Chen Zhuoyue, Wang Rongrong, He Jing, Liu Qian, Zhang Yifan, Wang Yijie, Liu Ling, Song Mingming, Chen Fulin
Provincial Key Laboratory of Biotechnology of Shaanxi, Key Laboratory of Resource Biology and Modern Biotechnology in Western China, Faculty of Life Science, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi Province 710069, China.
ACS Appl Bio Mater. 2025 Jan 20;8(1):236-251. doi: 10.1021/acsabm.4c01096. Epub 2024 Dec 26.
In this study, we designed a fusion protein, rhCR, by combining human collagen with the self-assembling peptide RADA-16 using genetic engineering technology. The rhCR protein was successfully expressed in . The rhCR can self-assemble into a three-dimensional nanofiber network under physiological conditions. The lyophilized rhCR sponge exhibited high elasticity modulus and stable swelling properties. experiments confirmed that the rhCR had good biocompatibility and could significantly promote the adhesion, proliferation, and migration of fibroblasts (L929), upregulating the expression of genes such as , , , and in L929 cells. When applied to a mouse liver hemorrhage model, rhCR hemostatic sponges rapidly formed nanofibers on the ruptured liver surface, activated platelet CD62P, and significantly reduced blood loss and bleeding duration compared to the recombinant human collagen (rhCol) alone. Furthermore, the rhCR application markedly accelerated wound healing in a mouse full-thickness skin defect model, with the wound healing rate in the rhCR group being 2.6 times that of the untreated group and 1.7 times that of the rhCol group on day 6 postinjury. Histological and immunofluorescence analyses revealed that the rhCR promoted collagen deposition and epidermal regeneration and improved the quality of skin tissue repair by stimulating tissue cells to produce cytokines, growth factors, and immune factors through immunological regulation. The rhCR fusion protein combines the advantages of collagen and RADA-16, overcoming the limitations of their separate use in hemostatic and tissue engineering applications. This biomaterial and its design idea hold promise for a variety of regenerative applications.
在本研究中,我们利用基因工程技术将人胶原蛋白与自组装肽RADA-16相结合,设计了一种融合蛋白rhCR。rhCR蛋白在 中成功表达。rhCR在生理条件下可自组装成三维纳米纤维网络。冻干的rhCR海绵表现出高弹性模量和稳定的溶胀性能。 实验证实,rhCR具有良好的生物相容性,可显著促进成纤维细胞(L929)的黏附、增殖和迁移,上调L929细胞中 、 、 和 等基因的表达。当应用于小鼠肝出血模型时,与单独的重组人胶原蛋白(rhCol)相比,rhCR止血海绵在破裂的肝脏表面迅速形成纳米纤维,激活血小板CD62P,并显著减少失血量和出血持续时间。此外,在小鼠全层皮肤缺损模型中,rhCR的应用显著加速了伤口愈合,在损伤后第6天,rhCR组的伤口愈合率是未治疗组的2.6倍,是rhCol组的1.7倍。组织学和免疫荧光分析表明,rhCR通过免疫调节刺激组织细胞产生细胞因子、生长因子和免疫因子,促进胶原蛋白沉积和表皮再生,提高皮肤组织修复质量。rhCR融合蛋白结合了胶原蛋白和RADA-16的优点,克服了它们在止血和组织工程应用中单独使用的局限性。这种生物材料及其设计理念在各种再生应用中具有广阔前景。