D. Santhi krupa, et al.

/International Journal of Advances in Pharmaceutical Research

IJAPR Research Paper

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www.ijapronline.org ISSN: 2230 – 7583

Comparative Evaluation of Wound Healing Activity of Combination

Ointment of Heliotropium indium (Boraginaceae) And Momordica
Charandia (Cucurbitaceae)
D. Santhi krupa*, Dr. K. Ravi Shankar, Ch. Pavan kumar, Y. Naga Prassanna,
N. Archana, A.Tirupathi Rao, M. Vinay kumar
Aditya Institute of Pharmaceutical Science and Research, ADB Road,
Surampalem - 533 43, A.P, India.
Received on 13 – 08 - 2013 Revised on 16 – 09- 2013 Accepted on 19– 09 – 2013
ABSTRACT
Herbal plants of Heliotropium indium and Momordium charantia have a potent pharmacological activity. In the
present study, we revealed the ethyl acetate extracts of both these herbs were studied for the wound healing activity
using wistar albino rats. The results were compared with standard Framycitin. Wound was made on the back of the
wistar albino ratsby excision wound method under anesthesia. Individual and combination herbal formulation
ointments (5% H.I.& 1%M.C. ) were prepared by using ethyl acetate solvent and was applied to the wound of the
animals. 98.8% of wound was cured in 9 days where as standard ointment produced only a 85.57%. It concluds that
combination of herbal formulated ointment had more prominent activity and decreased side effects.
Key Words; Heliotropium indium , Momordium charantia , framycetin , excision wound method ,wound healing.
INTRODUCTION
Herbs are being used in the traditional medicine in Many plant remedies have been used in wound
the treatment of disease is as old as mankind. The use healing since the days old but the report on their
of traditional medicine is even supported by World exact mechanism of action, toxicity and efficacy are
Health Organization provided it is delivered in a safer very sporadic. The present study is designed to
way and is efficient. In developing countries like validate the ethno-medicinal use of the leaves of
India people sometimes cannot afford for the costly Heliotropium indium belongs to family
treatment and in such a cases the herbal medicines Boraginaceae, mainly used for the treatment of
offer a better and efficient way of treatment at lesser tumors, asthma, wounds and leaves of Momordica
costs. In recent times, herbal medicines gained charantia belongs to the family Cucurbitaceae, used
importance as sources of lead compounds since they mainly to treat diabetes. Skin may be damaged by
are time tested, relatively safe for human use, eco- various reasons like radiations, microbes, chemical,
friendly, cheap, easily available and are affordable 1. extreme temperatures etc. A wound can be defined as
the break in the continuity of the soft tissues like
Author for Correspondence mucous membranes, skin and tissue surfaces2. A
Dasari santhi krupa, wound with various degree of damage of the tissue
Aditya Institute of Pharmaceutical Science and including that of the skin is termed an external wound
Research, and the wound which damages the underlying tissues
ADB Road, to varying leaving the skin intact is termed as an
Surampalem-533 437 . internal wound.
Mail id: shanthikrupa@gmail.com Wound healing is complex and dynamic
Ph. No: 9247679315 process of restoring the cell structure and tissue
layers and restores the functions of the damaged
tissue2. In this process the damaged tissue is replaced
by connective tissue which then forms a scar. Wound
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 D. Santhi krupa, et al. /International Journal of Advances in Pharmaceutical Research

healing occurs by the process of vascular responses
(homeostasis), inflammation, proliferation and
maturation, they can be grouped in two phases- early
phase and cellular phase3. The main constituent of
blood clot is fibrin which acts as mesh and traps the
blood cells, this gradually dries out acts as scab that
closes the wound temporarily4. During the
proliferation the wound is filled with a new
connective tissue and involves granulation,
contraction and epithelization5. Wound care, stress,
dehydration, age, nature of injury play a key role in
wound healing process. Our present studies involves
determination of the wound healing activity of the
combination of the herbal extracts, using framycetin
as standard drug.
Heliotropium indicum is a annual herb
commonly termed as Indian heliotrope is a native to
Asia and is a common weed in waste places. The
Infusion of the leaves and young shoots are used to
treat nettle rash. Infusion of the flowers taken in a
small dose regulates menstruation, where large doses
are abortive. Decoction of the leaves is used as a
vermifuge. Juice of the leaves acts as antiseptic and
anti-inflammatory and is also applied to wounds,
sores, boils and pimples on face. Boiled with castor
oil they are used to treat scorpion bites. It is also
employed locally for ophthalmic purpose when the
cornea is inflamed or excoriated.
Momordica charantia often called bitter
gourd and is widely grown in Asia, Africa, and
Caribbean, it is the most bitter of all edible fruits. It is
used for variety of purposes as Anti-cancer agent,
Anti helminthic agents, Anti-malarial, Anti-viral,
Cardio-protective agent and also as anti-diabetic
agent.
MATERIALS AND METHODS
The chemicals used include ethyl acetate,
framycetin (sanofi aventis), wool fat, hard paraffin,
cetostearyl alcohol and white soft paraffin all were of
laboratory grade obtained from Sigma-Aldrich.
The leafs of the herbs Heliotropium
indicum and Momordica charantia was collected
from surroundings of Surampalem, East Godavari
district (Andhra Pradesh).The collected leaves of
herbs are sun dried for about one week and are then
pulverized in to fine powder separately. The fine
powder is then subjected to maceration with ethyl
acetate as solvent for 5 days. The extract is filtered
through Whatmann’s filter paper No.1 and is
subjected to distillation. The extract thus obtained
was dried and is used in the preparation of ointments.

Table:1 Preparation of Extracts form Dried leaves of selected plants

Weight of dried Final weight of the
Volume of Ethyl-acetate
S.no: Plants species leaves powder extract
solvent (in ml)
(in grams) (in grams)
1. Momordica charantia 85 500 5
2. Heliotropium indicum 100 400 8

Preparation of ointment by fusion method

Ointment is prepared mixing the dried extracts of the leaves in the ointment base made from Wool fat, hard paraffin,
Cetoseryl alcohol, white soft paraffin.
Preparation of simple ointment:
Wool fat - 2 gm
Hard Paraffin-2 gm
Cetostearyl alcohol -2gm
White Soft Paraffin-34 gm.
Each ingredient was mixed and heated gently with stirring then cooled. The base was then packed in a wide
mouth container.
Preparation of Heliotropium indicum ointment
28.5gm of ointment base was taken and 1.5gm of Heliotropium indicum extract was added to it in a
motor and mixed slowly until a homogeneous product is formed. The ointment was then packed in a wide
mouth container.
Preparation of Momordica charantia ointment
26.7gm of ointment base was taken and 3.33gm of Momordica charantia extract was added to it in a
motor and mixed slowly until a homogeneous product is formed. The ointment was then packed in a wide
mouth container.
Preparation of combination ointment

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 D. Santhi krupa, et al. /International Journal of Advances in Pharmaceutical Research

25.2gms of ointment base was taken and 1.5gm of Heliotropium indicum extract and 3.3gm of
Momordica charantia extract were added to it in a motor and mixed thoroughly until the homogeneous product
is formed. The ointment was then packed in a wide mouth container.
Study Protocols
Wistar Albino Rats of either sex weighing between 130-150 g were purchased from the National Institute
of Nutrition, Hyderabad, India and are used for the test. The animals were numbered, weighed and then divided into
five groups with three animals in each.
Approximately 500mm2 full thickness of skin is removed from the dorsal thoracic central region of a
anaesthetized rat. And the wound were left undressed to the open environment. Then the ointments were applied
twice a day to the corresponding groups6.
GROUP-I: Received Standard Ointment (Framycetin Ointment).
GROUP-II: Received Heliotropium indicum Ointment.
GROUP-III: Received Momordica charantia Ointment
GROUP-IV: Receive a Combination Ointment.
GROUP-V: Control

RESULTS AND DISCUSSION

The treatment was done for different groups of animals with individual ointments and combination
ointment. The results of the wound healing activity of the ethyl acetate extract is presented in table 2 and figure 1 by
taking the epithelization time in to consideration for various treatment groups.

Table 2: Showing the Epithelization Rate for different treatment groups

Percentage of wound healing
3rd day 6th day 9th day
Standard ointment group 28.15 53.01 85.57
H.I ointment group 31.32 65.18 88.63
M.C ointment group 15.74 40.22 62.45
Combination ointment group 35.16 72.50 98.81

Figure: 1 Graph showing the Epithelization rate in different treatment groups

The epithelization period for standard ointment treated group is 16 days. The epithelization period for H.I treated
ointment group is 14 days. The epithelization period for M.C treated ointment group is 20 days. The epithelization
period for Control group is 23 days. The epithelization period for Combination (5% H.I &1% M.C) treated
ointment group is treated group is 12 days.
The epithelization period for Combination (5% H.I & 1% M.C) treated ointment group is rapid and
more effective than individual extract treated groups.
Though the advances in synthetic medicine had replaced the herbs usage, the herbs are still being used
among tribal people for curing various body ailments.

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Figure:2 Wister rat showing wound on its back at 0th day

Figure:3 Wister rat showing healing wound on its back at 3rd day

Figure:4 Wister rat showing healing wound on its back at 7th day

Figure:5 Wister rat showing healing wound on its back at 10th day

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 D. Santhi krupa, et al. /International Journal of Advances in Pharmaceutical Research

Figure:6 Wister rat showing completely healed wound on its back at 13th day

In our present study, we had taken the two easily Shokooeizadeh HR. Invitro antibacterial
available plants. One is grown near the fields i.e., activity of Rheum ribes extract obtained
Heliotropium indicum, which is having from various plant parts against clinical
phytochemical constituents like alkaloids, isolates of gram-negative pathogens. Iranian
triterpines, tannins, flavonoids, glycosides7 and J Pharm Res 2005, 2, 87.
other plant is Momordica charantia used as 2.
common vegetable consists of tri-terpenes, hristholm CM. Wound evaluation and
alkaloids, steroidal glycosides8, 9. cleaning. Emerg Med Clin North Am
These
1992,
two plants are hav
Momordica charantia is having a very mild 10, 665.
wound healing property, it has slow epithelization 3.
rate and has less wound contraction10. So to increase unt, T.K., “Basic principles of wound
the topical effectiveness of Momordica charantia healing”, J Trauma, 30, 122 (1990).
(1%), the combination ointment is prepared using 4.
Heliotropium indicum (5%). The constituents obson MC. Disturbances in wound healing.
were extracted by using ethyl acetate solvent 11, 12. Annals of Emergency Medicine 1994, 17,
Our observations had shown that percentage 1274.
wound reduction in combination treated groups are 5.
more prominent than that in standard treated group inter GD. Formulation of the scab and the
and individual extract treated groups. rate of epitheisation of superficial wounds
The usage of ointment preparation of extracts in the skin of the domestic pig. Nature 1962,
will decreases the unwanted systemic side effects 193, 293.
and makes the drug readily available at site of 6.
action. The combination ointment shows rapid andey, D.P., Singh, J.P., Roy, R., Singh,
epithelization time than individual ointments. V.P., Pandey, V.B (1996). Constituents of
Heliotropium indicum; OrientalJournalof
CONCLUSION Chemistry 12: 321-322.
The combination ointment prepared by using 7.
ethyl acetate extract of leaves of Heliotropium eddy, J.S., Rao, P.R., Reddy, M.S.
indicum (5%) and Momordica charantia (1%) (2002).Wound healing healing effects of
showed significant wound healing activity in Heliotropium indicum, activity of the
comparison with the ointments made from individual aqueous extract of Plumbago zeylanicum
extracts. The results offer the need for the further and Acalypha indica in rats. J.
investigation of these herbs to better assess their Ethnopharmacol. 79:249-251.
potentiality in the wound healing activity 8.
considering even the extracts obtained from other harma, S., Sharma, M.C., Kohli,
parts of these herbs. D.V.(2010). Wound healing activity of the
ether-chloroform extract of
REFERENCES momordica charantia fruits in rats.
1. DigestJournalof Nanomaterials and F
azly-Bazzaz BS. Khajehkaramadin M, Biostructure.5: 123 – 126.

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 D. Santhi krupa, et al. /International Journal of Advances in Pharmaceutical Research

9. Effect of Three Extracts from T

eoh, S.L., Latiff, A.A., Das,S. (2011). The Heliotropium indicum. Asian J Pharm
effect of topical extract of Momordica Biol.,75:183-185.
charantia (bitter gourd) on wound 12.
healing in non-diabetic rats and in rats odehe, Y., Barthelemy, A., Christine, L.,
with diabetes induced by streptozotocin. Metowogo, K., Jean David N’guessan,
Clin Exp Dermatol. 34, 815-22. Allico Joseph Djaman, Annelise Lobstein
10. and Nelly Frossard(2011). Isolation of R
ajaram Prashanthi, Natarajan Mohan and wound healing compounds from
Ganesan Vijayan Siva (2012). Wound Heliotropium indicum.Journalof
healing property of aqueous extract of seed Applied Pharmaceutical Science.10: 102-
and outer layer of Momordica charantia L. 106.
on albino rats. IndianJournalof Science and 13.
Technology.5:1936-1940. bayomi, S. (1993). Abstracts of the world
11. congress- medicinal and aromatic plants. D
odehe,Y., Amadou, F.C., Jean David Medicine in Africa. J. Altern. Complement.
N’guessan., Allico Joseph Djaman (2011). Med. 13,195-238.
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