人牙髓细胞对三氧化矿物凝聚体（MTA）和 MTA Plus 的反应：细胞毒性和基因表达分析。

Human dental pulp cells response to mineral trioxide aggregate (MTA) and MTA Plus: cytotoxicity and gene expression analysis.

机构信息

Department of Restorative Dentistry, Araraquara School of Dentistry, UNESP - Univ Estadual Paulista, Araraquara, São Paulo, Brazil.

Laboratory of Molecular Biology, Department of Genetic and Evolution, Federal University of São Carlos, São Carlos, São Paulo, Brazil.

出版信息

Int Endod J. 2017 Aug;50(8):780-789. doi: 10.1111/iej.12683. Epub 2016 Oct 3.

DOI:10.1111/iej.12683

PMID:27520288

Abstract

AIM

To investigate the cytotoxicity, osteogenic bioactivity and mRNA expression of osteogenic markers of bone morphogenetic protein 2 (BMP-2), osteocalcin (OC) and alkaline phosphatase (ALP) induced by the extracts of set MTA Plus (MTA P) (Avalon Biomed Inc. Bradenton, FL, USA) in comparison with MTA (Angelus, Londrina, PR, Brazil) on human dental pulp cells (hDPCs).

METHODOLOGY

Cell viability was assessed by mitochondrial dehydrogenase enzymatic (MTT) assay, and the mechanism of cell death was evaluated by flow cytometry. Bioactivity was evaluated by alkaline phosphatase (ALP) assay and detection of calcium deposits with alizarin red staining (ARS). The gene expression of BMP-2, OC and ALP was quantified with real-time PCR. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05).

RESULTS

MTA and MTA P were not cytotoxic and did not induce apoptosis. MTA P had significant higher ALP activity in relation to MTA and the control (P < 0.05). MTA had a significantly higher percentage of mineralized area than MTA P (P < 0.05). The expression of BMP2 and OC mRNA was significantly higher in cells exposed to MTA than MTA P after 1 day (P < 0.05). At day 3, the mRNA expression of ALP was significantly higher in MTA P compared with MTA (P < 0.05).

CONCLUSIONS

MTA and MTA Plus were noncytotoxic, increased mineralization processes in vitro and induced the expression of osteogenic markers.

摘要

目的

研究与普通矿化三氧化聚合物（MTA）相比，Set MTA Plus（MTA P）提取物对人牙髓细胞（hDPCs）的细胞毒性、成骨生物活性以及骨形态发生蛋白 2（BMP-2）、骨钙素（OC）和碱性磷酸酶（ALP）等成骨标志物的 mRNA 表达情况。

方法

通过线粒体脱氢酶酶促（MTT）测定法评估细胞活力，通过流式细胞术评估细胞死亡机制。通过碱性磷酸酶（ALP）测定法和茜素红染色（ARS）检测钙沉积来评估生物活性。采用实时 PCR 定量检测 BMP-2、OC 和 ALP 的基因表达。采用方差分析和 Bonferroni 或 Tukey 后检验（α=0.05）进行统计分析。

结果

MTA 和 MTA P 均无细胞毒性，也不会诱导细胞凋亡。与 MTA 和对照组相比，MTA P 具有更高的 ALP 活性（P<0.05）。MTA 具有比 MTA P 更高的矿化面积百分比（P<0.05）。与 MTA P 相比，暴露于 MTA 的细胞中 BMP2 和 OC mRNA 的表达在第 1 天显著更高（P<0.05）。在第 3 天，MTA P 中的 ALP mRNA 表达明显高于 MTA（P<0.05）。