海绵状的小肠粘膜下层促进成骨样细胞增殖分化  

Spongelike Small Intestinal Submucosa Matrix Can Promote Osteoblast-like Cells Proliferation and Differentiation

作　　者：房艳[1] 倪伟民[2] 单伟[1] 曾瑞霞[1] 刘学元[1] 

机构地区：[1]辽宁医学院基础医学院,锦州121000 [2]辽宁医学院附属第一医院,锦州121000

出　　处：《中国生物工程杂志》2013年第6期18-23,共6页China Biotechnology

摘　　要：目的:观察人脂肪干细胞(hADSCs)诱导分化的成骨样细胞在海绵状的猪小肠粘膜下层(SIS)表面的生长情况,探讨三维立体海绵状的SIS能否促进成骨样细胞的增殖和分化。方法:采用物理和化学结合的方法将猪近段空肠制备成脱细胞的SIS,再将薄膜状的SIS经液氮低温研磨制成微粒,交联后采用冷冻干燥技术重塑形为海绵状的SIS;原代培养hADSCs,流式术检测表面抗原,诱导其成骨、成软骨、成脂分化并染色鉴定;将诱导的成骨样细胞与海绵状SIS复合培养,扫描电镜观察细胞形态;应用SIS材料浸提液培养成骨样细胞,MTT法检测细胞增殖情况,ALP活度检测成骨分化情况。结果:脱细胞的SIS未见有核物质,海绵状的SIS呈三维立体状,具有大量均匀一致的孔隙;原代培养的hADSCs表达干细胞相关抗原,并可分化为成骨样细胞,茜素红将钙结节染成紫红色。成骨样细胞与海绵状SIS复合培养后,细胞生长旺盛增殖能力强,ALP表达量明显增加。结论:海绵状的SIS具有均匀的三维孔隙,细胞相容性好,能明显促进hADSCs来源的成骨样细胞的增殖及成骨分化,可成为骨组织工程新型的三维立体天然生物衍生材料。Objective： Observe the growth of osteoblast-like cells induced and differentiated by human adipose stem cells （hADSCs） in the spongelike porcine small intestinal matrix （SIS）. Explore ability of three- dimensional SIS promote osteoblast-like cells proliferation and differentiation. Methods： acellular SIS is prepared by combination of physical and chemical methods with porcine proximal jejunum. Particles was made with film- like SIS by milling-in liquid nitrogen cryogenic grinding. And the particles has been reshaped spongelike after crosslinked by freeze-drying technology, hADSCs is isolated and cultured by enzymatic digestion. And surface antigen has been identificated by flow cytometry, hADSCs has been induced and differentiated into osteoblast-like cells,chondroblast-like cells and adipocyte-like cells. The osteoblast-like cells is cultured in cavernous SIS. Cell morpholog is observed by scanning electron microscope. Osteoblast-like ceils is cultured in material extracts of SIS. The cell viability is evaluated with MTT. Osteogenic differentiation is detected by ALP activity. Results： Spongelike SIS is three-dimensional stereo-scaffold with a great of regular three-dimensional pore. Stem ceil- associated antigen is expressed by primary hADSCs, hADSCs can differentiated into osteoblast-like cells stained by alizarin red. Osteoblast-like cells can proliferate and express ALP obviously in cavernous SIS. Conclusion： spongelike SIS with three-dimensional pores presents superior cytocompatibility, and it can promote osteoblast- like cell differentiated by hADSCs proliferate and increase osteogenic activity significantly. It can became a novel three-dimensional natural biological materials for bone tissue engineering.

关 键 词：小肠粘膜下层 成骨样细胞 支架材料 增殖分化 

分 类 号：R318.08[医药卫生—生物医学工程]