摘要
目的探讨浓缩生长因子提取液(CGFe)对钛片表面MC3T3-E1细胞增殖分化的影响。方法实验组使用含CGFe的α-MEM培养液(含10%胎牛血清)培养MC3T3-E1细胞,对照组则使用不含CGFe的α-MEM培养液(含10%胎牛血清)培养细胞。采用MTT法测定培养1、3、5 d时的细胞增殖情况,碱性磷酸酶(ALP)活性测定法检测培养3、5 d时的细胞分化情况;通过扫描电子显微镜(SEM)观察培养12 h时细胞在钛片表面的形态;通过荧光实时定量聚合酶链反应(PCR)测定细胞培养3、7 d时核心结合蛋白因子2(Runx2)和成骨细胞特异性转录因子Osterix(Osx)基因的相对表达量。结果 MTT结果显示:随培养时间延长,细胞数量逐渐增加;各时间点实验组细胞的吸光度值均明显高于对照组(P<0.05)。ALP活性随着培养时间延长而增加,两个时间点实验组的吸光度值均明显高于对照组(P<0.05)。SEM观察:培养12 h时,实验组细胞在钛片表面的形态较对照组更加伸展。PCR结果显示:随着培养时间延长,两组细胞Runx2和Osx基因的表达量逐渐增加,两个时间点实验组的表达量均明显高于对照组(P<0.05)。结论 CGFe能有效地促进MC3T3-E1细胞的增殖、分化及在钛片表面的伸展。
Objective To evaluate the effect of concentrated growth factor extract （CGFe） on the proliferation and differen- tiation of MC3T3-E1 osteoblasts attached to sandblasted and acid etched titanium surfaces. Methods Trials were divided into experimental and control groups. The experimental group used α-MEM that contained CGFe （10% FBS）, whereas the control group only used α-MEM （10% FBS）. MTT assay was employed to detect the number of osteoblasts on the first, third, and fifth days. Alkaline phosphatase （ALP） activity and scanning electron microscope （SEM） were used to detect the osteoblast differentiations on the third and fifth days and to observe the osteoblast extensions on titanium surfaces for 12 h, respectively. Meanwhile, the levels of the osteogenetic biomarkers Runt-related transcription factor-2 （Runx2） and Osterix （Osx） on the third and seventh days were quantified via real-time polymerase chain reaction （PCR）. Results MTT assay indicated that on the first, third, and fifth days, the absorbance in the experimental group significantly increased than that in the control group （P〈0.05）. ALP activity： on the third and fifth days, the absorbance of the experimental group was significantly higher than that of the control group （P〈0.05）. SEM： at 12 h, the extension of the experimental group cells was larger than that of the control group. Real-time PCR： given the standardization in the group, the gene expression level of the control group on the third day was 1, and the Rtmx2 and Osx gene expressions in the experimental group were larger than those of the con- trol group. Conelusion CGFe can efficiently stimulate the proliferation, differentiation and extension of MC3T3-E1 cells.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2015年第1期84-87,共4页
West China Journal of Stomatology
关键词
浓缩生长因子
骨结合
成骨细胞
concentrated growth factor
osseointegration
osteoblast
