不同浓度TheraCal LC调控人牙髓干细胞成骨分化机制研究  

Mechanism of TheraCal LC at Different Concentrations Regulating Osteogenic Differentiation of Human Dental Pulp Stem Cells

作　　者：陈秋艳 张韦 嵇步云 CHEN Qiu-yan;ZHANG Wei;JI Bu-yun(Department of Stomatology,The First Affiliated Hospital of Wannan Medical College(Yijishan Hospital),Wuhu,Anhui 241000,China)

机构地区：[1]皖南医学院第一附属医院(弋矶山医院)口腔内科,安徽芜湖241000

出　　处：《河北北方学院学报(自然科学版)》2025年第8期7-12,16,共7页Journal of Hebei North University:Natural Science Edition

基　　金：皖南医学院校中青年科研基金(No.WK2022F29)。

摘　　要：目的探讨不同浓度TheraCal LC调控人牙髓干细胞成骨分化的机制。方法收集因正畸需拔除的恒牙,培养与鉴定人牙髓干细胞,取第3代进行拍照,观察人牙髓干细胞形态,分为成脂组(加入油红O染色液1.5 mL)与矿化组(加入茜素红染色液1.5 mL),分析人牙髓干细胞成脂、成骨分化能力。将对数生长人牙髓干细胞加入浓度为50、100、150、200μg·mL^(-1)的TheraCal LC,继续培养48h后CCK-8法检测细胞增殖活性、Western Blot法检测增殖相关蛋白[细胞周期蛋白D1(CyclinD1)、p21]及成骨分化相关蛋白[骨钙素(OCN)、骨桥蛋白(OPN)、Runt相关转录因子2(RUNX2)]表达量,检测碱性磷酸酶(ALP)活性、矿化结节水平。结果贴壁的人牙髓干细胞大部分为纤维细胞长梭形,形态一致;茜素红染色镜下可见深红色钙结节;油红O染色镜下可见橘色脂滴,表明人牙髓干细胞具有成脂、成骨分化能力。与50μg·mL^(-1)组相比,100、150、200μg·mL^(-1)组增殖活性、CyclinD1蛋白表达量、ALP活性、矿化结节水平、OCN、OPN、RUNX2蛋白表达量升高,p21蛋白表达量降低(P<0.05);与100μg·mL^(-1)组相比,150、200μg·mL^(-1)组增殖活性、CyclinD1蛋白表达量、ALP活性、矿化结节水平、OCN、OPN、RUNX2蛋白表达量降低,p21蛋白表达量升高(P<0.05);与150μg·mL^(-1)组相比,200μg·mL^(-1)组增殖活性、CyclinD1蛋白表达量、ALP活性、矿化结节水平、OCN、OPN、RUNX2蛋白表达量降低,p21蛋白表达量升高(P<0.05)。结论100μg·mL^(-1)TheraCal LC干预可明显促进人牙髓干细胞增殖、成骨分化,其机制可能与ALP活性、增殖及成骨分化相关蛋白表达量得到调控有关。Objective To investigate the mechanism of TheraCal LC at different concentrations regulating osteogenic differentiation of human dental pulp stem cells.Methods The permanent teeth that needed to be removed for orthodontic were collected,cultured and identified,and the 3rd generation of human dental pulp stem cells were photographed to observe the morphology of human dental pulp stem cells,which were divided into lipogenic group(adding oil red O staining solution 1.5 mL)and mineralized group(adding alizarin red staining solution 1.5 mL),and the adipogenic and osteogenic differentiation ability of human dental pulp stem cells was analyzed.The logarithmically grown human dental pulp stem cells were added to TheraCal LC with concentrations of 50,100,150,200μg·mL^(-1).After continued culture for 48h,cell proliferation activity was detected by CCK-8 assay and Western assay The expression of proliferation-related proteins(cyclin D1(CyclinD1),p21)and osteogenic differentiation related proteins(osteocalcin(OCN),osteopontin(OPN),RUNt-related transcription factor 2(RUNX2)),ALP activity and the level of mineralized nodles were detected by Blot assay.Results Most of the adherent human dental pulp stem cells were long fusiform fibrocytes with the same morphology.Alizarin red staining showed deep red calcium nodules.Orange-colored lipid droplets can be seen under the oil-red O staining microscope,indicating that human pulp stem cells have the ability of adipogenic and osteogenic differentiation.Compared with 50μg·mL^(-1)group,the proliferative activity,CyclinD1 protein expression,ALP activity,mineralized nodule level,OCN,OPN and RUNX2 protein expression in 100μg·mL^(-1),150μg·mL^(-1)and 200μg·mL^(-1)groups were increased,while the p21 protein expression was decreased(P<0.05).Compared with 100μg/mL group,the proliferative activity,CyclinD1 protein expression,ALP activity,mineralized nodule level,OCN,OPN,RUNX2 protein expression in 150μg·mL^(-1)and 200μg·mL^(-1)groups were decreased,while the p21 protein expression

关 键 词：TheraCal LC 人牙髓干细胞 成骨分化 矿化结节 增殖活性 

分 类 号：R781.3[医药卫生—口腔医学]