Research Journal of Pharmaceutical Dosage Forms and Technology.

7(2): April-June, 2015, 125-128

ISSN 0975-234X (Print) www.anvpublication.org

0975-4377 (online)

RESEARCH ARTICLE

Wound Healing Activity of Poly Herbal formulation

Syed Safiullah Ghori1* , Mohammed Gouse2, Niranjan Panda3, Shaik Khaled1,
Syed Basheeruddin1, Mirza Danish Baig1, Mohammed Yaqub1, Mohammed Mufaqqum1,
Arfa Nazneen1, Amtul Zehra Butul1
1
Pharmacology Research Lab, Anwarul Uloom College of Pharmacy, New Mallepally, Hyderabad 50001,
Telangana, India
2
Almocare Herbal and Ayurvedic Clinic, Chirag Ali lane, Abids, Hyderabad, Telangana, India
3
Department of Pharmaceutics, Anwarululoom College of Pharmacy, New Mallepally, Hyderabad 50001,
Telangana, India
*Corresponding Author E-mail: safiullahghori@gmail.com

ABSTRACT:
Aim: To evaluate the Wound healing potential of a Poly herbal formulation.
Material and Methods: In the present work a poly herbal Unani formulation; beeswax (base), sesame oil (10%),
olive oil (10%), honey (20%), psoraleac (5%), borax (10%), arnica (5%) and curcuma longa (1%) were used and
the wound healing potential was determined by both excision and incision wound healing models. Albino wistar
rats were used for this purpose. The wound area in excision wound model and tensile strength in incision wound
model were determined. Histopathological studies were also carried out to determine the phases of wound
healing.
Results: From the results obtained it was evident that the PUF has shown wound healing activity in both
excision and incision wound models. In the EWM the beneficial effect of the PUF was evident from the day of
first measurement as the progressive decrease in wound area was observed. On the 2 nd day of wounding, the
wound area shrinkage in 10% formulation treated group was more when compared to the other test groups. The
wound area was decreasing constantly and predominantly in all test treated groups as compared to the negative
control group. In the IWM there was decrease in tensile strength of the animals treated with the 10% formulation
and was comparable with the standard group. In the histopathological examination, the phases in the wound
healing process were observed and it showed that proliferation phase was the predominant stage of action of
PUF. The best re-modeling, particularly, re-epithelization were detected with the 10% formulation group On the
other hand; faster keratinization characterized with minor intraepithelial cornification was seen in 15% group.

KEYWORDS: Poly herbal Unani formulation (PUF), Excision wound model (EWM), Incision wound model
(IWM)

INTRODUCTION:
Several natural products which are composed of active
principles, like terpene alkaloids, flavonoids and
biomolecules have been reported to promote the process
of wound healing (Chitra et al., 1995). Herbs increase
the rate of tissue healing by providing different essential
Address for correspondence: substances, required at various steps of regeneration.
Syed Safiullah Ghori These herbs being cheaper and safer than allopathic
Anwarul Uloom College of Pharmacy, drugs may be useful in veterinary practice, especially in
New Mallepally, Hyderabad- 500001 Telangana, India India where these are found in plenty. Moreover the
current methods used to treat chronic wounds include
Received on 04.04.2015 Modified on 15.04.2015
Accepted on 28.04.2015 A&V Publications All right reserved
debridement, irrigation, antibiotics, tissue grafts and
Res. J. Pharm. Dosage Form. & Tech. 7(2): April-June, 2015; Page 125-128 proteolytic enzymes, which possess major drawbacks
DOI: 10.5958/0975-4377.2015.00018.X and unwanted side effects (Nayak et al, 2009).

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 Research Journal of Pharmaceutical Dosage Forms and Technology. 7(2): April-June, 2015, 125-128

The present study was designed the wound healing Experimental Design:
potential of a poly herbal formulation; beeswax (base), The animals were divided into 5 groups of 6 (n=6) rats
sesame oil (10%), olive oil (10%), honey (20%), each.
psoraleac (5%), borax (10%), arnica (5%), and Curcuma Group I- Negative Control: Ointment base
longa (1%). Group II- Standard: Soframycin
Group III- Test I: 5% w/w poly herbal formulation
MATERIALS AND PREPARATION: Group IV- Test II: 10% w/w poly herbal formulation
The formulation for assessment of wound healing Group V- Test III: 15% w/w poly herbal formulation
potential was procured from a local Ayurvedic
practitioner Dr. Mohammed Gouse. Excision Wound Model:
Animals were anesthetized prior to and during creation
Preparation of Test Formulation: of the wounds by diethyl ether inhalation chamber. The
Test formulation was taken in three different proportions rats were inflicted with excision wounds. The negative
(5%, 10% and 15%). Cetostearyl alcohol (2.5 g) was control group of animals was treated with ointment base,
melted on a water bath and liquid paraffin (25 g) was whereas the 5%, 10%, 15% extracts ointments are used
added and heated to 60oC. Test cream was dissolved in for 3 test groups, the reference drug (5% poly herbal
different proportions i.e. 5% (2.5 g), 10% (5 g), 15% (7.5 formulation) used for standard group and the wound area
g) in freshly boiled and cooled purified water and heated was determined on days 0, 2, 4, 8, 12 and 16 using
to 600C. This solution was added to the oily mixture and transparent paper and a marker. Change in wound area
stirred until cold. Cream was transferred to a suitable was calculated, giving an indication of the rate of wound
container and was stored for further use. contraction. The day of scar falling without any residual
raw wound were considered as period of epithelialization
Experimental Animals: (Rawat et al., 2011).
Adult male wistar rats weighing 150-200 gm were used
to evaluate wound healing activity. The animals were Experimental Design: The animals were divided into 5
maintained under standard laboratory conditions in groups of 6 (n=6) rats each.
polypropylene cages under 12 hours light/dark cycle, Group I- Negative Control: Ointment base
controlled temperature (242c), fed with commercial Group II- Standard: Soframycin
pellet diet and water ad-libitum in an animal house Group III- Test I: 5% w/w poly herbal formulation
approved by committee for the purpose and supervision Group IV- Test II: 10% w/w poly herbal formulation
on experiments on animals (1534/PO-/a/11/CPCSEA). Group V- Test III: 15% w/w poly herbal formulation
All the animals were acclimatized to the laboratory
environment for 10 hours before commencement of the Incision Wound Model
experiment. The experiments were carried out in All the animals were anaesthetized by chloroform
accordance with the instructions of Institutional Animal inhalation chamber and the back hair of the rats were
Ethical Committee, Anwarul Uloom College of shaved by using an electrical clipper. 5 cm long, two
Pharmacy, New Mallepally, Hyderabad 500001, linear-paravertebral incisions were made with a sterile
Telangana, India. surgical blade through the full thickness of the skin at a
distance of 1.5 cm from the midline of each side of the
METHODOLOGY: vertebral column (Ehrlich et al., 1968).
Draize test:
Male albino Wistar rats weighing (150-200 gm) with n= The ointments are applied once daily as per schedule i.e.
6 per group were used in skin irritation test. Two patches the negative control group of animals was treated with
each of two square inch area were prepared by shaving ointment base, whereas the 5%, 10%, 15% ointments
the dorsal surface of one rat. Patch made from two layers were used for 3 test groups, the reference drug
of light gauze was dipped in solutions containing (Safromycin) used for standard group, till 9 days. All the
different concentrations -0% (Control), 5%, 10% and sutures were removed on the 9th post wound day. On
15% of test extracts (PUF) prepared in PG: EtOH (7:3). Day 10 all the animals were killed under anesthesia. The
The animals were immobilized in the special holder Tensile Strength (in Newton) of one linear paravertebral
during the 24 hrs. patch exposure. Upon removal of the incised skin was measured using Tensiometer and
patches the animals were observed for any sign of average value was taken as the tensile strength and the
erythema or edema for a period of 72 hrs. The other paravertebral incised skin was taken carefully.
observations were repeated after 72 hrs. (Kaushal et al.,
2011).

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Experimental Design: RESULTS:

The animals were divided into 5 groups of 6 (n=6) rats Draize test:
each. No signs of allergy (allergic spots or redness of skin)
Group I- Negative Control: Ointment base were observed on rat's skin during the skin irritancy test.
Group II- Standard Safromycin There were no cases of wound infection in all the treated
Group III- Test I: 5% w/w poly herbal formulation groups.
Group IV- Test II: 10% w/w poly herbal formulation
Group V- Test III: 15% w/w poly herbal formulation Excision Wound Model
In the excision wound model the pharmacological
Statistical analysis: activity was observed and the wound healing potential of
The values were expressed as Mean SEM. P < 0.05 the poly herbal formulation has shown promising effects
was considered significant, denoted by symbol (*). as shown in the Table 1. The beneficial effect of poly
The data was analyzed by One-way Analysis of herbal formulation was evident from the day of first
Variance followed by Dunnetts multiple comparison measurement as the progressive decrease in wound area
post-hoc tests using Graph Pad Instat version 3.10 for was observed. On the 2nd day of wounding, the wound
Windows, Graph Pad Software, San Diego California area was decreasing constantly and predominantly in all
USA. test treated group as compared to the negative control
group. The P value was < 0.01, considered highly
significant. Variation among column means is
significantly greater than expected by chance.

Table 1: Effect Of PUF on wound healing activity using Excision wound model
Groups Day 2 Day 4 Day 6 Day 8 Period of Epithelization
-ve Control 468.4 7.30 200.7 14.43 159 2.93 192.1 20.78 18.3 1.14
Standard 496.9 3.15 268.0 7.50 221.7 2.52 136.4 3.94 12.8 0.94
Test I 297.2 0.48 267.6 0.50 191.1 0.32 213.3 0.25 18.1 0.94
Test II 296.3 0.96 254.1 0.34 174.9 0.48 165.48 0.36 20.5 0.76**
Test III 253.8 0.57 294.9 1.69 151.2 0.36 132.1 0.17** 18.3 0.88**
**P < 0.01

Figure 1: Effect of PUF on wound healing activity using Excision wound model

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Incision Wound Model ACKNOWLEDGMENT:

Poly herbal formulation has shown wound healing
activity in the incision wound model as shown in Table
2. The results of the measurements of tensile strength of
the animals treated with the 15% test were the next to the
highest value of the standard group. The P value of the
test treated groups was also P < 0.01, which is
considered as pharmacologically significant range.
Table 2: Effect of PUF on wound healing activity using Incision
wound model
Groups Tensile Strength
Negative Control 13.880.2587
Standard 28.760.2507**
Test I 21.050.3026**
Test II 27.440.3295**
Test III 27.500.2610**
**P<0.01
DISCUSSION:
Wound healing process begins with the restoration of a
damaged tissue as closely as possible to its natural state
and wound contraction is the course of shrinkage in
wounded area (Clark R.A 2001). The healing primarily
depends on the repairing ability of the tissue in addition
to type and degree of damage and general health status
of the tissue. The collagen is the main constituent of
extra cellular tissue, which is responsible for support and
strength. The poly herbal cream has shown considerably
good wound healing activity in Albino Wistar rats. The
poly herbal cream has been compared to that of standard
ointment and negative control groups. The ingredients
used in this poly herbal cream have been reported to
have wound healing activity and used traditionally.
The authors are thankful to the management of Anwarul
Uloom College of Pharmacy (affiliated to Jawaharlal
Nehru Technological University, Hyderabad) for
providing facilities to perform animal experimentation.
REFERENCES:
1. Chithra P, Suguna L, Chandrakasan G. (1995). Influence of
Arginine wound healing in rats. Journal of Clinical Biochemistry
and Nutrition 18, 111-117.
2. Nayak B.S, Vinutha B. Geetha B, and Sudha, B, Experimental
evaluation of Pentas lanceolata flowers for wound healing
activity in rats, Fitoterapia, vol. 76, no. 7-8, pp. 671675, 2005
3. Kaushal N, Naz S, Tiwary AK. (2011). Angelica archengelica
extract induced perturbation of rat skin and tight junctional
protein (ZO-1) of HaCaT cells. DARU: Journal of
Pharmaceutical Sciences 19, 1-11
4. Swati Rawat and Akhilesh Gupta. Development and study of
wound healing activity of an Ayurvedic formulation. Asian J.
Res. Pharma. Sciences 2011; Vol 1(1):26-28
5. Clark RA, Fibrin and wound healing. Ann N Y Acad Sci, 2001;
936: 355-67.
6. Ehrlich HP, Hunt TK. Effect of cortisone and vitamin A on
wound healing. Ann Surg. 1968; 167:3248

The measurements of the progression of wound healing

induced by the test cream, reference drug, negative
control groups were observed in this studies. In the
excision wound model, the 10% test cream to treated
groups of animals showed 56.5% contraction on the
wounds at Day 6. The same test cream demonstrated
80.6% contraction on the day 12, and healed completely
on day 14 which was close to contraction value of the
reference drug soframycin. However, the 5%
concentration of test cream presented no significant
results. Further, upon increasing the concentration to
15% of extract did not show any increase in contraction
value. In the incision wound model measurements of
tensile strength (in Newtons) showed significant results.
Tensile strength of the animals treated with the 10% test
cream demonstrated the highest value (38.9%) at day 10.
Topical application of the 10% test on the incision
wound model demonstrated a remarkable improvement
in wound tensile strength compared to other groups.

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