Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf e...Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf ethanol extracts on H2O2-induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms.Locally-grown Duzhong leaves were extracted with ethanol.MC3T3-E1 cells were treated with EuO (6.25,12.5,25,50,and 100 μg/ml) for 24 h,and then H2O2 (800 μmol/L) for an additional 24 h.Cell survival rate,percentage of apoptosis,and expressions of caspases 3,6,7,and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,microscopic analysis,Western blotting,and reverse transcription polymerase chain reaction (RT-PCR).The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC).EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 μg/ml,0.21;6.25 μg/ml,0.28;12.5 μg/ml,0.31;25 μg/ml,0.48;50 μg/ml,0.54;and 100 μg/ml,0.66 (P<0.05),with the half-effective concentration being around 25 μg/ml.MTT results were confirmed by microscopic analysis.Western blotting and RT-PCR analyses showed that the expressions of caspases 3,6,7,and 9 were significantly decreased in the EuO-treated cells compared with the control (EuOand H2O2-free) (P<0.05),with the half-effective concentration of EuO ranging from 12.5 to 25 μg/ml.We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells,and suppressed the H2O2-induced apoptosis in a rat MC3T3-E1 osteogenic cell model,likely due to the inhibition of caspases’ activities.The results indicate that EuO is a potent antioxidant,which may contribute to its many cellular protective functions,including the promotion of bone growth.展开更多
目的:探讨再矿化介质聚乙烯磷酸和聚丙烯酸对3种粘结剂(Fluoro Bond II、Xeno-III和i Bond)和牙本质所形成的树脂-牙本质界面再矿化程度的影响。创新点:(1)多数仿生再矿化研究集中于通过透射电子显微镜观察牙本质粘结剂界面的再矿化情况...目的:探讨再矿化介质聚乙烯磷酸和聚丙烯酸对3种粘结剂(Fluoro Bond II、Xeno-III和i Bond)和牙本质所形成的树脂-牙本质界面再矿化程度的影响。创新点:(1)多数仿生再矿化研究集中于通过透射电子显微镜观察牙本质粘结剂界面的再矿化情况,鲜有采用激光共聚焦显微镜(CLSM)作为观察手段的。虽有研究应用CLSM,但也仅是从定性的角度比较再矿化效应,没有定量分析比较,难以令人信服。而本项研究从定性和定量两方面分析对比。(2)本研究从当前牙本质粘结剂系统(共七代)常用的后四代中选择典型代表产品为研究对象,便于横向比较各类粘结剂。(3)本研究的定量结果除了比较同一时间实验组和对照组的矿化情况,还增加了纵向分析思路,进一步比较矿化程度随时间的变化以及不同牙本质粘结剂粘结界面的矿化情况。方法:将96颗健康前磨牙按照Fluoro Bond II、Xeno-III和i Bond粘结剂随机分为3组,每颗牙均暴露表层牙本质。3种粘结剂分别严格按照各自产品说明书处理牙本质表面,牙合面堆制5 mm厚的树脂。每颗牙沿牙合-龈向切成0.9 mm厚的薄片,获取中间两片样本用于矿化组和模拟组。矿化组采用含聚乙烯磷酸和聚丙烯酸的再矿化液浸泡;模拟组采用不含聚乙烯磷酸和聚丙烯酸的模拟体液浸泡。各组标本在储存1、2、3和4个月后,各取出8片,经苏丹明B荧光染料染色24 h,冲洗,吹干,置CLSM下观察渗入混合层及粘结层的荧光情况,测量荧光面积、平均荧光量及总荧光量。所有数据统计方法采用方差分析(ANOVA)和Tukey’s HSD检验分析。结论:本研究中聚丙烯酸和聚乙烯磷酸双仿生类似物分子对Fluoro Bond II、Xeno-III及i Bond粘结剂均显示不同程度的再矿化效应,其中对i Bond粘结剂再矿化效应最明显,Fluoro Bond II粘结剂次之,Xeno-III粘结剂再矿化效应较差,但能够起到抵制基底矿物继续丢失或阻止胶原降解的作用。CLSM结合应用苏丹明B是量化混合层再矿化的一项有效手段。因此,上述双仿生类似物分子应用于口腔粘结剂修复材料促使混合层再矿化,具有良好的应用前景,值得进一步深入研究。展开更多
研究目的:评价含有酪蛋白磷酸肽-无定形磷酸钙(CPP-ACP)的再矿化剂Tooth Mousse(TM)的使用、不同玷污层的处理以及样本储存时间对牙本质粘接微拉伸性能的影响。研究方法:将牙本质样本分成保留玷污层组和用15%乙二胺四乙酸(EDTA)处理90...研究目的:评价含有酪蛋白磷酸肽-无定形磷酸钙(CPP-ACP)的再矿化剂Tooth Mousse(TM)的使用、不同玷污层的处理以及样本储存时间对牙本质粘接微拉伸性能的影响。研究方法:将牙本质样本分成保留玷污层组和用15%乙二胺四乙酸(EDTA)处理90秒去除玷污层组。每组根据是否使用TM处理再分亚组。每个亚组分别用三种不同的粘结剂(两步法自酸蚀Clearfil SE Bond(CSE)、一步法自酸蚀G-Bond(GB)和全酸蚀Adper Single Bond 2(SB))与牙本质样本粘接,分别经过3天和6个月的去离子水储存。样本进行切割微拉伸测试并通过扫描电镜分析断裂界面模式。重要结论:经过再矿化剂TM预处理,可以减少牙齿的敏感性,并且对于这三种粘结系统经过长时间储存后的粘结性能没有影响。EDTA的处理对于长期储存的粘结性能没有显著影响。额外的TM和EDTA对短期(3天)粘接力会有效应,但对长期(6个月)的粘接力没有影响。展开更多
基金Project (No.2007C33030) supported by the Science and Technology Program of Zhejiang Province,China
文摘Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf ethanol extracts on H2O2-induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms.Locally-grown Duzhong leaves were extracted with ethanol.MC3T3-E1 cells were treated with EuO (6.25,12.5,25,50,and 100 μg/ml) for 24 h,and then H2O2 (800 μmol/L) for an additional 24 h.Cell survival rate,percentage of apoptosis,and expressions of caspases 3,6,7,and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,microscopic analysis,Western blotting,and reverse transcription polymerase chain reaction (RT-PCR).The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC).EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 μg/ml,0.21;6.25 μg/ml,0.28;12.5 μg/ml,0.31;25 μg/ml,0.48;50 μg/ml,0.54;and 100 μg/ml,0.66 (P<0.05),with the half-effective concentration being around 25 μg/ml.MTT results were confirmed by microscopic analysis.Western blotting and RT-PCR analyses showed that the expressions of caspases 3,6,7,and 9 were significantly decreased in the EuO-treated cells compared with the control (EuOand H2O2-free) (P<0.05),with the half-effective concentration of EuO ranging from 12.5 to 25 μg/ml.We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells,and suppressed the H2O2-induced apoptosis in a rat MC3T3-E1 osteogenic cell model,likely due to the inhibition of caspases’ activities.The results indicate that EuO is a potent antioxidant,which may contribute to its many cellular protective functions,including the promotion of bone growth.
基金Project supported by the National Natural Science Foundation of China(No.81271955)the Zhejiang Provincial Natural Science Foundation of China(No.Y2080338)
文摘目的:探讨再矿化介质聚乙烯磷酸和聚丙烯酸对3种粘结剂(Fluoro Bond II、Xeno-III和i Bond)和牙本质所形成的树脂-牙本质界面再矿化程度的影响。创新点:(1)多数仿生再矿化研究集中于通过透射电子显微镜观察牙本质粘结剂界面的再矿化情况,鲜有采用激光共聚焦显微镜(CLSM)作为观察手段的。虽有研究应用CLSM,但也仅是从定性的角度比较再矿化效应,没有定量分析比较,难以令人信服。而本项研究从定性和定量两方面分析对比。(2)本研究从当前牙本质粘结剂系统(共七代)常用的后四代中选择典型代表产品为研究对象,便于横向比较各类粘结剂。(3)本研究的定量结果除了比较同一时间实验组和对照组的矿化情况,还增加了纵向分析思路,进一步比较矿化程度随时间的变化以及不同牙本质粘结剂粘结界面的矿化情况。方法:将96颗健康前磨牙按照Fluoro Bond II、Xeno-III和i Bond粘结剂随机分为3组,每颗牙均暴露表层牙本质。3种粘结剂分别严格按照各自产品说明书处理牙本质表面,牙合面堆制5 mm厚的树脂。每颗牙沿牙合-龈向切成0.9 mm厚的薄片,获取中间两片样本用于矿化组和模拟组。矿化组采用含聚乙烯磷酸和聚丙烯酸的再矿化液浸泡;模拟组采用不含聚乙烯磷酸和聚丙烯酸的模拟体液浸泡。各组标本在储存1、2、3和4个月后,各取出8片,经苏丹明B荧光染料染色24 h,冲洗,吹干,置CLSM下观察渗入混合层及粘结层的荧光情况,测量荧光面积、平均荧光量及总荧光量。所有数据统计方法采用方差分析(ANOVA)和Tukey’s HSD检验分析。结论:本研究中聚丙烯酸和聚乙烯磷酸双仿生类似物分子对Fluoro Bond II、Xeno-III及i Bond粘结剂均显示不同程度的再矿化效应,其中对i Bond粘结剂再矿化效应最明显,Fluoro Bond II粘结剂次之,Xeno-III粘结剂再矿化效应较差,但能够起到抵制基底矿物继续丢失或阻止胶原降解的作用。CLSM结合应用苏丹明B是量化混合层再矿化的一项有效手段。因此,上述双仿生类似物分子应用于口腔粘结剂修复材料促使混合层再矿化,具有良好的应用前景,值得进一步深入研究。
基金Project supported by the National Natural Science Foundation of China(Nos.81271955 and 30973350)the Zhejiang Provincial Natural Science Foundation of China(No.Y2080338)
文摘研究目的:评价含有酪蛋白磷酸肽-无定形磷酸钙(CPP-ACP)的再矿化剂Tooth Mousse(TM)的使用、不同玷污层的处理以及样本储存时间对牙本质粘接微拉伸性能的影响。研究方法:将牙本质样本分成保留玷污层组和用15%乙二胺四乙酸(EDTA)处理90秒去除玷污层组。每组根据是否使用TM处理再分亚组。每个亚组分别用三种不同的粘结剂(两步法自酸蚀Clearfil SE Bond(CSE)、一步法自酸蚀G-Bond(GB)和全酸蚀Adper Single Bond 2(SB))与牙本质样本粘接,分别经过3天和6个月的去离子水储存。样本进行切割微拉伸测试并通过扫描电镜分析断裂界面模式。重要结论:经过再矿化剂TM预处理,可以减少牙齿的敏感性,并且对于这三种粘结系统经过长时间储存后的粘结性能没有影响。EDTA的处理对于长期储存的粘结性能没有显著影响。额外的TM和EDTA对短期(3天)粘接力会有效应,但对长期(6个月)的粘接力没有影响。
