Mechanisms of pNaSS-grafted poly(epsilon-caprolactone) scaffold-guided rAAV-mediated gene transfer for enhanced, safe human anterior cruciate ligament repair

膝关节前交叉韧带（ACL）损伤是骨科常见的关键问题和社会经济负担，可能导致骨关节炎，因为ACL具有有限的内在自我愈合能力。由于目前的临床（保守，手术）干预措施都不支持ACL病变的持久，完全修复，基于基因治疗的创新选择可能会提供强大的工具，以加强ACL修复的过程。在ACL病变中施用有效的、临床适应的重组腺相关病毒（rAAV）载体是在损伤部位暂时和空间转移修复基因以延长基因产物表达的有效方法。尽管如此，rAAV的安全临床使用仍然受到体内各种障碍的阻碍，特别是人体中存在针对AAV衣壳蛋白的天然中和抗体。为了解决这一具有挑战性的问题，本提案的目标是检验编码修复性腺相关病毒的治疗性rAAV载体能够抑制肿瘤细胞增殖的假设。（增殖，促合成代谢）基因（碱性成纤维细胞生长因子- FGF-2 -和转化生长因子β- TGF-β）可以通过功能化的聚苯乙烯磺酸钠接枝聚（ε-己内酯）（pNaSS接枝的PCL）膜作为保护性的受控递送平台，并且持久地增强人实验性ACL损伤离体的翻译前模型中ACL修复的相关过程和机制，包括在实验性体液中和的条件下，以及在临床相关的条件下，相对于直接应用无膜形式的载体，体内平移实验ACL损伤。该项目可能会提供新的，有效的治疗方法，以加强ACL修复患者在不久的将来。

Injuries of the knee anterior cruciate ligament (ACL) are prevalent, critical issues and a socioeconomical burden in orthopaedics, potentially leading to osteoarthritis, since the ACL has a limited intrinsic ability for self healing. As none of the current clinical (conservative, surgical) interventions support the durable, full repair of ACL lesions, innovative options based on gene therapy may provide strong tools to enhance the processes of ACL repair. Administration of the potent, clinically adapted recombinant adeno-associated virus (rAAV) vectors in ACL lesions is a potent approach to temporarily and spatially transfer reparative genes in sites of injury for a prolonged expression of the gene products. Still, the safe, clinical use of rAAV remains impeded by diverse obstacles in vivo, especially by the presence of natural neutralizing antibodies against the AAV capsid proteins in humans. To address this challenging issue, the goal of the present proposal is to test the hypothesis that therapeutic rAAV vectors coding for reparative (proliferative, pro-anabolic) genes (the basic fibroblast growth factor - FGF-2 - and the transforming growth factor beta - TGF-ß) may be delivered via functionalized poly(sodium styrene sulfonate)-grafted poly(epsilon-caprolactone) (pNaSS-grafted PCL) films as protective, controlled delivery platforms to safely, effectively, and durably enhance the processes and mechanisms relevant of ACL repair in a pre-translational model of human experimental ACL lesions ex vivo, including in conditions of experimental humoral neutralization, and in clinically relevant, translational experimental ACL lesions in vivo relative to direct application of the vectors in their film-free form. This project may offer new, effective therapies to enhance ACL repair in patients in a close future.