脱细胞兔髓核支架的形态学及体外细胞毒性研究

发布时间：2018-04-24 14:42

本文选题：支架 + 髓核　；参考：《实用医学杂志》2014年07期

【摘要】：目的:运用去污剂及核酸酶法制备兔脱细胞髓核支架,体外评价该支架的形态学及细胞毒性改变,探讨其作为组织工程髓核支架的可行性。方法:取健康成年新西兰大白兔髓核组织50枚,随机分为新鲜对照组(n=25)和脱细胞组(n=25),后者用0.3%TritonX-100+0.5%脱氧胆酸钠联合核酸酶进行脱细胞处理24 h,通过肉眼观、病理染色及扫描电镜观察髓核形态改变、细胞脱除和细胞外基质变化情况,并取兔骨髓间充质干细胞为对象,运用CCK-8法及死活细胞染色法在体外评价该支架的细胞毒性。结果:髓核脱细胞支架形态保持良好,细胞清除彻底,细胞外胶原基质结构破坏较小,与正常髓核相比,支架组织及其浸提液对兔骨髓间充质干细胞无明显毒副作用。结论:TritonX-100与脱氧胆酸钠联合核酸酶可有效脱除髓核细胞成分,较好地保留细胞外基质结构,且该支架对骨髓间充质干细胞无明显毒副作用,可以作为髓核组织工程研究的理想材料。
[Abstract]:Objective: to prepare rabbit nucleus pulposus scaffold with decontamination agent and nuclease method, to evaluate the morphology and cytotoxicity of the scaffold in vitro, and to explore the feasibility of the scaffold as tissue engineering nucleus pulposus. Method: 50 healthy adult New Zealand white rabbit nucleus pulposus were randomly divided into fresh control group (n=25) and acellular group (n=25), and the latter used 0.. 3%TritonX-100+0.5% deoxycholic acid sodium combined with nuclease for 24 h. The morphological changes of nucleus pulposus, cell removal and extracellular matrix change were observed by naked eye view, pathological staining and scanning electron microscope, and rabbit bone marrow mesenchymal stem cells were taken as the object. The cells of the scaffold were evaluated by CCK-8 method and dead cell staining method in vitro. Results: the morphology of the nucleus pulposus acellular scaffolds remained well, the cells were cleared thoroughly, the destruction of the extracellular matrix structure was small. Compared with the normal nucleus, the scaffold tissue and its extract had no toxic side effects on the rabbit bone marrow mesenchymal stem cells. Conclusion: the combined nuclease of TritonX-100 and sodium deoxycholate can effectively remove the components of the nucleus pulposus cells. This scaffold has no obvious toxic and side effects on bone marrow mesenchymal stem cells, and can be used as an ideal material for tissue engineering of nucleus pulposus.

【作者单位】：南方医科大学珠江医院骨科中心;
【基金】：国家自然科学基金面上项目(编号:81372869)
【分类号】：R318.08

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