摘要
目的:观察人骨髓间充质干细胞与纳米晶胶原基骨的复合培养,证实纳米晶胶原基骨是否可以成为组织工程的良好载体材料。方法:实验于2003-07/2004-08在清华大学材料科学与工程系完成纳米晶胶原基骨的制备及处理,在中日友好临床医学研究所免疫室完成纳米晶胶原基骨与骨髓间充质干细胞复合培养。健康自愿献髓者3人,在中日友好医院骨坏死与骨循环实验室体检。应用仿生原理制备纳米晶胶原基骨,体外培养骨髓间充质干细胞并与材料复合,通过共聚焦显微镜和环境扫描电镜观察骨髓间充质干细胞与材料的复合情况。结果:①共聚焦显微镜显示:在各层面扫描骨髓间充质干细胞均深入纳米晶胶原基骨的内部孔隙。②环境扫描电镜显示:第2天细胞贴附在多孔材料上,在纳米晶胶原基骨底部靠近培养板处细胞黏附明显较纳米晶胶原基骨上其他部位多,细胞也开始伸展,伸出伪足向材料微孔中长入,孔隙内见不到桥接的细胞连接细胞在整个材料及孔隙内表面贴附、伸展。第7天时细胞在材料上完全铺展。伪足向材料微孔中长入,在部分直径较小的孔隙中(70～150μm),细胞之间突起相互连接呈桥状。纳米晶胶原基骨临近培养板的一面有较多细胞生长,主要在材料的表面和材料的裂隙中,孔隙中较少。第14天,大量细胞在纳米晶胶原基骨表面和孔隙中生长,尤以孔隙中明显。细胞之间广泛存在突起连接,在孔隙中呈网状互相相连。结论:纳米晶胶原基骨是良好的组织工程支架材料,易与骨髓间充质干细胞复合,适合种子细胞的贴附、生长、增殖和分化。
AIM： To observe the combined cultivation of nano-collagen basal bone and human bone marrow derived mesenchymal stem cells, and verify whether the nano-collagen basal bone is an excellent carrier-material of tissue engineering.
METHODS： Preparation and disposal of nano-collagen basal bone was conducted in the Preserving and Reconstructive Center of Osteonecrosis and Joint, Tsinghua University from July 2003 to August 2004, and combined cultivation of nano-collagen basal bone and human bone marrow derived mesenchymal stem cells was done in the Research Institute of Clinical Medisine of China-Japan Friendship Hospital. Three health persons, who had donated their marrows voluntarily, received health examination in the Laboratory of Osteonecrosis and Joint, China-Japan Friendship Hospital. The nano-collagen basal bone was prepared with principles of bionics , and marrow derived mesenchymal stem cells in vitro were cultivated and combined with materials. Combination of bone marrow derived mesenchymal stem cells and materials were observed with confocal microscopy and enviremental scanning electron microscope.
RESULTS： ①Manifestation of confocal microscopy： Scanning of bone marrow derived mesenchymal stem cells bone in every slice was deep into the nano-collagen basal bone. ②Manifestation of enviremental scanning electron microscope： Stem cells adhered to the multiporous materials in the next day, and those in the bottom of nano-collagen basal bone and near the cultivating board were much more than those in other parts. Cells began to extend their parapodiums into the micropores of materials, while there were no bridging cells in the ventages, and knizocytes extended and adhered to the surface of materials as well as the inside of ventages. Cells extended completely on the 7^th day. Parapodiums growed into the micropores of materials, and there were bridge-like connections among cells in pores of small diameter （70-150μm）. There were much cells growing in the closed side of the cultivating board in nano-collagen basal bone, and mainly on the surface of materials as well as inside the leakages of materials, while there were little in the pores. Lots of cells growed on the surface of nanocollagen basal bone and the inside of pores on the 14^th day, especially inside the pores. There were extensive prominent connections among cells and reticulodromous connections in the pores.
CONCLUSION： Nano-cotlagen basal bone is a good scaffold materials of tissue engineering, which is easy to combined width bone marrow derived mesenchymal stem cells, and it is suitable for cells to adhere, grow, proliferate and differentiate.
出处
《中国临床康复》
CSCD
北大核心
2006年第13期41-43,共3页
Chinese Journal of Clinical Rehabilitation
基金
首都医学发展基金重大联合项目(2002-1007)
卫生部部属(管)临床学科重点项目(2004-2006)
中日友好医院2005年博士启动课题(20056150)~~
