Cucchiarini Magali, Schmidt Katharina, Frisch Janina, Kohn Dieter, Madry Henning
Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany
Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
Am J Sports Med. 2015 May;43(5):1197-205. doi: 10.1177/0363546514567063. Epub 2015 Feb 2.
Direct application of therapeutic gene vectors derived from the adeno-associated virus (AAV) might be beneficial to improve the healing of meniscal tears.
To test the ability of recombinant AAV (rAAV) to overexpress the potent transforming growth factor-β (TGF-β) in primary cultures of human meniscal fibrochondrocytes, in human meniscal explants, and in experimental human meniscal lesions as a new tool to enhance meniscal repair.
Controlled laboratory study.
The effects of the candidate treatment on the proliferative and metabolic activities of meniscal cells were monitored in vitro for up to 21 days and in situ in intact and injured human menisci for up to 15 days using biochemical, immunohistochemical, histological, and histomorphometric analyses.
Efficient production of TGF-β via rAAV was achieved in vitro and in situ, both in the intact and injured meniscus. Application of the rAAV TGF-β vector stimulated the levels of cell proliferation and matrix synthesis (type I collagen) compared with control gene transfer in all systems tested, especially in situ in regions of poor healing capacity and in sites of meniscal injury. No adverse effects of the candidate treatment were observed at the level of osseous differentiation, as tested by immunodetection of type X collagen. Most remarkably, a significant reduction of the amplitude of meniscal tears was noted after TGF-β treatment, an effect that was associated with increased expression levels of the α-smooth muscle actin contractile marker.
Overexpression of TGF-β via rAAV gene transfer is capable of modulating the reparative activities of human meniscal cells, allowing for the healing of meniscal lesions by convenient injection in sites of injury.
Direct gene-based approaches using rAAV have strong potential to develop new therapeutic options that aim at treating human meniscal defects.
直接应用源自腺相关病毒（AAV）的治疗性基因载体可能有助于改善半月板撕裂的愈合。
测试重组腺相关病毒（rAAV）在人半月板纤维软骨细胞原代培养物、人半月板外植体以及实验性人半月板损伤中过表达强效转化生长因子-β（TGF-β）的能力，作为增强半月板修复的新工具。
对照实验室研究。
使用生化、免疫组织化学、组织学和组织形态计量学分析，在体外长达21天以及在完整和损伤的人半月板原位长达15天监测候选治疗对半月板细胞增殖和代谢活性的影响。
在体外和原位，无论是完整半月板还是损伤半月板，均通过rAAV高效产生TGF-β。与所有测试系统中的对照基因转移相比，rAAV TGF-β载体的应用刺激了细胞增殖和基质合成（I型胶原蛋白）水平，尤其是在愈合能力差的区域原位以及半月板损伤部位。通过X型胶原蛋白免疫检测测试，在骨分化水平未观察到候选治疗的不良反应。最显著的是，TGF-β治疗后半月板撕裂幅度显著降低，这一效应与α-平滑肌肌动蛋白收缩标记物表达水平增加相关。
通过rAAV基因转移过表达TGF-β能够调节人半月板细胞的修复活性，通过在损伤部位方便地注射实现半月板损伤的愈合。
使用rAAV的基于基因的直接方法具有开发旨在治疗人半月板缺陷的新治疗选择的强大潜力。
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