生物蛋白胶双相接种法促进组织工程骨构建的观察

背景：很多学者采用生物蛋白胶修饰无机支架材料改善其细胞黏附能力，并证实该方法能有效促进骨髓间充质干细胞与支架材料黏附。目的：观察采用生物蛋白胶双相接种技术促进组织工程骨体外构建的可行性。设计、时间及地点：细胞生物学水平实验，2003-01/2008-10在解放军第三军医大学组织工程研发中心实验室完成。材料：医用生物蛋白胶为广州倍特生物技术有限公司产品，同种异体冻干脱钙骨基质由解放军第三军医大学西南医院骨组织库提供。方法：以生物蛋白胶为介质，用双相接种技术将羊骨髓间充质干细胞与同种异体冻干脱钙骨基质复合，在体外构建组织工程骨，以静置接种法作为对照。主要观察指标：通过细胞计数及MTT观察不同接种密度生物蛋白胶双相接种法所构建的组织工程骨细胞数量，并通过相差显微镜和扫描电微观察细胞在支架材料中的分布情况。结果：双相接种法在不同接种密度时都可保持细胞黏附率维持在80%左右，上架细胞的数量随接种细胞密度增加成比例增加；生物蛋白胶对细胞的增殖没有明显影响，提高接种密度使细胞生长曲线提前达到平台期，各组的细胞数量峰值接近。结论：双相接种技术可显著提高种子细胞与支架材料的复合数量、复合效率并且分布均匀，是一种工程骨快速高效组织的构建方法。

Enhancement of tissue-engineered bone construction by bioprotein biphasic seeding technique

Objective To initially study the feasibility of enhancing tissue engineering bone formatting in vitro with biphasic seeding technique. Methods Different densities of goats MSCs were seeded onto allogenic FDBM introduced by fibrin glue and cocultured in vitro. FDBM was soaked in cell suspension for 4 hours without fibrin glue in the control group. The adhesion and proliferation of MSCs were evaluated by cell counting, microscope, electronic scanning microscope and MTT test. Results With the biphasic seeding technique, the adhesive rates of different seeding densities were kept around 80 %, cells upon the scaffolds increased following the increase of seeding densities. In fibrin glue, MSCs lived and proliferated well. The higher seeding densities were, the earlier arrived platform phase of growth curves, and all peaks were similar. Conclusion Biphasic seeding technique can significantly increase the adhesive rates of MSCs seeding on the FDBM scaffolds, and maybe become a suitable method of formatting tissue engineering bone in vitro.