海绵状的小肠粘膜下层促进成骨样细胞增殖分化

中国生物工程杂志

2013, Vol. 33

Issue (6): 18-23

研究报告

海绵状的小肠粘膜下层促进成骨样细胞增殖分化

房艳¹, 倪伟民², 单伟¹, 曾瑞霞¹, 刘学元¹

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

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

FANG Yan¹, NI Wei-min², SHAN Wei¹, ZENG Rui-xia¹, LIU Xue-yuan¹

1. Department of Anatomy of Basic Medical College of LiaoNing Medical University, Jinzhou 121000, China;
2. Department of Neurosurgery of the First Affiliated Hospital of LiaoNing Medical University, Jinzhou 121000, China

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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。

关键词： 小肠粘膜下层; 成骨样细胞; 支架材料; 增殖分化

Abstract:

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 cells 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 cell-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.

Key words: Small intestinal submucosa matrix Osteoblast-like cells Scaffold Proliferation and differentiation

收稿日期: 2013-01-29 出版日期: 2013-06-25

ZTFLH:

R318.08

通讯作者: 倪伟民 E-mail: lnyxynwm@163.com

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引用本文:

房艳, 倪伟民, 单伟, 曾瑞霞, 刘学元. 海绵状的小肠粘膜下层促进成骨样细胞增殖分化[J]. 中国生物工程杂志, 2013, 33(6): 18-23.

FANG Yan, NI Wei-min, SHAN Wei, ZENG Rui-xia, LIU Xue-yuan. Spongelike Small Intestinal Submucosa Matrix Can Promote Osteoblast-like Cells Proliferation and Differentiation. China Biotechnology, 2013, 33(6): 18-23.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I6/18

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