羟基磷灰石对人牙髓细胞内游离钙的影响

Influence of hydroxyapatite on intracellular calcium ion level in human dental pulp cells observed by confocal laser scanning microscopy

目的:检测羟基磷灰石(hydroxyapatite,HA)对体外培养的人牙髓细胞内游离Ca2+的作用。方法:DMEM培养液体外培养的人牙髓细胞随机分为3组,2个实验组培养液中分别加入Ca(OH)2、HA共孵育24h。钙荧光指示剂Fluo-3/AM负载后,3组均加入HA干预,在激光扫描共聚焦显微镜下观测细胞内Ca2+的变化。结果:瞬时加入HA后,3组细胞的钙离子荧光强度在30s内均升高;空白对照组(A组)升高较为迅速,30s后即开始缓慢下降;Ca(OH)2组(B组)的升高幅度小于A组,60s时达到最高(230.63±5.24);HA组(C组)升高变化较小,30s时达到最高(161.25±3.81),随后小幅度下降。结论:HA可引起体外培养人牙髓细胞内[Ca2+]i的变化。

Objective： To investigate the effect of hydroxyapatite on intracellular calcium ion level in cultured human dental pulp cells in vitro. Methods： The fourth passage of cultured human dental pulp cells were maintained in DMEM medium and randomly divided into 3 groups. The blank group （group A） was kept in DMEM medium only, while the experimental groups were cultured in medium supplemented with Ca（OH） 2 （group B） and HA （group C） respectively for 24 h. The intracellular calcium level was measured with Fluo-3 probe by confoeal laser scanning microscope after re-stimulation with HA. Results： After HA stimulation, the intracellular calcium ion concentration in all groups increased quickly and reached the maximum level of （230.63 ± 5.24） in group B and （ 161.25 ± 3.81 ） in group C within 60 s and 30 s respectively, then decreased gradually. Conclusion： HA can cause the changes of intracellular calcium ion level in cultured human dental pulp cells.