Poloxamer 188对关节软骨加压负重后软骨细胞损伤的干预

目的 通过给正常人离体膝关节软骨体外加压负重,同时应用细胞表面活性剂Poloxamer 188的体外实验,探讨Poloxamer 188对钝性损伤时膝关节软骨的保护作用. 方法 将正常人离体膝关节软骨块样本分为正常对照组(n=6),非加压负重;非治疗组(n=12),加压负重;治疗组(n=12),加压负重后加Poloxamer 188.非治疗组和治疗组的样本加压负重570 N(20 MPa)的载荷.关节软骨加压负重后,各组中一半的标本在加压负重后1 h通过荧光染色观察,应用图像分析软件计算浅层软骨细胞(软骨全层的20%)的生存率.各组中剩下的一半标本用DMEM缓冲液依次漂洗3次,继续培养至加压负重后24 h.24 h后同样通过荧光染色观察,应用图像分析软件计算浅层软骨细胞(软骨全层的20%)的生存率. 结果 加压负重后软骨块中软骨细胞的存活率急剧下降.加压负重后软骨块中软骨细胞的坏死在浅层十分明显.非治疗组浅层软骨细胞存活率在第1小时和第24小时分别为55%和38%.与非治疗组相比,钝性损伤后治疗组的浅层细胞生存率在第1小时和第24小时显著增长为76%和57%(P<0.05). 结论 Poloxamer 188对浅层软骨细胞有一定的保护作用.利用Poloxamer 188早期干预软骨细胞的损伤,可能对延缓创伤后继发性关节炎的发病有一定的意义.

Intervention of Poloxamer 188 to chondrocyte damaged by blunt mechanical trauma in vitro

Objective To explore Poloxamer188, a non-ionic surfactant as a potential tool for early intervention into the chondrocyte damaged by blunt mechanical trauma in vitro. Methods Three groups were control group( n =6), no treatment group( n = 12) and Poloxamer 188 treatment group( n = 12). Two groups are then loaded to 20 MPa in unconfined compression. At 1 and 24 h the percentages of live and dead cells of superficial zone in compressed and control groups were determined with a cell viability stain. Results At 1 h post-trauma, specimens of Poloxamer 188 treatmet group ( 76% ) had a significantly increased percentage of live cells in the superficial zone versus the no treatment group(55% ). In 24 h the percentages of live cells in the superficial zone of the Poloxamer 188 treatment group (57%) were significantly greater than in the no treatment group (38%). Conclusions Poloxamer 188 surfactant could help restore the integrity of cell membranes in cartilage damaged by blunt mechanical trauma. Models of mechanical cartilage injury in vitro may explain aspect of the interactions between mechanical forces and degradative pathways which lead to osteoarthritis progression.