猪小肠黏膜下层海绵的构建与细胞黏附性观察  

Preparation of porcine small intestinal submucosa sponge and observation of cell adhesion

作　　者：孙慧哲 田伟[2] 曾亮[2] 裘锦云 张茜[3] 

机构地区：[1]沈阳医学院医学教育研究中心,辽宁省沈阳市110034 [2]沈阳医学院解剖教研室,辽宁省沈阳市110034 [3]沈阳医学院,辽宁省沈阳市110034

出　　处：《中国组织工程研究》2017年第34期5487-5492,共6页Chinese Journal of Tissue Engineering Research

基　　金：2014年省教育厅科学研究一般项目(L2014416)~~

摘　　要：背景:虽然小肠黏膜下层的结构与皮肤非常类似,但它的孔径大小与孔径率不如皮肤那样有利于种子细胞的长入。目的:利用EDC对于猪小肠黏膜下层进行化学改性,观察其改性后形态学结构及细胞相容性。方法:将小肠黏膜下层支架材料放入含0.2%胃蛋白酶的3%乙酸水溶液中,使小肠黏膜下层的质量浓度分别为1%、2%、3%、4%,磁力搅拌与冷冻干燥后获得小肠黏膜下层海绵,分别应用50,100,150 mmol/L的EDC进行交联改性,通过孔径及吸水率结果选取最佳质量浓度的小肠黏膜下层与EDC交联浓度,进行细胞培养。将第2代骨髓基质干细胞与小肠黏膜下层海绵共培养,1,2,3周后进行扫描电镜观察。结果与结论:①当小肠黏膜下层海绵质量浓度为1%时,在100 mmol/L EDC交联下的空间结构弹性好,结构规整,无空洞现象,孔径为100-150μm;②在100 mmol/L EDC交联下,质量浓度1%小肠黏膜下层海绵的吸水能力比2%高出0.35倍,质量浓度3%与4%小肠黏膜下层海绵的吸水能力相差不明显,质量浓度1%小肠黏膜下层海绵的结构更利于水分的流动与变化;③质量浓度1%的小肠黏膜下层海绵经100 mmol/L EDC交联后的结构、孔径及吸水率最好,可进行细胞实验。培养3周时,细胞变形相对完整且快,细胞形态在孔隙位置相对较大,外形接近长梭形,数目较多,在孔隙周围细胞形态较小,似圆盘形,数目相对较少,产生铺路石样改变,覆盖于大部分支架表面,并有大量颗粒样物质在周围出现,尤其在细胞集中位置更多见;④结果表明,EDC交联改性的小肠黏膜下层海绵具有良好细胞相容性。BACKGROUND： Although the porcine small intestinal submucosa is very similar to the skin in the structure, its pore sizeand porosity are not beneficial to the growth of seed cells as the skin does.OBJECTIVE： To observe the morphological and cytocompatibility of porcine small intestinal submucosa after chemicalmodification using 1-ethyl-3-（3-dimethylaminopropyl） carbodiimide （EDC）.METHODS： A porcine small intestinal submucosa sample was immersed in 3% acetic acid solution containing 0.2%pepsin to make 1%, 2%, 3%, 4% small intestine submucosa solutions. After magnetic stirring and freeze-drying, smallintestinal submucosa sponge was obtained and modified by cross-linking with 50, 100, 150 mmol/L EDC. Based on thedetection of pore size and water absorption, we selected the best concentrations of small intestinal submucosa and EDC,which were further used for cell culture. Passage 2 bone marrow mesenchymal stem cells were cocultured with the smallintestinal submucosa sponge, and observed under scanning electron microscope at 1, 2, 3 weeks after co-culture.RESULTS AND CONCLUSION： When cross-linked with 100 mmol/L EDC, the small intestinal submucosa sponge at amass concentration of 1% showed a reasonable structure and good elasticity with no appearance of voids, and the poresize ranged 100-150 μm. Moreover, the small intestinal submucosa sponge at a mass concentration of 1% showed a0.35-fold increase in the compared with that at a mass concentration of 2%, and its structure was more conducive towater flows and changes. The small intestinal submucosa sponges at a mass concentration of 3% and 4% showed nodifference in the water absorbing capacity. After cross-linked with 100 mmol/L EDC, the small intestinal submucosasponge at a mass concentration of 1% showed the best structure, pore size and water absorption, which were used forcell culture. At 3 weeks after cell culture, cell deformation was relatively intact and fast; there were many cells in the pores that were relatively large and approximately spindle

关 键 词：皮肤移植 细胞粘附 组织工程 生物材料 材料相容性 小肠黏膜下层 海绵 三维重建 细胞黏附 体外观察 

分 类 号：R318[医药卫生—生物医学工程]