Healing
Potential of Trichosanthes dioica
Roxb on Burn Wounds
Yogesh
Shivhare*, Priya Singh and
Dept. of Pharmacognosy, VNS Institute of
Pharmacy,
ABSTRACT
Trichosanthes
dioica Roxb is
a well-known plant in the traditional medicine. Based on its traditional
use, methanolic extract of the plant was selected for assessment of healing
potential in the form of simple ointment using full thickness burn wound model
in rats. The effect produced by the extract ointment showed significant healing
when compared with the control and standard groups. All parameters such as
wound contraction, epithelialization period, hydroxyproline content, and
histopathological studies were observed significant (P<0.01) in comparison to control group.
KEY
WORDS: T. dioica, Burn wound, Hydroxyproline.
INTRODUCTION
Wound can be defined as interruption of
cellular and anatomical or practical continuity of living tissues. It can be
produced by physical, chemical, electrical or microbiological rudeness to the
tissue1. Burn wound healing is a multifaceted course and does not
require much help but still cause distress and are prone to infection and other
complications. Infection is a major impediment of burn injury and is
responsible for 50-75% of hospital deaths2. In
Trichosanthes
dioica (Cucurbitaceae),
commonly known as palval, is distributed throughout the plains of
MATERIALS AND METHODS:
Plant material:
The fruits of Trichosanthes
dioica Roxb were collected from the local market of
Preparation of extract:
Fruits of Trichosanthes
dioica Roxb were dried in shade and powdered. About 125 g of powdered
fruits were extracted with 500ml methanol using soxhlet apparatus for 24 h at
75-80˚C. Solvent removal carried out under pressure afforded a semi solid
mass with a yield of 16.35g w/w with respect to dry powdered material.
Ointment formulation and standard used:
A 5% (w/w) simple ointment containing the methanolic
extract of Trichosanthes dioica was
prepared by trituration method in a ceramic mortar and pestle using white soft
paraffin base12, obtained from S.d. fine chemical,
Animals:
Healthy albino rats of either
sex (150-170g) were selected for all the present in vivo studies. The animals were fed on normal diet and water ad
libitum. The animals were used after an acclimatization period of 7 days to
the laboratory environment. Animal study was performed in division of
pharmacology, VNS Institute of Pharmacy,
Grouping of Animals:
Three groups of animals
containing six in each were used for burn wound model. The animals of group I,
II and III were considered as the control, reference standard and test
respectively.
Group I: Control
(treated with ointment base only)
Group II:
Standard (treated with 0.01% silver sulphadiazine)
Group III: Test
group (treated with 5% methanolic extract ointment of Trichosanthes dioica)
Burn wound model and drug treatment:
In burn wound
model13, three groups of animals containing six in each group were
anaesthetized by open mask method with anesthetic ether before wound creation.
The particular skin area was shaved one day prior to the experiment. Burn wounds
were created on dorsal part of shaved rats using a metal rod (1.5 cm diameter)
heated to 80- 85˚C and exposed for 20 seconds. After 24 h, dead tissues
were excised using sterile surgical blade. Control rats were dressed with
ointment base alone, while experimental rats were dressed with the 5% ointment
formulated with methanolic extract of Trichosanthes
dioica. All the rats were given regular dressing changes at every
alternative day.
Wound healing evaluation parameters:
The parameters
observed in the study were as follows:
Rate of wound contraction14 and Epithelialization period15
The progressive
changes in wound area were measured in mm2 by tracing the wound
margins on a transparent paper on every 2-day
interval. The wound areas in all groups were recorded on a graph paper. Wound contraction was expressed as reduction in percentage of the original wound size
by using the following formula:
% Wound
contraction =
Initial wound
size – Specific day wound size × 100
Initial wound size
Epithelialization
period was evaluated by noting the number of days mandatory for the escher to
fall off from the wound surface exclusive of leaving a raw wound behind
Hydroxyproline
estimation16
Hydroxyproline is an uncommon amino acid present in the collagen fibers of
granulation tissues. Its assessment helps clinically to realize the progress
rate at which the healing process is going on in the connective tissue of the
wound. Collected tissues were subjected to hydroxyproline estimation following
the method of Woessner.
Histopathological studies17
A sample of skin
tissues from control, standard and treated groups were excised from the healed
wound and preserved in 10% buffered formalin. Thin sections were cut and
stained with haematoxylin and eosin and observed under microscope for the
histological changes.
Statistical
Analysis
Experimental data
are expressed as mean ± standard error of mean (SEM). Statistical analysis was
performed using Dunnet vs. control test followed by one-way ANOVA to identify
the differences between pair of groups. Data were considered significant at p
< 0.01.
RESULTS:
The significant
increase in the wound-healing activity was observed in the animals treated with
the methanolic extract of Trichosanthes
dioica when compared with those
who received the ointment base treatments. In the experimental burn wound
model, the rate of wound contraction was significant (P< 0.01) when compared to control and standard groups. Contraction
of wound was promoted from 2nd day of treatment till 14th
day as shown in fig 1. The
epithelialization period of wounds, in case of animals treated with methanolic
extract of Trichosanthes dioica was
found to be quite earlier than control groups (P< 0.01) as depicted in fig 2.
It was also comparable with the standard drug silver sulphadiazine.
Hydroxyproline content was found to be increased significantly in the group
treated with methanolic extract of Trichosanthes
dioica than the control group which was shown in fig 3. Histopathological
study of the excised skin also evidenced the wound healing property of the
methanolic extract of Trichosanthes
dioica. The sections of the skin of treated animals showed significant
increase of blood vessels, fibroblasts and collagen fibers when compared to
animals of control groups. Photograph of skins are presented in fig 4.
Fig.4: Histopathology of skin in control (a)
and treated group (b). photographs are showing clear evidence for fibroblast
cells (F), blood vessels (B) & collagen fibers (C) in treated group than
the control group.
DISCUSSION:
Wound healing is
step by step process having different phases like haemostasis, inflammation,
proliferative and maturation. Hence, evaluation of the potentials of Trichosanthes dioica in burn wound
showed that the fruit extract exhibited wound healing activities and the
constituents of the fruit may play a useful role in the management of burn
wounds. In the present study, topical application of ointment formulated with
methanolic extract of Trichosanthes
dioica significantly showed the faster healing with earlier wound
contraction and rapid epithelialization which depends upon reparative abilities
of the tissues, type and extent of the damage and general state of the tissue.
Significant increase of hydroxyproline content implies more collagen deposition
in treated groups than the control group. Histopathological examination
revealed the formation of new blood vessels, fibroblasts and collagen fibers,
which supported the healing activity. Preliminary phytochemical analysis of
methanolic extract of Trichosanthes
dioica showed positive test to flavonoids, glycosides and tannins. Earlier
studies reported that flavonoids18, tannins19 and
triterpenoids20 are identified for wound healing action. Thus the
healing potential of Trichosanthes dioica
may be credited to the phytoconstituents present in it which may be either due
to their individual or additive effect that fastens the course of wound
healing.
CONCLUSION:
The results of
this study indicate that methanolic extract of Trichosanthes dioica have good potential for use in wound care and
further provide a justification for the use of the fruits of this plant in
wound management.
ACKNOWLEDGEMENT:
The authors great
fully acknowledge to Dr. Umesh K. Patil, Principal and Dr. R. S. Pawar Asst.
Professor VNS Institute of Pharmacy, for providing necessary facilities and
valuable suggestions to carry out this work.
REFERENCES:
1.
2.
Mokaddas
E, Rotimi VO and Sanyal SC. In vitro
activity of piperacillin/tazobactam versus other broad antibiotics against
nosocomial gram negative pathogens isolated from burn patients. Journal of
Chemotherapy. 1998; 10 (3): 208-214.
3.
Subrahmanyam
M. A prospective randomised clinical and histological study of superficial burn
wound healing with honey and silver sulfadiazine. Burns. 1998; 157-161.
4.
Diwan
PV, Tillo LD and Kulkarni DR. Influence of Tridax procumbens on wound
healing. Indian Journal of Medical Research. 1982; 75: 460–464.
5.
Nagappa
AN and Cheriyan B. Wound healing activity of the aqueous extract of Thespesia
populnea fruit. Fitoterapia 2001; 72: 503–506.
6.
Rasik
AM, Raghubir R, Gupta A, Shukla A, Dubey MP, Srivastava S, Jain HK and Park
Kulshrestha DK. Healing potential of Calotropis procera on dermal wounds
in guinea pigs. Journal of Ethnopharmacology. 1999; 68: 261–266.
7.
Suguna
L, Sivakumar P and Chandrakasan G. Effects of Centella asoatica extract
on dermal wound healing in rats. Indian Journal of Experimental Biology. 1996;
34: 1208–1211.
8.
Chaterjee
Asima and Pakrashi Satyesh Chandra. The Treatise of Indian Medicinal Plants,
Vol. 1, Publication and Information Directorate,
9.
Kirtikar
KR and Basu BD. Indian Medicinal Plants.
10.
11.
Sharma
G and Pant MC. Effect of feeding Trichosanthes
dioica (Parval) whole fruits on blood glucose, serum triglycerides,
phospholipid, cholesterol and high density lipoprotein-cholesterol levels in
the normal albino rabbits. Current Science. 1988; 57: 1085-87.
12.
Cooper.
Gunn’s Dispensing for pharmaceutical students. CBS Publisher and Distributors,
13.
K.
Shanmuga Priya, Gnanamani A, Radhakrishnan N and Mary Babu. Healing potential
of Datura alba on burn wounds in
albino rats. Journal of Ethanopharmacology. 2002; 83: 193-199.
14.
Sadaf
F, Saleem R, Ahmed M, Ahmad SI and Navaid-ul-Zafar. Healing potential of cream
containing extract of Sphaeranthus indicus on dermal wounds in Guinea
pigs. Journal of Ethnopharmacology. 2006; 107: 161–163.
15.
Rashed
AN, Afifi FU and Disi AM. Simple evaluation of wound healing activity of a
crude extract of Portuloca oleracea Linn. (growing in
16.
Woessner
JF. The determination of hydroxyproline in tissue and protein samples
containing small portion of this imino acid. Archives of Biochemistry and
Biophysics. 1961; 193: 440–447.
17.
McManus
JFA and Mowry RW. Staining Methods, Histological and Histochemical. Harper
& Row/Evanston, New York/London. 1965.
18.
Tsuchiya
H, Sato M, Miyazaki T, Fujiwara S, Tanigaki S and Ohyama M. Comparative study
on the antibacterial activity of phytochemical flavanones against methicillin-
resistant staphylococcus aureus. Journal of Ethanopharmacology. 1996; 50:
27-34.
19.
Scortichini
M. and Pia Rossi M. Preliminary in vitro
evaluation of the antimicrobial activity of terpenes and terpenoids towards Erwinia amylovora (Burrill). J. Appl
Bacteriol. 1991; 71: 109-12.
20.
Goren
N, Woerdenbag H and Bozok- Johanssan C. Cytotoxic and antibacterial activities
of sesquiterpene lactones isolated from Tanacetum
praeteritum. Planta medica. 1996. 62: 419-22.
Received on 03.12.2009
Accepted on 15.02.2010
© A&V Publication all right reserved
Research J. Pharmacology and
Pharmacodynamics 2(2): March –April 2010: 168 -171