CPC固化中海藻酸钙凝胶微球对细胞的保护及相关影响因素

Protective role of calcium alginate hydrogel beads in cells during calcium phosphate cement setting reaction and its influencing factors

目的:通过体外细胞实验,评价在磷酸钙骨水泥(calcium phosphate cement,CPC)浆料固化过程中海藻酸钙凝胶微球对小鼠成骨前体细胞(mouse preosteoblastic cell line,MC3T3-E1)的保护作用及相关影响因素。方法:将包封MC3T3-E1细胞的海藻酸钙凝胶微球与CPC浆料和固化块复合培养2 d,CCK-8检测MC3T3-E1细胞活性,钙黄绿素-AM(Calcein-AM)和碘化丙啶(PI)溶液分别对活细胞和死细胞染色。比较CPC浆料和CPC固化块、CPC固化后不同处理方式以及不同换液时间对MC3T3-E1细胞活性的影响。结果:CPC浆料组和固化块组的光密度值无显著差异(1 d:P=0.827;2 d:P=0.965),荧光图片见两组细胞数及形态无明显区别。CPC固化后的不同处理方式以及不同换液时间对细胞活性有不同影响。结论:在CPC浆料固化过程中,海藻酸钙凝胶微球对MC3T3-E1细胞有较好的保护作用。CPC浆料固化后可能仍持续影响细胞增殖,这种影响可能主要集中在固化后24 h内。CPC浆料固化后1 d内每6 h换液,有利于细胞的活性;2 d内每6 h换液可能对细胞活性产生不利影响。

Objective：To evaluate the protection of calcium alginate hydrogel beads for mouse preosteo- blastic cell line （MC3T3-E1） during the setting reaction of calcium phosphate cement （CPC） pastes and to analyze related influencing factors. Methods： The MC3T3-E1 cells encapsulated in alginate hydrogel beads were cultured in vitro with [3-tricalcium phosphate/calcium phosphate cement （[3-TCP/CPC） pastes and discs until 2 d. The cell viability and number of live/dead cells were tested using the CCK-8, Calcein-AM and PI, respectively. The effects of CPC paste and CPC disc, different treatment and time to renewal medium on cell viability were compared. Results： There were no significant differences in optical density and number of live cells between [3-TCP/CPC pastes and discs （1 d： P = 0. 827; 2 d： P = 0. 965 ）. There were different influences on cell viability between different treatment and time to renewal medium. Conclusion： The results indicated that the calcium alginate hydrogel beads had a good protec- tion to MC3T3-E1 cells during the setting reaction of CPC pastes. CPC paste might affect the cell proliferation even when solidification. After setting reaction, it may be favorable for cell viability to renew the medium every 6 h in 1 d.