成骨细胞在四种多孔医用材料内的灌注性三维动态接种研究  

Three-dimensional dynamic seeding of osteoblasts in flow perfusion bioreactors of four kinds of porousbiomedical material

作　　者：王林[1] 马真胜[1] 雷伟[1] 胡蕴玉[1] 王臻[1] 冯亚非[1] 张扬[1] 裴国献[1] 

机构地区：[1]西安,第四军医大学西京骨科医院,710032

出　　处：《中华创伤骨科杂志》2014年第3期242-248,共7页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（31170913）

摘　　要：目的探讨应用自行设计的灌注接种环路对4种医用多孔材料进行体外人胚成骨细胞动态接种的可行性，筛选较优的支架材料并初步探讨其作用机制。方法选取人同种多孔磷酸三钙（β-TCP）、异体松质骨粒、明胶海绵、胶原蛋白海绵4种医用材料作为支架载体，应用自行设计的灌注接种环路进行体外人胚成骨细胞动态接种（灌注接种组），以静态接种作为对照（静态接种组），通过检测支架上的细胞活力、活细胞接种率、细胞计数、细胞分布均匀度及组织学观察，比较不同接种方法和支架材料间细胞接种效果的差异。结果β-TCP、明胶海绵：灌注接种组细胞活力、活细胞接种率和细胞计数均优于静态接种组，差异行统计学意义（P〈0．05）；松质骨粒：两接种组间差异均无统计学意义（P〉0．05）；胶原蛋白海绵：灌注接种组细胞活力、活细胞接种率和细胞计数均低于静态接种组（P〈0．05），细胞分布均匀度蘑异无统计学意义（P〉0．05）。灌注接种：13-TCP、明胶海绵细胞接种率、细胞计数和细胞分布均匀度优于松质骨粒和胶原蛋白海绵，差异均有统计学意义（P〈0．05）；静态接种：胶原蛋白海绵的细胞接种率和细胞计数最高，差异均有统计学意义（P〈0．05）。结论灌注接种法可明显提高β-TCP和明胶海绵的细胞接种效率和细胞分布均匀性，可用于快速复合成骨细胞；支架材料显著影响细胞的灌注接种效果。Objective To determine optimal scaffold materials by evaluating the dynamic seeding of human fetal bone cells into 4 kinds of medical material in vitro using a self-designed perfusion loop. Methods A novel flow perfusion system with multi-loops was designed for dynamic cell seeding in vitro into three-dimensional （3D） porous scaffolds made of 4 different kinds of medical materials： β-tricaleium phos-phate（β-TCP）, human allogenic cancellous bone particles, gelatin sponge and collagen sponge. Static seeding scaffohts were used as controls. The seeding efficiency and spatial distribution of human fetal bone cells were compared between different seeding methods and scaffold materials using quantitative biochemical and image analyses. Results Perfusion seeding into β-TCP and gelatin sponge yielded significantly better seeding efficiency and more homogeneous distribution than static seeding （ P 〈 0. 05） . There were no significant differences regarding cell load and cell distribution in allogenic cancellous bone particles between the perfusion and static groups. Cell viability, cell seeding efficiency and cell counting in collagen sponge were significantly higher in the static group than in the perfusion group （ P 〈 0. 05）, but the uniformity of seeding was similar in both groups. In addition, perfusion seeding into β-TCP and gelatin sponge was much more effective than into allogenic cancellous particles and collagen sponge; static seeding into collagen sponge yields the highest cell seeding efficiency and cell counting. Conclusions Since perfusion seeding technique may yield higher seeding efficiency and more homogeneous distribution in β-TCP and gelatin sponge, it can be used to facilitate rapid cell seeding. Scaffold materials may have an obvious impact on the seeding efficiency and spatial dis- tribution of cells.

关 键 词：成骨细胞 组织支架 材料试验 细胞接种 灌注 

分 类 号：R318.08[医药卫生—生物医学工程]