Despite various clinical options, human anterior cruciate ligament (ACL) lesions do not fully heal. Biomaterial-guided gene therapy using recombinant adeno-associated virus (rAAV) vectors may improve the intrinsic mechanisms of ACL repair. Here, we examined whether poly(sodium styrene sulfonate)-grafted poly(ε-caprolactone) (pNaSS-grafted PCL) films can deliver rAAV vectors coding for the reparative basic fibroblast growth factor (FGF-2) and transforming growth factor beta (TGF-β) in human mesenchymal stromal cells (hMSCs) as a source of implantable cells in ACL lesions. Efficient and sustained rAAV-mediated reporter (red fluorescent protein) and therapeutic (FGF-2 and TGF-β) gene overexpression was achieved in the cells for at least 21 days in particular with pNaSS-grafted PCL films relative to all other conditions (up to 5.2-fold difference). Expression of FGF-2 and TGF-β mediated by rAAV using PCL films increased the levels of cell proliferation, the DNA contents, and the deposition of proteoglycans and of type-I and -III collagen (up to 2.9-fold difference) over time in the cells with higher levels of transcription factor expression (Mohawk, Scleraxis) (up to 1.9-fold difference), without activation of inflammatory tumor necrosis alpha especially when using pNaSS-grafted PCL films compared with the controls. Overall, the effects mediated by TGF-β were higher than those promoted by FGF-2, possibly due to higher levels of gene expression achieved upon rAAV gene transfer. This study shows the potential of using functionalized PCL films to apply rAAV vectors for ACL repair.

中文翻译：

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使用 pNaSS 移植的 PCL 膜，rAAV 介导的 FGF-2 和 TGF-β 过表达使人间充质基质细胞对 ACL 成纤维细胞分化的承诺


尽管有多种临床选择，人类前十字韧带 (ACL) 损伤仍无法完全愈合。使用重组腺相关病毒 (rAAV) 载体的生物材料引导的基因治疗可能会改善 ACL 修复的内在机制。在这里，我们检查了聚（苯乙烯磺酸钠）接枝的聚（ε-己内酯）（pNaSS 接枝的 PCL）薄膜是否可以传递编码修复性碱性成纤维细胞生长因子（FGF-2）和转化生长因子β（TGF）的 rAAV 载体。 -β) 存在于人间充质基质细胞 (hMSC) 中，作为 ACL 损伤中可植入细胞的来源。相对于所有其他条件（高达5.2 倍差异）。随着时间的推移，rAAV 使用 PCL 薄膜介导的 FGF-2 和 TGF-β 表达增加了细胞增殖水平、DNA 含量以及蛋白聚糖和 I 型和 III 型胶原蛋白的沉积（差异高达 2.9 倍）在转录因子表达水平较高的细胞（Mohawk、Scleraxis）中（差异高达 1.9 倍），而没有激活炎症性肿瘤坏死 α，特别是与对照组相比，使用 pNaSS 移植的 PCL 薄膜时。总体而言，TGF-β介导的作用高于FGF-2促进的作用，可能是由于rAAV基因转移后基因表达水平较高。这项研究显示了使用功能化 PCL 薄膜应用 rAAV 载体进行 ACL 修复的潜力。