Mouse dental papilla cells(mDPCs)are cranial neural crest-derived dental mesenchymal cells that give rise to dentin-secreting odontoblasts after the bell stage during odontogenesis.The odontoblastic differentiation of...Mouse dental papilla cells(mDPCs)are cranial neural crest-derived dental mesenchymal cells that give rise to dentin-secreting odontoblasts after the bell stage during odontogenesis.The odontoblastic differentiation of mDPCs is spatiotemporally regulated by transcription factors(TFs).Our previous work reveals that chromatin accessibility was correlated with the occupation of the basic leucine zipper TF family during odontoblastic differentiation.However,the detailed mechanism by which TFs regulate the initiation of odontoblastic differentiation remains elusive.Here,we report that phosphorylation of ATF2(p-ATF2)is particularly increased during odontoblastic differentiation in vivo and in vitro.ATAC-seq and p-ATF2 CUT&Tag experiments further demonstrate a high correlation between p-ATF2 localization and increased chromatin accessibility of regions near mineralization-related genes.Knockdown of Atf2 inhibits the odontoblastic differentiation of mDPCs,while overexpression of p-ATF2 promotes odontoblastic differentiation.ATAC-seq after overexpression of p-ATF2 reveals that p-ATF2 increases the chromatin accessibility of regions adjacent to genes associated with matrix mineralization.Furthermore,we find that p-ATF2 physically interacts with and promotes H2BK12 acetylation.Taken together,our findings reveal a mechanism that p-ATF2 promotes odontoblastic differentiation at initiation via remodeling chromatin accessibility and emphasize the role of the phosphoswitch model of TFs in cell fate transitions.展开更多
目的观察矿物三氧化物凝聚体(Mineral Trioxide Aggregate,MTA)和三种改良盖髓剂(nRoot、Vitapex、iRoot BP Plus)对人牙髓干细胞(hDPSCs)增殖、分化为成牙本质细胞的促进作用。方法①不同浓度四种盖髓剂对hDPSCs增殖的促进作用观察。取...目的观察矿物三氧化物凝聚体(Mineral Trioxide Aggregate,MTA)和三种改良盖髓剂(nRoot、Vitapex、iRoot BP Plus)对人牙髓干细胞(hDPSCs)增殖、分化为成牙本质细胞的促进作用。方法①不同浓度四种盖髓剂对hDPSCs增殖的促进作用观察。取第5代hDPSCs细胞分为MTA组、iRoot BP Plus组、nRoot组、Vitapex组及空白组,MTA组、iRoot BP Plus组、nRoot组、Vitapex组分别加入不同浓度(0.02、0.2、1、2 mg/mL)的MTA、iRoot BP Plus、nRoot和Vitapex培养液,空白组加入含10%FBS的DMEM/F12培养液。分别于培养24、48 h时,采用CCK-8法测算各组细胞增殖活性。②四种盖髓剂对hDPSCs分化为成牙本质细胞的促进作用观察。通过ALP活性检测筛选出四种材料对hDPSCs最适诱导浓度。取第4代hDPSCs分为五组:空白组、MTA组、iRoot BP Plus组、nRoot组和Vitapex组换为0.2 mg/mL的MTA、iRoot BP Plus、nRoot和Vitapex成骨培养基,空白组为正常成骨培养基。培养第21天时采用茜素红染色和半定量分析法观察各组细胞矿化结节形成情况,培养第7天时采用Western Blotting法检测细胞相关蛋白类核转录因子(Runt-related transcription factor 2,RUNX-2)、骨钙素(Osteocalcin,OCN)、牙本质涎磷蛋白(Dentin sialophosphoprotein,DSPP)以及牙本质基质蛋白1(Dentin matrix protein 1,DMP-1)。结果与空白组相比,培养第2天时0.02、0.2、1 mg/mL的MTA组、iRoot BP Plus在和nRoot组细胞增殖活性高(P均<0.05);与培养第1天时相比,培养第2天时2 mg/mL的MTA组和Vitapex组细胞增殖活性低(P均<0.05)。筛选0.2 mg/mL为后续受试浓度。与nRoot组和Vitapex组相比,iRoot BP Plus组和MTA组细胞钙沉积量高(P均<0.05)。与空白组相比,MTA组、iRoot BP Plus组、nRoot组和Vitapex组细胞OCN、RUNX-2、DSPP、DMP-1相对表达量均升高(P均<0.05);与nRoot组、Vitapex组相比,iRoot BP Plus组细胞OCN、RUNX-2、DSPP、DMP-1相对表达量均高(P均<0.05)。结论相比于MTA、Vitapex,nRoot和iRoot BP Plus更能促进hDPSCs的细胞增殖、诱导细胞分化为成牙本质细胞。展开更多
Aim: The oral cavity has the particularity to host multiple hard and soft tissues, in this paper, we will discuss the current therapies that lead to cell differentiation by regenerative therapies and the future altern...Aim: The oral cavity has the particularity to host multiple hard and soft tissues, in this paper, we will discuss the current therapies that lead to cell differentiation by regenerative therapies and the future alternatives proposed by medicinal plants and all the regenerative potential of these different tissues. Material and Methods: A detailed review of the literature through the various search engines: Scopus, PubMed, google scholar, Cochrane, etc., uses the selected keywords to explore the effect of the regenerative potential of several medicinal plants. Results: Through our research, we have proceeded to sort different medicinal plants, according to their repairing and regenerative potential on the different tissues of the oral cavity. Conclusion: Future studies are conceivable to explore the opportunities and potential provided by medicinal plants in the field of regenerative dentistry.展开更多
目的:探讨miR-20a-3p基因干扰对人牙髓干细胞(hDPSCs)成牙本质分化的影响及信号通路调控机制。方法:对hDPSCs转染miR-20a-3p-mimic和miR-20a-3p-inhibitor来上调或下调其表达,然后对hDPSCs进行成牙本质分化诱导培养。通过茜素红染色观...目的:探讨miR-20a-3p基因干扰对人牙髓干细胞(hDPSCs)成牙本质分化的影响及信号通路调控机制。方法:对hDPSCs转染miR-20a-3p-mimic和miR-20a-3p-inhibitor来上调或下调其表达,然后对hDPSCs进行成牙本质分化诱导培养。通过茜素红染色观察细胞钙基质的矿化程度并检测ALP活性。通过双荧光素酶报告基因测定实验验证miR-20a-3p与mothers against decapentaplegic homolog(SMAD)特异性E3泛素蛋白连接酶1(SMURF1)的靶向调节关系。通过RT-qPCR检测hDPSCs中miR-20a-3p以及SMURF1、Runt相关转录因子2(RUNX2)、骨钙蛋白(OCN)和牙本质涎磷蛋白(DSPP)mRNA水平。结果:与Control组相比,miR-20a-3p-mimic组的茜素红相对染色强度升高了67.09%,miR-20a-3p-inhibitor组降低了46.13%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的相对ALP活性升高了52.89%,miR-20a-3p-inhibitor组降低了53.32%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的RUNX2、OCN和DSPP mRNA相对表达量分别升高了2.19倍、1.86倍和2.35倍,miR-20a-3p-inhibitor组分别降低了63.26%、58.84%和68.12%(P<0.05)。双荧光素酶报告基因测定显示miR-20a-3p靶向抑制SMURF1(P<0.05)。过表达SMURF1抑制了miR-20a-3p对成牙本质分化的影响(P<0.05)。结论:miR-20a-3p通过靶向抑制SMURF1及其下游基因促进hDPSCs的成牙本质分化。展开更多
基金supported by the National Natural Science Foundation of China (No. 82071110 and No. 82230029) to Zhi Chenthe National Natural Science Foundation of China (No. 82071077 and No.82270948)+1 种基金“the Fundamental Research Funds for the Central Universities”“The Young Top-notch Talent Cultivation Program of Hubei Province” to Huan Liu
文摘Mouse dental papilla cells(mDPCs)are cranial neural crest-derived dental mesenchymal cells that give rise to dentin-secreting odontoblasts after the bell stage during odontogenesis.The odontoblastic differentiation of mDPCs is spatiotemporally regulated by transcription factors(TFs).Our previous work reveals that chromatin accessibility was correlated with the occupation of the basic leucine zipper TF family during odontoblastic differentiation.However,the detailed mechanism by which TFs regulate the initiation of odontoblastic differentiation remains elusive.Here,we report that phosphorylation of ATF2(p-ATF2)is particularly increased during odontoblastic differentiation in vivo and in vitro.ATAC-seq and p-ATF2 CUT&Tag experiments further demonstrate a high correlation between p-ATF2 localization and increased chromatin accessibility of regions near mineralization-related genes.Knockdown of Atf2 inhibits the odontoblastic differentiation of mDPCs,while overexpression of p-ATF2 promotes odontoblastic differentiation.ATAC-seq after overexpression of p-ATF2 reveals that p-ATF2 increases the chromatin accessibility of regions adjacent to genes associated with matrix mineralization.Furthermore,we find that p-ATF2 physically interacts with and promotes H2BK12 acetylation.Taken together,our findings reveal a mechanism that p-ATF2 promotes odontoblastic differentiation at initiation via remodeling chromatin accessibility and emphasize the role of the phosphoswitch model of TFs in cell fate transitions.
文摘目的观察矿物三氧化物凝聚体(Mineral Trioxide Aggregate,MTA)和三种改良盖髓剂(nRoot、Vitapex、iRoot BP Plus)对人牙髓干细胞(hDPSCs)增殖、分化为成牙本质细胞的促进作用。方法①不同浓度四种盖髓剂对hDPSCs增殖的促进作用观察。取第5代hDPSCs细胞分为MTA组、iRoot BP Plus组、nRoot组、Vitapex组及空白组,MTA组、iRoot BP Plus组、nRoot组、Vitapex组分别加入不同浓度(0.02、0.2、1、2 mg/mL)的MTA、iRoot BP Plus、nRoot和Vitapex培养液,空白组加入含10%FBS的DMEM/F12培养液。分别于培养24、48 h时,采用CCK-8法测算各组细胞增殖活性。②四种盖髓剂对hDPSCs分化为成牙本质细胞的促进作用观察。通过ALP活性检测筛选出四种材料对hDPSCs最适诱导浓度。取第4代hDPSCs分为五组:空白组、MTA组、iRoot BP Plus组、nRoot组和Vitapex组换为0.2 mg/mL的MTA、iRoot BP Plus、nRoot和Vitapex成骨培养基,空白组为正常成骨培养基。培养第21天时采用茜素红染色和半定量分析法观察各组细胞矿化结节形成情况,培养第7天时采用Western Blotting法检测细胞相关蛋白类核转录因子(Runt-related transcription factor 2,RUNX-2)、骨钙素(Osteocalcin,OCN)、牙本质涎磷蛋白(Dentin sialophosphoprotein,DSPP)以及牙本质基质蛋白1(Dentin matrix protein 1,DMP-1)。结果与空白组相比,培养第2天时0.02、0.2、1 mg/mL的MTA组、iRoot BP Plus在和nRoot组细胞增殖活性高(P均<0.05);与培养第1天时相比,培养第2天时2 mg/mL的MTA组和Vitapex组细胞增殖活性低(P均<0.05)。筛选0.2 mg/mL为后续受试浓度。与nRoot组和Vitapex组相比,iRoot BP Plus组和MTA组细胞钙沉积量高(P均<0.05)。与空白组相比,MTA组、iRoot BP Plus组、nRoot组和Vitapex组细胞OCN、RUNX-2、DSPP、DMP-1相对表达量均升高(P均<0.05);与nRoot组、Vitapex组相比,iRoot BP Plus组细胞OCN、RUNX-2、DSPP、DMP-1相对表达量均高(P均<0.05)。结论相比于MTA、Vitapex,nRoot和iRoot BP Plus更能促进hDPSCs的细胞增殖、诱导细胞分化为成牙本质细胞。
文摘Aim: The oral cavity has the particularity to host multiple hard and soft tissues, in this paper, we will discuss the current therapies that lead to cell differentiation by regenerative therapies and the future alternatives proposed by medicinal plants and all the regenerative potential of these different tissues. Material and Methods: A detailed review of the literature through the various search engines: Scopus, PubMed, google scholar, Cochrane, etc., uses the selected keywords to explore the effect of the regenerative potential of several medicinal plants. Results: Through our research, we have proceeded to sort different medicinal plants, according to their repairing and regenerative potential on the different tissues of the oral cavity. Conclusion: Future studies are conceivable to explore the opportunities and potential provided by medicinal plants in the field of regenerative dentistry.
文摘目的:探讨miR-20a-3p基因干扰对人牙髓干细胞(hDPSCs)成牙本质分化的影响及信号通路调控机制。方法:对hDPSCs转染miR-20a-3p-mimic和miR-20a-3p-inhibitor来上调或下调其表达,然后对hDPSCs进行成牙本质分化诱导培养。通过茜素红染色观察细胞钙基质的矿化程度并检测ALP活性。通过双荧光素酶报告基因测定实验验证miR-20a-3p与mothers against decapentaplegic homolog(SMAD)特异性E3泛素蛋白连接酶1(SMURF1)的靶向调节关系。通过RT-qPCR检测hDPSCs中miR-20a-3p以及SMURF1、Runt相关转录因子2(RUNX2)、骨钙蛋白(OCN)和牙本质涎磷蛋白(DSPP)mRNA水平。结果:与Control组相比,miR-20a-3p-mimic组的茜素红相对染色强度升高了67.09%,miR-20a-3p-inhibitor组降低了46.13%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的相对ALP活性升高了52.89%,miR-20a-3p-inhibitor组降低了53.32%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的RUNX2、OCN和DSPP mRNA相对表达量分别升高了2.19倍、1.86倍和2.35倍,miR-20a-3p-inhibitor组分别降低了63.26%、58.84%和68.12%(P<0.05)。双荧光素酶报告基因测定显示miR-20a-3p靶向抑制SMURF1(P<0.05)。过表达SMURF1抑制了miR-20a-3p对成牙本质分化的影响(P<0.05)。结论:miR-20a-3p通过靶向抑制SMURF1及其下游基因促进hDPSCs的成牙本质分化。
