table IV. It is evident
from Table IV that the degradation temperature of PCL is
lowered blending with PBAPCL.

Table IV. Thermal properties of PCL, PBAPCL and their blend.

Temperature ( C) for

Sample 10% Wt loss 50% Wt loss

PCL 358.2 386.9

PCL/PBAPCL (1:1 v/v) 304.8 354.3
Fig. 8. Optical micrograph of L-929 cells around 1:1 blend nanofiber
PBAPCL 300.6 358.4
mat.

36 J. Biomater. Tissue Eng. 1, 30–39, 2011

Balu et al. Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends

3.6. Enzymatic Degradation

The PCL, reported to undergo a two-stage degradation

process: first, the non-enzymatic hydrolytic cleavage of
ester groups followed by intracellular degradation as evi-
denced by observation of PCL fragments uptake in phago-
somes of macrophages and giant cells.26 Figure 7 shows
the weight retention profiles obtained during the enzymatic
degradation of the homopolymers and the blends. Nearly
30 Wt% of PCL degradation was observed in 5 weeks
with the lipase enzyme. The enzyme catalyzes the hydrol-
ysis of ester chemical bonds leading to degradation of the
polymer backbone.27 It can be seen that both PBAPCL
and the 1:1 blend showed higher enzymatic degradation
than the PCL. The 1:1 blend exhibited more susceptibil-
ity than PCL to enzymatic degradation as evident from a
weight loss of about 45% during the experiment. How-
ever, PBAPCL showed the highest weight loss of 51% for
5 weeks followed by disintegration into fragments in sub-
sequent weeks.

Fig. 9. MTT assay cell viability results. 3.7. In Vitro Cytotoxicity

Electrospun PCL/PBAPCL 1:1 blend tested by direct con-

PCL at molecular level. The improved wetability could tact method showed no cytotoxic reactivity to L-929
aid for better cell attachment, proliferation and might mouse fibroblast cells after 24 h of contact. Moreover,
affect the degradation of the blend by hydrolysis of ester the results of cytotoxicity test were observed and cer-
bonds.25 tified to be “0” grade (description given in Table III)

RESEARCH ARTICLE

Fig. 10. SEM micrograph of osteoblast cells on blend (1:1) nanofiber mat (A & B) after 2 days, (C & D) after 4 days.

J. Biomater. Tissue Eng. 1, 30–39, 2011 37

 Electrospun Polycaprolactone/Poly(1,4-butylene adipate-co-polycaprolactam) Blends
by SCTIMST, Trivandrum, India. Optical micrograph of
L-929 cells (Fig. 8) showed cell morphology and pheno-
type maintained around blend nanofiber mat indicating non
cytotoxic reactivity.
3.8. MTT Cell Viability
Cell viability for 1:1 blend fiber mat was quantitatively
evaluated to be about 90% using Eq. (1). Figure 9 shows
the cell viability test results by MTT assay. The results
showed the cells to be viable for the blend fiber environ-
ment. The MTT assay clearly indicate the cell viability as a
measure of living cells supporting the potential application
of the blend as a suitable scaffold for tissue engineering
applications.
3.9. Osteoblast Cell Adhesion and Proliferation
Figure 10 shows the SEM micrograph of osteoblast cells
cultured on electrospun 1:1 blend scaffold. It is evident
that the cells have attached well on the surface of the scaf-
fold and started proliferating on the 2nd day. Moreover, by
the 4th day the cell concentration was found to be higher
and the cells were well spread covering most of the areas
of scaffold surface. It was also observed that at the 4th day
most of the cells exhibited advanced stages of spreading,
that is cells exhibit cytoplasmic webbing and fully spread
with polygonal in shape. This clearly suggests that the
scaffold can be used in bone tissue engineering with min-
eralization by osteoblast cells upon cell culture for further
days.
4. CONCLUSIONS
A blend of PCL and PBAPCL has been electrospun to
form a porous scaffold. The blend was found to be semi-
crystalline in nature with molecular interactions observed
between PCL and PBAPCL. The scaffold showed bet-
ter physiological degradation property than PCL. In addi-
tion, the blend fiber mat was tested to be non-cytotoxic
after 24 h with fibroblast cells. The blend nanofibers
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showed cytoplasmic webbing of osteoblast cells in 4 days,
demonstrating the possibility for mineralization. Overall,
the PCL/PBAPCL blend fibrous mat provides a platform
from the well known biodegradable PCL polymer with a
tremendous potential as a scaffold material for guided bone
tissue regeneration.
Acknowledgments: This work was supported by
research grant (22(0447)/07/EMR-II) from Council for
Scientific and Industrial Research (CSIR), Government
of India. The authors would like to thank Dr. T. V.
Kumari, BMT wing, SCTIMST, Trivandrum, India for
direct contact in vitro cytotoxicity test and osteoblast cell
culture studies.
38
Balu et al.
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Received: 19 May 2011. Accepted: 10 June 2011.
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