半月板细胞自分泌的ColIV-RECM促进半月板植入物血管长入和组织再生的作用及机制研究

半月板撕裂后难以自我修复。人工半月板植入后早期血管长入困难，影响其组织再生和长期疗效。我们采用自主研制的特殊培养基制备出半月板细胞自分泌的富含Ⅳ型胶原及细胞迁移相关蛋白的细胞外基质（ColⅣ-RECM），前期验证其可明显促进细胞迁移和血管生成。本项目拟通过细胞迁移和成管实验研究这种ColⅣ-RECM启动内皮细胞及半月板细胞趋化链式反应的机制；通过上下调ColⅣ的表达及向普通培养基添加迁移相关蛋白研究上述成分对细胞迁移及血管形成的影响；通过检测FAK-actin迁移相关细胞内信号磷酸化变化、迁移和未迁移的细胞的基因表达差异,阐明上述成分促进血管再生的作用及分子机制；制作兔半月板部分切除模型，用3D复合打印的ColⅣ-RECM与聚己内酯植入物修补缺损，探索其体内促进血管长入、组织再生的作用及机制。本项目可建立一种通过ECM修饰促进血管化的人工半月板构建新模式，并具有良好的应用前景。

Meniscal tears are common sports injuries. The difficulty of early blood vessel growth in the artificial meniscus implant affects its tissue regeneration and long-term efficacy. This study intends to use a special medium with independent intellectual property rights to act on meniscus fibrochondrocytes to produce an extracellular matrix (ColIV-RECM) rich in type IV collagen and cell migration-related proteins, which can promote early vascularization of meniscus. The migration and tube formation experiments were used to study the role of ColⅣ-RECM in initiating chemotactic chain reaction of vascular endothelial cells and meniscus fibrochondrocytes, and to clarify the mechanism of promoting vascular growth and meniscal regeneration.By studying the effects of regulating the expression of ColⅣ and adding migration-related proteins to normal medium on cell migration and angiogenesis; by detecting FAK-actin migration-related intracellular signal phosphorylation changes and gene expression differences between migrating and non-migrating cells, clarify the role and molecular mechanism of the above components in promoting vascular regeneration, and then use 3D printing technology to produce ColIV-RECM composite scaffold with polycaprolactone was implanted in situ into rabbit knee meniscus defects to explore its role and mechanism in promoting vascular growth and tissue regeneration of meniscus grafts in vivo. This project can establish a new model of artificial meniscus for promoting vascularization through extracellular matrix modification, and has a good application prospect.