PRODUCTION AND in vitro EVALUATION OF RECOMBINANT HUMAN RHHB-EGF FOR WOUND HEALING AND TARGETED THERAPY I. Vovk, A., Didan, D. Zhukova, L. Dronko, A. Rebriev, A. Rybalko, E. Legach, O. Gorbatiuk, M. Usenko, A. Skvarchynskyi, T. Dovbymchuk, A.

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ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)

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Biotechnologia Acta Т. 18, No. 1 , 2025
P. 55-66, Bibliography 29, Engl.
UDC:: [615.454.1:577.112.6]+[616-08-031.84:616-001.4]
DOI: https://doi.org/10.15407/biotech18.01.055

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PRODUCTION AND in vitro EVALUATION OF RECOMBINANT HUMAN RHHB-EGF FOR WOUND HEALING AND TARGETED THERAPY

I. Vovk¹, A. Didan¹, D. Zhukova¹, L. Dronko², A. Rebriev¹, A. Rybalko¹, E. Legach³, O. Gorbatiuk⁴, M. Usenko⁵, A. Skvarchynskyi⁶, T. Dovbymchuk⁶, A. Siromolot¹, 6, S. Romaniuk¹, D. Kolybo₁

¹Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv
²National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”
³Institute for Problems of Cryobiology and Cryomedicine
⁴Institute of Genetic and Regenerative Medicine, M.D. Strazhesko National Scientific Center of Cardiology, Clinical and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv
⁵Institute of Molecular Biology and Genetics of the National Academy of Sciences of Ukraine, Kyiv
⁶Educational and Scientific Center “Institute of Biology and Medicine” Taras Shevchenko National University of Kyiv, Ukraine

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 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, 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.

Key words: human heparin-binding EGF-like growth factor (rhHB-EGF), recombinant protein, cell culture, fibroblasts, proliferation, migration, cytotoxicity, wound healing.

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