控释活性生物分子的氧化凝胶静电纺纤维及抑制肌腱粘连研究

Peritendinous adhesion is a common clinical disease which severely affects patients’ local body functions. Previously, we successfully employed electrospun fibrous membrane as drug carrier to prevent peritendinous adhesions caused by inflammation and cell adhesion, but were unable to prevent adhesions at later stage due to significant cell proliferation. It has been found that extracellular signal-regulated kinase 2 (ERK2) is a key factor for inhibiting adhesions caused by fibroblast proliferation. Therefore, in present study, we will synthesize polycationic nanoparticles for efficient delivery of ERK2-siRNA. Subsequently, an anti-adhesion film will be fabricated using oxide-gel electrospinning to incorporate polycationic-ERK2-siRNA nanoparticle complexes into electrospun fibers. In detail, the thiolated gelatin with oxidative crosslinking property, along with polycationic gelatin nanoparticles, will be compounded into the electrospun fibers in order to protect and control release the ERK2-siRNA. The in vitro study will include investigations of intracellular gene transfection and inhibition of fibroblasts adhesion and proliferation, whereas the in vivo study will examine the impact mechanism of tendon tissue adhesions. Importantly, this study will develop an oxide-gel electrospinning technology to protect water-soluble bioactive biomolecules, and will provide more insight for inhibiting tendon adhesion and promoting the rehabilitation of limb functions.

肌腱粘连是临床常见的疾病，严重影响患者局部功能。前期，我们通过携载药物的静电纺纤维膜有效的预防因炎症和细胞粘附导致的粘连，但是无法预防后期增生导致的再粘连。研究发现抑制增生导致粘连的重要靶点是细胞外信号调节激酶2（ERK2），在此基础上，本项目合成高效输送ERK2-siRNA的聚阳离子纳米颗粒，进一步设计氧化凝胶静电纺丝技术制备携载聚阳离子-ERK2-siRNA络合纳米颗粒的静电纺纤维膜作为防粘连膜研究。合成在氧气环境中可交联的巯基化明胶，构建携载聚阳离子络合颗粒的明胶纳米凝胶颗粒，再凝胶共纺法构建静电纺载药纤维，实现对活性生物分子的保护和控释。体外考察纤维膜力学、药物控释等理化性能、细胞内基因转染和成纤维细胞的抑制，体内分析肌腱粘连组织增生的影响机制。本研究将开发一种保护水溶性活性生物分子的氧化凝胶静电纺丝技术，并从分子机制上为防止肌腱粘连和促进患肢功能康复提供新的思路。

肌腱粘连是临床常见的疾病，严重影响患者局部功能。本项目合成了一种高效输送ERK2-siRNA的聚阳离子纳米颗粒，进一步设计氧化凝胶静电纺丝技术制备携载聚阳离子-ERK2-siRNA络合纳米颗粒的静电纺纤维膜作为防粘连膜研究。体外考察了该纤维膜细胞内基因转染和成纤维细胞的抑制，体内分析肌腱粘连组织增生的影响机制。解析了炎症、细胞粘附和增生与肌腱粘连之间的内在关系和作用机制。该复合支架被证实可以有效抑制成纤维细胞的增殖效率和细胞活性。ERK2-siRNA纳米粒子从复合防粘连屏障中释放出来后，仍具有很高的生物活性，可以下调细胞的Smad3和下游的胶原蛋白表达，抑制细胞增殖效率和细胞活性，并且植入体内后具有显著的抗粘连效果，同时不影响肌腱愈合后的力学强度。此外，针对肌腱粘连的临床需求，先后制备了复合MMP响应性水凝胶防粘连膜、水合高润滑的纳米纤维及免疫对冲的纤维防粘连膜。此类防粘连膜均在肌腱等软组织粘连方面获得较好的应用，为临床上防治肌腱粘连提供了新的思路和治疗策略。