不同频率周期性压力对组织工程软骨的影响

Effects of cyclic pressure at different frequendes on the construction of tissue-engineering cartilage

目的观察不同频率周期性压力对组织工程软骨的影响。方法将兔关节软骨细胞接种到PLGA支架上,随机分成4组。A、B、C组分别在频率为0.01、0.1、1 Hz的周期性压力(强度0～200 kPa)下培养;D组不加压,为对照组。2周后采用组织化学,免疫组织化学法观察四组工程软骨的细胞增殖及基质分泌。并定量检测硫酸糖氨多糖(GAG)含量。结果四组之间Ⅱ型胶原染色面积百分比及GAG含量差异均有统计学意义(分别F=478.379,P<0.01;F=67.402,P<0.01)。其中B组工程软骨细胞最多,排列规则,Ⅱ胶原染色面积百分比[(83.95±2.23)%]和GAG含量[(476.33±25.66)μg]最高(P<0.01)。结论周期性压力对软骨细胞的新陈代谢具有促进作用,其中0～200 kPa、0.1 Hz频率的周期性压力能最好的促进软骨细胞增殖,合成Ⅱ胶原、GAG等细胞外基质。

Objective To explore the effects of cyclic pressure at different frequencies on the construction of tissue-engineering cartilage. Methods Chondrocytes were obtained from articular cartilages of freshly killed 3-day-old rabbits. Three-dimensional cartilage tissue was produced using PLGA scaffold as a carrier. All samples were cultured in bioreactor （ Celdyne HFB-40）. Four different culture conditions were examined：group A （0-200 kPa,0.01 Hz） ,group B （0-200 kPa,0.1 Hz） ,group C （0-200 kPa, 1 Hz） ,group D （ no pressure）. After culture for 2 weeks in each group,the level of GAG in each sample was measured. Macroscopical, histological and immunohistochemical analyses were performed. Results There were significant differences in type Ⅱ collagen and GAG among the 4 groups （ F = 478. 379, P 〈 0.01;F = 67. 402, P 〈 0.01 respectively）. In the group B, tissue-engineering chondrocytes were the most, and produced the most type Ⅱ collagen [ （ 83.95 ± 2.23 ） % ） ] and GAG [ （476.33 ± 25.66） μg ] among 4 groups （ P 〈 0.01 ）. Conclusion The cyclic pressure （0-200 kPa,0.1 Hz） effectively promotes chondroeytes to synthesize and secrete extracellular matrix just as GAG and type Ⅱ collagen in 4 groups, suggesting that cyclic pressure at different frequencies have important effects on metabolism of tissue-engineering cartilage.