基于拉曼光谱的具有不同浓度梯度的生物玻璃/胶原人工韧带涂层对腱骨愈合的效应及其机制研究

The graft choice in ACLR (Anterior Cruciate Ligament Reconstruction) is always placed in hot and difficult issues in sports medicine. Currently, more and more researchers focus on the study of artificial ligament. However, drawbacks such as low graft-bone healing, synovitis and unfirmed interference fixation were reported in the use of LARS(Ligament Advanced Reinforcement System). The preliminary results showed that some bioactive materials coating on the surface of PET artificial ligament in its inter-tunnel segment could promote the tendon-bone healing, however, the transition of the ligament-bone healing is complicate due to less knowledge of this structure. To address these limitations, a tissue engineering approach aimed at recreating the complex enthesis through biomimetic design has significant potential to improve the clinical outcome of ACL reconstruction using synthetic grafts. We hypothesize that incorporation of a biomimetic Bioglass-collagen gradient that based on the Raman spectrum to mimic a compositionally, mechanically and biologically native ACL interface. Furthermore, the Biomechanical tests, cellular compatibility tests and in vivo experiments on animal will be carried out to approve the advantages of the coated ligament on biocompatibility, tendon-bone healing, choose the best coating method, to further understand the mechanism of the signal pathways, MAPK and c-Jun pathways, and to test some relative osteostimulative factors and VEGF. Our research will provide theoretical and experimental foundation for the mimicked native artificial ligament.

膝关节前交叉韧带损伤重建手术的移植物选择是一直是运动医学研究的热点及难点。目前人工韧带的研究在国内外受到高度关注，但其存在腱骨愈合不佳、并发滑膜炎、挤压螺钉固定易松动等缺陷。前期研究表明对现有材料表面进行生物玻璃涂层处理可有效促进其在体内与骨的愈合，但是在腱骨结合部的愈合一直是难点。本项目旨在通过拉曼光谱分析正常将腱骨结合部构成成分及变化规律，在此基础上运用胶原压制成型法（Plastically Compressed Collagen, PCC）对现有材料（聚对苯二甲酸乙二醇酯）进行仿生涂层处理，并通过材料表征、体外细胞相容性及成骨性能试验以及体内动物实验包括病理检测及生物力学测试，验证其体内外优势。同时对生物玻璃涂层后MAPK及c-Jun信号通路的相关分子检测以及体内成骨成血管因子的检测，探索生物玻璃涂层促材料体内外生物性能的机制，为仿生新型生物型人工韧带在临床应用提供理论和实验依据。

膝关节前交叉韧带损伤重建手术的移植物选择一直是运动医学研究的热点及难点。目前，人工韧带的研究在国内外受到高度关注，但其存在腱骨愈合不佳、并发滑膜炎、挤压螺钉固定易松动等缺陷。前期研究表明对现有材料表面进行生物玻璃涂层处理可有效促进其在体内与骨的愈合，但是在腱骨结合部的愈合一直是难点。本项目拟在通过拉曼光谱分析正常将腱骨结合部构成成分及变化规律，此基础上运用胶原压制成型法(Plastically Compressed Collagen, PCC)对现有材料(聚对苯二甲酸乙二醇酯)进行仿生涂层处理，并进行相应验证。但计划开展后发现聚多巴胺（PDA）通过共价键的结合与传统意义上的简单物理涂层譬如：自然沉积、等离子喷涂、胶原压制成型法等等，有着显著差别。它主要的作用方式有共价成键、静电相互作用、π-π堆栈作用等，其中居于主导地位的作用方式与被界面的性质有关，如金属表面能够与聚多巴胺通过金属有机共轭结构发生成键反应，聚乙烯表面的聚多巴胺修饰可能以静电相互作用为主导，而含苯环类聚合物则可能通过π-π堆栈作用与聚多巴胺结合。这样的结合方式更有利于涂附材料的粘附，从而避免涂层材料在运用中的丢失。本研究达到预期目标，将渐变的涂层方法应用于制备人工韧带表面涂层以促进移植物骨愈合，通过可控制的修饰移植物表面，提高移植物对细胞粘附和诱导成骨的能力，实现人工韧带移植物骨界面成骨，为解决组织工程研究领域中“人工韧带移植物骨愈合不良”等关键性问题提供新方法和新思路。