适宜于构建组织工程化脂肪的蚕丝蛋白支架：最佳孔径筛选

背景：既往组织工程脂肪的研究中，支架材料的孔径往往被忽略。如孔径过大，细胞会顺着过大的孔隙流走，难以保留在支架中；孔径过小，细胞则主要分布于支架材料的表面，不易进入支架中，同时也不利于新生血管生长。 目的：筛选用于构建组织工程脂肪的蚕丝蛋白支架的适宜孔径。 方法：在保持蚕丝蛋白浓度不变条件下，通过改变冷冻-干燥温度与时间，制备出6批次不同孔径的蚕丝蛋白支架；贴壁法分离脐带间充质干细胞，化学染色检测其体外诱导成骨和成脂的能力；电镜下测定6批次蚕丝蛋白支架的孔径；观察脐带间充质干细胞在不同孔径蚕丝蛋白支架上黏附、增殖的能力。 结果与结论：6批次蚕丝蛋白支架的孔径分别为：(39.94±17.27)，(53.51±16.18)，(63.97±19.76)，(71.08±18.07)，(87.33±21.78)，(121.97±44.10) µm。脐带间充质干细胞在2号支架材料上黏附良好，并充分伸展，而第1，3，4，5，6号支架材料，除第1，3号支架上偶见细胞外，其余支架材料上均未见细胞生长。由此说明人脐带间充质干细胞与蚕丝蛋白支架构建组织工程脂肪时，支架材料的最佳孔径为50 µm。 关键词：蚕丝蛋白支架；孔径；组织工程脂肪；脐带间充质干细胞；支架材料 doi:10.3969/j.issn.1673-8225.2010.08.008

Suitable construction of tissue-engineered adipose with silk fibroin porous scaffolds: A choice of optimal pore

BACKGROUND: In previous studies on tissue-engineered adipose, the pore size of scaffolds is easily neglected. If the pore size was large, cells would flow away along the pore and could not maintain in the scaffold. If the pore size was small, cells would distribute on the surface of the scaffold, and could not enter the scaffold. Simultaneously, it would be not contribute to growth of new vessels. OBJECTIVE: To screen appropriate silk fibroin porous scaffolds to construct tissue-engineered adipose. METHODS: Under unchanged concentration of fibroin, six kinds of silk fibroin porous scaffolds in different aperture were prepared by changing freezing and drying temperature and time. Human umbilical cord mesenchymal stem cells (hUCMSCs) were harvested using attachment method. The potentiality of its osteogenic and adipogenic differentiation was detected by chemistry staining. The apertures of six kinds of silk fibroin porous scaffolds were measured by scanning electron microscope. The adhesion and proliferation of hUCMSCs on silk fibroin porous scaffolds in different aperture were observed. RESULTS AND CONCLUSION: The apertures of six silk fibroin porous scaffolds were (39.94±17.27), (53.51±16.18), (63.97±19.76), (71.08±18.07), (87.33±21.78), (121.97±44.10) µm, respectively. The strong ability of attachment of hUCMSCs appeared in the number two scaffold, and in other materials (number 1,3,4,5,6), there were not any cell in them except number one and three. Therefore, 50 µm might be a suitable aperture to construct tissue-engineered adipose with silk fibroin porous scaffolds using hUCMSCs.