培养基加氟后渗透压的变化及其对成釉细胞形态影响的观察
The effects of additional fluoride on the osmotic pressure of DMEM and its influence on the morphology of ameloblasts
张乐 1曹伟靖 1高江红 2董宁 2阮建平2
作者信息
- 1. 延安大学附属医院,陕西延安716000
- 2. 西安交通大学口腔医院,陕西西安710004
- 折叠
摘要
目的:观察加入不同浓度的NaF后DMEM培养基的渗透压变化情况及其对成釉细胞形态的影响.方法:配置含NaF 0(对照组)、10.5、21、42、63、84 mg/L的DMEM培养液,并用冰点渗透压仪检测其渗透压值.取含NaF 0、42、84 mg/L的3组DMEM用于实验,同时设6组不含NaF的DMEM,并分别用58.5 mg/LNaCl、0.05 mg/L甘油、1mg/mL山梨醇将其中3组的渗透压调至与42 mg/L NaF组的渗透压值(323.3 mOsm/kg)接近;用117 mg/L NaCl、0.09 mg/L甘油、1.5 mg/mL山梨醇将其余3组的渗透压调至与84 mg/L NaF组的渗透压值(324.1 mOsm/kg)接近.最后取纯化后的成釉细胞,并将其随机分为9组,分别换用上述含不同浓度NaF、NaCl、甘油、山梨醇的DMEM培养基,并继续培养48 h后,用倒置显微镜和HE染色法观察各组成釉细胞形态变化.结果:42 mg/L NaF组的渗透压均低于对照组,而84 mg/L NaF组的渗透压值均高于对照组.倒置显微镜观察显示:对照组细胞主要呈铺路石样分布,且形态规整,仅有个别细胞脱壁悬浮;42 mg/L NaF组的细胞体积减小,部分细胞皱缩明显,少数细胞脱壁悬浮;84 mg/L NaF组的大部分细胞脱壁漂浮,仅残余个别贴壁细胞,且胞膜破裂,形态紊乱;不同浓度NaCl、甘油、山梨醇各组的细胞形态均与对照组相似,无明显差异.HE染色结果显示:对照组细胞成簇分布,形态不一,多呈圆形、椭圆形及不规则形,胞浆呈紫粉色,胞核为紫蓝色,胞浆内无空泡现象;42 mg/L NaF组的细胞胞浆内出现较多的空泡样变;84 mg/L NaF组仅可见少量细胞,且细胞皱缩成团,其余各组的细胞形态均与对照组无明显差别.结论:染氟成釉细胞的形态变化与氟离子浓度有关,培养基的渗透压升高对成釉细胞的形态无明显影响.
Abstract
AIM:To study the effects of additional fluoride on the osmotic pressure of DMEM and its influence on the morphology of ameloblasts.METHODS:The osmotic pressure of DMEM containing 10％ fetal bovine serum and NaF at 0(control),10.5,21,42,63 and 84 mg/L respectively was measured.Three groups of DMEM containing NaF of 0,42 and 84 mg/L,six sets of DMEM without NaF and with additional 58.5 mg/L of NaCl,0.05 mg/L of glycerol,1 mg/mL of sorbitol and with the same penetrating hydraulic data as with that of 42 mg/L NaF;and with 117 mg/L of NaCl,0.09 mg/L of glycerol,1.5 mg/mL of sorbitol and with the same penetrating hydraulic data as with that of 84 mg/L of NaF,respectively,were prepared for the experiments.Ameloblasts were cultured in the prepared the medium for 48 h.The morphology of the cells was respectively observed by HE staining and inverted microscope.RESULTS:The osmotic pressure of DMEM with additional NaF ＜ 42 mg/L was lower than than that of the control,The osmotic pressure of DMEM with additional NaF≥42 mg/L was higher than that of the control.T.In the 42 mg/L NaF group,a few cells were wrinkled,dwindled and suspended.In the 84 mg/L NaF group almost all the cells suspended and only a few cells attached,and whose cytomembrane were broken.The cellular morphology in NaCl group,glycerol group and sorbitol group were similar to that of the control group.HE staining showed that in 42 mg/L NaF group,most of the cells had vacuolation.In 84 mg/L NaF group,few cells were attached and wrinkled and dwindled into a mass.The ameloblasts in other groups were similar with those in the control group.CONCLUSION:The osmotic pressure of DMEM varys with the different concentration NaF.The osmotic pressure change of medium did not influnce ameloblasts morphology.
关键词
渗透压/氟牙症/成釉细胞/氟化物/渗透压调节剂Key words
osmotic pressure/fluorosis/ameloblast/fluoride/osmotic regulatory agent引用本文复制引用
出版年
2017
NETL
NSTL
维普
万方数据