中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2012年
6期
364-368
,共5页
王钰莹%运慧媛%缪宏颖%胡伟平
王鈺瑩%運慧媛%繆宏穎%鬍偉平
왕옥형%운혜원%무굉영%호위평
干细胞%牙髓%成骨细胞%细胞分化
榦細胞%牙髓%成骨細胞%細胞分化
간세포%아수%성골세포%세포분화
Stem cells%Dental pulp%Osteoblasts%Cell differentiation
目的 比较成骨细胞和矿化液对牙髓干细胞(dental pulp stem cells,DPSC)向成骨细胞分化的作用,选择理想的诱导方法.方法 利用插入式小室将成骨细胞与DPSC共培养作为共培养组,矿化液培养DPSC作为矿化液组.在光学显微镜和透射电镜下观察DPSC形态学变化；应用茜素红染色法检测矿化结节；采用反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-RCR)分别榆测DPSC骨涎蛋白、Runt相关转录因子2、骨钙蛋白、Ⅰ型胶原等骨源性基因表达情况.结果 茜素红染色显示培养28 d后共培养组DPSC矿化结节明显多于矿化液组.培养15 d后共培养组骨涎蛋白和Ⅰ型胶原基因阳性表达水平(分别为9.807±1.135和2.913±0.310)显著高于矿化液组(分别为6.478±0.781和1.703±0.184,P＜0.05).结论 成骨细胞分泌大量细胞因子或可溶性蛋白,对DPSC的诱导作用较矿化液明显.
目的 比較成骨細胞和礦化液對牙髓榦細胞(dental pulp stem cells,DPSC)嚮成骨細胞分化的作用,選擇理想的誘導方法.方法 利用插入式小室將成骨細胞與DPSC共培養作為共培養組,礦化液培養DPSC作為礦化液組.在光學顯微鏡和透射電鏡下觀察DPSC形態學變化；應用茜素紅染色法檢測礦化結節；採用反轉錄聚閤酶鏈反應(reverse transcription polymerase chain reaction,RT-RCR)分彆榆測DPSC骨涎蛋白、Runt相關轉錄因子2、骨鈣蛋白、Ⅰ型膠原等骨源性基因錶達情況.結果 茜素紅染色顯示培養28 d後共培養組DPSC礦化結節明顯多于礦化液組.培養15 d後共培養組骨涎蛋白和Ⅰ型膠原基因暘性錶達水平(分彆為9.807±1.135和2.913±0.310)顯著高于礦化液組(分彆為6.478±0.781和1.703±0.184,P＜0.05).結論 成骨細胞分泌大量細胞因子或可溶性蛋白,對DPSC的誘導作用較礦化液明顯.
목적 비교성골세포화광화액대아수간세포(dental pulp stem cells,DPSC)향성골세포분화적작용,선택이상적유도방법.방법 이용삽입식소실장성골세포여DPSC공배양작위공배양조,광화액배양DPSC작위광화액조.재광학현미경화투사전경하관찰DPSC형태학변화；응용천소홍염색법검측광화결절；채용반전록취합매련반응(reverse transcription polymerase chain reaction,RT-RCR)분별유측DPSC골연단백、Runt상관전록인자2、골개단백、Ⅰ형효원등골원성기인표체정황.결과 천소홍염색현시배양28 d후공배양조DPSC광화결절명현다우광화액조.배양15 d후공배양조골연단백화Ⅰ형효원기인양성표체수평(분별위9.807±1.135화2.913±0.310)현저고우광화액조(분별위6.478±0.781화1.703±0.184,P＜0.05).결론 성골세포분비대량세포인자혹가용성단백,대DPSC적유도작용교광화액명현.
Objective To find an ideal method inducing dental pulp stem cells(DPSC) osteogenic differentiation.To compare the effect of co-culture method and that of mineralizing culture medium.Methods DPSC were co-cultured with osteoblasts using cell culture inserts system as experiment group,and DPSC were cultured in mineralizing culture medium as control group. The cell morphology and ultrastructure and mineralized nodes were analyzed under phase contrast microscope,transmission electron microscope,and alizarin red S staning. Bone sialoprotein (BSP),Runx-2,osteocalcin,and collagen-1( Col-1 ) osteoblastic genes expressions of DPSC cultivated in special niche of osteoblasts were assayed by reverse transcription polymerase chain reaction (RT-PCR). Results The mineralization nudoles of experiment group were more than control group.Fifteen days later,BSP and Col-1 genes in the DPSC of cocultures were 9.807 ± 1.135 and 2.913 ± 0.310,respectively. And those in the DPSC of mineralizing culture medium were 6.478 ± 0.781 and 1.703 ± 0.184,respectively.Co-cultures and mineralizing were significantly different( P ＜0.05 ).Conclusions As osteoblasts can secret lots of osteogenic cell eytokines,they have more significant effect than mineralizing culture medium on osteogenesis of DPSC.
