猪小肠黏膜下层支架复合兔骨髓间充质干细胞构建组织工程骨膜对T淋巴细胞增殖的影响  

Influence of tissue-engineered periosteum constructed by porcine small intestinal submucosa scaffold combined with rabbit mesenchymal stem cells on T lymphocyte proliferation

作　　者：刘佳[1] 徐宏文[1] 王栓科[2] 伏雯[1] 

机构地区：[1]广州市儿童医院外七科,广东省广州市510120 [2]兰州大学第二附属医院骨科,甘肃省兰州市730000

出　　处：《中国组织工程研究与临床康复》2008年第27期5239-5242,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:小肠黏膜下层为无细胞胶原层,含有多种生长因子,主要由Ⅰ、Ⅲ型纤维胶原蛋白构成,与骨膜相似。目的:观察猪小肠黏膜下层支架构建兔组织工程骨膜对兔外周血T淋巴细胞增殖的影响,探讨兔组织工程骨膜的免疫学特性。设计、时间及地点:体外细胞学对比观察,于2005-09/2007-03在兰州大学附属第二医院骨科研究所完成。材料:健康家猪小肠,出生20d的新西兰纯种兔4只。方法:采用沉淀法以猪小肠黏膜下层作为支架材料,复合培养兔骨髓间充质干细胞成骨诱导细胞构建兔组织工程骨膜。分离兔外周血T淋巴细胞作为单向混合淋巴细胞反应中的效应细胞。实验分3个刺激组:成骨诱导细胞组(间充质干细胞成骨诱导14d)、组织工程骨膜组(间充质干细胞成骨诱导14d复合小肠黏膜下层支架)、单纯小肠黏膜下层组(单纯小肠黏膜下层支架),分别加入单相混合淋巴细胞反应液中。主要观察指标:MTT法体外检测各组对外周血T淋巴细胞增殖水平的影响。结果:成骨诱导细胞组、组织工程骨膜组、单纯小肠黏膜下层组的淋巴细胞增殖抑制率分别为(11±3)%,(23±5)%,(48±10)%。成骨诱导细胞组和组织工程骨膜组淋巴细胞增殖抑制率低于单纯小肠黏膜下层组,差异有显著性意义(P<0.01),成骨诱导细胞组与兔组织工程骨膜组比较差异无显著性意义(P>0.05)。结论:构建的组织工程骨膜可抑制兔外周血T淋巴细胞增殖,组织工程骨膜与单纯间充质干细胞成骨诱导细胞同样具有调节免疫作用。BACKGROUND： Small intestinal submucosa is an acellular collagen layer, which contains many growth factors and is similar to the periosteum, comprising type I and III fibrous collagen proteins. OBJECTIVE： To investigate the influence of rabbit tissue-engineered periosteum constructed by porcine small intestinal submucosa （PSIS） scaffold on the proliferation of peripheral blood T lymphocytes, and to approach immunological property of rabbit tissue-engineered periosteum. DESIGN, TIME AND SETTING： A controlled observation of in vitro cytology was performed in the Institute of Orthopaedics, the Second Affiliated Hospital of Lanzhou University （Lanzhou, Gansu, China） between September 2005 and March 2007. MATERIALS： Healthy porcine small intestine and four New Zealand purebred rabbits of 20 days old were used in this study. METHODS： By means of precipitation method, rabbit tissue-engineered periosteum was constructed using co-cultured osteoblasts derived from mesenchymal stem cells as seed cells and PSIS as scaffolds. Peripheral blood T lymphocytes were used as the effector cells in mixed lymphocyte reactions, and then cultured with three stimulus groups, namely osteoblastic induction group （induction of mesenchymal stem cells for 14 days）, tissue-engineered periosteum group （induction of mesenchymal stem cells for 14 days combined with PSIS scaffold） and PSIS group （PSIS scaffold alone）. Afterwards, they were added to the ongoing one-way mixed lymphocyte reactions. MAIN OUTCOME MEASURES： The proliferation ofT lymphocytes in peripheral blood was tested using MTT method in vitro. RESULTS： The inhibition ratio of T lymphocytes in the osteoblastic induction, tissue-engineered periosteum and PSIS groups was （11 ＋3）%, （23 ＋5）% and （48＋ 10）%, respectively. The T lymphocytes inhibition ratio was significantly different among three stimulus groups （P 〈 0.01）. The osteoblastic induction group was not significantly different from the tissue-engineered periosteum group （

关 键 词：组织工程 骨膜 混合淋巴细胞反应 猪小肠黏膜下层 骨髓间充质干细胞 生物材料 

分 类 号：R318[医药卫生—生物医学工程]