Center of Experimental Orthopaedics , Saarland University Medical Center , Homburg D-66421 , Germany.
Centro de Investigacións Científicas Avanzadas (CICA) , Universidade da Coruña , Campus de A Coruña , 15071 A Coruña , Spain.
Mol Pharm. 2018 Jul 2;15(7):2816-2826. doi: 10.1021/acs.molpharmaceut.8b00331. Epub 2018 May 24.
Recombinant adeno-associated virus (rAAV) vectors are well suited carriers to provide durable treatments for human osteoarthritis (OA). Controlled release of rAAV from polymeric micelles was already shown to increase both the stability and bioactivity of the vectors while overcoming barriers, precluding effective gene transfer. In the present study, we examined the convenience of delivering rAAV vectors via poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) polymeric (PEO-PPO-PEO) micelles to transfer and overexpress the transcription factor SOX9 in monolayers of human OA chondrocytes and in experimentally created human osteochondral defects. Human osteoarthritic (OA) chondrocytes and human osteochondral defect models were produced using human OA cartilage obtained from patients subjected to total knee arthroplasty. Samples were genetically modified by adding a rAAV-FLAG-h sox9 vector in its free form or via polymeric micelles for 10 days relative to control conditions (unmodified cells). The effects of sox9 overexpression in human OA cartilage samples were monitored by biochemical, histological, and immunohistochemical analyses. Delivery of rAAV-FLAG-h sox9 via polymeric micelles enhanced the levels of sox9 expression compared with free vector administration, resulting in increased proteoglycan deposition and in a stimulated cell proliferation index in OA chondrocytes. Moreover, higher production of type II collagen and decreased hypertrophic events were noted in osteochondral defect cultures when compared with control conditions. Controlled therapeutic rAAV sox9 gene delivery using PEO-PPO-PEO micelles is a promising, efficient tool to promote the remodelling of human OA cartilage.
重组腺相关病毒（rAAV）载体非常适合提供持久的治疗人类骨关节炎（OA）。已经证明，聚合物胶束中 rAAV 的控制释放可以增加载体的稳定性和生物活性，同时克服障碍，阻止有效的基因转移。在本研究中，我们研究了通过聚（环氧乙烷）（PEO）和聚（环氧丙烷）（PPO）聚合物（PEO-PPO-PEO）胶束递送 rAAV 载体以转导和过表达转录因子 SOX9 在单层人 OA 软骨细胞中和在实验性创建的人骨软骨缺陷中的便利性。使用从接受全膝关节置换术的患者中获得的人 OA 软骨来产生人 OA（OA）软骨细胞和人骨软骨缺陷模型。与对照条件（未经修饰的细胞）相比，通过添加游离形式或通过聚合物胶束添加 rAAV-FLAG-h sox9 载体来遗传修饰样品 10 天。通过生化、组织学和免疫组织化学分析监测 sox9 过表达对人 OA 软骨样本的影响。与游离载体给药相比，通过聚合物胶束递送 rAAV-FLAG-h sox9 增强了 sox9 表达水平，导致 OA 软骨细胞中糖胺聚糖沉积增加和细胞增殖指数增加。此外，与对照条件相比，在骨软骨缺陷培养物中观察到 II 型胶原产量增加和肥大事件减少。使用 PEO-PPO-PEO 胶束进行受控治疗性 rAAV sox9 基因传递是一种有前途的、有效的工具，可以促进人 OA 软骨的重塑。
