口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (3): 249-253.DOI: 10.13701/j.cnki.kqyxyj.2018.03.009

• 颞下颌关节研究 • 上一篇    下一篇

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

赵萤1, 陈慧2, 张旻1*   

  1. 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
  • 收稿日期:2018-01-24 出版日期:2018-03-30 发布日期:2018-03-27
  • 通讯作者: *张旻,E-mail:zhangmin@fmmu.edu.cn
  • 作者简介:赵萤(1984~ ),女,青海西宁人,硕士,助理研究员,主要从事口腔生物力学方面研究。
  • 基金资助:
    国家自然科学基金(编号:31570951)

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

ZHAO Ying1, CHEN Hui2, ZHANG Min1*   

  1. 1. 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;
    2. Tongren Hospital of WuHan University, Wuhan 430060, China
  • Received:2018-01-24 Online:2018-03-30 Published:2018-03-27

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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。

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

Abstract: 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.

Key words: Hydrostatic pressure, BMSCs, PRF, Chondrogenesis, Elasticity moduli