中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (2): 261-266.doi: 10.3969/j.issn.2095-4344.0016

• 材料生物相容性 material biocompatibility • 上一篇    下一篇

SD大鼠乳鼠脱细胞心脏支架的细胞相容性评价

韦雅淑1,徐亦辰2,赵文婧1,王慧丰1,刘红静1,谢丹尼1,2,陈维平1 
  

  1. 1广西医科大学组织学与胚胎学教研室,广西壮族自治区南宁市  530021;2广西壮族自治区计划生育研究中心,广西壮族自治区南宁市  530021
  • 收稿日期:2017-12-13 出版日期:2018-01-18 发布日期:2018-01-18
  • 通讯作者: 谢丹尼,主任医师,广西壮族自治区计划生育研究中心,广西壮族自治区南宁市 530021 陈维平,教授,广西医科大学组织学与胚胎学教研室,广西壮族自治区南宁市 530021
  • 作者简介:韦雅淑,女,1989年生,广西壮族自治区河池市大化县人,壮族,广西医科大学在读硕士,主要从事组织工程化心肌模型的建立研究。
  • 基金资助:

    广西自然科学基金(2012GXNSFAA053126);广西科技厅资助项目(桂科自0832157、桂科攻0592007-1G);广西急诊与医学救援人才小高地•广西高校急诊医学重点实验室开放课题(GXJZ201505);广西医科大学青年基金 (GXMUYSF201528);2017年度广西高校中青年教师基础能力提升项目(2017KY0096);广西再生医学重点实验室开放课题资助项目(桂再重开201703);广西壮族自治区人口和计划生育委员会科研基金(桂人口计生研1108)

Cell biocompatibility of an acellular heart scaffold of Sprague-Dawley neonatal rats

Wei Ya-shu1, Xu Yi-chen2, Zhao Wen-jing1, Wang Hui-feng1, Liu Hong-jing1, Xie Dan-ni1, 2, Chen Wei-ping1
  

  1. 1Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; 2Guangxi Zhuang Autonomous Region Family Planning Research Center, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Received:2017-12-13 Online:2018-01-18 Published:2018-01-18
  • Contact: Xie Dan-ni, Chief physician, Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; Guangxi Zhuang Autonomous Region Family Planning Research Center, Nanning 530021, Guangxi Zhuang Autonomous Region, China Chen Wei-ping, Professor, Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Wei Ya-shu, Studying for master’s degree, Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:

    the Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 2012GXNSFAA053126; the Funded Project of Guangxi Department of Science and Technology, No. 0832157, 0592007-1G; Emergency and Medical Rescue Talent Small Highland in Guangxi Zhuang Autonomous Region • Open Topics of Key Laboratory for Emergency Medicine in Guangxi Universities, No. GXJZ201505; the Science Foundation for the Youth in the Guangxi Medical University, No. GXMUYSF201528; 2017 Basic Ability Promotion Project for Young and Middle-Aged Teachers in Guangxi Universities, No. 2017KY0096; Open Project of Guangxi Key Laboratory of Regenerative Medicine, No. 201703; Research Foundation of the Population and Family Planning Commission of Guangxi Zhuang Autonomous Region, No. 1108

摘要:

文章快速阅读:

 

文题释义:
脱细胞支架:采用各种脱细胞方法,去除同种异体组织或器官中原有的细胞和引起免疫排斥反应的抗原成分,并尽可能保留细胞外基质特定成分,维持组织的三维空间结构。理想的支架材料应具备良好的生物相容性和细胞亲和性,可支持细胞黏附生长,植入体内不会引起机体的的炎症及毒性反应;可以模仿天然的组织结构和功能,具有适当的抗拉伸能力;支架被机体吸收的速率与种子细胞增殖、分化速率相似。
细胞相容性:种子细胞与生物材料之间的相互作用后产生的各种生物、物理、化学反应,细胞生物相容性是评价生物材料生物相容性的重要内容之一。一般来讲,良好的细胞相容性是首先要求支架不破坏细胞的生长环境,安全、无毒性,种子细胞能在材料上能黏附、伸展和增殖,并在特定的环境和相应条件诱导作用下能定向分化。
 
背景:脱细胞心脏支架材料具有低致敏性和天然的三维结构、具有良好的生物相容性和细胞亲和性等优点,成为心肌组织工程研究的热点。
目的:探究SD大鼠乳鼠脱细胞心脏支架的细胞相容性。
方法:选择第3代SD大鼠乳鼠骨髓间充质干细胞与SD大鼠乳鼠脱细胞心脏支架共培养7,14 d,构建细胞-支架复合物,采用苏木精-伊红染色、扫描电镜观察骨髓间充质干细胞在支架内的生长情况。采用心肌组织裂解液对细胞-支架复合物进行定向分化诱导,连续培养14 d,以平面定向分化诱导连续培养14,20 d的骨髓间充质干细胞为对照组,采用RT-PCR方法检测骨髓间充质干细胞内肌球蛋白重链α-MHC、锌指转录因子GATA-4的表达。
结果与结论:①苏木精-伊红染色显示,细胞-支架复合物中存在大量嗜碱性细胞核,细胞-支架复合物共培养14 d的细胞数量明显多于共培养7 d;扫描电镜下观察显示,共培养14 d的细胞-支架复合物纤维表面有大量细胞黏附;②平面定向分化诱导连续培养14,20 d的骨髓间充质干细胞分别呈竹节样、肌管样结构。在定向分化诱导连续培养14 d的细胞-支架复合物中可检测到α-MHC、GATA-4的表达,α-MHC、GATA-4的表达量分别与竹节样细胞和肌管样结构阶段相当;③结果表明脱细胞心脏支架的细胞相容性良好。

关键词: 生物材料, 组织工程, 骨髓间充质干细胞, 细胞-支架复合物, 共培养, 细胞相容性, 诱导分化, GATA-4, α-MHC

Abstract:

BACKGROUND: Owing to the advantages of low sensitization and natural three-dimensional structure, good biocompatibility and cell affinity, acellular heart scaffold materials are of great current interest in cardiac tissue engineering.
OBJECTIVE: To investigate the cytocompatibility of an acellular heart scaffold of neonatal rats.
METHODS: In order to construct the seed cell-scaffold complex, passage 3 bone marrow mesenchymal stem cells (BMSCs) of Sprague-Dawley neonatal rats were cultured with an acellular heart scaffold of Sprague-Dawley neonatal rats for 7 and 14 days. Hematoxylin eosin staining and scanning electron microscopy were used to observe the growth of BMSCs in the scaffold. The cell-scaffold complex was induced in myocardial tissue lysate for 14 days. BMSCs with planar orientation differentiation for 14 and 20 days were used as control group. RT-PCR was used to detect the expression of myosin heavy chain α-MHC and zinc finger transcription factor GATA-4 in BMSCs.
RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining showed the acellular heart scaffold contained a large number of eosinophilic fibrous structures, and the cell number of cell-scaffold complex after co-culture for 14 days was higher than that after co-culture for 7 days. Under the scanning electron microscope, a large amount of cells adhered to the fiber surface of the acellular scaffold at 14 days of co-culture. (2) BMSCs with planar orientation differentiation for 14 and 20 days had the bamboo-like and myotube-like structures. In the cell-scaffold complex with planar orientation differentiation for 14 days, the expression of α-MHC and GATA-4 could be detected, and their expression levels fulfilled the requirement for the presence of bamboo-like cells and myotube-like structure. These results indicate that the acellular heart scaffold exhibits good cytocompatibility.

Key words: Stem Cells, Myocytes, Cardiac, Myosin Heavy Chains, Tissue Engineering

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