Wound Healing Activity of Sida cordifolia in rats
S. Akilandeswari*, R. Senthamarai, S. Shakila Banu
Department of Pharmaceutical Chemistry and Pharmacognosy, Periyar College of Pharmaceutical Sciences Tiruchirappalli – 620021.
*Corresponding Author E-mail: akipcog@yahoo.co.in
ABSTRACT:
Sida cordifolia (Malvaceae), Shrubby, branched, softly hairy and with much stellate hair nearly all over and subpersistant. Leaves 2.5-5 cm, long, coordinate, ovate-oblong, crenate, obtuse or subacute, not acuminate; petioles 1.2-3.8 cm long.1 The mucilaginous leaves are used as a demulcent and their infusion is given in fever as a refrigerant. They are also used against dysentery and for poulticing ulcers. A decoction of the leaves is said to possess emollient and diuretic properties. Fresh infusion they are bruised and applied to boils to promote suppuration. Leaves are cooked and eaten in cases of bleeding in piles. The leaves are cooling, aphrodisiac, unctuous, strengthening and promote sexual vigor and vital factor. They are good for rheumatism, flatulence, colic, haemothermia, emaciation, vitiated conditions of tridosa, seminal weakness and arthritis. However, there were no reports on both ethnobotanical and pharmacological profile of this plant. Hence, the present study was made to evaluate the wound healing potential.2
KEYWORDS: Wound healing, Sida cordifolia, Methanolic extract, Nitrofurazone ointment.
INTRODUCTION:
Sida cordifolia (Malvaceae), Shrubby, branched, softly hairy and with much stellate hair nearly all over and subpersistant. Leaves 2.5-5cm, long, coordinate, ovate-oblong, crenate, obtuse or subacute, not acuminate; petioles 1.2-3.8cm long.1 The mucilaginous leaves are used as a demulcent and their infusion is given in fever as a refrigerant. They are also used against dysentery and for poulticing ulcers. A decoction of the leaves is said to possess emollient and diuretic properties. Fresh infusion they are bruised and applied to boils to promote suppuration. Leaves are cooked and eaten in cases of bleeding in piles. The leaves are cooling, aphrodisiac, unctuous, strengthening and promote sexual vigor and vital factor. They are good for rheumatism, flatulence, colic, haemothermia, emaciation, vitiated conditions of tridosa, seminal weakness and arthritis. However, there were no reports on both ethnobotanical and pharmacological profile of this plant. Hence, the present study was made to evaluate the wound healing potential.2
MATERIAL AND METHODS:
Plant Extract and standard used:
The dried plant materials were, pulverized by a mechanical grinder, sieved through 40 mesh. The powdered materials were extracted with methanol using soxhlet extraction apparatus. This methanol extract was then concentrated and dried under reduced pressure. The methanol free semisolid mass thus obtained was used for the experiment. Two types of ointment formulation were prepared from the extract; 5% (w/w) and 10% (w/w), where 5g and 10g of the extract were incorporated in 100 g of simple ointment base B.P3 respectively. Nitrofurazone ointment (0.2% w/w, Smithkline – Beecham) was used as a standard drug for comparing the wound healing potential of the extract.
Animals used:
Wistar Albino rats (150 – 180gms) were selected for these studies. Six rats were taken for each group. The rats were used after an acclimatizationperiod of 7 days to the laboratory environment. They were provided with food and water ad libitum.
Excision wound Model4:
Four groups with six animals in each group were anaesthetized with ether. The rats were depilated on the back. One excision wound was inflicted by cutting away a 500mm2 full thickness of skin from the depilated area, the wound was left undressed to open environment. Then the drugs, i.e., the reference standard, (0.2% w/w) Nitrofurazone (NFZ) ointment, simple ointment B.P., Sida cordifolia extract ointment (5% w/w), and Extract ointment (10% w/w) were applied once daily till the wound was completely healed5. This model was used to monitor wound contraction and wound closure time. Wound contraction was calculated as percent reduction in wound area. The progressive changes in wound area were monitored planimetrically by tracing the wound margin on graph paper every alternate day.
Incision wound model:
Four groups with six animals in each group were anaesthetized and two Para vertebral long incisions were made through the skin and cutaneous muscles at a distance of about 1.5cm from the midline on each side of the depilated back of the rat. Full aseptic measures were not taken and no local or systemic antimicrobials were used throughout the experiment6. No ligature was used for stitching. After the incision was made, the parted skin was kept together and stitched with black silk at 0.5 cm intervals; surgical threads (No.000) and a curved needle (No.11) were used for stitching. The continuous threads on both wound edges were tightened in the same manner as has already described above. The extract ointments and the NFZ ointment were administered once daily for 9 days; when wound were cured thoroughly the sutures were removed on the ninth day and tensile strength was measured with a tensiometer7.
Measurement of Healing:
Tensile strength, the force required to open a healing skin wound, was used to measure healing. The instrument for this measurement is called tensiometer was designed on the same principle as the thread tester used in the textile industry. It consisted of a 6 x 12 inch board with one post of 4 inch long fixed on each side of the longer ends. The board was placed at the end of a table. A pulley with bearing was mounted on the top of one of the posts. An alligator clamp with 1cm width, was tied on the tip of the post without pulley by a piece of fishing line (20-lb test monofilament), so that the clamp could reach the middle of the board. Another alligator clamp was tied on a piece of fishing line with a 1 – L polyethylene bottle tied on the other end. Before testing, the animal was anaesthetized with ether in an open mask. The sutures of the wound were cut out with a pair of scissors. The animal was then placed on a stack of paper towels that could be adjusted so that the wound was on the same level of the tips of the posts. The clamps were then carefully clamped on the skin of the opposite sides of the wound at a distance of 0.5cm away from the wound. The longer piece of fishing line was placed on the pulley and the position of the board was adjusted so that the polyethylene bottle was freely hanging in the air. Water was added to the polyethylene bottle at a rapid but constant rate by siphon from a large reservoir (20-L Bottle) until the wound began to open up. The amount of water in the polyethylene bottle was weighed and considered as the tensile strength of the wound.
Statistical analysis:
Data are expressed as mean ± SEM and subjected to student’s t- test by comparing with the control.
RESULTS AND DISCUSSION:
The measurements of the progress of the wound healing induced by the NFZ ointment (0.2% w/w), Sida cordifolia ointment (5% w/w and 10% w/w) the respective control groups (i.e. simple ointment treated groups) in the excision wound method are shown in Table. 1 and Fig. 1, 2 and 3. It is observed that the wounds contracting ability of the Sida cordifolia ointment (5% w/w) significantly greater than that of the control. In the case of NFZ ointment and Sida cordifolia ointment treated groups it was found to be 18±2 d.
In the incision wound studies, there was a significant increase in tensile strength of the 10-d old wound due to treatment with Sida cordifolia ointment and the reference standard NFZ ointment when compared with the respective control. Measurements of the tensile strength are shown in Table.2 and Fig. 4, 5 and 6. The tensile strength of the NFZ ointment-and the Sida cordifolia ointment treated groups were almost the same.
The process of wound healing occurs in four phases (i) coagulation, which prevents blood loss, (ii) inflammation and debridement of wound, (iii) repair, including cellular proliferation and (iv) tissue remodeling and collagen deposition8. Any agent, which accelerates the above process is a promoter of wound healing. Plant products have been show to possess good therapeutic potential as anti-inflammatory agents and promoter of wound healing due to the presence of active terpenes, alkaloids and flavonoids9,10. An essential oil from the leaves had broad-spectrum antifungal activity comparable to that of 0.5% hamycin a glycosidal mixture extract of centella asiatica has been reported to be responsible for enhanced repair only in incised wounds11 and in stimulating collagen in human skin fibroblast cells12. The wound healing property of Sida cordifolia appears to be due to the presence of its active principles which accelerates the healing process and confers breaking strength to the healed wound.
Table 1: Effect of Plant Extract and Nitrofurazone Ointment on Wound Healing by Excision Wound Method
|
Post wounding days |
Wound Area (mm2) mean ± S.E. and percentage of wound contraction |
|||
|
Simple ointment |
Nitrofurazone ointment (0.2% w/w) |
Sida cordifolia Extract ointment (5% w/w) |
Sida cordifolia Extract ointment (10% w/w) |
|
|
0 |
529±19.3 (0.0) |
518±17.4 (0.0) |
523±24.0(0) |
536±18.4 (0.0) |
|
2 |
467±18.4 (11.7) |
417±21.6 (19.4) |
468±19.8(10.5) |
446±31.4 (16.8) |
|
4 |
403±21.5 (23.8) |
321±29.4*(38.0) |
377±17.9*(29) |
409±21.3*(23.7) |
|
6 |
371±14.6 (29.8) |
231±23.4**(55.4) |
228±13.3*(56.4) |
326±19.3**(39.2) |
|
8 |
313±13.9 (40.8) |
173±17.6**(66.7) |
151±14.5**(71.1) |
208±17.1**(61.2) |
|
10 |
297±14.6 (45.2) |
129±11.6**(75.0) |
89±10.1**(82.9) |
156±16.9**(70.9) |
|
12 |
276±11.9 (47.8) |
75±6.9**(85.5) |
53±6.4**(89.8) |
94±11.3**(82.5) |
|
14 |
259±14.3 (51.0) |
34±2.4**(93.4) |
32±4.8**(93.8) |
57±3.1**(89.4) |
|
16 |
231±16.7 (56.3) |
9±0.8**(98.2) |
21±1.2**(96) |
28±1.7**(94.7) |
|
18 |
211±15.3 (60.1) |
0.0**(100.0) |
0.0**(100) |
12±0.6 (97.8) |
|
20 |
179±11.8 (66.1) |
0.0**(100.0) |
- |
0.0 (100) |
*P<0.01, **P<0.001 Vs Control by Students‘t’-test (n=6)
Figures in parenthesis represent percentage of wound contraction.
Fig. 1, 2 and 3 Effect of Plant Extract and Nitrofurazone Ointment on Wound Healing By Excision Wound Method
Table 2 : Effect of Plant Extracts and Nitrofurazone Ointment on Incision Wound Model In Rats
|
Group |
Treatment |
Tensile strength (g) |
|
1 |
Simple Ointment |
363 ± 12.9 |
|
2 |
Sida cordifolia Extract Ointment (5% w/w) |
620 ± 26.1* |
|
3 |
Sida cordifolia Extract Ointment (10% w/w) |
623 ± 15.9* |
|
4 |
Nitrofurazone ointment (0.2% w/w) |
659 ± 27.1* |
*p<0.001 Vs control by student‘t’-test. Data are expressed as mean ± S.E
Fig. 4,5 and 6 - Effect of Plant Extracts and Nitrofurazone Ointment on Incision Wound Model In Rats
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Received on 06.12.2019 Modified on 18.02.2020
Accepted on 29.03.2020 © RJPT All right reserved
Research J. Pharm. and Tech. 2020; 13(12):6147-6149.
DOI: 10.5958/0974-360X.2020.01072.0