Sriwidodo Sriwidodo, Maksum Iman Permana, Subroto Toto, Wathoni Nasrul, Subarnas Anas, Umar Abd Kakhar
Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, Indonesia.
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jatinangor 45363, Indonesia.
J Exp Pharmacol. 2020 Sep 29;12:339-348. doi: 10.2147/JEP.S265727. eCollection 2020.
Human epidermal growth factor (hEGF) has biological activities and can be used in medicines and cosmetics. A high level of effectiveness of hEGF can be obtained when three disulfide bonds fold perfectly. Extracellular secretion from BL21 using the PelB signal peptide is a new way to obtain hEGF with a structure that folds appropriately.
This study aimed to determine the activity and effectiveness of recombinant hEGF excreted by BL21 on wound healing in induced diabetic mice.
Cell proliferation and migration tests were performed on NIH3T3 cells, followed by wound healing tests in induced diabetic mice, along with histological and endotoxin test at various hEGF concentrations (25, 50, and 75 µg/mL).
Based on the results, hEGF at a level of 50 μg/mL showed optimal proliferation and migration activities. Wound healing in induced diabetic mice showed faster-wound closure within 12 days at hEGF 50 and 75 µg/mL with a percentage wound closure of 95% and 98.5%, respectively, which was significant versus control. In the histology test, the number of fibroblasts showed an increase and was significant at hEGF 75 µg/mL compared to the control group. The single test vial (STV) showed that hEGF solution was free of endotoxin.
Recombinant hEGF produced by extracellular secretion using BL21 has optimal diabetic wound healing activity through increased fibroblast proliferation.
人表皮生长因子(hEGF)具有生物活性,可用于医药和化妆品领域。当三个二硫键完美折叠时,hEGF可获得较高的有效性。利用PelB信号肽从BL21中进行细胞外分泌是获得具有适当折叠结构的hEGF的一种新方法。
本研究旨在确定BL21分泌的重组hEGF对诱导型糖尿病小鼠伤口愈合的活性和有效性。
对NIH3T3细胞进行细胞增殖和迁移试验,随后在诱导型糖尿病小鼠中进行伤口愈合试验,并在不同hEGF浓度(25、50和75μg/mL)下进行组织学和内毒素检测。
基于结果,50μg/mL水平的hEGF显示出最佳的增殖和迁移活性。在诱导型糖尿病小鼠中,hEGF 50和75μg/mL组在12天内伤口愈合更快,伤口闭合百分比分别为95%和98.5%,与对照组相比具有显著差异。在组织学检测中,成纤维细胞数量增加,与对照组相比,hEGF 75μg/mL组差异显著。单测试瓶(STV)显示hEGF溶液无内毒素。
利用BL21通过细胞外分泌产生的重组hEGF通过增加成纤维细胞增殖具有最佳的糖尿病伤口愈合活性。