Comparative effects of Pistacia atlantica leaf and phenytoin ointment on rat skin wound healing: histopathological changes and bioactive compounds.

Abstract

Plant-derived bioactive compounds are increasingly considered as safer alternatives to synthetic drugs in wound management. This study aimed to comparatively evaluate the wound-healing efficacy of Pistacia atlantica (bene tree) leaf extract ointment versus phenytoin in a rat skin wound model. This experimental interventional study was conducted using Pistacia atlantica leaf extract. The extract was first analyzed by gas chromatography-mass spectrometry (GC-MS) and subsequently formulated into herbal ointments. Thirty-five rats were anesthetized using ketamine and xylazine, and standardized excisional skin wounds were created on the dorsal region. Animals were randomly assigned into four groups: control (C), 5% Pistacia ointment (P5), 25% Pistacia ointment (P25), and phenytoin (F). Treatments were applied for 21 days. Wound photographs were taken on days 1, 3, 5, 7, 14, and 21, and wound areas were measured using ImageJ software. At the end of the study, tissue samples were collected in 10% formalin for histopathological analysis with hematoxylin-eosin and Masson's trichrome staining. A significant decrease in body weight was observed in the phenytoin group compared with all other groups (p&#x2009;<&#x2009;0.05). Wound contraction and healing percentages were highest in the P25 and F groups, followed by the P5 and control groups. Histopathological analysis of P25-treated wounds, consistent with the F group, revealed a significant enhancement in epithelialization, angiogenesis, granulation tissue formation, fibroblast proliferation, collagen deposition, and hair follicle regeneration compared with the C group (p&#x2009;<&#x2009;0.001). Furthermore, inflammatory cell infiltration during the healing process was markedly and significantly reduced in the P25 group compared to the other study groups (p&#x2009;<&#x2009;0.001). GC-MS analysis revealed that 1,2,3-benzenetriol (antibacterial), cis-13-octadecenoic acid (anti-inflammatory), and phytol (antimicrobial, anti-inflammatory, antioxidant) were the predominant compounds in the extract. The results indicated that wound healing using an ointment containing Pistacia atlantica leaf extract occurred in a dose-dependent manner and accelerated the healing process through histopathological mechanism without adverse effects compared with phenytoin. The findings support its potential as a safe and effective phytotherapeutic alternative for wound management.