|Year : 2015 | Volume
| Issue : 1 | Page : 27-30
Healing effect of phenytoin on excisional wound in experimental albino rats
Mohua Sengupta1, Prithwijit Banerjee2, Suhrita Paul3, Janmejoy Sengupta4, Mainak Ghosh5
1 Department of Pharmacology, National Medical College, Baharampur, Kolkata, West Bengal, India
2 Department of Pharmacology, Sagore Dutta Medical College, Baharampur, Kolkata, West Bengal, India
3 Department of Pharmacology, Medical College, Baharampur, Kolkata, West Bengal, India
4 Department of Anaesthesiology, KPC Medical College, Baharampur, Kolkata, West Bengal, India
5 Department of Pharmacology, Murshidabad Medical College, Baharampur, Kolkata, West Bengal, India
|Date of Web Publication||8-Dec-2014|
HIG-Q 1, Niva Park Phase II', PO - Brahmapur, Kolkata - 700 096, West Bengal
Source of Support: None, Conflict of Interest: None
Background and Objective: A common side effect with phenytoin is gingival hyperplasia. This apparent stimulatory effect of phenytoin on connective tissue suggested the possibility for its use in wound healing. This study aims to evaluate the effect of 1% and 2% phenytoin ointment on the excisional wound healing process in experimental albino rats, in comparison to 5% betadine ointment. Materials and Methods: Albino rats of Wistar strain (150200 g) were divided randomly into four groups (n = 6). The animals were anesthetized with ether, shaved on the back, and the skin was disinfected using cotton and alcohol wipes. Excisional round, full-thickness skin wounds of diameter 15 mm were created in the paravertebral area, at 5 mm from the midline on the back of all the animals, using sterile disposable acuderm biopsy needle. The following medications were applied topically to the wound twice daily for a maximum of 20 days. Group A rats served as negative control (untreated). Group B rats were applied 5% betadine ointment (positive control). Group C and group D rats had 1% and 2% phenytoin powder applied on them, respectively. Wound healing was measured on days 0, 4, 8, 12, 16, and 20 of the experiment. Number of days taken for complete epithelization of wound was also noted. Statistical analysis was done using analysis of variance (ANOVA) followed by TukeyKramer test and P < 0.05 was considered significant. Results: Both 1% and 2% phenytoin powder hastened healing and reduced the time taken for complete epithelization of excisional wound in albino rats, when compared to negative control (P < 0.05). While 2% phenytoin powder was superior (P < 0.05) to both 1% phenytoin powder and 5% betadine, the latter two were comparable (P > 0.05). Conclusion: 2% phenytoin may be considered as an important agent for wound healing, but its role in the healing of infected wound needs to be explored further.
Keywords: Phenytoin powder, time for complete epithelization, wound healing
|How to cite this article:|
Sengupta M, Banerjee P, Paul S, Sengupta J, Ghosh M. Healing effect of phenytoin on excisional wound in experimental albino rats. Muller J Med Sci Res 2015;6:27-30
|How to cite this URL:|
Sengupta M, Banerjee P, Paul S, Sengupta J, Ghosh M. Healing effect of phenytoin on excisional wound in experimental albino rats. Muller J Med Sci Res [serial online] 2015 [cited 2019 Oct 16];6:27-30. Available from: http://www.mjmsr.net/text.asp?2015/6/1/27/146419
| Introduction|| |
Widespread existence of unhealed wounds, ulcers, and burns has a great impact on public health and economy. Non-healing and chronic wounds are a significant healthcare problem in today's medical practice.  Appropriate treatment and wound care accelerate the healing process and prevent infection and chronicity of the wound.  Despite extensive efforts to improve wound healing, the outcomes of existing methods are far from optimal.  Many interventions, including new medications and technologies, are being used to help achieve significant wound healing and to eliminate infections.
Phenytoin (diphenylhydantoin) was introduced into therapy in 1937 for the effective control of convulsive disorders.  A common side effect with phenytoin is the development of fibrous overgrowth of gingiva.  This apparent stimulatory effect of phenytoin on connective tissue suggested an exciting possibility for its use in wound healing.  Studies have shown topical phenytoin to promote healing of decubitus ulcers, venous stasis ulcers, diabetic ulcers, traumatic wounds, burns, and leprosy trophic ulcers, ,, but the efficacy of phenytoin in treatment of such conditions is still controversial. The present literature available indicates that topical phenytoin deserves further investigation as a wound healing agent in preclinical studies in a controlled environment.  The present study was conducted to evaluate the effect of phenytoin on the wound healing process compared to standard therapy betadine on laboratory rats.
| Materials and Methods|| |
The experiment was conducted on 24 albino rats of Wistar strain of either sex weighing 150200 g. The animals were caged individually (to avoid licking of wound) in a controlled environment (temperature 25°C ± 2°C) with a 12-h lightdark cycle. Food and water were available ad libitum to the rats. Permission of Institutional Animal Ethics Committee was duly obtained. Prior to creating excisional wounds, the rats were anesthetized by ether anesthesia, shaved on the back, and skin was disinfected using cotton and alcohol wipes. Using sterile surgical instrument, round, full-thickness skin wounds of 15 mm diameter (178 mm 2 ) were created in the paravertebral area, at approximately 5 mm from the midline on the back of rats.  The animals were randomly divided into four groups (n = 6). Group A served as negative control. To group B rats, 5% betadine ointment (positive control) was applied. Rats of groups C and Dhad 1% and 2% phenytoin powder  applied on them, respectively. Wound healing was measured on days 0, 4, 8, 12, 16, and 20 of the experiment. 
To measure the contracture of the wound, a transparent plastic paper was placed on the wound and its shape was drawn on the same paper with a marker and then matched with the graph paper (mm 2 ) for finding the area of the wound to the nearest square millimeter. 
Time taken for complete epithelization (fall of scab without any raw area) was also noted in all the groups. 
Statistical analysis was performed with Graph Pad InStat 3 version. One-way analysis of variance (ANOVA) followed by Tukey-Kramer multiple comparisons test was done. P < 0.05 was considered significant and P < 0.001 was considered highly significant.
| Results|| |
Compared to group A rats, rats of all other groups had significantly reduced wound surface area over a period of 20 days [Table 1]. Contracture of wound surface area was noted as early as on day 4 with 2% phenytoin (group D) (P < 0.05) in comparison with all the other groups.
On the 8 th day of treatment, there was statistically significant reduction (P < 0.05) in wound area in treatment groups B (5% betadine) and C (1% phenytoin) and a highly significant reduction (P < 0.01) in group D (2% phenytoin), when compared to group A (control). This difference widened throughout with P value gradually becoming very highly significant (P < 0.001) in group D in the later part of the study.
In Between-group comparison showed that throughout the study, group B was comparable to group C and the difference was not statistically significant (P > 0.05), while the difference of group B in comparison to group D was statistically significant as early as on the 12 th day and highly significant on the 16 th and 20 th days. Group D showed greater efficacy in healing of wound when compared to group C, which was statistically significant (P < 0.05) on 16 th day onward [Table 1].
The time taken for initiation of granulation tissue formation and the average number of days required for complete epithelization were significantly less (P < 0.05) in all the treatment groups B, C, and D, compared to Group A. Between-group comparison showed that groups B and C were comparable (P > 0.05), whereas group D was superior to groups C and B (P < 0.05) [Table 2].
|Table 2: Number of days required for initiation|
of granulation tissue formation and complete
epithelization of wound (n = 6)
Click here to view
[Figure 1] depicts the comparative reduction of wound surface area in square millimeter (y axis) plotted against the number of days taken for the same (x axis).
| Discussion|| |
Wound healing is the process of restoration of physical integrity of internal or external body structures and involves complex interactions between the cells and various other factors. The healing process consists of a sequence of overlapping events including inflammatory responses, regeneration of the epidermis, shrinkage of the wound, and finally connective tissue formation and remodeling.  On day 4 after injury, new granulation tissue begins to invade the wound gap and numerous new capillaries grow through the new stroma with its granular appearance. After migrating into wounds, fibroblasts begin the synthesis of the extracellular matrix. 
The mechanism by which phenytoin accelerates the wound healing process is unknown. Clinical studies suggest the following as some of the ways by which topical phenytoin hastens the healing process: Stimulation of fibroblastic proliferation, enhancing the formation of granulation tissue, decreasing collagenase activity (by reducing its production or secretion or both), promoting deposition of collagen and other connective tissue components, decreasing bacterial contamination, and decreasing the formation of wound exudate. ,, Biopsies of phenytoin-treated open wounds show neovascularization and collagenization.  Local pain relief due to membrane-stabilizing action of topical phenytoin therapy has been observed in several studies. , Pendse et al., reported the antibacterial activity of phenytoin in the wound healing process.  Lipid peroxidation is an important process in several types of injuries like burns, wounds, and skin ulcers. Therefore, a drug like phenytoin, which inhibits lipid peroxidation, may hasten the process of wound healing. 
Topical application of phenytoin results in direct access of the drug to the target site and avoids the risk of getting systemic side effects.  The findings of the above-mentioned study were taken into consideration for selecting the route of administration of phenytoin for wound healing in the present study. Modaghegh et al.,  compared four topical phenytoin formulations (gel, cream, phenytoin sodium powder, and phenytoin powder) in a rat model of wound healing and concluded that the phenytoin powder showed the most favorable results. Thus, we considered phenytoin as powder formulation in our study to avoid contamination.
| Conclusion|| |
In the present study, it is seen that both 1% and 2% phenytoin powder hasten healing and the time taken for complete epithelization of excisional wound in albino rats when compared to negative control. While 2% phenytoin powder was superior to both 1% phenytoin powder and 5% betadine, the latter two were comparable. These findings support the findings of earlier studies, but its systemic side effects and its action in contaminated wounds need further evaluation.
| Acknowledgment|| |
The authors would like to thank all the members of Department of Pharmacology, Medical College, Kolkata, for their constant support to carry out this experiment.
| References|| |
Alizadeh A, Mohagheghi M, Khaneki M, Saeed PK. A study of the effect of magnesium hydroxide on the wound healing process in rats. Med J Islamic World Acad Sci 2007;16:165-70.
Clark RA. Basis of cutaneous wound repair. J Dermatol Surg Oncol 1993;19:693-706.
Anstead GM, Hart LM, Sunahara JF, Liter ME. Phenytoin in wound healing. Ann Pharmacother 1996;30:768-75.
Bhatia A, Prakash S. Topical phenytoin for wound healing. Dermatol Online J 2004;10:5.
Tripathi KD. Antiepileptic drugs. In: Tripathi KD, editor. Essentials of Medical Pharmacology. 6 th
ed. New Delhi, India: Jaypee Brothers; 2008. p. 382-5.
Simpson GM, Kunz E, Slafta J. Use of sodium diphenylhydantoin in treatment of leg ulcers. N Y State J Med 1965;65:886-8.
Bansal NK, Mukul. Comparison of topical phenytoin with normal saline in the treatment of chronic trophic ulcers in leprosy. Int J Dermatol 1993;32:210-3.
Malhotra YK, Amin SS. Role of topical phenytoin in trophic ulcers of leprosy in India. Int J Lepr Other Mycobact Dis 1991;59:337-8.
Modaghegh S, Salehian B, Tavassoli M, Djamshidi A, Rezai AS. Use of phenytoin in healing of war and non-war wounds. A pilot study of 25 cases. Int J Dermatol 1989;28:347-50.
Iftikhar F, Arshad M, Rasheed F, Amraiz D, Anwar P, Gulfraz M. Effects of acacia honey on wound healing in various rat models. Phytother Res 2010;24:583-6.
Rathi B, Badri N. Wound healing effects of beetle leaf in albino rats. Orissa J Pharm 2006;3:21-5.
Clark RA, Nielsen LD, Welch MP, McPherson JM. Collagen matrices attenuate the collagen-synthetic response of cultured fibroblasts to TGF-beta. J Cell Sci 1995;108:1251-61.
Rhodes RS, Heyneman CA, Culbertson VL, Wilson SE, Phatak HM. Topical phenytoin treatment of stage II decubitus ulcers in the elderly. Ann Pharmacother 2001;35:675-81.
McAnally LE, Thompson D. Use of phenytoin for wound healing. Hosp Pharm 1992;27:649-50.
Talas G, Brown RA, McGrouther DA. Role of phenytoin in wound healing - a wound pharmacology perspective. Biochem Pharmacol 1999;57:1085-94.
Pendse AK, Sharma A, Sodani A, Hada S. Topical phenytoin in wound healing. Int J Dermatol 1993;32:214-7.
Senel O, Cetinkale O, Ozbay G, Ahçioðlu F, Bulan R. Oxygen free radicals impair wound healing in ischemic rat skin. Ann Plas Surg 1997;39:516-23.
[Table 1], [Table 2]
|This article has been cited by|
||Evaluation of wound healing activity of plumbagin in diabetic rats
| ||Yi Shao,Minyan Dang,Yukiat Lin,Feng Xue |
| ||Life Sciences. 2019; |
|[Pubmed] | [DOI]|
||Antimicrobials tethering on suture surface through a hydrogel: a novel strategy to combat postoperative wound infections
| ||Himadri Kalita,Ankita Hazarika,Sanjeeb Kalita,Raghuram Kandimalla,Rajlakshmi Devi |
| ||RSC Adv.. 2017; 7(52): 32637 |
|[Pubmed] | [DOI]|
||The influence of growth factors on skin wound healing in rats
| ||Elen Carolina David Joćo De Masi,Antonio Carlos Ligocki Campos,Flavia David Joćo De Masi,Marco Aurelio Soatti Ratti,Isabela Shin Ike,Roberta David Joćo De Mais |
| ||Brazilian Journal of Otorhinolaryngology. 2016; |
|[Pubmed] | [DOI]|