PLGA/CPC支架材料复合骨髓基质干细胞构建组织工程骨的体外效果评价  

Evaluation of tissue engineering bone constructed with PLGA/CPC scaffold material and bone marrow stromal cells in vitro

作　　者：于祥茹[1] 韩晓谦[1,2] 袁浩天 董志恒[2] 程梁[1] 吴哲[1] 

机构地区：[1]吉林大学口腔医院口腔修复科,吉林长春130021 [2]滨州医学院附属医院口腔科,山东滨州256600

出　　处：《吉林大学学报（医学版）》2014年第2期294-299,I0003,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅自然科学基金资助课题(201215052)

摘　　要：目的：探讨聚乳酸聚羟基乙酸／磷酸钙骨水泥（PLGA／CPC）复合骨髓基质干细胞（BMSCs）构建组织工程骨的可行性，为预防剩余牙槽嵴萎缩和促进骨再生提供科学理论指导。方法：采用溶剂浇铸一粒子沥滤技术结合相分离法制备PLGA／CPC支架材料；体外分离、培养、纯化大鼠BMSCs；第3代BMSCs经Dil染色后接种于复合支架材料。实验分为实验组（PLGA／CPC＋BMSCs）和对照组（单纯BMSCs）。扫描电镜和激光共聚焦观察支架材料的孔隙率及BMScs在支架上的黏附情况；CCK-8和碱性磷酸酶（AKP）法检测2组细胞增殖和分化。结果：支架材料孔隙率达90％以上，孔径平均为200300μm，支架材料与细胞黏附性较好；原代BMSCs前3d增殖较慢，3～6d增殖较快，6d后增殖较慢；BMSCs表面抗原CD44和CD105均呈阳性表达；茜素红和Ⅰ型胶原染色，BMSCs有良好的成骨活性；Dil染色，细胞标记率达90％以上，标记物对细胞形态无明显影响。CCK-8和AKP法检测，实验组和对照组BMSCs增殖率和AKP活性差异无统计学意义（P〉0．05）。结论：PLGA／CPC是理想的组织工程支架材料。Objective To study the feasibility of tissue engineering bone constructed with poly lactic-co-glycolic acid/ calcium phosphate cement（PLGA/CPC） and bone marrow stromal cells （BMSCs）, and to provide scientific and theoretical and guiding for preventing residual ridge resorption and promoting bone regeneration. Methods Solvent casting - particle leaching technology combined with the phase separation process was used to prepare PLGA/CPC scaffold material; the BMSCs of rats were isolated, cultured, and purified in vitro ; the third generation of BMSCs after Dil staining were planted on the composite scaffold material. The experiment was divided into experiment group（ PLGA/CPC ＋ BMSCs） and control group （BMSCs）. Scanning electron microscope and laser confocal microscope were used to observe the adhesion of BMSCs on the PLGA/CPC material; CCK-8 and alkaline phosphatase （AKP） methods were used to detect the proliferation and differentiation of BMSCs of two materials. Results The porosity of PLGA/CPC scaffold was above 90%, and the average pore diameter was 200-300 μm and the BMSCs were combined with the scaffolds very well; the proliferation of original generation of BMSCs was low during the first 3 d, and it became fast during 3-6 d, but it became low after 6 d; the expressions of surface antigens CD44 and CD105 in BMSCs were positive; the alizarin red and typeⅠ collagen staining results showed that BMSCs had good osteogenic activity; the Dil staining results showed that the rate of marked cells was more than 90%, and the markers had no significant effect on the morphology of BMSCs. The CCK-8 and AKP results showed there were no significant differences of proliferation rate of BMSCs and AKP activities between experiment group and control group（P〉0.05）. Gonclusion PLGA/CPC is an ideal tissue engineering scaffold material.

关 键 词：骨髓基质干细胞 骨组织工程 聚乳酸羟基乙酸 磷酸钙骨水泥 茜素红染色 

分 类 号：R783.1[医药卫生—口腔医学]