流体剪切力影响人关节软骨细胞合成Ⅱ型胶原的研究

Effects of fluid shearing force on type lI collagen synthesis by human chondrocytes in vitro

目的检测体外培养的人关节软骨细胞在不同大小的间断剪切力作用下合成Ⅱ型胶原的能力。方法选取培养的软骨组织，按摇床作用的转速分为：空白对照组和低、中等、高转速组（转速分别为20、40、60转／min）；通过免疫组化法检测关节软骨细胞Ⅱ型胶原蛋白的表达，RT—PCR法检测Ⅱ型胶原mRNA表达。结果中等转速组和高转速组Ⅱ型胶原免疫组化平均光密度（OD）值分别为（0．3061±0．0530）和（0．3777±0．0397），均高于低转速组平均OD值（0．2184±0．0135）和空白对照组平均OD值（0．1846±0．0363），差异均有统计学意义（P〈0．05）；低、中等、高转速组Ⅱ型胶原mRNAOD比值分别为（0．120±0．003）、（0．150±0．005）、（0．210±0．006），均高于空白对照组OD比值（0．080±0．006），差异均有统计学意义（P〈0．05）。结论一定强度间断剪切力能够影响人软骨细胞的代谢、增殖活动，使其合成Ⅱ型胶原增多。

Objective To detect type II collagen synthesis by cultured human chondrocytes under the influence of different intermittent fluid shearing forces. Methods Second passage human monolayer chondrocytes were divided into low-speed （20 rpm/min）, mid-speed （40 rpm/min） and high-speed （60 rpm/min） groups according to the rotation speed of the rocking bed. The expression of type Ⅱ collagen was analyzed using immunohistochemistry and RT-PCRs. Results The absorbance of the mid-speed and high-speed groups was significantly greater than that of the low-speed group and a control group. The average optical density of type Ⅱ collagen by RT-PCR was significantly higher in all three sheared groups than in the control group. Conclusions Intermittent shearing can enhance cellular proliferation and the metabolism of human chondrocytes in vitro and promote the expression of type Ⅱ collagen.