Chinese Medicine, 2011, 2, 138-142
doi:10.4236/cm.2011.24023 Published Online December 2011 (
Copyright © 2011 SciRes. CM
Comparative Efficacy of Tagetes erecta and Centella
asiatica Extracts on Wound Healing in Albino Rats
Sandipan Chatterjee1, Tigari Prakash1*, Dupadahalli Kotrsha2, Nadendla Rama Rao3, Divakar Goli1
1Department of Phar macology, Acharya & B M Reddy College of Pharmacy, Bangalore, India
2Department of Parasi t ology, Faculty of Medicine, University of Malaysia, Kuala Lumpur, Malaysia
3Department of Pharmaceutical Chemistry, Chalapathi Institute of Pharmaceutical Science,
Guntur, India
Received July 20, 2011; revised August 22, 2011; accepted August 29, 2011
The study was undertaken to evaluate the comparative efficacy of hydroalcoholic extracts of leaves of Ta-
getes erecta (T. erecta) and aerial parts of Centella asiatica (C. asiatica) on Excision, Incision and Dead
space wound models in albino rats. Extract of T. erecta and C. asiatica (P < 0.001) showed significant in-
crease in rate of wound contraction, epithelization and formation of scar faster in excision wound model
compare to control group. T. erecta extract (250 and 500 mg/kg) showed significantly increased the wound
breaking strength in incision wound model and wet and dry granulation tissue weights, breaking strength in a
dead space wound model compare to control and C. asiatica treated group (P < 0.001). In this investigation
the work is conclude that the comparison made between the above two plants, the T. erecta extract showed
potent wound healing activity then the reported C. asiatica in different wound parameters.
Keywords: Centella asiatica, Tagetes erecta, Excision, Incision, Dead Space Wound Model
1. Introduction
Wound healing or wound repair, is the body’s natural pr-
ocess. Have regenerating dermal and epidermal tissues.
When an individual is wounded, a set of events takes
place in a predictable fashion to repair the damages. Wo-
unds were probably the first medical problem faced by
human race. The field of pharmacologic modulation in
wound healing is still in its infancy. Indians were acqu-
ainted with a far larger number of medicinal plants than
the natives of other country. In fact ancient Indian sur-
geon sushruta has enumerated large number of herbal dr-
ug in the management of wounds singly or in combina-
tion. Some Ayurvedic medicinal plants, namely, Centella
asiatica, Curcuma longa, Aloe vera, and Tagetes erecta
Linn, were found to be effective on wound healing in ex-
perimental models.
Tagetes erecta (T. erecta) is commonly known as Ma-
rigold. The plant is widely distributed throughout India,
Asia, Central Europe, USA and Africa. The leaves are
reported to be effective against piles, kidney troubles,
muscular pain, ulcers, wounds and earache. The pounded
leaves are used as an external application to boils and
carbuncles. The steam distillation of fresh leaves offer
0.3% of essential oil with a strong, sweet lasting odor
and contains d-limonene, ocimene, l-linalyl acetate, l-li-
nalool tagetone, n-nonyl aldehyde, lutein [1].
Centella asiatica (C. asiatica) is commonly known as
Indian pennywort. The plant is widely distributed in Asia,
Africa, and North and South America [2]. This plant is
reportedly useful in wound healing and protecting ag-
ainst ulcer formation and is reported to have antimicro-
bial properties. Extracts of the plant have been shown to
accelerate wound healing through topical and systemic
routes [3]. The extracts of C. asiatica plant are reported
to have sedative, antidepressant, analgesic, and anticon-
vulsive effects [4]. It is also reputed within the Indian
system of medicine for its use in treating chronic and ob-
stinate eczema, psoriasis, syphilis and leprosy [5]. The
titrated extract of C. asiatica contains three principal
ingredients like asiaticoside, asiaticacid, and madecassic
acid. All these ingredients are known to be clinically
effective in the treatment of systemic scleroderma, ab-
normal scar formation [6] and wound-healing activity [7,
8]. The present study was planned to evaluate the com-
parative efficacy of the hydroalcoholic extract of aerial
parts of leaves of T. erecta and C. asiatica on different
wound models.
2. Materials and Methods
2.1. Plant Materials
Leaves of T. erecta and arial part of C. asiatica was col-
lected during the month of November from the local ar-
eas of Burdwan District, West Bengal, India and both the
plants were authenticated by Professor Udayan Sarkar,
HOD, Dept of Botany, Sonamukhi College, Bankura, In-
dia. A voucher specimen (Reference no-SC341/06) of
plants has been deposited in the department.
2.2. Preparation of Hydro Alcoholic Extracts
The plants material was cleaned in water and air dried,
powder was made using an electric mixer. The powder
(75 g) was packed into a Soxhlet apparatus and was ex-
tracted with 500 ml of hydro alcoholic solvent (350 ml
alcohol-95% and 150 ml water) at 65˚C to 70˚C for 24 h.
The extracts were dried under vacuum and weights of the
dried mass were recorded (Yield: C. asiatica-10% and T.
2.3. Animals
Healthy Wistar albino rats of either sex weighing be-
tween 180 - 250 g were used for the wound healing study
and albino mice weighing between 25 - 30 g were used
for the oral acute toxicity studies. The animals were
housed individually in Poly propylene cages and main-
tained at 24˚C ± 2˚C, a 12 h light/dark cycle fed with
standard pellet rat chow (Pranav agro industries Ltd.
Sangli, India) and water ad libitum. The experimental
protocol and procedures used in this study were ap-
proved by the Institutional Animal Ethics Committee
(Protocol no: IAEC /PP/06/07).
2.4. Toxicity Studies
The female albino mice weighing 20 - 25 g were used for
the study. The acute oral toxicity studies of hydro alco-
holic extracts of T. erecta and C. asiatica was carried out
by using fixed-dose method according to OECD [9]
guideline No. 401. Animals were kept fasting overnight
prior to the experimentation.
2.5. Surgical Procedures and Treatment
2.5.1. Excision Wound Model
In the present experimental model two different types of
creams were formulated of each extract (5% and 10%
w/w of T. erecta and the other having 5% and 10% w/w
of C. asia tica) and the wounds were medicated with one
local application (5 - 10 mg) of each cream for every day
until the day of epithelization and their excision wound
healing effect on rats was observed. The animals were
randomly allocated into five groups of six animals each.
Group 1: Wounds untreated served as control; Group 2:
Wounds treated with cream having T. erecta (5% w/w);
Group 3: Wounds treated with cream having C.asiatica
(5% w/w); Group 4: Wounds treated with cream having
T. erecta (10% w/w); and Group 5: Wounds treated with
cream having C. asiati ca (10% w/w).
This surgical procedure was performed according to
Panchatcharam. The wounding procedures were carried
out under anesthetized condition by thiopentone sodium
(40 mg/kg, ip), the backs of animals were shaved and
sterilized with 70% ethyl alcohol. Excision wounds sized
2.5 cm diameter in average were made after leaving at
least 5 mm space from the ears and treated topically
every day until the day of epithelization (Figure 1). The
physical attributes of wound healing viz wound closure
Excision wound Incision wound Dead space wound
Figure 1. Different surgical w ound models on r ats.
Copyright © 2011 SciRes. CM
(contraction), epithelization and scar features were recor-
ded. The wound contraction was studied by traced 1 mm2
graph paper on the day of wounding and subsequently on
the alternate days, until healing was complete and calculated
as percentage reduction of initial wound area [10].
2.5.2. I n cision Wound Model
The animals were randomly allocated into five groups of
six animals each. The extracts were dissolved in water
and administered orally, daily for 10 days. Group 1: was
assigned as vehicle control group and received no treat-
ment; Group 2: received extract of T. erecta 250 mg/kg;
Group 3: received extract of C. asiatica 250 mg/kg;
Group 4: received extract of T. erecta 500 mg/kg; Group
5: received extract of C. asiatica 500 mg/kg, orally.
Under light ether anesthesia, the animals were secured
to operation table in its natural position. Two Para verte-
bral straight incision of 6 cm each was made through the
entire thickness of the skin, on either side of the vertebral
column of the rat with help of sharp blade [11]. Wounds
were closed with interrupted sutures, 1 cm apart and
showed in Figure 1. The sutures were removed on the
8th day. The tensile strength of wound was determined
on the 10th post wounding day by continuous constant
water flow technique [12].
2.5.3. Dead Spac e Wound Model
In the case of dead space wound model the animals were
randomly divided into five groups of six animals each.
The extracts were dissolved in water and administered
orally, daily for 10 days. Group 1: was assigned as con-
trol group and received no treatment; Group 2: received
extract of T. erecta 250 mg/kg; Group 3: received extract
of C. asia tica 250 mg/kg; Group 4: received extract of T.
erecta 500 mg/kg; Group 5: received extract of C. asiat-
ica 500 mg/kg, orally.
Physical and mechanical changes in the granulation
changes in the granuloma tissues were studies in this
model. These wounds were created by implanting two
polypropylene tubes (0.5 × 2.5 cm each), one on either
side in the lumber region on the dorsal surface of each rat
and it showed in Figure 1. The wounds were sutured and
mopped with cotton swabs in 70% alcohol. Animals were
placed independently in cages after recovery from anes-
thesia. On the 10th post wounding day, the granulation
tissue formed on the implanted tubes was carefully dis-
sected out under anesthetic condition. The wet weight of
granulation tissue was noted. The breaking strength of
granulation tissue was measured by the method of Lee.
Later, these granulation tissues were collected, dried at
60˚C for 24 h. The weight of the dried granulation tissue
was noted and expressed as mg/100g body weight.
2.6. Statistical Analysis
The data are represented as mean ± S.E.M, and statistical
significance between treated and control groups was
analyzed by using one way ANOVA followed by Tukey
multiple comparison test using Graph Pad Instate soft-
ware (GPIS) (Version 1.13). P < 0.05 was considered
statistically significant.
3. Results
The acute toxicity studies of the hydroalcoholic extract
of leaves of T. erecta and aerial part of C. asiatica were
found to be non-lethal up to dose of 2 g/kg body weight.
Hence 1/8th and 1/4th of 2 g/kg dose (i.e. 250 and 500
mg/kg, orally) was selected for incision and dead space
wound model. In the excision wound model, the T.
erecta extract treated animals showed a significant in-
creased percentage of wound contraction, decreased in
the epithelization period and formation of scar, compare
to C. asiatica extract treated animals. Data are presented
in Table 1 and Figure 2. In the incision wound model, a
significant difference (P < 0.001) was observed with the
treatment of C. asiatica and T. erecta in wound breaking
strength group has compared to control group. Results
are represented in Table 2. The effect of C. asiatica (500
mg/k g ) and T. erecta extract (250 & 500 mg/kg) signifi-
cantly increased the granulation tissue weight (wet and
dry) and granulation breaking strength (P < 0.001) in the
dead space wound model. By comparison, a significant
decrease was observed in the T. erecta 250 mg/kg (P <
0.05) and non significant in C. asiatica 250 mg/kg
treated group, data are presented in Table 2.
4. Discussion
The objective in wound management is to heal the wound in
the shortest time possible, with minimal pain, discomfort,
812 16
Da y s
Control C. asiatica 5%C . a sia tic a 10
T. erecta 5% T . erecta 10%
Figure 2. Effect of hydroalcoholic extracts of Centella asiat-
ica and Tagetes erecta for percentage of wound contraction
(in days), in excision wound model.
Copyright © 2011 SciRes. CM
Table 1. Effect of hydroalcoholic extracts of Centella asiatica and Tagetes erecta for percentage of wound contract ion (in days),
epithelization (in days) and formation of scar in exci sion wound model.
% Wound contraction in days
4 8 12 16
Period of
( in days)
Mean size
of scar area
(in mm2)
Control 36.29 ± 1.52 54.59 ± 1.90 83.94 ± 1.40 90.41 ± 0.80 21.80 ± 0.65 55.50 ± 1.78
C. asiatica
5% (w/w) 47.95 ± 2.06** 67.80 ± 1.40** 88.28 ± 0.70* 96.77 ± 0.64*** 19.33 ± 0.76 48.33 ± 2.20
T. erecta
5% (w/w) 49.74 ± 1.49** 71.45 ± 2.30** 92.79 ± 0.74*** 98.82 ± 0.39*** 18.00 ± 0.51* 44.66 ± 2.50*
C. asiatica
10% (w/w) 55.11 ± 2.30*** 76.50 ± 2.22*** 94.12 ± 1.16*** 99.48 ± 0.21*** 17.66 ± 0.71* 43.33 ± 2.54*
T. erecta
10% (w/w) 59.59 ± 3.08*** 83.73 ± 4.01*** 99.72 ± 0.12*** 100.00 ± 0.0*** 14.16 ± 0.75*** 29.00 ± 2.80***
The value are expressed as mean ± SEM, (n = 6). If *P < 0.05, **P < 0.01, ***P < 0.001 vs. control.
Table 2. Effect of hydroalcoholic ext ract of Centella asiatica and Tagetes erecta in incision and dead space wound models.
Incision wound Dead space wound
Treatment Breaking Strength, (g) Wet tissue weight mg/100 g ratDry tissue weight mg/100 g rat Braking Strength, (g)
Vehicle control 259.20 ± 7.45 152.30 ± 4.20 14.20 ± 1.04 256.10 ± 9.14
C. asiatica 250 mg/kg 281.20 ± 9.20* 175.60 ± 5.00 21.20 ± 0.99 280.60 ± 9.50
T. erecta 250 mg/kg 300.70 ± 13.60** 185.30 ± 4.20 22.28 ± 2.20* 297.10 ± 12.90
C. asiatica 500 mg/kg 324.50 ± 11.30*** 193.50 ± 5.30** 25.27 ± 2.24** 331.70 ± 14.70***
T. erecta 500 mg/kg 351.40 ± 11.07*** 214.50 ± 8.30*** 27.21 ± 2.30*** 353.60 ± 5.90***
The value are expressed as mean ± SEM, (n = 6). If *P < 0.05, **P < 0.01, ***P < 0.001 vs. Vehicle control.
and scarring to the patient. At the site of wound closure a
flexible and fine scar with high tensile strength is desired
[13,14]. The healing process begins with tissue reposi-
tion by cell proliferation presented in connective tissue.
The main cells that trigger the wound healing process are
the macrophage cells that remove foreign bodies and
direct granular tissue development. Sequentially fibro-
blast and endothelial cells migrate towards the injured
area increasing tissue permeability and collagen fiber
production and it decreases vascularization by a contrac-
tion process by the scar process [15,16].
The ability of the wound is to heal is a biological
process, which follows a definite pattern of cellular and
molecular events, ultimately leading to complete repair
of the injured tissues. High order animal’s posses very
limited regenerative capabilities except for those of some
organs e.g. liver, parenchyma and outer surface of the
skin. The healing rate differs among species and tissues
and even between sites of the same tissue. However, in
general the pattern of wound healing in domestic rumi-
nants appears to be similar. The initial events of wound
healing mechanism occurred smoothly under favorable
condition. The normal process of wound healing may be
disrupted. During this condition before using drugs,
many herbal preparations are extensively used in the
rural area [17].
The result suggest that treatment with hydroalcoholic
extract of C. asiatica and T. erecta have beneficial in-
fluence various phase of wound healing fibroplasias, col-
lagen synthesis and wound contraction result in faster
In the present study the significant increase in the
breaking strength was observed in the animals treated
with extract of T. erecta when compared to extract of C.
asiatica on 10th post wound healing day.
In addition to excision and incision wound model,
dead space wound model also revealed a significant in-
crease in the dry granulation tissue weight indicating hi-
gher protein content and significant in breaking strength
of granulation tissue both resutured incision wound was
observed in the hydroalcoholic extract of T. erecta
treated group compare to C. asiatica treated group.
The observation and result obtained in this study indi-
cated that crude extract of T. erecta significantly stimu-
lated wound contraction, breaking strength of the inci-
sion wound and increased in the dry granulation weight
in the treated group compared with extract of C. asiatica.
In this study one more attempt has been made to inclu-
Copyright © 2011 SciRes. CM
sion T. erecta plant in the management of wound healing
in folk medicine is justifiable.
5. Conclusions
In the present work it can be concluded that the com-
parison made between the above plants of hydroalcoholic
extract of T. erecta showed promising wound healing
activity then the reported C. asiatica in different wound
6. Acknowledgements
The authors are grateful to Premnath Reddy, Chairman
Acharya Institute, and Principal, Acharya & B. M. Re-
ddy College of Pharmacy, Bangalore, India, for the con-
stant support and facilities to this study.
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