PLGA微囊支架材料的制备及对成骨细胞粘附和增殖的影响

摘要]目的：制备一种新型结构的聚乳酸-羟基乙酸共聚物poly（1actic—co—glycolicacid，PLGA]微囊支架，对其进行表征，并检测其对成骨细胞增殖与粘附的影响。方法：采用复乳法制备PLGA微囊，以激光粒度仪分析其粒径分布，筛选出合适的粒径大小，通过二氯甲烷蒸汽熏蒸使微囊结合，再经过烧结最终形成三维支架，通过扫描电子显微镜观察微囊及微囊支架形态。将MC3T3-E1成骨细胞接种到该支架上，通过扫描电镜观察细胞在支架上的粘附形态，通过CCK-8法检测细胞增殖的情况，并与无支架对照组进行比较。结果：制备的PLGA微囊支架具有一定孔径和孔隙率，具有三维连通的多孔结构，且孔隙结构均匀。细胞可在支架表面粘附生长。微囊支架上的细胞增殖活性高于对照组。结论：本方法制备的PLGA微囊支架具有一定的孔隙率和内部连通性，有利于细胞的粘附和增殖，在骨修复中有应用前景。

Study on preparation of a PLGA microcapsule scaffold and its influence on adhesion and proliferation of osteoblast cells

Objective ：To prepare a novel poly （ lactic-co-glycolic acid） （PLGA） microcapsule scaffold for bone tissue engineering, characterize the scaffold,and detect its influence on the proliferation and adhesion of osteoblasts, nethods：PLGA was used to prepare microcapsules by double emulsion method. After analyzing the size distribution of PLGA microcapsules by the Malvern Mastersizer 2000, we sieved the microspheres into different sizes. Then we used ethylene chloride vapor to make the random packing of PLGA micro- capsules with appropriate size combined. Further sintering helped us to create a three-dimensional microcapsule scaffold. The structure of microcapsule scaffolds were examined by scanning electron microscopy. Osteoblast cell line MC3T3-E1 was chosen to seed on the scaffold,and its adhesive status was observed by SEM. CCK-8 kit was used to analyze the cell proliferation of the group with scaffolds, and to compare with the group without scaffolds. Results：SEM showed that PLGA scaffolds has porous structure,with a certain pore size distribution and porosity. The in vitro study demonstrated that excellent cellular attachment throughout the microcapsule scaffolds. The results of CCK-8 demonstrated the cell viability of cells cultured on PLGA microcapsule scaffolds was higher than the control group after 9 days. Conclusions： From the above results we can conclude that the microcapsule-based scaffold has certain porosity and internal connectivity,which contributes to cell adhesion and proliferation. That means it may be promising as polymeric substitutes for bone re- pair.