骨髓基质细胞与快速成形仿生人工骨支架载体的体外附着实验

Attachment of marrow stromal cells on poly-DL-lactideco-glycolide/tircalcium phosphate scaffold in vitro

目的:探讨具有良好生物相容性的聚乳酸-羟基乙酸与磷酸三钙人工合成仿生复合材料骨移植支架材料的可行性。方法:实验于2004-11在温州医学院附属第二医院完成。新西兰兔,雌性,体质量2.5～3.0kg。仿生人工骨材料为清华大学机械学院快速成形实验室提供,将其剪成2m m×4m m×1mm大小,750mL/L乙醇浸泡消毒30m in,紫外灯照射2h灭菌,烘干备用。在新西兰兔股骨上端,抽取约1.5mL骨髓,进行骨髓基质细胞的分离和培养。将第3代骨髓基质细胞以4×1010L-1细胞密度接种于仿生人工骨材料中,第7天取材,透射显微镜及其扫描电镜下观察细胞粘附情况。结果:仿生人工骨复合物表面及其孔隙内长满细胞,透射显微光镜下可见细胞覆盖的光照影。扫描电镜下观察:3d时,仿生人工骨复合物载体表面细胞生长密集,胞体呈不规则的球形、梭形或多角形,从胞体生出的细胞突长短不一,有的与其他细胞相连。第7天时细胞在孔洞中长满,有大量的胶原纤维分泌。结论:快速成形仿生人工骨复合材料与骨髓基质细胞有较好的生物相容性,具备作为细胞支架骨移植材料的性质。

AIM： To investigate the feasibility of poly-DL-lactideco-glycolide/ tircalcium phosphate （PLGA/TCP）, which has good biocompatibility, scaffold as bone graft material. METHODS： The experiment was carried out in the Second Affiliated Hospital of Wenzhou Medical College in November 2004. Female New Zealand rabbits of 2.5-3.0 kg were used. The artificial bone material, which was provided by the laboratory of rapid formation, School of Mechanical Engineering, Tsinghua University, was sheared into blocks with the size of 2mm×4mm×1mm, and then soaked into 750 mL/L ethanol to disinfected for 30 minutes and sterilized under ultraviolet radiation for 2 hours, and then dried by fire for later use. About 1.5 mL bone marrow was extracted from upper femur to isolate and culture MSCs. The third generation MSCs were inoculated into the PLGA,/TCP scaffold with the density of 4×10^10L^-1, and the material was removed at 7 days, and the attachment of cells was observed under transmission microscope and the scanning electron microscope.RESULTS： There were a great quantity of cells on the surface and aperture of scaffold under the transmission microscope. At 3 days, too many cells gathered on the surface of the material, the irregular cell bodies showed globular shape, fusiform shape or multi-angles, the cell process grew from the cell bodies were not of the same length, and some were connected with other cellsl. The cell numbers was increased fat 7 days. At the same time, collagen fibers also were observed. CONCLUSION： The rapidly formed PLGA,/TCP has better biocompatibility with MSCs, and can be used as bone graft material.