磷酸钙骨水泥材料的孔径大小与大鼠骨髓基质干细胞黏附能力之间的关系  

Effect of pore size on cell adhesion in calcium phosphate cement scaffolds

作　　者：董鑫[1] 孟国林[1] 白峰[1] 刘建[1] 刘昌胜[2] 郝赋[1] 张金康[1] 贾帅军[1] 阚成双[1] 唐鹏[1] 张建彬[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西发710032 [2]华东理工大学材料科学与工程学院生物材料研究所

出　　处：《中华创伤骨科杂志》2011年第6期571-575,共5页Chinese Journal of Orthopaedic Trauma

基　　金：围家科技支撑计划（2008BA116B02）

摘　　要：目的评价小同大小孔径的磷酸钙骨水泥（CPC）材料对大鼠骨髓基质干细胞（BMSCs）黏附能力的影响。方法采用盐析法制备3种不同孔径（200～300μm、300～450μm、450～600μm）的CPC材料，利用Micro—CT测董3种材料的平均孔径、孔隙率。无菌条件下取新生大鼠BMSCs原代培养并传代；将3组材料分别放置于24孔板内，每种材料接种2×10^5个细胞后。于接种后12、24h收集细胞并计数，计算细胞黏附率；并对24h收集细胞行Annexin V FTTC／PI双标记，流式细胞仪检测细胞捌亡、坏死检测，并利用扫描电镜观察细胞形态及黏附情况。结果Micro—CT测量结果显示：3种CPC材料孔径间相瓦连通，孔隙率均〉67％，平均孔径分别为240、410、510μm。接种后12h，3种材料表面细胞黏附率及坏死牢比较差异均无统计学意义（P〉0．05）；200—300μm组在接种后24h细胞黏附草明显少于300～450μm组和450～600μm组，差异有统计学意义（P〈0．05）；200～300μm组材料衣面细胞坏死数革叫显多于450～600μm组，差异有统计学意义（P〈0．05）。结论孔径大小可影响大鼠BMSCs存多孔CPC材料上的黏附能力，随着孔径增大，细胞黏附率逐渐增高，细胞坏死率逐渐降低，为进一步研究孔径结构对细胞的影响提供了实验依据。Objective To study the effect of different pore sizes of calcium phosphate cement （CPC） scaffolds on cell adhesion. Methods Rat bone mesenchymal stein cells （BMSCs） primarily cultured were used in the present study. CPC scaffolds of 3 pore sizes （200 to 300 μm, 300 to 450 μm, 450 to 600 μm） were respectively groups A, B and C. Micro-CT was used to analyze the average pore size and porosity of each group. The 3 kinds of CPC were placed in 24-well plates and 2 ×10^5 cells were incubated per well. In each group, the cells were harvested 12 and 24 hours after incubation to count cell numbers and adhesion rate. Cell apoptosis and necrosis were detected by flow cytometry and cell morphology and adhesion were observed by scanning electron microscope （SEM） 24 hours after incubation. Results Micro-CT showed that pores of the CPC used in this study were interconnected and the porosity of each group was greater than 67%. The average pore sizes of the 3 groups were 240 μm, 410 μm and 510 μm respectively. At 12 hours, there were no significant differences between the 3 groups in cell adhesion and necrosis （ P 〉 0. 05）. At 24 hours, cell adhesion in group A was significantly less than in groups B and C （ P 〈 O. 05）, and cell necrosis in group A was significantly more than in group C （ P 〈 0.05）. Conclusion In CPC scaffolds for culture of BMSCs, cell adhesion rate may increase gradually and cell necrosis may decrease gradually with the increase of pore size.

关 键 词：磷酸钙骨水泥 骨髓 间充质干细胞 细胞黏附 大鼠 

分 类 号：R318.08[医药卫生—生物医学工程]