中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2013年
1期
27-31
,共5页
王燕萍%吴锦涛%王子露%郑阳玉%张光东%于金华
王燕萍%吳錦濤%王子露%鄭暘玉%張光東%于金華
왕연평%오금도%왕자로%정양옥%장광동%우금화
干细胞研究%磷酸二氢钾%细胞分化
榦細胞研究%燐痠二氫鉀%細胞分化
간세포연구%린산이경갑%세포분화
Stem cells research%Potassium dihydrogen phosphate%Cell differentiation
目的 探讨磷酸二氢钾(KH2 PO4)对根尖牙乳头干细胞(stem cells from apical papilla,SCAP)成牙与成骨向分化能力的影响,以期为组织工程牙再生、骨再生及临床运用KH2PO4提供实验依据.方法 通过酶消化法分离培养人SCAP,将SCAP分别培养于常规培养液α-最低基础培养基及含1.8 mmol/L KH2PO4的α-最低基础培养基常规培养液中,分为空白对照组和实验组(KH2PO4刺激组).通过碱性磷酸酶(alkaline phosphotase,ALP)活性检测、茜素红染色实验及实时定量反转录聚合酶链反应(reverse transcription PCR,RT-PCR)和蛋白质印迹法检测SCAP体外成牙及成骨向分化的能力.结果 ALP活性检测结果显示,3d时实验组ALP活性[(0.370 ±0.013) Sigma单位·min-1 ·mg-1]显著高于空白对照组[(0.2845±0.0084) Sigma单位·min-1·mg-1],差异有统计学意义(P＜0.01)；培养5、7d后茜素红染色结果显示实验组形成了明显的矿化结节,实验组5、7d时钙离子浓度[分别为(0.539 ±0.007)、(1.617 ±0.042) μg/g]均显著高于空白对照组[(0.138 ±0.037) μg/g],P＜0.01.RT-PCR及蛋白质印迹法检测结果显示:实验组SCAP培养3、7d后ALP、核心结合蛋白因子2、成骨相关转录因子、骨钙蛋白及牙本质涎磷蛋白的基因及蛋白表达均显著高于空白对照组(P＜0.01,P＜0.05).结论 1.8 mmol/L KH2P04可以明显促进人SCAP体外成牙及成骨能力.
目的 探討燐痠二氫鉀(KH2 PO4)對根尖牙乳頭榦細胞(stem cells from apical papilla,SCAP)成牙與成骨嚮分化能力的影響,以期為組織工程牙再生、骨再生及臨床運用KH2PO4提供實驗依據.方法 通過酶消化法分離培養人SCAP,將SCAP分彆培養于常規培養液α-最低基礎培養基及含1.8 mmol/L KH2PO4的α-最低基礎培養基常規培養液中,分為空白對照組和實驗組(KH2PO4刺激組).通過堿性燐痠酶(alkaline phosphotase,ALP)活性檢測、茜素紅染色實驗及實時定量反轉錄聚閤酶鏈反應(reverse transcription PCR,RT-PCR)和蛋白質印跡法檢測SCAP體外成牙及成骨嚮分化的能力.結果 ALP活性檢測結果顯示,3d時實驗組ALP活性[(0.370 ±0.013) Sigma單位·min-1 ·mg-1]顯著高于空白對照組[(0.2845±0.0084) Sigma單位·min-1·mg-1],差異有統計學意義(P＜0.01)；培養5、7d後茜素紅染色結果顯示實驗組形成瞭明顯的礦化結節,實驗組5、7d時鈣離子濃度[分彆為(0.539 ±0.007)、(1.617 ±0.042) μg/g]均顯著高于空白對照組[(0.138 ±0.037) μg/g],P＜0.01.RT-PCR及蛋白質印跡法檢測結果顯示:實驗組SCAP培養3、7d後ALP、覈心結閤蛋白因子2、成骨相關轉錄因子、骨鈣蛋白及牙本質涎燐蛋白的基因及蛋白錶達均顯著高于空白對照組(P＜0.01,P＜0.05).結論 1.8 mmol/L KH2P04可以明顯促進人SCAP體外成牙及成骨能力.
목적 탐토린산이경갑(KH2 PO4)대근첨아유두간세포(stem cells from apical papilla,SCAP)성아여성골향분화능력적영향,이기위조직공정아재생、골재생급림상운용KH2PO4제공실험의거.방법 통과매소화법분리배양인SCAP,장SCAP분별배양우상규배양액α-최저기출배양기급함1.8 mmol/L KH2PO4적α-최저기출배양기상규배양액중,분위공백대조조화실험조(KH2PO4자격조).통과감성린산매(alkaline phosphotase,ALP)활성검측、천소홍염색실험급실시정량반전록취합매련반응(reverse transcription PCR,RT-PCR)화단백질인적법검측SCAP체외성아급성골향분화적능력.결과 ALP활성검측결과현시,3d시실험조ALP활성[(0.370 ±0.013) Sigma단위·min-1 ·mg-1]현저고우공백대조조[(0.2845±0.0084) Sigma단위·min-1·mg-1],차이유통계학의의(P＜0.01)；배양5、7d후천소홍염색결과현시실험조형성료명현적광화결절,실험조5、7d시개리자농도[분별위(0.539 ±0.007)、(1.617 ±0.042) μg/g]균현저고우공백대조조[(0.138 ±0.037) μg/g],P＜0.01.RT-PCR급단백질인적법검측결과현시:실험조SCAP배양3、7d후ALP、핵심결합단백인자2、성골상관전록인자、골개단백급아본질연린단백적기인급단백표체균현저고우공백대조조(P＜0.01,P＜0.05).결론 1.8 mmol/L KH2P04가이명현촉진인SCAP체외성아급성골능력.
Objective To determine the effects of KH2PO4 on the odonto-and osteogenic differentiation potential of human stem cells from apical papillae (SCAP) in vitro.Methods SCAP were isolated and cultured respectively in alpha minimum essential medium (α-MEM) or α-MEM containing 1.8 mmol/L KH2PO4.Alkaline phosphotase(ALP) activity,alizarin red staining,real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to examine the odonto and osteogenic potential of SCAP in the two media.Results SCAP cultured in α-MEM containing 1.8 mmol/L KH2PO4 exhibited a higher ALP activity [(0.370 ± 0.013) Sigma unit · min-1 · mg-1] at day 3 than control group [(0.285±0.008) Sigma unit · min-1 · mg-1] and KH2PO4-treated SCAP formed more calcified nodules at day 5 [(0.539 ± 0.007) μg/g] and day 7 [(1.617 ± 0.042) μg/g] than those in normal medium[(0.138 ±0.037) μg/g,P ＜0.01].The expression of odonto-and osteogenic markers were significantly up-regulated after the stimulation of KH2P04 at day 3 and 7 respectively,as compared with control group.Conclusions 1.8 mmol/L KH2PO4 can promote the odonto and osteogenic differentiation potential of human SCAP.
