Effects of calcium silicate endodontic cements on biocompatibility and mineralization-inducing potentials in human dental pulp cells.

The objective of this study was to evaluate the biocompatibility, inflammatory response, and odontoblastic potential of Biodentine (Septodont, Saint Maur des Fosses, France), Ortho-MTA (OMTA; BioMTA, Seoul, Korea), Angelus-MTA (AMTA; Angelus, Londrina, Brazil), and IRM (Dentsply Tulsa Dental, Tulsa, OK) in human dental pulp cells. The underlying signaling mechanisms were also investigated. Biocompatibilities were examined by the 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide assay. Differentiation was assessed by alkaline phosphatase activity, alizarin red S staining, and reverse-transcription polymerase chain reaction for marker genes. The levels of inflammatory mediators and cytokines were measured by reverse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Signal transduction analysis was performed by Western blotting. Biodentine, OMTA, and AMTA showed favorable cell proliferation, alkaline phosphatase activity, formation of mineralized nodules, and expression of odontoblastic marker genes that were similar to those of IRM. The levels of proinflammatory mediators including nitric oxide, prostaglandin E2, inducible nitric oxide synthase, and cyclooxygenase-2 were lower for Biodentine, OMTA, and AMTA compared with the IRM group. All test materials induced reactive oxygen species production and the expression of hemeoxygenase-1, nuclear factor-E2-related factor-2, and mitogen-activated protein kinases. These data indicate for the first time that the biocompatibility, inflammatory response, and odontoblastic differentiation of Biodentine were similar to that of OMTA and AMTA in HDPCs, which suggests that Biodentine could be good alternative pulp capping agent.

Chang SW ，Lee SY ，Ann HJ ，Kum KY ，Kim EC ... -  《-》

Effects of ProRoot MTA, Bioaggregate, and Micromega MTA on odontoblastic differentiation in human dental pulp cells.

The aim of this study was to compare the biocompatibility and odontogenic potential of newly developed Bioaggregate (BA) and Micromega MTA (MMTA) with ProRoot MTA (PMTA) and intermediate restorative material (IRM) by using human dental pulp cells. Biocompatibility was assessed by an 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide assay and scanning electron microscopy. Differentiation was evaluated by alkaline phosphatase (ALP) activity, alizarin red staining, and reverse transcriptase-polymerase chain reaction for the maker genes. The levels of inflammatory mediators and cytokines were measured by reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay. PMTA, BA, and MMTA exhibited equally good biocompatibility, whereas IRM showed cytotoxicity compared with these materials. PMTA, BA, and MMTA increased the ALP activity, promoted mineralization nodule formation, and enhanced the mRNA expression level of the osteogenic/odontogenic markers (ALP, osteopontin, osteocalcin, dentin sialophosphoprotein, and dentin matrix protein-1) compared with IRM. The levels of proinflammatory mediators and proinflammatory cytokines were lower in PMTA, BA, and MMTA compared with the IRM group. Collectively, the biocompatibility, odontogenic potentials, and inflammatory response of BA and MMTA are equal to those of PMTA and superior to those of IRM.

Chang SW ，Lee SY ，Kum KY ，Kim EC ... -  《-》

Odontogenic effect of a fast-setting pozzolan-based pulp capping material.

Mineral trioxide aggregate (MTA) is widely used as a pulp capping material. Recently, a MTA-derived fast-setting pozzolan cement (Endocem; Maruchi, Wonju, Korea) was introduced in the endodontic field. Our aim in this study was to investigate the odontogenic effects of this cement in vitro and in vivo. Human dental pulp cells (hDPCs) were cultured, and the effects of Endocem and a previously marketed MTA (ProRoot; Dentsply, Tulsa, OK) on biocompatibility were evaluated by assessing cell morphology and performing a cell viability test. Chemical composition of each material was analyzed by energy-dispersive X-ray spectroscopic analysis. Odontoblastic differentiation was analyzed by alkaline phosphatase activity and alizarin red S staining. The expression of odontogenic-related markers, namely dentin sialophosphoprotein, dentin matrix protein 1, and osteonectin, was evaluated by real-time polymerase chain reaction, Western blotting, and immunofluorescence analysis. Pinpoint pulp exposures were made on rat teeth and then capped with ProRoot or Endocem. After 4 weeks, reparative tertiary dentin formation and inflammatory responses were investigated histologically. The biocompatibility of Endocem was similar to that of ProRoot. Energy-dispersive X-ray spectroscopic analysis showed that ProRoot and Endocem contained similar elemental constituents such as calcium, oxygen, and silicon. Alkaline phosphatase activity and mineralized nodule formation increased in ProRoot- and Endocem-treated cells compared with medium only-treated cells in the control group (P < .05). The expression of odontogenic-related markers was significantly higher in the ProRoot- and Endocem-treated groups than the control group (P < .05), but there was no significant difference in the expression of these markers between the 2 experimental groups (P > .05). Four weeks after the pulp capping procedure, continuous tertiary dentin had formed directly underneath the capping materials and the pulp exposure area in all samples in the 2 treated groups. Furthermore, most specimens either had no inflammation or minor pulpal inflammation. Our results indicate that ProRoot and Endocem have similar biocompatibility and odontogenic effects. Therefore, Endocem is as effective a pulp capping material as ProRoot.

Park SJ ，Heo SM ，Hong SO ，Hwang YC ，Lee KW ，Min KS ... -  《-》

Effects of simvastain and enamel matrix derivative on Portland cement with bismuth oxide-induced growth and odontoblastic differentiation in human dental pulp cells.

We previously reported that bismuth oxide containing Portland cement (BPC) showed similar biocompatibility to Portland cement (PC) in periodontal ligament cells. However, the bioactivity of simvastatin and Emdogain (Biora AB, Malmö, Sweden) on BPC was not reported. The aim of this study was to evaluate the effects of simvastatin and Emdogain on BPC compared with mineral trioxide aggregate (MTA) in human dental pulp cells (HDPCs). Cell growth was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) assay. Differentiation was evaluated by alkaline phosphatase (ALP) activity, alizarin red staining, and reverse-transcriptase polymerase chain reaction. The cell growth of HDPCs exposed to Emdogain and simvastatin plus BPC was superior to those administered BPC alone and similar to those that received MTA for 14 days. The simvastatin and Emdogain groups increased the odontogenic potential of the BPC group with respect to ALP activity, mineralization nodules, messenger RNA expression of ALP, osteopontin, osteocalcin, Runx2, and osterix. These results suggest that simvastatin and Emdogain improved cell growth and the differentiation of the BPC group in HDPCs and may be useful ingredients in BPC as pulp-capping material.

Lee SY ，Min KS ，Choi GW ，Park JH ，Park SH ，Lee SI ，Kim EC ... -  《-》

Biodentine induces human dental pulp stem cell differentiation through mitogen-activated protein kinase and calcium-/calmodulin-dependent protein kinase II pathways.

Biodentine (Septodont, Saint-Maur-des-Fossès, France), a new tricalcium silicate cement formulation, has been introduced as a bioactive dentine substitute to be used in direct contact with pulp tissue. The aim of this study was to investigate the response of human dental pulp stem cells (hDPSCs) to the material and whether mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-κB), and calcium-/calmodulin-dependent protein kinase II (CaMKII) signal pathways played a regulatory role in Biodentine-induced odontoblast differentiation. hDPCs obtained from impacted third molars were incubated with Biodentine. Odontoblastic differentiation was evaluated by alkaline phosphatase activity, alizarin red staining, and quantitative real-time reverse-transcriptase polymerase chain reaction for the analysis of messenger RNA expression of the following differentiation gene markers: osteocalcin (OCN), dentin sialophosprotein (DSPP), dentin matrix protein 1 (DMP1), and bone sialoprotein (BSP). Cell cultures in the presence of Biodentine were exposed to specific inhibitors of MAPK (U0126, SB203580, and SP600125), NF-κB (pyrrolidine dithiocarbamate), and CaMKII (KN-93) pathways to evaluate the regulatory effect on the expression of these markers and mineralization assay. Biodentine significantly increased alkaline phosphatase activity and mineralized nodule formation and the expression of OCN, DSPP, DMP1, and BSP. The MAPK inhibitor for extracellular signal-regulated kinase 1/2 (U0126) and Jun N-terminal kinase (SP600125) significantly decreased the Biodentine-induced mineralized differentiation of hDPSCs and OCN, DSPP, DMP1, and BSP messenger RNA expression, whereas p38 MAPK inhibitors (SB203580) had no effect. The CaMKII inhibitor KN-93 significantly attenuated and the NF-κB inhibitor pyrrolidine dithiocarbamate further enhanced the up-regulation of Biodentine-induced gene expression and mineralization. Biodentine is a bioactive and biocompatible material capable of inducing odontoblast differentiation of hDPSCs. Our results indicate that this induction is regulated via MAPK and CaMKII pathways.

Luo Z ，Kohli MR ，Yu Q ，Kim S ，Qu T ，He WX ... -  《-》