Department of Orthopaedic Surgery of the Affiliated Hospitals, Chongqing Medical University, Chongqing 400016, China; Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, Illinois 60637.
J Biomed Mater Res A. 2013 Dec;101(12):3542-50. doi: 10.1002/jbm.a.34661. Epub 2013 Apr 29.
Articular cartilage lesions in the knee are common injuries. Chondrocyte transplant represents a promising therapeutic modality for articular cartilage injuries. Here, we characterize the viability and transgene expression of articular chondrocytes cultured in three-dimensional scaffolds provided by four types of carriers. Articular chondrocytes are isolated from rabbit knees and cultured in four types of scaffolds: type I collagen sponge, fibrin glue, hyaluronan, and open-cell polylactic acid (OPLA). The cultured cells are transduced with adenovirus expressing green fluorescence protein (AdGFP) and luciferase (AdGL3-Luc). The viability and gene expression in the chondrocytes are determined with fluorescence microscopy and luciferase assay. Cartilage matrix production is assessed by Alcian blue staining. Rabbit articular chondrocytes are effectively infected by AdGFP and exhibited sustained GFP expression. All tested scaffolds support the survival and gene expression of the infected chondrocytes. However, the highest transgene expression is observed in the OPLA carrier. At 4 weeks, Alcian blue-positive matrix materials are readily detected in OPLA cultures. Thus, our results indicate that, while all tested carriers can support the survival of chondrocytes, OPLA supports the highest transgene expression and is the most conductive scaffold for matrix production, suggesting that OPLA may be a suitable scaffold for cell-based gene therapy of articular cartilage repairs.
膝关节软骨损伤是常见的损伤。软骨细胞移植是治疗关节软骨损伤的一种很有前途的治疗方法。在这里，我们描述了在四种载体提供的三维支架中培养的关节软骨细胞的活力和转基因表达。从兔膝关节中分离出关节软骨细胞，并在四种支架中培养：I 型胶原海绵、纤维蛋白胶、透明质酸和开孔聚乳酸（OPLA）。用表达绿色荧光蛋白（AdGFP）和荧光素酶（AdGL3-Luc）的腺病毒转导培养的细胞。通过荧光显微镜和荧光素酶测定法测定软骨细胞的活力和基因表达。通过阿利新蓝染色评估软骨基质的产生。兔关节软骨细胞可被 AdGFP 有效感染，并表现出持续的 GFP 表达。所有测试的支架都支持感染的软骨细胞的存活和基因表达。然而，在 OPLA 载体中观察到最高的转基因表达。在 4 周时，在 OPLA 培养物中很容易检测到阿利新蓝阳性基质材料。因此，我们的结果表明，虽然所有测试的载体都可以支持软骨细胞的存活，但 OPLA 支持最高的转基因表达，并且是最有利于基质产生的支架，这表明 OPLA 可能是关节软骨修复细胞基因治疗的合适支架。
