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1. Research Journal of Pharmaceutical Dosage Forms and Technology. 7(2): April-June, 2015, 125-128
125
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 2nd
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)
Address for correspondence:
Syed Safiullah Ghori
Anwarul Uloom College of Pharmacy,
New Mallepally, Hyderabad- 500001 Telangana, India
Received on 04.04.2015 Modified on 15.04.2015
Accepted on 28.04.2015 ©A&V Publications All right reserved
Res. J. Pharm. Dosage Form. & Tech. 7(2): April-June, 2015; Page 125-128
DOI: 10.5958/0975-4377.2015.00018.X
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
substances, required at various steps of regeneration.
These herbs being cheaper and safer than allopathic
drugs may be useful in veterinary practice, especially in
India where these are found in plenty. Moreover the
current methods used to treat chronic wounds include
debridement, irrigation, antibiotics, tissue grafts and
proteolytic enzymes, which possess major drawbacks
and unwanted side effects (Nayak et al, 2009).

2. Research Journal of Pharmaceutical Dosage Forms and Technology. 7(2): April-June, 2015, 125-128
126
The present study was designed the wound healing
potential of a poly herbal formulation; beeswax (base),
sesame oil (10%), olive oil (10%), honey (20%),
psoraleac (5%), borax (10%), arnica (5%), and Curcuma
longa (1%).
MATERIALS AND PREPARATION:
The formulation for assessment of wound healing
potential was procured from a local Ayurvedic
practitioner Dr. Mohammed Gouse.
Preparation of Test Formulation:
Test formulation was taken in three different proportions
(5%, 10% and 15%). Cetostearyl alcohol (2.5 g) was
melted on a water bath and liquid paraffin (25 g) was
added and heated to 60o
C. Test cream was dissolved in
different proportions i.e. 5% (2.5 g), 10% (5 g), 15% (7.5
g) in freshly boiled and cooled purified water and heated
to 600
C. This solution was added to the oily mixture and
stirred until cold. Cream was transferred to a suitable
container and was stored for further use.
Experimental Animals:
Adult male wistar rats weighing 150-200 gm were used
to evaluate wound healing activity. The animals were
maintained under standard laboratory conditions in
polypropylene cages under 12 hours light/dark cycle,
controlled temperature (24±2˚c), fed with commercial
pellet diet and water ad-libitum in an animal house
approved by committee for the purpose and supervision
on experiments on animals (1534/PO-/a/11/CPCSEA).
All the animals were acclimatized to the laboratory
environment for 10 hours before commencement of the
experiment. The experiments were carried out in
accordance with the instructions of Institutional Animal
Ethical Committee, Anwarul Uloom College of
Pharmacy, New Mallepally, Hyderabad 500001,
Telangana, India.
METHODOLOGY:
Draize test:
Male albino Wistar rats weighing (150-200 gm) with n=
6 per group were used in skin irritation test. Two patches
each of two square inch area were prepared by shaving
the dorsal surface of one rat. Patch made from two layers
of light gauze was dipped in solutions containing
different concentrations -0% (Control), 5%, 10% and
15% of test extracts (PUF) prepared in PG: EtOH (7:3).
The animals were immobilized in the special holder
during the 24 hrs. patch exposure. Upon removal of the
patches the animals were observed for any sign of
erythema or edema for a period of 72 hrs. The
observations were repeated after 72 hrs. (Kaushal et al.,
2011).
Experimental Design:
The animals were divided into 5 groups of 6 (n=6) rats
each.
Group I- Negative Control: Ointment base
Group II- Standard: Soframycin
Group III- Test I: 5% w/w poly herbal formulation
Group IV- Test II: 10% w/w poly herbal formulation
Group V- Test III: 15% w/w poly herbal formulation
Excision Wound Model:
Animals were anesthetized prior to and during creation
of the wounds by diethyl ether inhalation chamber. The
rats were inflicted with excision wounds. The negative
control group of animals was treated with ointment base,
whereas the 5%, 10%, 15% extracts ointments are used
for 3 test groups, the reference drug (5% poly herbal
formulation) used for standard group and the wound area
was determined on days 0, 2, 4, 8, 12 and 16 using
transparent paper and a marker. Change in wound area
was calculated, giving an indication of the rate of wound
contraction. The day of scar falling without any residual
raw wound were considered as period of epithelialization
(Rawat et al., 2011).
Experimental Design: The animals were divided into 5
groups of 6 (n=6) rats each.
Group I- Negative Control: Ointment base
Group II- Standard: Soframycin
Group III- Test I: 5% w/w poly herbal formulation
Group IV- Test II: 10% w/w poly herbal formulation
Group V- Test III: 15% w/w poly herbal formulation
Incision Wound Model
All the animals were anaesthetized by chloroform
inhalation chamber and the back hair of the rats were
shaved by using an electrical clipper. 5 cm long, two
linear-paravertebral incisions were made with a sterile
surgical blade through the full thickness of the skin at a
distance of 1.5 cm from the midline of each side of the
vertebral column (Ehrlich et al., 1968).
The ointments are applied once daily as per schedule i.e.
the negative control group of animals was treated with
ointment base, whereas the 5%, 10%, 15% ointments
were used for 3 test groups, the reference drug
(Safromycin) used for standard group, till 9 days. All the
sutures were removed on the 9th post wound day. On
Day 10 all the animals were killed under anesthesia. The
Tensile Strength (in Newton) of one linear paravertebral
incised skin was measured using Tensiometer and
average value was taken as the tensile strength and the
other paravertebral incised skin was taken carefully.

3. Research Journal of Pharmaceutical Dosage Forms and Technology. 7(2): April-June, 2015, 125-128
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Experimental Design:
The animals were divided into 5 groups of 6 (n=6) rats
each.
Group I- Negative Control: Ointment base
Group II- Standard Safromycin
Group III- Test I: 5% w/w poly herbal formulation
Group IV- Test II: 10% w/w poly herbal formulation
Group V- Test III: 15% w/w poly herbal formulation
Statistical analysis:
The values were expressed as Mean ± SEM. P < 0.05
was considered significant, denoted by symbol (*).
The data was analyzed by One-way Analysis of
Variance followed by Dunnett’s multiple comparison
post-hoc tests using Graph Pad Instat version 3.10 for
Windows, Graph Pad Software, San Diego California
USA.
RESULTS:
Draize test:
No signs of allergy (allergic spots or redness of skin)
were observed on rat's skin during the skin irritancy test.
There were no cases of wound infection in all the treated
groups.
Excision Wound Model
In the excision wound model the pharmacological
activity was observed and the wound healing potential of
the poly herbal formulation has shown promising effects
as shown in the Table 1. The beneficial effect of poly
herbal formulation was evident from the day of first
measurement as the progressive decrease in wound area
was observed. On the 2nd
day of wounding, the wound
area was decreasing constantly and predominantly in all
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

4. Research Journal of Pharmaceutical Dosage Forms and Technology. 7(2): April-June, 2015, 125-128
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Incision Wound Model
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 13.88±0.2587
Standard 28.76±0.2507**
Test I 21.05±0.3026**
Test II 27.44±0.3295**
Test III 27.50±0.2610**
**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 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 Newton’s) 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.
ACKNOWLEDGMENT:
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.
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