关节骨软骨仿生支架浸提液对种子细胞生物学特性的影响  

Effects of the extract solution of bionic articular osteochondral scaffold on the biological characteristics of seed cells

作　　者：刘伟[1] 王晨昱[1] 常旭东 欧阳成伟 董燕[4] 戴刚 李敏[1] LIU Wei;WANG Chen-yu;CHANG Xu-dong;OUYANG Cheng-wei;DONG Yan;DAI Gang;LI Min(Gansu Provincial Key Laboratory of Preclinical Study for New Drug Development,Institute of Pathology,School of Basic Medical Sciences,Lanzhou University,Lanzhou 730000,China;Second Department of Joint Surgery,Gansu Provincial Hospital of Traditional Chinese Medicine and First Affiliated Hospital of Gansu University of Traditional Chinese Medicine,Lanzhou 730050,China;School of Material Science and Engineering,Lanzhou Jiaotong University,Lanzhou 730070,China;Second Department of Obstetrics,Gansu Provincial Maternity and Child-care Hospital,Lanzhou 730050,China;Department of Trauma Joint Surgery,First Affiliated Hospital of Army Medical University,Chongqing 400038,China)

机构地区：[1]兰州大学基础医学院病理学研究所,甘肃省新药临床前研究重点实验室,兰州730000 [2]甘肃省中医院暨甘肃中医药大学第一附属医院关节骨二科,兰州730050 [3]兰州交通大学材料科学与工程学院,兰州730070 [4]甘肃省妇幼保健院产二科,兰州730050 [5]陆军军医大学第一附属医院创伤关节外科,重庆400038

出　　处：《兰州大学学报（自然科学版）》2023年第6期838-845,共8页Journal of Lanzhou University(Natural Sciences)

基　　金：国家自然科学地区基金项目(31360230);甘肃省科技重大专项(1203FKDA036)。

摘　　要：为了观测自制胶原涂层羟基磷灰石纳米晶/聚磷酸钙纤维/L-聚乳酸基含“界层结构”仿生一体化关节骨软骨复合组织工程支架材料对种子细胞增殖活性及相关分化功能的影响,采用CCK-8、流式细胞仪检测及体外分化诱导培养,分别检测新型支架材料浸提液对人骨髓间充质干细胞(hBMSCs)及人脐带间充质干细胞(hUCMSCs)的增殖活性、凋亡及成骨、成软骨分化能力的影响;通过实时荧光定量PCR和Western blot检测细胞成骨及成软骨分化诱导后ALP、Osterix、Runx2、COL1A1及COL2A1的mRNA和蛋白表达水平.结果显示,新型支架材料浸提液对hBMSCs及hUCMSCs的增殖活性及凋亡均无显著影响;体外分化诱导后均可呈现成骨及成软骨分化表型,5种基因的mRNA和蛋白表达均显著上调;支架材料浸提液处理后,其成骨及成软骨分化能力更明显,5种基因的mRNA和蛋白表达上调更显著.提示自制新型仿生一体化支架材料可保持hBMSCs及hUCMSCs种子细胞良好的增殖活性及成骨、成软骨分化功能.To investigate the effects of collagen-coated nano-hydroxyapatite/calcium polyphosphate fibers/poly-L-lactic acid bionic integrated articular bone and cartilage composite tissue engineered scaffold material with a boundary layer structure on the growth,proliferation and related differentiation function of seed cells in vitro.We examined the effects of novel scaffold material extract solution on the proliferation,apoptosis and osteogenic or chondrogenic differentiation capacity of human bone marrow mesenchymal stem cells(hBMSCs)and human umbilical cord mesenchymal stem cells(hUCMSCs)using cell counting kit-8,flow cytometry,and differentiation induction cultures.The mRNA and protein expression levels of alkaline phosphatase(ALP),Osterix,Runx2,COL1A1 and COL2A1 in two kinds of mesenchymal stem cells were measured by real-time fluorescence quantitative PCR and Western blot after osteogenic and chondrogenic differentiation induction.The results indicated that there were no significant effects on the proliferation and apoptosis of hBMSCs and hUCMSCs after treatment with the novel scaffold material extract solution.hBMSCs and hUCMSCs showed osteogenic or chondrogenic differentiation phenotypes after the induction of osteogenic or chondrogenic differentiation in vitro.The mRNA and protein expression of ALP,Osterix,Runx2,COL1A1 and COL2A1 were significantly upregulated.The differentiation of osteogenesis and chondrogenesis were more obvious when cells were treated with the scaffold material extract solution,and the mRNA and protein expression of ALP,Osterix,Runx2,COL1A1 and COL2A1 were upregulated more significantly.The results suggested that the selfmade novel bionic integrated scaffolds could maintain the proliferative viability and the osteogenic and chondrogenic differentiation functions of both hBMSCs and hUCMSCs in vitro.

关 键 词：复合组织工程 关节软骨 仿生支架材料 人间充质干细胞 增殖分化 

分 类 号：R318.08[医药卫生—生物医学工程]