Evaluation of Wound Healing Activity of Ethanolic Extract of Aerva tomentosa Forsk
Deepak Kumar*1,DN
Prasad1, JS Dua1 and SP Bhatnagar2
*1Dept.
of Pharmacognosy,
2Department of Pharmaceutical Sciences, B.I.T., Mesra, Ranchi,
835215,(INDIA).
ABSTRACT
Herbal drugs are already in vouge as therapeutic agents for thousands of
years but in present times there is a need to re-valuate their clinical
efficacy. Aerva tomentosa Forsk. is recognized by its white
axillary bunches of small woolly flowers and traditionaly claimed for its wound
healing activity. Wound healing activity of ethanolic
extract of Aerva
tomentosa Forsk. was
evaluated on excision and incision wound using Wistar
rats of either sex. Percentage of wound contraction was measured in excision
wound, whereas tensile strength of healed wound was measured in incision wound.
In excision wound, wound contraction was 93.8 % on 16th day for Aerva tomentosa
treated groups. In incision model, significant increase in tensile strength of
healed wound was observed in the concentrated ethanolic
extract.
KEYWORDS: Excision wound,
incision wound, framycetin sulphate.
INTRODUCTION
Aerva tomentosa Forsk. (A.t) (amaranthaceae)
is a common wayside weed grown mostly on waste place and found on the plains of
the warmer parts of
MATERIALS AND METHODS:
Plant
material collection:
The aerial parts of Aerva tomentosa were collected from wild
source of the state of Punjab during flowering season in the month of November
– December and authenticated by Dr.
Sumer Chandra, Scientist D, F.R.I. Deemed University, I.P.E., Kaulagarh Road, Dehradun, 248195,
Utranchal as Specimen voucher no. SCP- 302.
Extraction of plant materials:
The aerial
parts were shade dried, powdered and
passed through 80 mesh sieve. The powdered material was extracted with ethanol
using Soxhlet’s apparatus. The extract obtained was
dried in rotary vacuum evaporator at 400 C yielding a yellowish
green colored, viscous mass (1.2%). The phytochemical
tests of ethanolic extracts of Aerva
tomentosa showed the presence of alkaloids, saponin, flavonoid and tannin.
Table
1 Wound healing activity of ethanolic extract of Aerva tomentosa on
excision wounds using topical application method
|
Post wounding days /
% contraction |
Control |
Framycetin sulfate
(Stand.gp.)1% |
Conc.ethanolic extract
of Aerva toment. |
|
Day 4 |
12.1±0.30 |
20.8±0.31 |
17.1±0.27 |
|
Day 8 |
31.2±0.89 |
59.8±0.90 |
44.6±0.88 |
|
Day 12 |
72.6±1.05 |
89.5±0.76٭٭ |
82.5±0.99٭٭ |
|
Day 16 |
88.6±0.88 |
97.33±0.66٭٭٭ |
93.8±1.07٭٭٭ |
One way ANOVA.
Table
2 Wound breaking strength of Incision wound on 10th day of Ethanolic extract of Aerva tomentosa (gm)
|
Control |
Standard
group Framycetin sulfate |
Ethanolic extract of Aerva tomentosa |
|
230.5±0.99 |
512.3±1.22٭٭٭ |
478.0±1.41٭٭٭ |
N= 6, values (mean±
SEM), ***p< 0.001 considered significant. all p values calculated with
respect to control.
Animals:
Healthy Wister albino
rats of either sex weighing about 150-180g were used for wound healing
activity. All animal experiments were carried out under the CPCSEA norms, after
obtaining the approval of Institutional Animal Ethical Committee
(1080/C/07/CPCSEA/2007). All animals were procured from the Institute of
Microbial Technology (IMTECH), Sector 39-A,
Excision
Wound Model7:
The hair on the skin
from dorsal thoracic region were removed by using a suitable depilatory (Anne
French hair removing cream).Circular wounds of approximately 10mm diameter were
inflicted on the cleared skin by cutting under mild ether anesthesia, 5 cm away
from the ears on the depilated part of skin and the entire thickness of skin
from the demarcated area was excised to get a wound. For this a round seal of
10mm diameter was impressed on dorsal thoracic region. Area of the wounds were
measured immediately in sq. mm by tracing the wounds on to tracing paper and
there after on graph paper by using carbon paper, on day of wounding and this
was taken as the initial wound area reading.
Standard and test sample were applied once daily topically till complete
contraction of the skin takes places directly on wound, starting from day of
excision. The wound area was measured subsequently on 4th, 8th, 12th
and 16th days post wounding8. The percentage of wound
contraction was calculated from the days of measurements of wound area.
Incision
wound model:
For incision wound
model each animal was secured to operation table in its natural position under
light ether anesthesia and hairs were removed from back portion using depilator. Light par vertebral straight incisions were made
on the cleared surface by cutting the skin of the animals through the entire
thickness of skin with a sterilized sharp blade. The wounds were created at a
length of about 1.5 cm. After the incision, the parted skin was kept together
and sutured with absorbable sterilized surgical needled suture USP, Vicryl (synthetic, 0.5 matric,
coated polyglactin 910, made by Ethicon, Baddi, H.P.) thread as interrupted sutures at 0.5cm apart
with curved needle (no.11)9. The concentrated ethanolic
extract of A.t. was applied once a day for 10 days in
a similar manner as control and standard drug. Wound tensile strength was
measured on the 10th day by continuous constant water flow technique
of Lee10.
Measurement
of Wound Breaking Strength of Incised Wounds:
Measurement of wound
breaking strength was performed following Lee with certain modifications. A
board was placed on a raised platform on the table, on which the anaesthetized
rat was made to lie on its abdomen. Two clamps were clamped on either sides of
healed wound at a distance of 0.5cm.The left clamp was fastened tightly to a
stand by means of a thread. The right clamp was connected to a leak proof
polythene bag through a pulley, by means of a thread. A reservoir containing
water was placed at a suitable height and connected to a polythene bag by means
of a rubber tube. The position of the board was adjusted so that, the polythene
bag was hanging freely. Water was added to polythene bag rapidly at constant
rate from the reservoir until the wound opened. Amount of water in polythene
bag was measured in ml and was considered as tensile strength of the wound11.All
results are expressed as mean ± S.E.M. The results obtained were compared with
the control and analysed using statistical method.
RESULTS:
In excision wounds,
wound contraction was 97.33 and 93.8 % respectively on 16th day for framycetin SO4 and ethanolic
extract of A.t.treated groups. Significant wound
contraction was observed on 16th day for A.t.
treated groups in comparison with the control group.
In incision model,
significant increase in tensile strength of healed wounds was observed in Aerva tomentosa ethanolic extract.
DISCUSSION:
Wound healing comprises
of different phases such as contraction, granulation and collagenation12.It
normally involves an initial inflammatory phase followed by fibroblast
proliferation, formation of collagen fibers and shrinking, occurring concurrently
but independent of one another13.Phytoconstituents such as flavonoid and tannin are reported to be responsible for
wound healing activity. Wound healing effect is also attributed to free radical
scavenging activity of flavanoids14.Flavonoids are known to reduce
lipid peroxidation not only by preventing or
slowing onset of cell
necrosis, but also by improving vascularity. Lipid peroxidation is an important process in several types of
injuries like burns, infected wounds and skin ulcers. Hence any drug that
inhibits lipid peroxidation is believed to increase
strength of collagen fibers, by increasing circulation or by preventing cell
damage or by promoting DNA synthesis15.Flavonoid and tannin are
known to promote wound healing process mainly by their astringent and
antimicrobial property. Hence presence of flavonoid
and tannin in the ethanolic extract of A.t. may be responsible for its wound healing activity.
Hence, the results of present study support the traditional use of Aerva tomentosa in
the treatment of wounds. Further, work may be undertaken to isolate and
identify the bio-active constituents responsible for this activity.
REFERENCE:
1. Annonymous.”The Wealth of India”, Raw materials,vol IA.Pub.andInformation
Directorate,CSIR,N.Delhi.revised edn,1988; 91-92.
2.
Aiyar VN,
Narayanan V, Sheshadri TR, Vydeeswaran
S. Chemical Components of some Indian Medicinal Plants. Indian Journal of
Chemistry.1973; 11; 89.
3.
Maher SMA, Valhari
MU and Khatri LM. Identification and Quantification
of Ecdysteroids from Aerva
tomentosa and Pandiaka
involucrate. Pakistan Journal Science and Industrial research.1995; 38; 2;
91-93.
4.
Jaswant B, Ragunathan V and Sulochana N. A
rare flavonol glycoside from Aerva tomentosa Forsk
as antimicrobial and hepatoprotective agent. Indian
Journal of Chemistry.2003; 42B; 956-958.
5.
Dymock A, Warden
CJ and Hooper DH.”Pharmacographica Indica”, 1, Thacker Spints and
company,
6.
Radwan HM, Nazif NM.The Flavonoidal
constituents of the Ethyl fraction of Aerva bovei webb in Hook.F.and its antimicrobial activity. Phytomedicine,
Supplement II, 2000; 53.
7.
Morton JP,
8.
Madhavan V,
9.
Ghosh T, Dash
GK, Bose A and Panda BR.Wound healing properties of Argemone Mexicana. Indian J.Nat.Prod.2004; 20(4);
3-5.
10. Lee KH.
Studies on the mechanism of action of salicylate II:
Retardation of wound healing by Aspirin.J. Pharmacol. Sci.1968; 57; 1042.
11.
Madhavan V, Yadav DK, Murali A and Yoganarsimhan SN, Wound healing activity of Aqueous and
alcoholic extracts of leaves of Colebrookea oppositifolia Smith. Indian Drugs.2009; 46(3); 209-213.
12.
Vidyas SM,
Krishna V, Manjunath BK,
13.
Ganachari MS, Kumar
S and Patel A. Wound healing activity of Saussurea lappa roots.Indian
Drugs.2005; 42(5); 295.
14.
Lara OO, Fakoya FA,
Agbani EO and Lwalewa EO.
Vascular permeability-Increase effect of the leaf essential oil of Ocimum gratissimum
Linn. as a mechanism for its wound healing property.Afr.J.Trad
Compl. Alt Med, 2005 ;( 2); 253.
15.