体外构建人工仿生骨膜

In vitro constructing artificial biomimetic periosteum

背景:小肠黏膜下层既具有良好的生物相容性和降解性,又富含多种生长因子,能明显促进细胞的黏附、增殖及分化,在国外已被广泛应用于骨与软骨、血管、皮肤、膀胱、平滑肌及胰岛等组织的修复,且已表现出良好的组织工程化细胞支架性能。目的:探讨兔骨髓间充质干细胞经体外诱导成成骨细胞与猪小肠黏膜下层复合构建组织工程骨膜的可行性。方法:采用贴壁筛选法分离2周龄健康新西兰大白兔骨髓间充质干细胞,并进行体外扩增培养、诱导分化及鉴定。将经成骨诱导分化的骨髓间充质干细胞与猪小肠黏膜下层复合构建组织工程骨膜,观察细胞在生物材料上的附着、生长、增殖情况。结果与结论:接种5 d后,细胞散在附着于小肠黏膜下层材料上,细胞形态呈圆形,细胞之间无连接;10 d后细胞之间形成桥粒连接,成骨细胞伸出突起,与小肠黏膜下层贴附;15 d后细胞增殖,分泌基质,在小肠黏膜下层表面形成多层细胞组成的复层膜样结构。表明将骨髓间充质干细胞诱导成成骨细胞后与猪小肠黏膜下层复合可构建组织工程骨膜,有可能成为理想的组织工程支架材料。

BACKGROUND：The smal intestinal submucosa has good biocompatibility and biodegradability, and also contains a variety of growth factors that can significantly promote celladhesion, proliferation and differentiation. Currently, the smal intestinal submucosa has been widely used in bone and cartilage, blood vessels, skin, bladder, smooth muscle and pancreatic tissue repair, showing good performance as a tissue-engineered cellscaffold. OBJECTIVE：To investigate the in vitro feasibility of tissue engineered periosteum constructed by porcine smal intestinal submucosa and osteoblasts differentiated from bone marrow mesenchymal stem cells. METHODS：Bone marrow mesenchymal stem cells were harvested from 2-week-old healthy New Zealand rabbits by using adherent method, and then cells were cultured, induced, differentiated and identified in vitro. Fol owing induced differentiation and identification, the bone marrow mesenchymal stem cells were compounded with porcine smal intestinal submucosa to fabricate tissue engineered periosteum. The adhesion, growth, and proliferation of cells on the materials were observed. RESULTS AND CONCLUSION：At 5 days after inoculation, the cells receiving osteogenic induction could quickly adhere and proliferate on the surface of porcine smal intestinal submucosa and be interconnected;at 10 days, the desmosomes formed among the cells, cellprocesses from osteoblasts were visible and attached to the smal intestine submucosa;at 15 days, cellproliferation and secretion of matrix appeared, and multi-layer membrane-like structure formed on the surface of the smal intestine submucosa. These findings indicate that after osteogenic induction, the bone marrow mesenchymal stem cells can be combined with porcine smal intestinal submucosa to construct a tissue engineered periosteum, which is hoped to be an ideal scaffold for tissue engineering.