Fabricating a novel HLC-hBMP2 fusion protein for the treatment of bone defects.
Fabricating a novel HLC-hBMP2 fusion protein for the treatment of bone defects.
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DOI：
10.1016/j.jconrel.2020.11.058
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发表时间：
2020-12
期刊：
影响因子：
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通讯作者：
Zhuoyue Chen;Zhen Zhang;Zhaoyue Wang;Jiawei Wu;Yihang Wang;He Si;Xin Xie;L. Shang;Daidi Fan;Fulin Chen
中科院分区：
文献类型：
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作者：
Zhuoyue Chen;Zhen Zhang;Zhaoyue Wang;Jiawei Wu;Yihang Wang;He Si;Xin Xie;L. Shang;Daidi Fan;Fulin Chen
骨形成蛋白（BMPs）等骨诱导剂经胶原蛋白海绵（CS）输送治疗严重骨创伤被认为是一种最佳选择。先前的研究表明，BMP的浓度和释放速率难以精确控制。在这里，我们提出了重组人样胶原（HLC）和人BMP-2 （hBMP2）的重组融合蛋白的制备。融合蛋白保留了高效液相色谱的特性，使得重组蛋白可以在酵母（如asppichia pastorisGS115）中表达，且甲醇诱导后纯化快速，易于批量生产。在体液环境中保持了HLC和hBMP2的稳定性质，具有良好的生物相容性和无细胞毒性。此外，重组融合蛋白制备了垂直通孔结构，提高了力学性能，从而促进了骨髓间充质干细胞（MSCs）向融合材料的迁移。此外，融合蛋白在体内降解并释放hBMP2，从而提高了hBMP2的诱导活性，提高了hBMP2的利用率，并精确控制了hBMP2的释放。该融合蛋白用于大鼠颅骨缺损具有骨诱导活性，促进骨修复，修复率分别比单独使用hplc和对照提高3.5倍和4.2倍。植入部位周围未见明显炎症反应、感染或挤压。我们的数据强烈表明，这种新的重组融合蛋白可能比hBMP2/胶原海绵的简单叠加更有利于骨缺损的治疗。
Treating serious bone trauma with an osteo-inductive agent such as bone morphogenetic proteins (BMPs) has been considered as an optimized option when deliveredviaa collagen sponge (CS). Previous works have shown that the BMP concentration and release rate from approved CS carriers is difficult to control with precision. Here we presented the fabrication of a recombinant fusion protein from recombinant human-like collagen (HLC) and human BMP-2 (hBMP2). The fusion protein preserved the characteristic of HLC allowing the recombinant protein to be expressed in Yeast (such asPichia pastorisGS115) and purified rapidly and easily with mass production after methanol induction. It also kept the stable properties of HLC and hBMP2 in the body fluid environment with good biocompatibility and no cytotoxicity. Moreover, the recombinant fusion protein fabricated a vertical through-hole structure with improved mechanical properties, and thus facilitated migration of bone marrow mesenchymal stem cells (MSCs) into the fusion materials. Furthermore, the fusion protein degraded and released hBMP-2in vivoallowing osteoinductive activity and the enhancement of utilization rate and the precise control of the hBMP2 release. This fusion protein when applied to cranial defects in rats was osteoinductively active and improved bone repairing enhancing the repairing rate 3.5- fold and 4.2- fold when compared to the HLC alone and the control, respectively. There were no visible inflammatory reactions, infections or extrusions around the implantation sites observed. Our data strongly suggests that this novel recombinant fusion protein could be more beneficial in the treatment of bone defects than the simple superposition of the hBMP2/collagen sponge.