连续低强度超声对骨软骨体外保存的影响  

Effect of continuous low-intensity ultrasound on the preservation of osteochondral tissue in vitro

作　　者：华真 李众 张学芬 亓建洪 宋洪强 周路 熊藉培 王凯声 耿彩云 张艺 HUA Zhen;LI Zhong;ZHANG Xue-fen;QI Jian-hong;SONG Hong-qiang;ZHOU Lu;XIONG Jie-pei;WANG Kai-sheng;GENG Cai-yun;ZHANG Yi(College of Sports Medicine and Rehabilitation,Shandong First Medical University,Tai'an 271000,China)

机构地区：[1]山东第一医科大学运动医学与康复学院,山东泰安271000

出　　处：《中国矫形外科杂志》2023年第22期2072-2077,共6页Orthopedic Journal of China

基　　金：山东省自然科学基金(编号:ZR2017LH018);山东省中医药科技项目(编号:2021M148)。

摘　　要：[目的]探究连续低强度超声(continuous low intensity ultrasound,cLIUS)对骨软骨体外保存的影响。[方法]6个月龄猪股骨髁获取骨软骨标本60个,随机分为两组,均于DMEM培养液在4℃保存。静态组不做超声处理。超声组每天给予20min超声刺激,频率42 kHz,电功率50 W。于第14、28 d检测细胞存活率、蛋白多糖表达、组织形态学、生物力学性能。cLIUS刺激2 h后检测ERK蛋白表达情况。[结果]第14 d,超声组细胞存活率显著高于静态组[(75.5±1.8)%vs(65.6±1.5)%,P<0.05];随时间推移,两组细胞存活率均显著下降(P<0.05);第28 d两组差异无统计学意义(P>0.05)。组织学染色显示,第14 d,超声组组织形态结构优于静态组,且番红O染色IOD[(224.7±4.1)vs(166.5±3.7),P<0.05]、甲苯胺蓝染色IOD[(223.7±6.0)vs(166.7±5.0),P<0.05]均显著高于静态组;第28 d两组上述指标差异无统计学意义(P>0.05)。随时间推移,两组组织形态结构紊乱,蛋白多糖显著流失(P<0.05)。第14 d,超声组杨氏模量显著优于静态组[(8.7±0.3)MPa vs(7.0±0.0)MPa,P<0.05];第28 d两组差异无统计学意义(P>0.05)。随时间推移,两组杨氏模量显著下降(P<0.05)。超声刺激2 h后,超声组ERK蛋白表达水平显著高于静态组[(0.9±0.1)vs(0.5±0.2),P<0.05]。[结论]cLIUS可以提高猪骨软骨的体外保存效果,延长保存时间,上调ERK蛋白的表达水平。[Objective]To investigate the effect of continuous low-intensity ultrasound(cLIUS)on the preservation of osteochondral tis-sue in vitro.[Methods]Sixty osteochondral specimens were harvested from a 6-month-old pig femur condyles and were randomly dividedinto two groups,which were stored in DMEM medium at 4℃,those in the static group was not treated with ultrasound,while the samples inthe ultrasonic group was stimulated for 20 min every day at a frequency of 42 kHz and an electric power of 50W.The survival rate,proteo-glycan expression,histomorphology and biomechanical properties were detected at the 14 th and 28 th days.The expression of extracellularregulatory protein kinase(ERK)was detected 2 hours after cLIUS stimulation.[Results]The ultrasound group got significantly higher cellsurvival rate than the static group at 14 days[(75.5±1.8)%vs(65.6±1.5)%,P<0.05],whereas which became not statistically significant be-tween the two group at 28 days.As time went by,the cell survival rate in both groups decreased significantly(P<0.05).Histological stainingshowed that the ultrasound group was better than the static group,the former had higher the saffron O staining IOD[(224.7±4.1)vs(166.5±3.7),P<0.05],toluidine blue staining IOD[(223.7±6.0)vs(166.7±5.0),P<0.05]than the latter at 14 days,while which returned to be not sig-nificant difference between the two groups at 28 days(P>0.05).The tissue morphology and structure of the two groups were disordered andproteoglycan was significantly lost over time(P<0.05).In addition,the ultrasound group got significantly higher Young's modulus than thestatic group 14 days[(8.7±0.3)MPa vs(7.0±0.0)MPa,P<0.05],which became not significantly different either at 28 days between the twogroups(P>0.05).The Young's modulus of the two groups decreased significantly with time(P<0.05).At 2 hours after ultrasonic stimulation,the ultrasonic group had significantly higher expression of ERK protein than the static group[(0.9±0.1)vs(0.5±0.2),P<0.05].[Conclusion]The cLIUS improves the preserv

关 键 词：同种异体骨软骨移植 关节软骨 低强度超声 力学刺激 细胞外调节蛋白激酶 

分 类 号：R68[医药卫生—骨科学]