Cellular response to Mineral Trioxide Aggregate

Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 2008

The objective of this study was to determine the effect of MTA on the expression of cytokines in mouse pulp tissue. Study design. Pulp tissue was exposed to MTA, and the expression of CCL5/RANTES, CCL2/MCP1, IL-1␣, IFN-␥, TNF-␣, IL-4, and IL-6 was evaluated by RT-PCR at 10 and 20 days after exposure. Control groups were not exposed to MTA. Results. We found no detectable expression of CCL2, IL-4, and IL-6 in the tissue from either group, while TNF-␣ was expressed at high levels 20 days after exposure (P Ͻ .05). CCL5 and IL-1␣ mRNA expression was lower in the MTAtreated group 10 days after treatment (P Ͻ .05). At 20 days after the surgical procedure, IFN-␥ mRNA expression was also lower in the MTA-treated group (P Ͻ .05). Conclusions. These findings suggest that MTA down-regulates the inflammatory cytokines CCL5, IL-1␣ and IFN-␥ and may have an anti-inflammatory effect.

Journal of Endodontics, 2012

Introduction: The purpose of this study was to compare the cytotoxicity and cytokine expression profiles of EndoSequence Root Repair Material (ERRM; Brasseler, Savannah, GA) putty, ERRM flowable, and ProRoot mineral trioxide aggregate (MTA; Dentsply Tulsa Dental, Johnson City, TN) using osteoblast cells (MG-63). Methods: Four millimeters in diameter of each material was placed in the center of a 6-well culture plate, and a 2-mL suspension (10 5 cells/mL) of human osteoblasts was seeded in each well. Photomicrograph images were used to evaluate cytotoxicity as evidenced by the lack of osteoblast cell growth in relation to the materials with AH-26 (Dentsply Tulsa Dental) as the positive control. In addition, reverse-transcriptase polymerase chain reaction (RT-PCR) was used to evaluate the expression of interleukin (IL)-1b, IL-6, IL-8, and tumor necrosis factor-a (TNF-a). Cytokine expression of MG-63 cells upon lipopolysaccharide treatment was used as controls. RT-PCR results were normalized by the expression of the housekeeping gene b-actin and were used to measure cytokine expression. Statistical analysis was performed using analysis of variance. Results: Results showed that ERRM putty and MTA exhibited minimal levels of cytotoxicity; however, ERRM was slightly more cytotoxic although not statistically significant. The expression of IL-1b, IL-6, and IL-8 was detected in all samples with minimal TNF-a expression. Conclusions: We concluded that ERRM and MTA showed similar cytotoxicity and cytokine expressions.

Journal of Endodontics, 2009

The purpose of this study was to investigate the effects of mineral trioxide aggregate (MTA) on survival, mineralization, and expression of mineralization-related genes of cementoblasts. Immortalized cementoblasts (OCCM) were maintained with Dulbecco modified Eagle medium containing 10% fetal bovine serum. Methyl-thiazol-diphenyltetrazolium experiments were performed at 24 and 72 hours to evaluate bioactive components released by MTA (0.002-20 mg/mL) on the cell survival of OCCM. Von Kossa staining was used to evaluate biomineralization of OCCM cells. Images of cementoblasts were taken on day 3 by using inverted microscopy. Gene transcripts for bone sialoprotein (BSP), OCN, collagen type I (COL I), and osteopontin (OPN) were evaluated on days 3 and 5 by using semiquantitative reverse transcriptase polymerase chain reaction. The 20 mg/mL concentration of MTA was toxic for OCCM cells, whereas other concentrations of MTA tested exhibited similar cell numbers when compared with control group, and the 0.02 mg/mL concentration of MTA increased OCCM cell survival at 72 hours. Although an apparent decrease in mineralization was observed in the highest 3 concentrations of MTA used, 0.02 and 0.002 mg/mL concentrations of MTA induced greater biomineralization of OCCM cells than seen in the control. Moreover, increased BSP and COL I mRNA expression was observed at 0.02 and 0.002 mg/mL concentrations of MTA. MTA did not have a negative effect on the viability and morphology of cementoblasts and induced biomineralization of cementoblasts at the concentrations of 0.02 and 0.002 mg/mL. Based on these results MTA can be considered as a favorable material regarding cell-material interaction. (J Endod 2009;35:513-519)

Bone Reports, 2015

Mineral trioxide aggregate (MTA) has been recommended for various uses in endodontics. To understand the effects of MTA on alveolar bone, we examined whether MTA induces osteoblastic differentiation using MC3T3-E1 cells. MTA enhanced mineralization concomitant with alkaline phosphatase activity in a dose-and timedependent manner. MTA increased production of collagens (Type I and Type III) and matrix metalloproteinases (MMP-9 and MMP-13), suggesting that MTA affects bone matrix remodeling. MTA also induced Bglap (osteocalcin) but not Bmp2 (bone morphogenetic protein-2) mRNA expression. We observed induction of Atf6 (activating transcription factor 6, an endoplasmic reticulum (ER) stress response transcription factor) mRNA expression and activation of Atf6 by MTA treatment. Forced expression of p50Atf6 (active form of Atf6) markedly enhanced Bglap mRNA expression. Chromatin immunoprecipitation assay was performed to investigate the increase in p50Atf6 binding to the Bglap promoter region by MTA treatment. Furthermore, knockdown of Atf6 gene expression by introduction of Tet-on Atf6 shRNA expression vector abrogated MTA-induced mineralization. These results suggest that MTA induces in vitro osteoblastogenesis through the Atf6-osteocalcin axis as ER stress signaling. Therefore, MTA in endodontic treatment may affect alveolar bone healing in the resorbed region caused by pulpal infection.

World Journal of Dentistry

hydroxide [Ca(OH) 2 ] and Ca silicate hydrate. 4 Additionally, bismuth oxide is included as a radiopacifier. 5 As the material sets, the Ca from Ca silicate precipitates, forming Ca(OH) 2. The dicalcium silicate and tricalcium silicate formed or the tricalcium aluminate IntroductIon The placement of a filling material in either an orthograde or retrograde manner necessitates the prevention of communication pathways between the root canal and adjacent tissues. This objective requires the use of a nontoxic, biocompatible, stable, and insoluble material that is noncarcinogenic and nongenotoxic in nature. 1 In restorative dentistry, a number of materials such as dental amalgam, interim restorative material, super-ethoxy benzoic acid, and glass ionomers have been employed. However, the aforementioned materials are subject to various limitations including corrosion susceptibility, electrolysis, staining and expansion upon contact with moisture, poor marginal adaptability leading to leakage, sensitivity to moisture, and adverse toxic effects on tissues. 2 Since its introduction in 1993 by Torabinejad et al. as a root-filling material, mineral trioxide aggregate (MTA) has become a valuable tool in operative dentistry and endodontics. MTA is commonly used for various procedures, including vital pulp therapy, root end closure, root perforation repairs, and surgical and regenerative procedures. MTA is a bioactive Ca silicate-based material that is placed in close proximity to tissues to elicit a local immune response and stimulate the repair of both hard and soft periradicular tissues. 3 MTA's properties of being osteoconductive, osteoinductive, and biocompatible contribute to its effectiveness. The MTA's patented composition includes CaO and silica. The fine hydrophilic particles of these components constitute 70-95% of the material. These components form tricalcium silicate, dicalcium silicate, tricalcium aluminate, and tetra-Ca aluminoferrite, which in the presence of moisture, form a colloidal gel made of calcium

Journal of Endodontics, 2010

Introduction: Recently, some studies have compared mineral trioxide aggregate (MTA) with Portland cements, concluding that the principal ingredients of Portland cements are similar to those of MTA. The purpose of the present study was to evaluate the effect of gray MTA, white MTA, and gray and white Portland cements on inflammatory cells in rats. Methods: Fresh mixtures mixed with distilled water were placed in polyethylene tubes, which were implanted in the dorsal subcutaneous connective tissue of 60 Sprague-Dawley rats along with empty tubes as controls. Tissue specimens were collected after the rats were sacrificed after 7, 15, 30, 60, and 90 days. The specimens were fixed, stained, processed, and histologically evaluated under a light microscope. Inflammatory reactions were classified as grade 0: without inflammatory cells, grade I: sporadic infiltration of inflammatory cells, grade II: moderate infiltration (<25 cells), grade III: dense and severe infiltration (25-125 cells), and grade IV: very dense and severe infiltration (>125 cells). Data were analyzed with the nonparametric (two factor) analysis of variance and Kruskal-Wallis H-test. Results: All the groups showed grade III inflammation after 7 and 15 days; there was a decrease in the inflammatory process after 30, 60, and 90 days. After 90 days, gray MTA, white MTA, and control groups had grade 0 inflammatory process, but gray Portland cement and white Portland cement groups showed grade 0 to grade I inflammatory processes. Conclusion: MTAs were more biocompatible; however, more studies are required.

Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics, 2003

Mineral trioxide aggregate (MTA) is being widely used for root-end fillings, pulp capping, perforation repairs, and other endodontic procedures. MTA and Portland cement (PC) have many similar physical, chemical, and biologic properties. PC cement has shown promising potential as an endodontic material in several studies in vitro and in vivo. The purpose of this study was to compare the cytotoxic effect in vitro and the tissue reaction of MTA and Portland cement in bone implantation in the mandibles of guinea pigs. Millipore culture plate inserts with freshly mixed or set material were placed into the culture plates with already attached L929 cells. After an incubation period of 3 days, the cell morphology and cell counts were studied. Adult male guinea pigs under strict asepsis were anesthetized, during which a submandibular incision was made to expose the symphysis of the mandible. Bilateral bone cavities were prepared and Teflon applicators with freshly mixed materials were insert...

Journal of Materials Science: Materials in Medicine, 2004

Some endodontic sealers have been shown to cause local and systemic effects, mainly due to microleakage of chemicals from the sealer. To avoid the risk of toxic effects in vivo, the biological compatibility of ®lling materials has to be assessed. In vitro compatibility of Proroot 2 MTA cement in comparison with two different ®llers used in clinical practice, was examined by testing the adherence, viability, proliferation and secretion of collagen of osteoblast-like cells. In our experimental system, Saos-2 cells challenged with Proroot 2 MTA for 24 and 72 h showed a better behaviour than the cells exposed to the other compounds under assay. We found that the cells attached to the rough surface of Proroot 2 MTA cement and spread onto the rough surface. Moreover, the cells on Proroot 2 MTA were viable, grew, and released some collagen even at 72 h, while cell metabolism and growth was dramatically reduced onto sEBA and amalgam surfaces. A parallel behaviour was found after the cells were challenged with extracts of the different ®llers. In conclusion, according to our in vitro study, Proroot 2 MTA showed a good interaction with bone-forming cells: such behaviour may partially account for its satisfying clinical performance.

Journal of Orthopaedic Research, 2006

The purpose of the current study was to evaluate the effects of alumina particles on secretion of several cytokines involved in bone resorption in cocultures of macrophages and osteoblasts. To distinguish the contribution of each individual cell type, we have established a heterologous in vitro system that makes use of mouse J774 cells and primary cultured human osteoblasts. J744 cells decreased the production of TNF-a when they were cocultured with osteoblasts. Treatment of J744 cells with alumina particles increased TNF-a secretion, but the induction was lower when cells were cocultured with osteoblasts. Secretion of IL-6 by J744 cells was very low, and increased in the presence of osteoblasts. Alumina particles were only able to stimulate the release of IL-6 by J744 cells when cells were cocultured with osteoblasts. On the other hand, incubation of osteoblasts with alumina particles enhanced the release of IL-6 and GM-CSF. Coculturing osteoblasts with J744 cells induced them to release IL-6 and GM-CSF, and treatment with alumina further increased the secretion of both mediators by osteoblasts. According to these in vitro results, it seems rather plausible that alumina particles are able to initiate an inflammatory response in vivo. ß

International Endodontic Journal, 2005

Rezende TMB, Vargas DL, Cardoso FP, Sobrinho APR, Vieira LQ. Effect of mineral trioxide aggregate on cytokine production by peritoneal macrophages. International Endodontic Journal, 38, 896-903, 2005. Aim To test the effect of two commercial brands of grey mineral trioxide aggregate (ProRoot Ò and MTA-Â ngelus Ò ) on cytokine production by M1 and M2 inflammatory macrophages. Methodology M1 (from C57BL/6 mice) and M2 peritoneal inflammatory macrophages (from C57BL/6 IL12p40 )/) mice) were obtained and cultured in vitro in the presence of MTA. The cellular viability and the production of tumour necrosis factor-a, interleukin (IL)-12 and IL-10 in response to stimulation with interferon-c and Fusobacterium nucleatum or Pepto-streptococcus anaerobius were evaluated. Data were analysed by Mann-Whitney, Kruskal-Wallis and anova tests. Results The cements did not interfere with cellular viability or with cytokine production by either type of macrophage. However, M2 macrophages produced higher levels of IL-10 when stimulated with F. nucleatum than M1 macrophages (P < 0.05).