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	不同浓度骨碎补总黄酮复合可注射骨修复材料对MC3T3-E1成骨细胞作用的实验研究
	李晋玉 ; 徐林 ; 谭荣伟 ; 金合 ; 李志勇 ; 俞兴 ; Li Jinyu ; Xu Lin ; Tan Rongwei
	2016-03-30 ; 2016-03-30
关键词	骨碎补总黄酮 MC3T3-E1细胞 海藻酸基可注射骨修复 四甲基偶氮唑盐比色法 体外细胞实验 MC3T3-E1 Alginate-based injectable bone repair materials Assemble Flavone of Drynaria Rhizome(AFDR) In vitro experiments MTT R318.08
其他题名	Analysis on the effect of the application of AFDR in combination with injectable bone repair materials on MC3T3-E1 osteoblasts in rats
中文摘要	目的本实验旨在观察中药骨碎补提取物骨碎补总黄酮(AFDR)复合海藻酸基可注射骨修复材料对成骨细胞MC3T3-E1体外培养的影响。方法将成骨细胞株MC3T3-E1随机分为空白对照组、单纯使用海藻酸基可注射骨修复材料组、AFDR0.04mg/L、0.16mg/L、0.64mg/L、1.28mg/L、5.12mg/L 7组,体外复合海藻酸基可注射骨修复材料,以24h和48h为观察时间点,倒置显微镜观察MC3T3-E1细胞形态,台盼蓝拒染法检测MC3T3-E1存活率,MTT法观察细胞增殖效应,碱性磷酸酶(ALP)活性和骨钙素定量检测分别观察不同浓度骨碎补总黄酮促进MC3T3-E1分化作用,采用Von kossa钙化染色法观察其促MC3T3-E1细胞钙化作用。结果①倒置显微镜下观察复合海藻酸基可注射骨修复材料的MC3T3-E1细胞形态,细胞形态呈三角形、多边形、长梭形等,呈集落样生长,无接触抑制,有伪足伸出,胞液透亮,核圆形。AFDR各组不同程度促进成骨细胞的增殖、分化:7组能不同程度的促进成骨细胞的增殖、分化,与空白对照组比较,以0.16 mg/L和0.64mg/L剂量组的成骨细胞增殖数量最多,1.28mg/L,5.12mg/L的AFDR对细胞增殖作用不明显;②MC3T3-E1平均存活率≥90%;③骨碎补总黄酮(AFDR)0.16 mg/L和0.64mg/L剂量组在培养24小时和48小时,其OD值与空白对照组及组间均有显著性差异,可促进MC3T3-E1细胞增殖(P<0.05和P<0.01);④AFDR 0.16 mg/L和0.64mg/L剂量组能使MC3T3-E1的ALP活性升高(P<0.05);⑤AFDR 0.64mg/L剂量组能促进MC3T3-E1骨钙素合成(P<0.05);⑥AFDR 0.16 mg/L和0.64mg/L剂量组均可促进细胞钙化(P<0.05)。结论 AFDR可显著促进在海藻酸基可注射骨修复材料上MC3T3-E1的增殖、分化和钙化,其作用具有时间依赖和剂量依赖关系。; Objective This study was designed to observe the effect of AFDR in combination with alginate-based injectable bone repair materials on osteoblast MC3T3-E1 cultured in vitro.Methods MC3T3-E1 cells were randomly divided into control group,pure materials group,AFDR0.04mg/L,0.16mg/L,0.64mg/L,1.28mg/L,5.12mg/L 6 group.The observation time point:24h and 48h,inverted microscope MC3T3-E1 cell morphology,trypan blue staining MC3T3-E1 cell viability.THE effects were observed by MTT cell proliferation.All data are expressed as mean ± standard deviation,using SPSS 13.0 software for analysis of variance.The effect of different concentrations of Total Flavonoids promoting differentiation of MC3T3-E1 was observed by ALP activity and osteocalcin quantitative detection.The effect of promoting calcification of MC3T3-E1 cells were observed by Von kossa calcification staining.Results ①inverted microscope composite alginate-based injectable bone repair materials MC3T3-E1 cell morphology,cell morphology triangular,polygonal,fusiform,etc,were colony-like growth,no contact inhibition,extending pseudopodia,cytosol translucent,round nuclei.Different sets of degrees AFDR promote osteoblast proliferation and differentiation;②MC3T3-E1 average survival rate≥90%;③ AFDR 0.64mg / L dose group was cultured for 24h and 48h,its OD value with the blank control group and the groups were significantly different,with statistical significance(P<0.05 and P<0.01);④ AFDR 0.16 mg / L and 0.64mg/L dose can increase activity of ALP of MC3T3-E1(P<0.05);⑤ AFDR 0.64mg/ml dose can promote the synthesis of osteocalcin of MC3T3-E1(P<0.05);⑥AFDR 0.16 mg / L and 0.64mg/L dose groups can promote cell calcification(P<0.05).Conclusion AFDR can significantly facilitate the alginate-based injectable bone repair material on the proliferation、differentiation and calcificationof MC3T3-E1,its role has a time-dependent and dose-dependent manner.
语种	中文 ; 中文
内容类型	期刊论文
源URL	[http://ir.lib.tsinghua.edu.cn/ir/item.do?handle=123456789/141697]
专题	清华大学
推荐引用方式 GB/T 7714	李晋玉,徐林,谭荣伟,等. 不同浓度骨碎补总黄酮复合可注射骨修复材料对MC3T3-E1成骨细胞作用的实验研究[J],2016, 2016.
APA	李晋玉.,徐林.,谭荣伟.,金合.,李志勇.,...&Tan Rongwei.(2016).不同浓度骨碎补总黄酮复合可注射骨修复材料对MC3T3-E1成骨细胞作用的实验研究..
MLA	李晋玉,et al."不同浓度骨碎补总黄酮复合可注射骨修复材料对MC3T3-E1成骨细胞作用的实验研究".(2016).

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