低强度脉冲超声对兔关节软骨细胞增殖及Ⅱ型胶原表达的影响  

Effect of low intensity pulsed ultrasound on the cell proliferation and type Ⅱ collagen expression of rabbit articular chondrocytes

作　　者：郭杨[1] 马勇[1,2] 董睿[1] 刘尚仑[1] 屠娟[3] 黄桂成[1] 

机构地区：[1]南京中医药大学骨伤研究所,南京210023 [2]南京中医药大学附属医院骨伤科,南京210029 [3]南京大学声学研究所

出　　处：《安徽医科大学学报》2014年第12期1739-1742,共4页Acta Universitatis Medicinalis Anhui

基　　金：江苏省"六大人才高峰"项目(编号:2010-WS-082);江苏省自然科学基金(编号:BK2011812)

摘　　要：目的探讨低强度脉冲超声(LIPUS)对体外培养兔关节软骨细胞增殖及其合成Ⅱ型胶原的影响。方法选取生长状况良好的第3代兔膝关节软骨细胞,随机分为空白组及3种强度的LIPUS干预组,其超声强度分别为40、50、80m W/cm2,其余作用参数相同,中心频率:1.0 MHz,脉冲重复频率:1.0 KHz,脉冲宽度:200μs,作用周期:50。各LIPUS干预组每日照射20 min,连续照射3 d后,采用CCK-8比色法检测软骨细胞增殖情况,免疫组化染色分析Ⅱ型胶原表达量的变化。结果强度50、80 m W/cm2的LIPUS干预组软骨细胞增殖较空白组显著增多(P<0.01),且50 m W/cm2LIPUS的作用最为明显,而40 m W/cm2LIPUS促软骨细胞增殖效应与空白组相比差异无统计学意义(P=0.249);各强度LIPUS组Ⅱ型胶原表达量均高于空白组(P<0.01),尤以50 m W/cm2LIPUS组Ⅱ型胶原表达量增加最为显著,优于40、80 m W/cm2LIPUS的效应(P<0.01)。结论 LIPUS能够促进体外培养关节软骨细胞增殖,增加软骨细胞Ⅱ型胶原的分泌,且强度为50 m W/cm2时效应较为明显。Objective To investigate the effect of low intensity pulsed ultrasound （LIPUS）on the cell proliferation and type Ⅱ collagen expression of rabbit articular chondrocytes cultured in vitro.Methods The third generation of rabbit knee articular chondrocytes in good condition were randomly divided into 4 groups：control group （without LIPUS）,low-intensity LIPUS group,mid-intensity LIPUS group and high-intensity LIPUS group.The cells in LIPUS groups were stimulated with an intensity of 40 mW/cm2,50 mW/cm2 and 80 mW/cm2 respectively.The frequency was 1.0 MHz with a sine wave of 200 μs tone burst repeated at 1.0 KHz and 50 cycles.The ultrasound exposure time was 20 minutes per day in LIPUS groups.After 3 days,the cell proliferation was determined by Cell Counting Kit-8 assay.The immunocytochemical staining was used to analyze the expression of type Ⅱ collagen.Results Compared with control group,an intensity of 50 mW/cm2 and 80 mW/cm2 LIPUS both could significantly promote chondrocytes proliferation（P ＜0.01）.The effect of 50 mW/cm2 LIPUS was more obvious.There was no statistical difference between 40 mW/cm2 LIPUS group and control group（P =0.249）.The expression of type Ⅱ collagen in each LIPUS group was significantly higher than that of control group（P ＜0.01）.The effect of 50 mW/cm2 LIPUS group was the most significant and superior to that of the other two LIPUS groups （P ＜ 0.01）.Conclusion LIPUS can promote the cell proliferation and type Ⅱ collagen expression of articular chondrocytes,which has better biological effect at 50 mW/cm2 intensity.

关 键 词：低强度脉冲超声 软骨细胞 细胞增殖 Ⅱ型胶原 

分 类 号：R274.3[医药卫生—中医骨伤科学]