关节腔内骨髓间充质干细胞与同种异体骨的共培养

发布时间：2017-12-27 15:20

本文关键词：关节腔内骨髓间充质干细胞与同种异体骨的共培养　出处：《中国组织工程研究》2016年19期 　论文类型：期刊论文

【摘要】：背景:构建组织工程软骨的时候,同种异体骨与骨髓间充质干细胞是常用的支架材料和种子细胞,以往大多采用体外培养的方式。膝关节内游离体可以长期存在于关节腔中,并维持一定的软骨组织学特性。因此,关节腔可能为软骨细胞的生长和发育提供良好的环境条件。目的:探讨在关节腔内骨髓间充质干细胞与同种异体骨复合培养的效果。方法:纳入5只新西兰新生白兔进行骨髓间充质干细胞分离与培养,利用1只成年新西兰白兔制备同种异体骨,进行骨髓间充质干细胞与同种异体骨复合。实验分组:腔内培养组将骨髓间充质干细胞与同种异体骨复合物培养于动物关节腔内,正常对照组设为同腔内正常软骨,体外培养组实施常规体外培养。培养4,8,12周进行组织学苏木精-伊红染色观察及Ⅱ型胶原免疫组织化学染色观察。结果与结论:1培养12周进行苏木精-伊红染色和观察,正常对照组细胞质和软骨基质出现红染,细胞核出现蓝染,软骨细胞按照一定的方向呈紧密状有序排列。腔内培养组细胞质和软骨基质出现红染,细胞核出现蓝染,支架材料基本吸收。软骨细胞长入支架之中,细胞按照一定应力方向排列,且形态变小。体外培养组软骨细胞出现大量增殖,但呈无序状排列;2经免疫组织化学染色和观察,计数可得,随着培养时间的推移,腔内培养组的A值呈现出不断上升的情况,体外培养组和正常对照组均未出现明显的改变。且培养4,8,12周,正常对照组和腔内培养组的A值均显著高于体外培养组(P0.05)。培养4,8周,腔内培养组的A值均显著低于正常对照组(P0.05)。但培养12周,腔内培养组与正常对照组A值差异无显著性意义(P0.05);3实验结果表明,在体外和关节腔内环境下,均可以利用同种异体骨与骨髓间充质干细胞复合培养获得组织工程软骨,且关节腔内培养可以获得更好的培养效果。
[Abstract]:Background: when constructing tissue-engineered cartilage, allogeneic bone and bone marrow mesenchymal stem cells are commonly used scaffold materials and seed cells. In the past, most of them were cultured in vitro. The free body in the knee joint can exist in the articular cavity for a long time and maintain a certain histological characteristics of the cartilage. Therefore, the articular cavity may provide good environmental conditions for the growth and development of chondrocytes. Objective: To investigate the effect of bone marrow mesenchymal stem cells (MSCs) combined with allograft bone in the articular cavity. Methods: a total of 5 New Zealand white rabbits were enrolled in this study. Bone marrow mesenchymal stem cells were isolated and cultured. 1 adult New Zealand white rabbits were used to prepare allogenic bone, and bone marrow mesenchymal stem cells were combined with allogeneic bone. Experimental group: in vivo culture group, bone marrow mesenchymal stem cells and allograft bone complex were cultured in animal joint cavity, normal control group was set as normal cartilage in same cavity, and in vitro culture group was cultured in vitro. The 4,8,12 weeks were observed and observed by hematoxylin eosin staining and collagen immunohistochemical staining. Results and conclusion: 1 after 12 weeks of hematoxylin eosin staining and observation, the normal control group and the cytoplasm of cartilage matrix red staining, nuclei appear blue, cartilage cells in a certain direction is closely aligned. Intracavitary culture group, cytoplasm and cartilage matrix red staining, nuclei appeared blue, the basic scaffold material absorption. The chondrocytes grow into the scaffold, and the cells are arranged in a certain direction of stress, and the morphology becomes smaller. In vitro culture of chondrocytes was the emergence of a large number of proliferation, but in a disordered arrangement; 2 by immunohistochemical staining and counting can be observed, with the increase of culture time, intracavitary culture group A values showing a rising situation, group and normal control group were cultured in vitro without noticeable change. The A values in the normal control group and the intracavitary culture group were significantly higher than those in the culture group (P0.05) for 4,8,12 weeks. For 4,8 weeks, the A value of the intracavitary culture group was significantly lower than that of the normal control group (P0.05). But 12 weeks of culture, intracavitary culture group and normal control group A value had no significant difference (P0.05; 3) the experimental results show that in vitro and joint cavity environment, can use bone allograft and bone marrow mesenchymal stem cells co cultured for tissue engineering cartilage, articular cavity and culture can get training effect better.
【作者单位】：滨州医学院烟台附属医院骨关节科;滨州医学院烟台附属医院创伤科;
【分类号】：R68;R318.08
【正文快照】： 文章快速阅读:文题释义:局部微环境:在将构建的组织工程软骨植入局部进行修复治疗的过程中,会受到局部微环境的较大影响。局部的实际理化环境与各种生长因子,以及力学刺激等均会对组织工程软骨产生极大的影响,并直接影响到软骨修复效果。为此,在进行组织工程软骨构建的时候,可

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