Pak. j. sci. ind. res. Ser. B: biol. sci.

2022 65B(1) 55-61

Wound Healing Potential/Activity of Polyherbal Ointment Containing

Salvadora persica, Azadirachta indica and Calendula officinalis
Extracts: An Experimental Study
Hina Imran*, Atiq-ur-Rehman, Tehmina Sohail, Sohail Shaukat and Ayesha Khokar
Pharmaceutical Research Centre, PCSIR Laboratories Complex, Karachi, Pakistan

(received December 14, 2018; revised July 16, 2019; accepted September 3, 2019)

Abstract. The present study explore the wound healing potential of a novel herbal ointment formulation,
fast wound healing ointment (FWHO) composed of 10% w/w crude ethanolic extracts of Salvadora persica
(sticks), Azadirachta indica (leaves) and Calendula officinalis (whole plant) on excision wound model in
albino rats. The FWHO was applied topically once daily on the wound to evaluate the rate of contraction
and period of epithelialization in rats. Complete contraction and epithelialization of wound was observed
at 19 day (P<0.0005) in test group animals compared with standard (polyfax skin ointment) at 23 day
(P<0.0005) and control group (blank petroleum jelly) at 29 day respectively. It was observed that 10%
FWHO has potent wound healing capability as evidenced by decrease in the period of epithelialization
and increase in the rate of wound contraction in test group as compared to the standard and control groups.
The test group illustrates 34.4%, while the standard group shows 20.6% decrease in the time of epithelialization
as compare to control group. The study revealed that test ointment (10% FWHO) has a potential to promote
wound healing by accelerating the rate of epithelialization and scar formation as compare to standard
(polyfax skin ointment) and control (blank petroleum jelly) groups.
Keywords: extracts of S. persica, A. indica, C. officinalis, excision wound model

Introduction ancient times due to their widespread availability,

Wounds and healing of wounds are two vital events in minimal toxicity and their effectiveness as crude
the life of animals and human. Wound is a physical preparations (Gautam et al., 2014).
trauma, where the skin is torn, cut, burnt or punctured Although many traditional healer have successfully
due to disruption of normal anatomic structure and used hundreds of medicinal plants to manage various
function. The most common symptoms of wounds are forms of health conditions (Wambugu and Waweru,
bleeding, loss of function, heat, redness around the 2016). Henceforth it was aimed to find out wound
wound, painful or throbbing sensation, swelling of local healing effects of FWHO which was composed of 10%
tissue and oozing from wound. The healing process is w/w crude extract of Salvadora persica (sticks),
the survival mechanism representing an attempt to Azadirachta indica (leaves) and Calendula officinalis
maintain normal anatomical structure and function. It (whole plant) to establish the scientific base for its use
is a natural body reaction against the injury which starts (Maan et al., 2017; Imran et al., 2015; Emeka et al.,
immediately after wounding and occurs in four phases 2013).
i.e. hemostasis, inflammation, proliferation and
maturation (Kumaran et al., 2016; Gupta et al., 2015). Salvadora persica (Salvadoracea) commonly known
A curative agent should ideally improve one or more as Miswak abundantly found in Asian countries, contains
phases of healing process without causing harmful important phyto-constituents such as vitamin C,
effects. The side effects of synthetic medicine are salvadorine, salvadourea, alkaloids, trimethylamine,
alarming and produce to numerous undesirable effects cyanogenic glycosides, tannins, saponins and salts
despite their strong pharmacological action (Parasuraman mostly as chlorides etc. Medicinally possesses anti-
et al., 2014). Today there is widespread interest is oxidant, antiseptic, antimicrobial, hemostatic, anti-
developed in producing plant derived drugs. The use plaque, analgesic, anti-inflammatory, anti-pyretic,
of plants as a whole or extracts of their certain parts to astringent and diuretic properties. It is well documented
accelerate the process of healing has been in use since that the promising wound healing activity of S. persica
*Author for correspondence; is due to the synergistic effect of both antimicrobial and
E-mail: [email protected] antioxidant activities which increase wound contraction

55
56 Hina Imran et al.

and accelerate the wound-healing process (Tatke et al., cut into small pieces and then finally chopped to make
2017; Imran et al., 2015; Ahmad et al., 2011). Calendula coarse powder. The extract was prepared by soaking
officinalis L. (Asteraceae) known as pot marigold or the equal weights i.e. 2 Kg of each material into 6 liters
common marigold is the herb of ancient time with of commercial ethanol and caped in screw tight separate
medicinal repute used in traditional and homeopathic bottles for one week with occasional shaking. After
medicine. Commonly it is used in first-aid treatment of filtration through muslin cloth and through Whatman
wound for reducing inflammation, as antiseptic to clean no. 1 filter paper, the filtrates were then evapourated
wound, as disinfectant to prevent infection and helps under reduce pressure (-760 mmHg) at 35-40 °C in a
to promote wound healing (Parente et al., 2012; Paul rotary evapourator. The thick semi-solid crude extracts
et al., 2017). It also increases blood flow and oxygen were collected in Amber glass bottles and stored in
to wounds and affected areas which in turns helps the refrigerator till use.
body to grow new tissue and healing of wound more Preparation of fast wound healing ointment (FWHO).
rapidly. The high content of flavonoids acts as The test sample (FWHO) was prepared by incorporating
antioxidants and protects the body cells from damage the active ingredients (ethanolic extracts of S. pesica,
caused by oxidation that may suppress the immune C. officinalis and A. indica) in the base petroleum jelly
function (Bernatoniene et al., 2011). Azadirachta indica by trituration using mortar and pestle in a concentration
(Meliaceae) known as Neem is a well reputed useful of 10% (w/w). The prepared ointment was filled in
medicinal plant with wide spectrum of biological sterilized airtight containers and stored at room
activities. The active ingredients like nimbidin, nimbin temperature.
and nimbidol are sufficiently found with anti-
inflammatory, antibacterial, antifungal and antiviral Selection of animal. Eighteen albino male rats (180-
properties (Alzohairy, 2016; Chhibber and Sharma, 200 g) reared at animal house of PCSIR Labs Complex,
2014). It is considered valuable in various skin problems Karachi were selected and grouped accordingly for
like itching, burning and skin ulcer. A. indica has an wound healing activity. Animals were housed and
excellent amount of amino acids, vitamins and minerals maintained under standard conditions and fed with
that are playing very important role in the proliferation standard diet and water ad libitum. They were kept
phase of wound healing process (Chundran et al., 2015; carefully following acclimatization period of 7 days to
Kumari and Keshri, 2015). At present more than 50% ensure their suitability for the study. Animals showing
of clinically used drugs are plant origin (Vamsi et al., any signs of illness were excluded from the study.
2014). This interest primarily stems from the belief that Wound healing activity. Excision wound model was
green medicines are commonly safe and dependable as used for the study of rate of contraction and
compared to costly synthetic drugs. These herbal drugs epithelialization of wound. Animals were anaesthetized
are considered as a therapeutic weapon to fight against with diethyl ether and a circular wound of about 150
various ailments/diseases in birds, humans and animals mm2 was made on depilated dorsal thoracic region of
with less or without any side effects. Herbal drugs are rats. Wounded areas were measured immediately by
boon to our society. This present study is majorly aimed means of Vernier Caliper and consider as initial wound
to determine the effect of FWHO on the rate of wound area reading. Treatment with the ointments started
healing (percent wound contraction and period of immediately after wound creation. Animals of Group
epithelialization) in excision wounds in Wistar albino I were treated with thin layer of (10% FWHO) as test
rat model. drug, Group II was treated with thin layer of (polyfax
skin ointment) as reference drug, while Group III was
Material and Methods treated with thin layer of blank petroleum jelly as control
Preparation of extracts. The specified portion of the respectively. The entire wound was left open. All samples
plants i.e. S. persica (twigs) was purchased from local were applied topically once daily until the achieving of
medicinal market of Karachi, whereas A. indica (leaves) complete healing of wound. The observation was made
and C. officinalis (whole plant) were collected from every 3 rd day of post wounding day. The wound
garden of PCSIR Labs. Complex, Karachi, Pakistan. contraction was calculated as percentage reduction in
All parts were air dried under the shade at room wound area with respect to initial wound area, while
temperature in controlled room environment until the the epithelialization time was noted as the number of
material gets completely dried. Each plant sample was days after wound required for scar to fall off leaving
Wound Healing Potential: An Experimental Study 57

no raw wound behind. Rate of wound contraction was of standard and control groups (Table 1, Fig. 2). FWHO
calculated by the formula given below (Nguyen et al., also demonstrated the better healing capacity as it
2017; Imran et al., 2015; Kodati et al., 2011); completely healed the wound within the period of 19
days (P<0.0005) in animals of test group whereas
Percent wound contraction = wounds treated with Polyfax skin ointment showed
wound area at day 0 - wound area at day n
___________________________________ similar effects within 23 days (P<0.0005) in standard
x100
wound area at day 0 group as compared to control group which showed
complete wound healing and epithelialization within
Statistical analysis. All the results obtained were
29 days. During the excision wound healing study, it
analyzed statistically by using Student's t test and P<0.05
was observed that FWHO not only effectively and
were considered significant.
promptly heals wounds by inducing fast epithelialization
but also prevents infection because of its antiseptic
Results and Discussion effects like with polyfax skin ointment.
The studies on excision wound healing model reveals
Wound healing is a complex process aiming at the re-
that all three groups showed decrease in wound area
construction of damage tissue which requires precise
from day one to last day. Rate of wound contraction in
coordination of connective tissue repair, re-
control rats was 33.3% to 68.8% from day 3 to day 9,
epithelialization and angiogenesis. Healing of wound
77.7% to 92.4% from day 12 to day 18 and 96% to
and generation of new tissue require the proliferation
98.2% from day 21 to day 27. The mean time for
of fibroblasts not only to increase cell numbers but also
complete normalization of skin in control group animals
to produce several extra cellular matrix proteins and
was noted on day 29 shown in Fig. 1. FWHO showed
growth factors (Jettanacheawchankit et al., 2009). Plants
time dependent healing effect on wound surface area.
are more potent healers as they promote the repair
Increase in wound contraction in treated group was
mechanism in the natural way and function as efficacious
43.1% to 84% from day 3 to day 9 and 94.6% to 100%
natural antibiotics. The plant base materials are used in
from day 12 to day 18, whereas in standard group 41.7%
first aid for wound wash. They act as antiseptic,
to 81.3% from day 3 to day 9 and 92% to 97.7% from
antibiotics, coagulants, vasodilators and antioxidants
day 12 to day 18, while 100% contraction was noted
by enhancing the healing of acute as well as chronic
on day 19 (P<0.0005) respectively as compared to
wounds due to the presence of phytochemical agents
control group. The data obtained indicates that there
and vitamins (Pereira and Bártolo, 2016). Properly
is significant promotion in wound-healing activity in
prepared herbal formulations/products act as potent and
the animals of test group treated with FWHO which
effective medicines and generate the desired effects
showed faster rate of epithelialization than in the animals
without numerous adverse side effects generally
associated with synthetic antibiotics (Wachtel-Galor
and Benzie, 2011). The aim of present study is to
formulate and evaluate a new successful poly herbal
remedy containing necessary natural supplements and
other physiological active compounds that are essential
for healthy skin, for its nourishment and to promote
prompt wound healing in short period of time (Ahmad
et al., 2014; Meena et al., 2009). Fast wound healing
ointment (FWHO) is a poly herbal preparation containing
plant extracts of S. persica, A. indica and C. officinalis
as active agents, while petrolatum jelly is used as a
base. Petroleum jelly is a mixture of natural
hydrocarbons and mineral oils obtained from
petroleum. It locks skin moisture and act as lubricant
that softens and soothes the skin (Park and Song, 2010).
Fig. 1. Excision wound on back of neck of albino The preparation formulated by using these herbs because
rat on day 1st. they have strong evidences to have anti-inflammatory,
58 Hina Imran et al.

immuno-modulatory, antimicrobial, anti-oxidantand Table 1. Percent wound healing activity of FWHO in

and analgesic activities respectively which are excision wound model
complementary to wound healing process (Talha Bin Animal % of wound healing
Emran, 2015; Khatkar et al., 2014; Ahmad et al., 2011; groups 3rd 6th 9th 12th 15th 18th 21st 24th 27th 30th
Mariod et al., 2009; Biswas et al., 2002). Polyfax skin day day day day day day day day day day
ointment was used as a standard ointment, contains (1 Test 43.5 64 85 95 98.9 99.7 100 100 100 100
g contains: polymyxin B 10000 IU, bacitracin 500 IU). group
Polymyxin and bacitracin ointment (Polyfax®) is used Standard 42.1 67.1 81.3 91.9 95.7 98.2 99.1 100 100 100
group
to treat skin infections caused by bacteria. It act as a
Control 30.3 49.4 65.1 76.5 85.3 92 96 98.5 99.7 100
topical antibiotic commonly used to treat wounds, burns, group
skin grafts, skin ulcers, itching and rashes
(Glaxosmithkline Pakistan Limited). Number of animals=6

In the excision wound model, animals treated with

FWHO showed better and fast healing as compared to 29 days***
the standard and control groups (Fig. 1). Also there was 23 days***
significant decrease in the epithelialization period. It
19days
was observed that FWHO treated group showed
considerable wound healing activity from the third day
onwards, which was comparable to the standard drug
polyfax skin ointment and blank petroleum jelly for
control groups. The percentage of wound contraction
was found much more in FWHO treated group (19 days
for 100% contraction) which was more than that of
standard group and control group animals i.e. 23 and
Test group Standard group Control group
29 days for 100% contraction respectively (Table 1,
Fig. 2). The healing time of test and standard group
Fig. 2. Mean healing time in days of FWHO in
was also compared with control group and it was
excision wound model.
observed that the test group had 34.4% ability to decrease
the time of epithelialization, while 20.6% decrease time
of epithelialization was observed in standard verses epithelialization in shortest period of time (Talekar et
control group. On the basis of the result obtained in the al., 2017; Imran et al., 2015; Khatak et al., 2010). A.
present investigation, we can conclude easily that the Indica is reported for fibroblast proliferation, collagen
FWHO has remarkable wound healing activity. The synthesis and neo-vascularization that resulted in an
better activity of polyherbal formulation may be due to increased wound tensile strength and accelerate wound
the synergistic action of the plants constituents present healing (Rajasree et al., 2012). The amino acids, vitamins
in the formulation like alkaloids, saponins, tannins, and minerals present in A. indca also play important
flavonoids, curcumin and triterpinoids (Vamsi et al., role in proliferation phase of wound healing processes
2014). S. persica is a well reputed plant with properties (Emeka et al., 2013). Its antimicrobial, antioxidant and
that are needed for wound healing like analgesic, anti- anti-inflammatory properties accelerates healing process
inflammatory, antioxidant, antiseptic and antimicrobial by preventing the infection and speeding up the
along with hemostatic activities. Hemostasis plays a proliferation phase in the entire wound healing process
vital role in healing process as it arrest the bleeding (Chundran et al., 2015; Raina et al., 2008). A study
from damaged blood vessel, initiate tissue repair and conducted by Shafeie et al. (2015) reported that topical
prevents tissue death through hemorrhage (Imran et al., application of Calendula officinal is gel contributions
2015; Ahmed et al., 2011). It is also reported that to the reduction of edema and the regulation of micro
phytochemicals (flavonoids, saponin, sterol, tannins) circulation. The polysaccharides in Calendula flowers
and mineral salts (copper, manganese, zinc, vitamins also demonstrate strong bio adhesive or mucilaginous
and amino acids etc.) found in S. persica which also effect may not only help to decrease local inflammation
help to promote wound repair by increasing the rate of by shielding tissues from irritants, but also facilitate
Wound Healing Potential: An Experimental Study 59

tissue hydration (Ahmed et al., 2003). Calendula may present formulation is found very effective in wound
also facilitate wound healing by increasing wound healing by showing quick response following its
angiogenesis and collagen, nucleoprotein and application on wounds. It can be concluded that the
glycoprotein metabolism leading to improvements in FWHO is a valuable herbal product which may be
both local circulation and in the formation of granulation effectively used for the treatment and management of
tissue. Several other experimental studies also support wounds due to its promising quick healing property.
to these claims and demonstrate that the daily application
of calendula cream or a 1:10 alcoholic extract of Conflict of Interest. The authors declare no conflict
Calendula to paravertebral incisions in rats facilitates of interest.
collagen maturation and epithelialization within 10 to
References
25 days (Leach, 2008). All these studies strongly support
our findings. Ahmad, M., Imran, H., Yaqeen, Z., Zakir-ur-Rehman,
Atiq-ur-Rahman, Fatima, N., Sohail, T. 2011.
The results of this study indicate that daily topical Pharmacological profile of Salvadora persica.
application of FWHO significantly accelerated the rate Pakistan Journal of Pharmaceutical Sciences, 24:
of wound healing. The potency and powerful healing 323-330.
action of FWHO could be due to the combined action Ahmad, S., Bibi, A., Ishaq, M.S., Afridi, M.S., Kanwal,
of phyto-constituents that accelerates cell reproduction, F., Zakir, M., Fatima, F. 2014. Phytochemical
tissue regeneration and simultaneously depressing the analysis, antioxidant activity, fatty acids composition
inflammatory process along with providing local and functional group analysis of Heliotropium
analgesic, antibacterial, antifungal and astringent effects bacciferum. Scientific World Journal, 2014: 1-8.
(Thangapazham et al., 2016; Manjunatha et al., 2006). Ahmed, S., Rahman, A., Qadiruddin, M., Qureshi, S.
Furthermore, the wound healing effect can be attributed 2003. Elemental analysis of Calendula officinalis
to free radical scavenging activity of flavonoids and plant and its probable therapeutic role in health.
triterpenoids. They are known to reduce lipid Pakistan Journal of Scientific and Industrial
peroxidation, not only by preventing or slowing the Research, 46: 283-287.
onset of cell necrosis, but also improving vascularity. Alzohairy, M.A. 2016. Therapeutics role of Azadirachta
It is documented that any drug that inhibits lipid indica (Neem) and their active constituents in
peroxidation can increase the viability of collagen fibrils, diseases prevention and treatment. Evidence-Based
which in turn results in increase in the strength of Complementary and Alternative Medicine, 2016:
collagen fiber by increasing the circulation, preventing 1-11.
the cell damage and promoting DNA synthesis (Rekha Biswas, K., Chattopadhyaym, R.K., Banerjee, U.,
et al., 2015). Roy et al. (2012) reported a clinical study Bandyopadhyay, U. 2002. Biological activities and
that topical application of compounds with free radical medicinal properties of neem (Azadirachta indica).
scavenging properties showed improve wound healing Current Science, 82: 1336-1345.
significantly and protect tissues from oxidative damage. Bernatoniene, J., Masteikova, R., Davalgiene, J., Peciura,
In epithelialization of wound and chemotaxis in fibrosis R., Gauryliene, R., Bernatoniene, R., Majiene, D.,
alkaloids are also known to play a pivotal role (Fetse Lazauskas, R., Civinskiene, G., Velziene, S.,
et al., 2014). All these reports strengthen our results. Muselik, M., Chalupova, Z. 2011. Topical
On the basis of wound healing studies it can be application of Calendula officinalis (L.): formulation
summarized that prepared ointment possesses superior and evaluation of hydrophilic cream with
wound healing contracting ability than the standard and antioxidant activity. Journal of Medicinal Plants
control groups. Thus, the prepared topical ointment is Research, 5: 868-877.
effective in terms of acceptability and possesses a Chhibber, S., Sharma, N. 2014. Medicinal and
versatile approach in healing the wound within short therapeutical potential of neem (Azadirachta
period of time. indica): a review. International Journal of
Scientific and Research Publications, 4: 1-5.
Conclusion Chundran, N.K., Husen, I.R., Rubianti, I. 2015. Effect
The aim of present investigation is to formulate a natural, of neem leaves extract (Azadirachta indica) on
safe, indigenous herbal wound healing remedy. The wound healing. Althea Medical Journal, 2: 199-
60 Hina Imran et al.

203. Khatak, M., Khatak, S., Siddqui, A.A., Vasudeva, N.,

Emeka, A.O., Emamoke, J.O., Theodore, A.A., Julius, Aggarwal, A., Aggarwal, P. 2010. Salvadora persica
C.O. 2013. The wound healing effects of aqueous (short review). Pharmacognosy Reviews, 4: 209-
leave extracts of Azadirachta indica on Wistar rats. 214.
Journal of Natural Sciences Research, 3: 181-186. Leach, M.J. 2008. Calendula officinalis and wound
Fetse, J.P., Kyekyeku, J.O., Dueve, E., Mensah, K.B. healing: a systematic review. Wounds, 20: 236-43.
2014. Wound healing activity of total alkaloidal Meena, A.K., Bansal, P., Kumar, S. 2009. Plants-herbal
extract of the root bark of Alstonia boonei wealth as a potential source of ayurvedic drugs.
(Apocynacea). British Journal of Pharmaceutical Asian Journal of Traditional Medicines, 4: 152-
Research, 4: 2642-2652. 70.
Gautam, M.K., Purohit, V., Agarwal, M., Singh, A., Mariod, A.A., Mattaus, B., Hussein, I.H. 2009. Chemical
Goel, R.K. 2014. In vivo healing potential of Aegle characterization of the seed and antioxident activity
marmelos in excision, incision and dead space of various parts of S. persica. Journal of the
wound models. The Scientific World Journal, 2014: American Oil Chemists’ Society, 86: 857-865.
1-9. Maan, P., Yadav, K.S., Yadav, N.P. 2017. Wound healing
Gupta, A., Verma, S., Gupta, A., Jangra, M., Pratap, R. activity of Azadirachta indica A. Juss stem bark
2015. Evaluation of Prosopis cineraria (Linn.) in mice. Pharmacognosy Magazine, 13: 316-320.
Druce leaves for wound healing activity in rats. Manjunatha, B.K., Vidhya, S.M., Krishna, V., Mankani,
Annals of Pharma Research, 3: 70-74. K.L. 2006. Wound healing activity of Leucas hirta.
Imran, H., Ahmad, M., Rahman, A., Yaqeen, Z., Sohail, Indian Journal of Pharmaceutical Sciences, 68:
T., Fatima, N., Iqbal, W., Yaqeen, S.S. 2015. 380-384.
Evaluation of wound healing effects between Nguyen, M.C., Le, D.T., Kamei, K., Dang, T.P.T. 2017.
Salvadora persica ointment and Solcoseryl jelly Wound healing activity of Streptocaulon juventas
in animal model. Pakistan Journal of Pharma- root ethanolic extract. Wound Repair and
ceutical Sciences, 28: 1777-1780. Regeneration, 25: 956-963.
Jettanacheawchankit, S., Sasithanasate, S., Sangvanich, Parente, L.M.L., Junior, R.S.L., Tresvenzol, L.M.F.,
P., Banlunara, W., Thunyakitpisal, P. 2009. Vinaud, M.C., Paula, J.R., Paulo, N.M. 2012.
Acemannan stimulates gingival fibroblast Wound healing and anti-inflammatory effect in
proliferation; expressions of keratinocyte growth animal models of Calendula officinalis L. growing
factor-1, vascular endothelial growth factor, and in Brazil. Evidence-Based Complementary and
type I collagen; and wound healing. Journal of Alternative Medicine, 2012: 1-7.
Pharmacological Sciences, 109: 525-531. Park, E.-K., Song, K.-W. 2010. Rheological evaluation
Kumaran, P.S., Sumathi, A., Shanthini, K.R., Mallika, of petroleum jelly as a base material in ointment
J. 2016. Wound healing activity of methanolic and cream formulations: steady shear flow
extract of the peel of Abelmoschus esculents. behaviour. Archives of Pharmacal Research, 33:
European Journal of Pharmaceutical and Medical 141-150.
Research, 3: 208-212. Paul, S., Rahman, A.N.M.A., Al Mahmud, M.A., Uzzal,
Kumari, M., Keshri, U.S.P. 2015. Wound healing effect A.R., Das, Z.C. 2017. Wound healing by marigold
of Azadirachta indica and Curcuma longa in guinea (Calendula officinalis) and turmeric (Curcuma
pigs. Scholars Bulletin, 1: 271-275. longa) paste: a comparative approach. Journal of
Kodati, D.R., Goud, P.K., Burra, S., Galipelly, S.K. Advance Veterinary and Animal Research, 4: 333-
2011. Evaluation of wound healing activity of 342.
methanolic root extract of Plumbago zeylanica L. Parasuraman, S., Thing, G.S., Dhanaraj, S.A. 2014.
in wistar albino rats. Pelagia Research Library, 2: Polyherbal formulation: concept of ayurveda.
239-248. Pharmacognosy Review, 8: 73-80.
Khatkar, S., Dhiman, P., Sachin, M.N., Khatkar, A., Pereira, R.F., Bártolo, P.J. 2016. Traditional therapies
Redhu, N. 2014. Biological and medicinal properties for skin wound healing. Advances in Wound Care,
of Azadirachta indica: a review. International 5: 208-229.
Journal Pharma Professional’s Research, 5: 1054- Rekha, T.R., Kashmira, J.G., Poonam, S., Surendra, S.
1060. 2015. Development of wound healing herbal
Wound Healing Potential: An Experimental Study 61

formulation herbal wound guard. International Parab, P.B. 2017. Studies on wound healing
Journal of Scientific and Research Publications, potential of polyherbal formulation using in vitro
5: 1-5. and in vivo assays. Journal of Ayurveda and
Rajasree, P.H., Vishwanad, V., Cherian, M., Eldhose, Integrative Medicine, 8: 73-81.
J., Singh, R. 2012. Formulation and evaluation of Tatke, P., Nehete, M., Gabhe, S. 2017. Evaluation of
antiseptic polyherbal ointment. International antioxidant, antimicrobial and wound healing
Journal of Pharmacy and Life Sciences, 3: 2021- potential of Salvadora persica twig extracts. World
2031. Journal of Pharmaceutical Research, 6: 1186-
Roy, P., Amdekar, S., Kumar, A., Singh, R., Sharma, 1199.
P., Singh, V. 2012. In vivo antioxidative property, Thangapazham, R.L., Sharad, S., Maheshwari, R.K.
antimicrobial and wound healing activity of flower 2016. Role of phytochemicals in skin regeneration.
extracts of Pyrostegia venusta (Ker Gawl) Miers. Advances in Wound Care, 5: 230-241.
Journal of Ethnopharmacology, 140: 186-192. Wachtel-Galor, S., Benzie, I.F.F. 2011. An introduction
Raina, R., Prawez, S., Verma, P.K., Pankaj, N.K. 2008. to its history, usage, regulation, current trends and
Medicinal plants and their role in wound healing. research needs. In: Herbal Medicine: Biomolecular
Online Veterinary Journal, 3: 1-7. and Clinical Aspects, Benzie, I.F.F., Wachtel-Galor,
Shafeie, N., Naini, A.T., Jahromi, H.K. 2015. S., 2nd edition, CRC Press/Taylor & Francis, Boca
Comparison of different concentrations of Raton, Florida, USA.
Calendula officinalis gel on cutaneous wound Vamsi, S., Satish, C., Nagaveni, K., Jyothi, M.J., Latha
healing. Biomedical and Pharmacology Journal, P. 2014. Formulation and evaluation of polyherbal
8: 979-992. wound healing ointment. International Journal of
Talha Bin Emran, Uddin, M.M.N., Rahman, M.A., Pharma Research & Review, 3: 66-73.
Uddin, M.Z., Islam, M.M. 2015. Phytochemical, Wambugu, F.K., Waweru, W.R. 2016. Evaluation of
antimicrobial, cytotoxic, analgesic and anti- wound healing activity of ethanolic extract of leaves
inflammatory properties of Azadirachta indica: a of croton megalocarpus using excision wound
therapeutic study. Journal of Bioanalysis and model on Wistar albino rats. International Journal
Biomedicine, 12: 1-7. of Science and Research Methodology, 4: 182-
Talekar, Y.P., Apte, K.G., Paygude, S.V., Tondare, P.R., 194.