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EVALUATION OF WOUND HEALING ACTIVITY OF ETHANOLIC EXTRACT OF OCIMUM BASILICUM LEAVES IN MALE ALBINO RATS
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Review Article - (2021) Volume 10, Issue 3

EVALUATION OF WOUND HEALING ACTIVITY OF ETHANOLIC EXTRACT OF OCIMUM BASILICUM LEAVES IN MALE ALBINO RATS

Renu Solanki1*, Shiv Kumar Purohit2, Vipin Mathur1 and Manoj Mathur2
*Correspondence: Renu Solanki, Lachoo Memorial College of Science and Technology, Pharmacy Wing, Jodhpur, Rajasthan, India,
1Lachoo Memorial College of Science and Technology, Pharmacy Wing, Jodhpur, Rajasthan, India
2College of Veterinary and Animal Science, Rajasthan Agricultural University, Rajasthan, India

Abstract

Wounds are major cause of physical disabilities. Wound healing consists of orderly progression of series of events that establish the integrity of the damaged tissues. Recent pharmacological studies have established various pharmacological properties of Ocimum basilicum (Ban-tulsi) like anabolic, hypoglycemic, smooth muscle relaxant, cardiac depressant, antifertility, adaptogenic, antifertility, antidiabetic, antimicrobial, antiallergic and immunomodulator properties. The present study was undertaken on ethanolic extract of leaves of Ocimum basilicum on wound healing activity through topical route on excision wound model. The activity was compared with standard drug Povidone Iodine ointment (5% w/w) respectively. The leaves of Ocimum basilicum ethanolic extract showed faster rate of healing when compared with standard topical application.

Keywords

Ocimum basilicum, Wound healing activity, Excision wound model.

Introduction

Wounds are major cause of physical disabilities. They arise due to physical, chemical or microbial agents. Healing is a survival mechanism and represents an attempt to maintain normal anatomical structure and junction.[1] Many immunosuppressant, cytotoxins and non-steroidal anti-inflammatory drugs suppress the wound healing. Management of wound healing is a complicated and expensive step.[2] Restoration of damaged tissue (wound) is an important process which plays vital role in survival of life. It is imminent for the basis of all surgical manipulations.[3] Many plants have proved to possess significant healing properties in different types of wounds. Using certain plants, possessing antiseptic, astringent, anti-inflammatory, antimicrobial property the rate of wound healing can be enhanced.[4] Such plant can increase the rate of tissue healing by providing different essential substances, required at various steps of wound healing. Plants being cheaper and safer than allopathic drugs, so the treatment by natural ways may be useful in veterinary practice, especially in India where these are found in plenty[5] Ocimum basilicum is a plant commonly known as Ban-tulsi, belongs to the family Lamiaceae [6-7] Recent pharmacological studies have established various pharmacological properties of Ocimum basilicum (Ban-tulsi) like anabolic, hypoglycemic, smooth muscle relaxant, cardiac depressant, antifertility, adaptogenic, antifertility, antidiabetic, antimicrobial, antiallergic and immunomodulator properties.[8-10] Based on the above source of information, the present study aimed to evaluate the wound healing activity of leaves extract of Ocimum basilicum.

Materials and Methods

Collection of Plant Materials

The fresh leaves of Ocimum basilicum were purchased from local nursery garden during the month of November 2007. The plant material was identified and authenticated at Botanical Survey of India, Jodhpur by Dr. P. J. Parmar. The fresh plant material was dried under shade. Dried plant material was powdered using mechanical grinder and passed through sieve no.60 to get the powder of desired coarseness. Powdered material was preserved in an air tight container.

Extraction and Drug Formulation

800 g of the coarsely powdered, dried leaves of Ocimum basilicum was mixed with 4600 ml of ethanol, allowed to stand for 2 days. It was filtered and distilled under vacuum to get concentrated ethanolic extract. The ethanolic extract of Ocimum basilicum (EtObEx) was stored under desiccators for further phytochemical and pharmacological screening. A 5% w/w ointment formulation was made by incorporating the extracts EtObEx separately with simple ointment base I.P10 for external application of the drugs in the excision wound model.

Preliminary Phytochemical Analysis

A preliminary phytochemical screening was carried out for the extract employing the standard procedure to reveal the presence of alkaloids, steroids, terpenoids, flavonoids, saponins, tannins, glycosides, carbohydrates, phytosterols and proteins[11].

Animals

Adult albino rats (wistar strain) of either sex weighing 180-200 g were taken. They were placed in polypropylene cages in a controlled room environment (25°C±2°C) at a natural day night cycle and they were provided with standard laboratory food and water ad libitum. All the experimental procedures and protocol used in this study were reviewed and approved by the Institutional Animal Ethical Committee (IAEC), Utkal University.

Wound Healing Activity

Animals were assigned into 4 groups containing 6 animals in each group. Group I was called as control group, was kept untreated, group II was called as standard group, was treated with market formulation (Povidone Iodine ointment), group III was called as test group III, was treated with ointment of ethanolic leaves extract of Ocimum basilicum (EtObEx). Excision wound model was selected for assessing wound healing activity of Ocimum basilicum (EtObEx) on topical route of administration in a form of a 5%w/w extract in simple ointment base I.P.10 respectively for 15 days. Animals were anaesthetized with ether and shaved on part to be exposed. A circular piece (500 mm2 area) was impressed on the dorsal thoracic region 5cm away from ears and 1 cm away from the vertebral column. The animals were individually housed in separate cages. The test and standard preparation were topically applied once a day till epithelialisation was complete, starting from the day of the operation. The wounds were traced on 1 mm2 graph paper on the day of wounding and then subsequently on the 4th, 8th, 12th and 16th post wound days and thereafter daily until healing was complete. The parameters studied were percentage of wound contraction or wound closure and period of epithelialisation.[12]

Results and Discussion

Preliminary Studies

The results of extractive value and preliminary phytochemical screenings studies for EtObEx are shown in the table1. From this it was found that the 6.53% yield of Ocimum basilicum leaves was obtained. The Ocimum basilicum extract showed the presence of phytoconstituents like β sitosterol, caempferol, rutin, quercetin, carbohydrate, proteins, calcium, magnesium etc.

Pharmacological Screening

Wound healing involves a highly dynamic integrated series of cellular physiological and biochemical processes that occurs in living organisms.[13-14] The majority of world population relies on traditional medicine for their health care.[15] This is also the case in the treatment of wounds. Many research proposed that wound healing can be improved by herbal drugs having antiseptic, antibacterial, antioxidant and anti- inflammatory properties.[16-17] In the present study, the excision wound model is considered for evaluation of wound healing activity of ethanolic extract of Ocimum basilicum (EtObEx) as topical application. The rate of wound contraction in mm2 was taken as a measure of wound healing.

During the initiation of the study from the day 0 there was not much difference in the healing of wounds in all 3 groups. But after day 9, the healing process was faster in the group treated with test formulation of ethanolic extract of Ocimum basilicum as compared to the standard group which was treated with Povidone Iodine ointment as shown in table 2. Statistical analysis was done by ANOVA and Dunnet’s multiple comparision tests. Results are expressed as mean ± SE. n=6 in each group. Significant difference was compared to control group at p<0.01. From the results, it was observed that in the topical route, test group III shows faster wound closure and wound contraction and the results are significant (P<0.01) when compared with standard drug Povidone iodine ointment.

Conclusion

p>Thus, it can be concluded that when the ethanolic extracts of leaves of Ocimum basilicum. was separately applied externally on male albino rats by topical route through excision wound model then it showed faster as well as better wound closure and wound contraction as compared to standard marketed formulation called as Povidone Iodine ointment. Therefore, we may interpret that the presence of such response may be due to the presence of various phytoconstituents which were found to be present in the leaves of Ocimum basilicum. However further studies can be performed to isolate the particular component responsible for generating wound healing activity.

Table 1: The percent yield and presence of phytoconstituents in ethanolic extract of Ocimum basilicum

Extract Percentage yield (w/w) Presence of phytoconstituents
Ocimum basilicum 6.53 Carbohydrate, b-sitosterol, caempferol, rutin, quercetin, proteins

Table 2: Effect of topical administration of ethanolic extract of Ocimum basilicum on excision wound model.

Comparative mean wound area of different groups (in mm2)
Post wounding days Group I (control) Group II (standard) Group III (test group III)
0 226.85± 1.78 226.86± 1.98 221.56± 1.68
3 166.62± 1.96 153.86± 1.86 132.06± 1.27
6 143.04± 1.20 113.04± 1.47 63.58± 1.56
9 94.88± 1.07 63.58± 1.56 21.62± 0.69
12 62.58± 1.46 7.08± 1.76 06.09± 0.54
15 37.46± 1.22 0.00± 0.00 0.00± 0.00

References

  1. Harshmohan, M (2005), “A Text Book of Pathology”, 5, Jaypee Brothers Medical Publishers, New Delhi, 252.
  2. Gupta, N and Gupta, SK (2004), “An Indian community based epidemiological study of wound”,Journal of Wound Care, Vol. 22 (2), 323-25.
  3. Majumdar, M and Kamath, JV (2005), “Herbal concept on wound healing”, Journal of Pharmaceutical Research, Vol. 4 (1), 01-03.
  4. Jaiswal, S and Singh, SV (2004), “Plants used for tissue healing of animals”, Natural Product Radiance, Vol.3 (4), 284-90.
  5. Hahn, DR (1985), “Modern Pharmacognosy”, 4, Handbook Teaching Material Co, Seoul, Korea, 332.
  6. Chopra, RN; Nayar, SL and Chopra, IC (1956), “Glossary of Indian Medicinal Plants”, 1, National Institute of Science and Communication, New Delhi, 495.
  7. Nadkarni, KM (1954), “Indian Materia Medica”, 3, Popular Prakashan Book Depot, New Delhi, 248.
  8. Rastogi, RP and Mehrotra, BN (1984), “Compendium of Indian Medicinal Plants”, CDRI, Lucknow, 494.
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  10. Anonymous (1996), “Indian Pharmacopoeia”, Information Directorate, Ministry of Health and Family Welfare, New Delhi, 931.
  11. Rangari, V (2002),   “Pharmacognosy   and   Phytochemistry”,   1,   Career   Publication, Nashik, 53.
  12. Ghosh, MN (2005), “Fundamentals of Experimental Pharmacology”, 3, Scientific Book Agency, Kolkata, 680.
  13. Srinivas, RB; Kirankumar, R and Naidu, VM (2008), “Evaluation of antimicrobial, antioxidant and wound healing potentials of Haloptela integrifolia”, Journal of Ethnopharmacology, Vol 11 (5), 249-56.
  14. Mukherjee, PK (2002), “Quality Control of Herbal Drugs-An approach to Evaluation of Botanicals”, 1, Business Horizons Pharmaceuticals Publishers, Bombay, 541.
  15. Zhang, X (1996), “Traditional medicines used in wound healing”, Hamdard Medicus, Vol. 39 (3), 102-03.
  16. Somashekar, S; Saraswati, U and Laxinarayana, V (2006), “Wound healing activity of Ocimum sanctum. Linn with supportive role of antioxidant enzymes”, International Journal of Physiology and .Pharmacology, Vol. 50 (2), 163-68.
  17. Sunil, SJ; Agrawal, N and Patil MB (2008). “Antimicrobial and wound healing activities of leaves of Alternanthere sessilis”, International Journal of Greenpharmacy, Vol. 14 (2), 141-44.

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