流体静压力对BMSCs/PRF复合构建的组织工程软骨移植物力学性能的影响  

Effects of Hydrostatic Pressure on Mechanical Property of Tissue Engineering Cartilage Constructed by BMSCs/PRF Compound Membranes

作　　者：赵萤[1] 陈慧[2] 张旻 ZHAO Ying;CHEN Hui;ZHANG Min(State Key Laboratory of Military Stomatology , National Center for Oral Diseases and Clinical Medicine, Department of General Dentistry ＆ Emergency, School of Stomatology , The Fourth Military Medical University, Xi＇an 710032, China;Tongren Hospital of WuHan University, Wuhan 430060, China)

机构地区：[1]军事口腔医学国家重点实验室国家口腔疾病临床医学研究中心陕西省口腔疾病国际联合研究中心第四军医大学急诊与综合临床科,陕西西安710032 [2]武汉大学附属同仁医院,湖北武汉430060

出　　处：《口腔医学研究》2018年第3期249-253,共5页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:31570951)

摘　　要：目的:观察流体静压力对骨髓间充质干细胞(bone marrow stem cells,BMSCs)复合富血小板纤维蛋白(platelet-rich fibrin,PRF)构建的组织工程软骨力学性能的影响。方法:采用密度梯度离心法分离培养兔BMSCs并进行表面标志物鉴定,并制备细胞膜片。采用全血离心的方法分离兔PRF,构建BMSCs/PRF双膜复合体。将复合体随机分为3组:对照组、诱导组以及压力+诱导组。Real-time PCR检测成软骨标志基因Sox-9、Aggrecan及Col-II的表达量。HE染色、甲苯胺蓝染色观察复合体组织学改变。ElectroForce Systems3200力学测量仪测试BMSCs/PRF双膜复合体的弹性模量。结果:诱导组及压力+成软骨分化诱导组中成软骨标志基因Sox-9、Aggrecan及Col-II的表达量显著高于对照组,且压力+诱导组中Aggrecan和Col-II的表达量显著高于压力组。HE染色发现,压力+诱导组中软骨细胞的数量较对照组及诱导组显著增高。甲苯胺蓝染色可见压力+诱导组中大量蓝紫色异染的细胞外基质。弹性模量检测发现,压力+诱导组的弹性模量显著高于对照组及诱导组。结论:流体静压力可提高BMSCs/PRF双膜复合体构建组织工程软骨的质量及力学性能。Objective：To evaluate the effects of hydrostatic pressure on mechanical property of tissue engineering cartilage which was constructed by BMSCs/PRF compound membranes.Methods：BMSCs were obtained by density gradient centrifugation.BMSCs cell sheet was obtained after cell surface markers detection.After PRF membrane was obtained by whole blood centrifuge,BMSCs/PRF compound membranes were constructed.BMSCs/PRF constructs were randomized into 3 groups.BMSCs/PRF construct cultured without any mechanical stimulation or chemical induction was adopted as control group.BMSCs/PRF compound membranes treated with chondrogenic medium was induction group.BMSCs/PRF compound membranes treated with chondrogenic medium and hydrostatic pressure was defined as pressure＋induction group.Gene expression of Sox-9,aggrecan,and Col-II were analyzed by real-time PCR for all groups.HE staining and toluidine blue staining were performed to observe the histomorphology.Elasticity moduli of BMSCs/PRF compound membranes of each group were measured by ElectroForce System3200.Results：The mRNA expressions of Sox-9,aggrecan,and Col-II in induction group and pressure＋induction group were significantly higher than those of control group.Besides,the expressions of aggrecan and Col-II in pressure＋induction group were significantly higher than those of the induction group.HE staining showed that the density population of the chondrocytes in pressure＋induction group increased greatly compared with control and induction group.The metachromatic particles could be found by toluidine blue staining in pressure＋induction group.Elasticity moduli of tissue engineering cartilage constructed by BMSCs/PRF compound membranes inpressure＋induction group was also enhanced compared with that of control and induction group.Conclusion： Hydrostatic pressure promotes the quality and mechanical property of tissue engineering cartilage constructed by BMSCs/PRF compound membranes.

关 键 词：流体静压力 骨髓间充质干细胞 富血小板纤维蛋白 成软骨分化 弹性模量 

分 类 号：R782[医药卫生—口腔医学]