Development of chondro-protective and regenerative strategy using alginate-recovered-chondrocyte tissue

背景与目的：关节软骨组织的退行性变是由于关节软骨细胞合成代谢和分解代谢因子失衡所致。我们进行了这项研究，以了解有效的软骨保护方法，使用最近开发的藻酸盐回收软骨细胞组织和腺病毒基因transfer technology.Methods：蛋白多糖（PG）的合成和降解进行了评估，使用35硫酸盐和阿尔新蓝快速过滤法。采用二甲基亚甲基蓝染料结合法定量PG。结果：腺病毒载体（Adv）能有效地将目的基因导入ARC组织中。Adv/BMP-7转导的ARC组织分泌大量的BMP-7长达三周。Adv/BMP-7可增加ARC组织PG的合成和含量。IL-1增加ARC组织PG的降解和释放，增加MMP-3的产生。结论：腺病毒介导的基因转染是一种在ARC组织中表达目的基因的有效方法。Adv/BMP-7对IL-1b刺激的ARC组织显示出显著的软骨保护作用。这些结果表明，在受影响的关节中基因转移或诱导BMP-7将在体内提供软骨保护。合作外国研究者：Koichi增田，拉什医学院骨科

Background and purpose: Degeneration of articular cartilage tissue is derived from imbalance between anabolic and catabolic factors for articular chondrocytes. We performed this study to obtain insight into efficient chondroprotective methods using recently developed alginate-recovered-chondrocyte tissue and adenoviral gene transfer technique.Methods : Synthesis and degradation of proteoglycan (PG) were assessed using 35sulfate and alcian blue rapid filtration assay. PG was quantified using dimethylmethylene blue dye binding method. Bone morphogenic protein-7 (BMP-7) and matrix metalloproteinase-3 (MMP-3) were measured using ELISA.Results : Adenoviral vector (Adv) efficiently transduced target genes into ARC tissue. A significant amount of BMP-7 was secreted by Adv/BMP-7-transduced ARC tissue up to three weeks. Adv/BMP-7 increased synthesis and content of PG of ARC tissue. IL-1□ increased degradation and release of PG as well as production of MMP-3 of ARC tissue. Adv/BMP-7 canceled these anabolic effects of IL-lb on ARC tissue.Conclusion : Adenoviral gene transfer is an efficient method to express target genes in ARC tissue. Adv/BMP-7 showed significant chondroprotective effects on IL-1b-stimulated ARC tissue. These results suggest that gene-transfer or induction of BMP-7 in affected joints would provide chondroprotection in vivo.Collaborating foreign investigator : Koichi Masuda, Department of Orthopedics, Rush Medical College