Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice

Abstract

Context

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 E. coli BL21 using the PelB signal peptide is a new way to obtain hEGF with a structure that folds appropriately.

Object

This study aimed to determine the activity and effectiveness of recombinant hEGF excreted by E. coli BL21 on wound healing in induced diabetic mice.

Methods

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).

Results

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.

Conclusion

Recombinant hEGF produced by extracellular secretion using E. coli BL21 has optimal diabetic wound healing activity through increased fibroblast proliferation.

Keywords:

• recombinant hEGF
• fibroblast proliferation
• diabetic wound healing

Abbreviations

EGF, epidermal growth factor; GFs, growth factors; hEGF, human epidermal growth factor; VEGF, vascular endothelial growth factor; FGF, fibroblast growth factor; STV, single test vial; LAL, limulus amebocyte lysate; WST-8, 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; H&E, hematoxylin and eosin; bFGF, basic fibroblast growth factor; TGF-β, transforming growth factor-β.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This work was supported by the Ministry of Research and Technology, Penelitian Terapan Unggulan Perguruan Tinggi (PTUPT), Research Grants of the Ministry of Research and Technology / National Research and Innovation Agency (1827/UN6.3.1/LT/2020).