ISSN: 0975-8585

Research Journal of Pharmaceutical, Biological and Chemical

Sciences

Wound Healing Activity of Oil Extract of Tectona Grandis Leaves in Wistar

Rats.

Prathibha M1 *, Tatiyana M1, Bairy Kl1, Adiga S2, and Sunitha K3.
1
Department of Physiology, Melaka Manipal Medical College, Manipal University, Manipal 576104, Karnataka, India.
2
Department of Ayurveda, Kasturba Medical College , Manipal 576104, Karnataka, India.
3
Department of Pharmacology, Jawaharlal Institute of Medical Education and Research, Puducherry, India

ABSTRACT

Frontal leaves of Tectona grandis are used traditionally for the treatment of variety of wounds.
Scientific data is not available regarding the wound healing activity of oil extract of frontal leaves of Tectona
grandis. Hence this study was undertaken to investigate the effect of topical application of oil extract of
frontal leaves of Tectona grandis on incision and excision wound models in rats. 4 groups (control, vehicle
control - coconut oil, standard - aloe vera, test - oil extract of Tectona grandis leaves) of wistar rats were used
each for excision and incision wound model. Parameters studied were wound breaking strength in incision
wound model and percentage of wound contraction and period of epithelisation in excision wound model.
Results showed a significant increase in the breaking strength (p<0.001) in incision wound model; decrease in
period of epithelisation (p<0.01) and increase in wound contraction rate (p<0.001) in excision wound model
in the test group when compared to control group. Therefore, this study ascertains the wound healing activity
of Tectona grandis leaves and it substantiates the use of frontal leaves of Tectona grandis in Ayurveda for
treatment of wounds.
Keywords: excision and incision wound model, wistar rats, Tectona Grandis leaves, oil extract

*Corresponding author

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INTRODUCTION

Wound is a discontinuity in a normal tissue structure that results in a variety of cellular and molecular
changes. Wound healing is a complex process that includes inflammation, granulation tissue formation,
epithelisation, collagen synthesis and tissue remodelling [1, 2].

Many dietary modification and nutritional and herbal supplements have proved to improve quality of
wound healing by influencing repairative processes or by limiting the damaging effects of inflammation. Some
of these factors [3] help in reversing delayed wound healing due to drugs like corticosteroids [4], anticancer [5]
and non-steroidal anti-inflammatory drugs [6].

Various medicinal plants have been used for centuries as a remedy for treating human diseases [7,8].
The plant, Tectona grandis (family verabinaceae) is grown throughout India and is also known as Indian teak.
Tectona grandis (TG) is a large deciduous tree 10-12 meter tall. Leaves of TG are opposite, elliptic or obovate in
shape, rough and glabrous above [9]. According to ancient Indian literatures, leaves of TG are having cooling
property and are used commonly as hemostatic. They are useful in treatment of inflammations, leprosy, skin
diseases, pruritus, stomatitis, ulcers and hemorrhages.

The phytochemical screening of alcoholic extract of TG leaves reveals the presence of alkaloids,
flavonoids, tannins [10]. It is also shown to have nitric oxide scavenging activity [11]. Hydro alcoholic extract of
TG leaves improves incision, excision and burn wound healing in rats [12]. It has been found that topical
application of methanolic extract of this leaves have antifungal and antibacterial activity [13,14] .

In ayurvda practice, oils are used as one of the mode of application in treatment of the wounds [15].
Traditionally prepared oils are used as a base for external applications of medicine. There is no scientific data
available on the wound healing activity of oil extract of frontal leaves of Tectona grandis which is prepared
according to traditional methods. Hence this study was undertaken to evaluate the effect of topical
application of oil extract of leaves of TG on incision and excision wound models in wistar rats.

MATERIALS AND METHODS

Experimental animals

Healthy inbred wistar rats of either sex, weighing between 200 - 250 g were used. The experimental
protocol was approved by Institutional Animal Ethics Committee and animals were maintained under standard
environmental condition of temperature 23±2C, humidity 50± 5% and 10 -14 hours light and dark cycle
respectively in the animal house approved by Committee for Purpose of Control and Supervision on
Experiments on Animals (CPCSEA). The animals were provided with standard rat feed and water; ad libitum.
The animals were housed individually in polyprophylene cages containing sterile husk as bedding after making
wounds till the completion of wound healing.

Drugs used

Injection ketamine, aloe vera gel and coconut oil were obtained from the pharmacy of Kasturba
Hospital. Fresh frontal leaves of TG were collected locally from Udupi district, Karnataka and they were
authenticated by department of Ayurveda, KMC, Manipal.

Traditional method of preparation of oil extract

Procedure followed in this study for the preparation of medicated oil extract is recognized by Drugs and
Cosmetic Act [16] and it is also included in the Ayurvedic Formulary of India (AFI) [17]. Tender leaves of TG
plant were collected in the month of December and dried under shade for 15 days and powdered. One part
of coarse powder leaves was added to sixteen parts of water and boiled to reduce the volume to one forth.
The decoction is strained using a muslin cloth. Some fresh tender leaves were grounded to make a fine paste.
Coconut oil is taken in a vessel and heated for some time. Then one part fine paste of leaves were added to
four parts of coconut oil and sixteen parts of above prepared decoction. This mixture is boiled on mild fire with
frequent stirring to avoid paste to adhere to the vessel and boiling continued till all the water evaporates.

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Well-cooked oil should not have any residual moisture. The oil is strained while warm through muslin cloth and
allowed to cool [15].

Wound Induction

Incision wound model

Animals were fasted for twelve hours prior to the experiment. Rats were anaesthetized by using
intraperitoneal injection of ketamine (50mg/ kg body wt) [18,19]. Two Para vertebral straight incisions of 6 cm
were made through the entire thickness of the shaved skin one on each side of vertebral column with the help
of a sharp blade. After mopping the wounds dry, wounds were closed by using 4-0 silk thread and straight
round body needle. The interrupted sutures were place at equidistance points of 1 cm each [20]. Wounds were
then mopped with cotton swabs soaked in 70% alcohol. The animals were housed individually.

Excision wound model

After fasting for 12 hours rats were anaesthetized by using intraperitoneal injection of ketamine (50
mg/kg body wt). A round seal of 2.5cm diameter was impressed on the dorsal of thoracic region 5 cm away
from the ears on shaved back of the rat. Full thickness skin from demarked area was excised to get a wound
2
measuring 500 mm [21]. After achieving full haemostasis by mopping the wound with cotton swab soaked in
warm saline, animals were placed in their individual cages.

Drug treatment

4 groups of animals (n=6) were used each for incision and excision wound models. All the drugs were
applied topically. Control groups received distilled water, vehicle control groups received coconut oil, standard
groups received aloe vera gel and test groups received oil extract of TG leaves. Day of wounding is considered
as day 0. In case of incision wound model, drugs were applied once daily from the day 0 till day 9 post
wounding day. In excision wound model, animals were treated once daily from 1 to 21 post wounding day
[18].

Parameters assessed:

Incision wound model

th
Removal of the sutures was done on 7 post wounding day. The wound breaking strength was
estimated on 10 day by continuous, constant water flow technique. The breaking strength was expressed as
minimum weight of water necessary to bring about gaping of wound [6].

Excision wound model

Two physical attributes of healing namely wound contraction rate and epithelisation period were
studied . To monitor wound contraction the progressive changes in wound area will be followed
planimetrically .Wound area were traced on a transparent paper on day 0, 4, 8,12 and 16. The tracings were
then transferred to 1mm graph sheets and the wound area was measured. Percentage of wound contraction
was calculated using the following equation:

2
Initial wound size (500 mm ) – Specific day wound size
% of wound contraction= X 100
Initial wound size

Epithelisation period was monitored by noting the number of days required for the eschar to fall off
leaving no raw wound behind [21,22].

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Stastical Analysis

Data was analyzed using SPSS 16. The results were expressed as mean ± SEM. Differences between
the experimental groups were compared using one-way ANOVA followed by Bonferroni’s post hoc test.
Values of p<0.05 were considered statistically significant.

RESULTS

In incision wound model (Table 1), topical application of oil extract of TG showed significant increase
in the wound breaking strength (p<0.001) when compared to control group and vehicle control group.
Standard group also showed significant improvement and test group results were comparable to that of
standard group.

Table 1: Mean wound breaking strength in incision wound model

Group Wound breaking strength (gms)

(n=6)
Control( C) 172±13.15
Vehicle control (VC) 183.23± 6.94
Standard (SD) 258.35 ± 20.75 *
Oil extract of TG 308.33 ± 14.70***
*
Control vs SD p< 0.05; Control vs TG,*** p<0.001

In excision wound model (Table 2), TG treated animals showed significant reduction in wound area
when compared to control and vehicle control group on day 8, 12 and 16 (p<0.001) and there was no
significant difference between test group and standard.

Table 2: Mean wound contraction rate in excision wound model

Groups Percentage of wound contraction

(n=6)
Day 4 Day 8 Day 12 Day 16
Control 13.72 ±2.77 58.96 ±2.63 81.66 ±1.656 91.33 ±0.571
Vehicle 18.88 ± 2.66 70.36 ± 3.14 8 1.55 ±1.679 93. 55 ± 0.899
control
Standard 22.36±3.77 75.28± 1.64* 88.23 ± 1.514* 98.4 ±0.280*
Oil extract of 24.8 ± 2.83 76.36 ±1.37*** 92.23 ±1.19*** 96.76 ± 1.06***
TG
th th th th th th
Control vs TG on 8 , 12 , 16 day ***p<0.001; Control vs SD on 8 , 12 , 16 day ***p<0.05

Period of epithelisation (Table 3) was shorter in test group when compared to that of control and
vehicle control group (p<0.05) but there was no significant difference between test and standard group.

Table 3: Mean Period of epithelisation for excision wound model

Groups Period of epithelisation(days)

Control 19.33 ± 0.49
Vehicle control 19 ± 0.36
Standard 15 ± 1.09**
Oil extract of TG 15.33 ± 0.61**
Control vs TG ***p<0.05; Control vs SD **p<0.05

DISCUSSION AND CONCLUSION

Wound healing is a process of restoration of damaged tissue to its normal state and wound
contraction is the process of shrinkage of the area of the wound. The results of the present study showed that
the oil extract of Tectona grandis leaves when applied topically improved the wound breaking strength and
wound contraction rate in incision and excision wound models respectively.

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The above mentioned prohealing actions of the extract maybe due to the presence of
phytoconstituents like alkaloids, carbohydrates, glycosides phytosterols, saponins, proteins aminoacids and
flavonoids. The flavonoids [23], and triterpenoids[24] are known to promote the wound-healing process
mainly due to their astringent and antimicrobial properties, which could be responsible for improved wound
contraction rate and shortened period of epithelisation.

Traditionally prepared oil extracts have principally three components namely, a liquid which may be
aqueous decoction of one or more herbs, a fine paste of the herbs and a vegetable oil. Normally crude sesame
oil (SO) is used but occasionally castor oil and coconut oil is also used.

In the present study coconut oil is used as a vehicle because it is also used in treating wounds
traditionally. Various studies proved that coconut oil itself has wound healing activity [25]. Therefore, using
coconut oil as base to prepare oil extract potentiates the wound healing activity of TG. Moreover, properly
prepared oils have long expiry (about one year) without any separate preservatives which helps in the easy
storage of the medicinal extract. The oil preparation by using above mentioned procedure is home based,
economical and convenient. In this study, aloe vera gel is used as a standard drug for wound healing as it is
known to have wound healing and anti- inflammatory activity [26].

This study ascertains that oil extract of TG leaf extract possess good wound healing activity when
applied topically in incision and excision wound models and it substantiates the use of TG leaves in folklore
medicine for treatment of wounds. This study also support that oil prepared in traditional method can be
used as a wound healing agent. Further phytochemical studies are suggested to isolate, characterize and
identify the specific active compounds in this plant responsible for wound healing activity.

ACKNOWLEDGEMENT

The authors thank Manipal University for supporting the research.

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