中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2015年
1期
38-42
,共5页
刘红春%郭晓恒%刘骁%陈伟建%赵娜如%廖义东
劉紅春%郭曉恆%劉驍%陳偉建%趙娜如%廖義東
류홍춘%곽효항%류효%진위건%조나여%료의동
羟基磷灰石类%氟化物%细胞增殖%细胞分化
羥基燐灰石類%氟化物%細胞增殖%細胞分化
간기린회석류%불화물%세포증식%세포분화
Hydroxyapatites%Fluorides%Cell proliferation%Cell differentiation
目的 评价氟取代比例为20％的氟-羟基磷灰石(fluor-hydroxyapatite,FHA)对骨肉瘤MG63细胞增殖和成骨分化的影响,以期为FHA的临床应用提供依据.方法 采用化学沉淀法合成FHA,扫描电镜、X射线衍射仪、傅里叶红外光谱仪观察结构并表征.设置FHA组(培养基中加入FHA)、羟基磷灰石(hydroxyapatite,HA)组(培养基中加入HA)和空白对照组(单纯培养基),每组样本量为3,共培养骨肉瘤MG63细胞.甲基噻唑基四唑法检测3组细胞增殖能力,检测3组细胞碱性磷酸酶(alkaline phosphatese,ALP)活性.反转录PCR检测3组细胞成骨分化相关基因(Ⅰ型胶原、ALP、骨钙素和核心结合因子)mRNA表达情况.结果 FHA的X射线衍射图谱主晶相与HA标准图谱一致.培养3d后FHA组细胞增殖(A值为1.87±0.06)显著高于空白对照组(1.25±0.02)(P＜0.05),FHA组与HA组(1.84±0.03)差异无统计学意义(P＞0.05).培养3d后FHA组ALP活性(4.62±0.09)显著高于空白对照组(1.92±0.05)(P＜0.05).与空白对照组相比,FHA上调细胞Ⅰ型胶原、ALP、骨钙素mRNA表达,下调核心结合因子mRNA表达.结论 FHA对MG63细胞增殖和成骨分化相关基因mRNA的表达有促进作用,具有良好的生物相容性.
目的 評價氟取代比例為20％的氟-羥基燐灰石(fluor-hydroxyapatite,FHA)對骨肉瘤MG63細胞增殖和成骨分化的影響,以期為FHA的臨床應用提供依據.方法 採用化學沉澱法閤成FHA,掃描電鏡、X射線衍射儀、傅裏葉紅外光譜儀觀察結構併錶徵.設置FHA組(培養基中加入FHA)、羥基燐灰石(hydroxyapatite,HA)組(培養基中加入HA)和空白對照組(單純培養基),每組樣本量為3,共培養骨肉瘤MG63細胞.甲基噻唑基四唑法檢測3組細胞增殖能力,檢測3組細胞堿性燐痠酶(alkaline phosphatese,ALP)活性.反轉錄PCR檢測3組細胞成骨分化相關基因(Ⅰ型膠原、ALP、骨鈣素和覈心結閤因子)mRNA錶達情況.結果 FHA的X射線衍射圖譜主晶相與HA標準圖譜一緻.培養3d後FHA組細胞增殖(A值為1.87±0.06)顯著高于空白對照組(1.25±0.02)(P＜0.05),FHA組與HA組(1.84±0.03)差異無統計學意義(P＞0.05).培養3d後FHA組ALP活性(4.62±0.09)顯著高于空白對照組(1.92±0.05)(P＜0.05).與空白對照組相比,FHA上調細胞Ⅰ型膠原、ALP、骨鈣素mRNA錶達,下調覈心結閤因子mRNA錶達.結論 FHA對MG63細胞增殖和成骨分化相關基因mRNA的錶達有促進作用,具有良好的生物相容性.
목적 평개불취대비례위20％적불-간기린회석(fluor-hydroxyapatite,FHA)대골육류MG63세포증식화성골분화적영향,이기위FHA적림상응용제공의거.방법 채용화학침정법합성FHA,소묘전경、X사선연사의、부리협홍외광보의관찰결구병표정.설치FHA조(배양기중가입FHA)、간기린회석(hydroxyapatite,HA)조(배양기중가입HA)화공백대조조(단순배양기),매조양본량위3,공배양골육류MG63세포.갑기새서기사서법검측3조세포증식능력,검측3조세포감성린산매(alkaline phosphatese,ALP)활성.반전록PCR검측3조세포성골분화상관기인(Ⅰ형효원、ALP、골개소화핵심결합인자)mRNA표체정황.결과 FHA적X사선연사도보주정상여HA표준도보일치.배양3d후FHA조세포증식(A치위1.87±0.06)현저고우공백대조조(1.25±0.02)(P＜0.05),FHA조여HA조(1.84±0.03)차이무통계학의의(P＞0.05).배양3d후FHA조ALP활성(4.62±0.09)현저고우공백대조조(1.92±0.05)(P＜0.05).여공백대조조상비,FHA상조세포Ⅰ형효원、ALP、골개소mRNA표체,하조핵심결합인자mRNA표체.결론 FHA대MG63세포증식화성골분화상관기인mRNA적표체유촉진작용,구유량호적생물상용성.
Objective To investigate the effects of 20％ fluor-hydroxyapatite(FHA) on proliferation and osteogenic differentiation of human MG63 osteosarcoma cells.Methods FHA was prepared by chemical precipitation method,and its structure and surface features were tested by scanning microscope,Xray diffraction(XRD) and Fourier transform infrared spectroscopy.MG63 cells were cultured and divided into FHA,hydroxyapatite(HA) and control groups(n=3).The proliferation of the cells was evaluated using methylthiazol tetrazolium(MTT) assay.ALP activity of the cells was assessed.Osteogenic differentiation was evaluated based on the reverse transcription PCR(RT-PCR) of differentiation-related genes,namely,collagen type Ⅰ(Col Ⅰ),alkaline phosphatase(ALP),osteocalcin(OCN) and core binding factor α1(Cbfα1).The data were analyzed statistically by one-way analysis of variance using SPSS 13.0 software.Results XRD test showed that the main crystalline phase of FHA was similar to that of HA.Absorptance value of cells exposed to FHA(1.87±0.06) measured by MTT was higher than that of the control(1.25±0.02) on the third day(P＜ 0.05),and there was no statistically significant difference between the cells exposed to FHA and HA(1.84± 0.03)(P＞0.05).ALP activity of the cells exposed to FHA (4.62±0.09) was higher than that of the control (1.92 ± 0.05)(P＜0.05).RT-PCR tests showed that compared with the control,FHA up-regulated the expression of Col Ⅰ,ALP and OCN mRNA,down-regulated the expression of Cbfα1 mRNA.Conclusions FHA enhances the proliferation and osteogenic differentiation-related gene expression,and has good biocompatibility.
