Production and in vitro evaluation of recombinant human rhHB-EGF for wound healing and targeted therapy

Aim. The goal of the study was to evaluate the biological activity of recombinant human heparinbinding EGF-like growth factor (rhHB-EGF) on mouse fibroblasts in vitro and the possibility of its use as a potential agent for promoting wound healing and tissue regeneration. Methods. The study employed a scratch assay to evaluate the migration of mouse fibroblasts (L929 and NIH-3T3), the MTT test to assess cell proliferation and/or metabolic activity, MALDI-TOF mass spectrometry for protein identification, and flow cytometry to determine cell viability. Results. In the concentration range of 500–1000 ng/ml rhHB-EGF, no cytotoxic effect was recorded, but an increase in proliferation and/or metabolic activity, as well as migration of fibroblasts, was detected, with a maximum effect at 500 ng/ml rhHB-EGF in the cell incubation medium. A 30% overgrowth of the wound surface of fibroblasts was demonstrated in the scratch assay test under the influence of rhHB-EGF compared to the corresponding control. Conclusions. rhHB-EGF at a concentration of 500 ng/ml can be used in preparations to stimulate wound healing and tissue regeneration due to its ability to stimulate proliferation/metabolic activity and migration of fibroblasts, as well as the lack of cytotoxicity. Further, in vivo studies are needed for a comprehensive evaluation of this possibility.