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加速矿化三氧化物凝聚体对牙髓干细胞龛中牙本质细胞分化的影响。

The effect of accelerated mineral trioxide aggregate on odontoblastic differentiation in dental pulp stem cell niches.

机构信息

Department of Pediatric Dentistry, Faculty of Dentistry, Marmara University, Istanbul, Turkey.

Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Istanbul, Turkey.

出版信息

Int Endod J. 2018 Jul;51(7):758-766. doi: 10.1111/iej.12747. Epub 2017 Feb 22.

Abstract

AIM

To investigate the effect of accelerated-set mineral trioxide aggregate (MTA) on the proliferation and odontoblastic differentiation of human dental pulp cell niches (DPSC).

METHODOLOGY

ProRoot White MTA (WMTA; Dentsply Tulsa Dental, Johnson City, TN, USA) was mixed with various additives, which included distilled water, 2.5% disodium hydrogen phosphate (Na HPO ; Merck, Darmstadt, Germany) and 5% calcium chloride (CaCl ; Merck). DPSC niches extracted from third molars were cultured directly on MTA in the culture medium. Cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4- sulphophenyl)-2H-tetrazolium (MTS) assay. Cell growth and expression of odontoblastic differentiation markers (dentine sialophosphoprotein (DSPP) and collagen type 1 (COL1)) were determined using Real-Time Polymerase Chain Reaction analysis. Osteo-/odontogenic differentiation of DPSC niches was evaluated by measurement of alkaline phosphatase activity (ALP). Calcium deposition was assessed using von Kossa staining. The results were analysed statistically using Mann-Whitney tests and Kruskal-Wallis tests.

RESULTS

MTA mixed with 5% CaCl and 2.5% Na HPO exhibited optimal cell viability (P < 0.05) compared to MTA mixed with distilled water. MTA mixed with 5% CaCl and 2.5% Na HPO significantly increased ALP activity (P < 0.05), significantly promoted mineralization nodule formation (P < 0.05) and significantly enhanced the mRNA expression level of the osteogenic/odontogenic markers (P < 0.05; DSPP and COL1) compared with MTA mixed with distilled water.

CONCLUSIONS

MTA mixed with 5% CaCl and 2.5% Na HPO was biocompatible with dental pulp stem cell niches. Accelerated-set MTA promoted better differentiation in DPSC niches than conventional MTA. The accelerators could provide an alternative to MTA mixed with distilled water.

摘要

目的

研究加速凝固型矿化三氧化物凝聚体(MTA)对人牙髓细胞龛(DPSC)增殖和成牙本质分化的影响。

方法

ProRoot White MTA(WMTA;登士柏天津牙科有限公司,美国田纳西州约翰逊市)与不同添加剂混合,包括蒸馏水、2.5% 磷酸氢二钠(NaHPO4;默克公司,德国达姆施塔特)和 5% 氯化钙(CaCl2;默克公司)。从第三磨牙中提取的 DPSC 巢直接培养在培养基中的 MTA 上。通过 3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺苯基)-2H-四唑(MTS)法评估细胞活力。通过实时聚合酶链反应分析测定细胞生长和牙本质分化标志物(牙本质涎磷蛋白(DSPP)和胶原蛋白 1(COL1))的表达。通过碱性磷酸酶活性(ALP)测定评估 DPSC 巢的成骨/成牙本质分化。通过 von Kossa 染色评估钙沉积。使用曼-惠特尼检验和克鲁斯卡尔-沃利斯检验对结果进行统计学分析。

结果

与蒸馏水混合的 MTA 相比,与 5%CaCl2 和 2.5%NaHPO4 混合的 MTA 表现出最佳的细胞活力(P<0.05)。与蒸馏水混合的 MTA 相比,与 5%CaCl2 和 2.5%NaHPO4 混合的 MTA 显著增加了 ALP 活性(P<0.05),显著促进了矿化结节的形成(P<0.05),并显著增强了成骨/成牙本质标志物的 mRNA 表达水平(P<0.05;DSPP 和 COL1)。

结论

与蒸馏水混合的 MTA 与牙髓干细胞巢具有生物相容性。加速凝固型 MTA 促进了 DPSC 巢更好的分化,优于传统 MTA。加速剂可以为与蒸馏水混合的 MTA 提供替代方案。

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