Histological Studies on a Newly Isolated Bacillus subtilis D10 Protease in the Debridement of Burn Wound Eschars Using Mouse Model

Abstract

Background: Proteases are among the most important industrial enzymes, playing a critical role in the physiological, biochemical, and regulatory processes of all living organisms. This study evaluated the histological effects of a Bacillus subtilis D10 protease in combination with the antibacterial ointment silver sulfadiazine (SSD) on the burned skin of mice. Materials and Methods: The bacterial proteolytic enzyme was produced and purified through DEAE-Sepharose CL-6B and Sephadex G-100 FF. The in vitro protease specificity was then determined. The dorsal skin of albino mice was burned with 80% HCl solution, then treated under three conditions: cold cream, SSD, and SSD combined with the tested protease. After 15 days of daily treatment, the mice were sacrificed and skin tissue samples were histopathologically examined using hematoxylin eosin, and Masson trichrome staining. Results: The D10 protease hydrolyzed the proteinaceous components of eschars (fibrin, normal collagen, and denatured collagen) in vitro. Mice skins treated with protease and SSD mixture showed promising results, with more rapid healing than the other treatments. This group regenerated epidermis and dermis with newly formed granulated follicles, fibroblasts and blood capillaries in the dermis, and collagen fibers in the hypodermis. Conclusions: These results suggest that the serine protease produced by B. subtilis D10 promotes wound healing of mice skin burnt with HCl and restores the normal architectural pattern in a shorter time than the standard treatments.

Saccharomyces cerevisiae OS303 expression of an alkaline protease from a newly isolated Bacillus subtilis D9

“…The use of them as detergent additives represent the largest application among the industrial enzymes. Another industrial process that has received attention is the enzyme aided dehairing of animal hides and skin in the leather industry (Al-Dhuayan et al, 2021). The goal of this study was to clone the local Bacillus subtilis D9 alkaline protease gene in the yeast system for high expression level of D9 alkaline protease.…”

“…Different reports have focused on the characterization and applications of alkaline proteases produced by B. subtilis group, recovered from Saudi Arabia, because of their industrial importance in feather degradation, wound healing, and detergents (Al-Dhuayan et al, 2021;Mahmoud et al, 2021;Almahasheer et al, 2022).…”

The Use of Menthol in Skin Wound Healing—Anti-Inflammatory Potential, Antioxidant Defense System Stimulation and Increased Epithelialization

“…Our results are in accordance with several studies reported in the literature. Such studies showed that the inflammatory phase restores homeostasis, and that, in the proliferative phase, fibroblasts and other cells from the connective tissue infiltrate the wound site and secrete cytokines, attracting keratinocytes and leading to re-epithelialization [26,46]. Preventing prolonged inflammation by suppressing the production of inflammatory cytokines is a desirable target for wound healing products; excessive inflammation results in chronic wounds and scar formation [47].…”

In vitro and in silico characterization of alkaline serine protease from Bacillus subtilis D9 recovered from Saudi Arabia

“…are mostly the microbial source to generate the proteases. Bacillus -derived alkaline proteases have numerous applications in industry, including organic synthesis, food, pharmacology, leather, and bioremediation ( Tarhriz et al., 2014 ; Al-Dhuayan et al., 2021 ).…”