School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea.
BioMAX/N-Bio Institute, Institute of BioEngineering, Seoul National University, 1 Gwanakro, Gwanak-gu, Seoul, 08826, Republic of Korea.
Adv Biol (Weinh). 2021 Jan;5(1):e2000176. doi: 10.1002/adbi.202000176. Epub 2021 Jan 4.
The instability of recombinant basic fibroblast growth factor (bFGF) is a major disadvantage for its therapeutic use and means frequent applications to cells or tissues are required for sustained effects. Originating from silkworm hemolymph, 30Kc19α is a cell-penetrating protein that also has protein stabilization properties. Herein, it is investigated whether fusing 30Kc19α to bFGF can enhance the stability and skin penetration properties of bFGF, which may consequently increase its therapeutic efficacy. The fusion of 30Kc19α to bFGF protein increases protein stability, as confirmed by ELISA. 30Kc19α-bFGF also retains the biological activity of bFGF as it facilitates the migration and proliferation of fibroblasts and angiogenesis of endothelial cells. It is discovered that 30Kc19α can improve the transdermal delivery of a small molecular fluorophore through the skin of hairless mice. Importantly, it increases the accumulation of bFGF and further facilitates its translocation into the skin through follicular routes. Finally, when applied to a skin wound model in vivo, 30Kc19α-bFGF penetrates the dermis layer effectively, which promotes cell proliferation, tissue granulation, angiogenesis, and tissue remodeling. Consequently, the findings suggest that 30Kc19α improves the therapeutic functionalities of bFGF, and would be useful as a protein stabilizer and/or a delivery vehicle in therapeutic applications.
重组碱性成纤维细胞生长因子(bFGF)的不稳定性是其治疗应用的主要缺点,这意味着需要频繁地应用于细胞或组织,以维持其持续效果。30Kc19α 来源于家蚕血液,是一种具有细胞穿透能力的蛋白质,同时也具有蛋白质稳定特性。本文研究了将 30Kc19α 与 bFGF 融合是否可以增强 bFGF 的稳定性和皮肤穿透特性,从而提高其治疗效果。ELISA 结果证实,30Kc19α 与 bFGF 融合蛋白增加了蛋白质稳定性。30Kc19α-bFGF 还保留了 bFGF 的生物学活性,因为它促进了成纤维细胞的迁移和增殖以及内皮细胞的血管生成。研究发现,30Kc19α 可以改善通过无毛小鼠皮肤的小分子荧光染料的透皮传递。重要的是,它增加了 bFGF 的积累,并通过毛囊途径进一步促进其向皮肤的转运。最后,当在体内皮肤创伤模型中应用时,30Kc19α-bFGF 有效地穿透真皮层,促进细胞增殖、组织肉芽形成、血管生成和组织重塑。因此,这些发现表明 30Kc19α 改善了 bFGF 的治疗功能,并且作为一种蛋白质稳定剂和/或治疗应用中的输送载体将是有用的。