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

背景和目的：关节软骨组织退变是由于关节软骨细胞合成代谢因子和分解代谢因子失衡所致。本研究利用最新发展的海藻酸盐回收软骨细胞组织和腺病毒基因转移技术，探讨有效的软骨保护方法。方法：采用35硫酸盐和阿新蓝快速渗滤法检测蛋白多糖的合成和降解。用二甲基亚甲基蓝结合法对PG进行定量。用ELISA法检测骨形态发生蛋白-7(BMP-7)和基质金属蛋白酶-3(MMP-3)的表达。结果：腺病毒载体(ADV)能有效地将靶基因导入ARC组织。经Adv/BMP-7转导的ARC组织在3周内可分泌大量BMP-7。Adv/BMP-7可增加ARC组织PG的合成和含量。IL-1促进ARC组织PG的降解和释放以及MMP3的产生。Adv/BMP-7可阻断IL-1b对ARC组织的这些合成代谢作用。结论：腺病毒基因转移是ARC组织表达靶基因的有效方法。Adv/BMP-7对IL-1b刺激的ARC组织具有明显的软骨保护作用。这些结果表明，在受影响的关节中进行BMP-7的基因转移或诱导将提供活体软骨保护。合作外国研究员：拉什医学院骨科正田光一

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