Cytotoxicity of Different Concentrations of Three Root Canal Sealers on Human Mesenchymal Stem Cells.

This study assessed the dose-dependent effect on the cytotoxicity of BioRoot RCS (BR) and Endosequence BC (BC) sealers in human bone marrow mesenchymal stem cells (hMSCs) compared to those of the AH Plus sealer. Cells were exposed to different dilutions of extracts from freshly prepared sealers (1:2, 1:8, 1:32). Unexposed cells acted as the negative control. Cytotoxicity was evaluated by an alamar blue assay. Cell morphology was analyzed by using scanning electron microscopy after exposure to the different sealers' extracts. Statistical analysis was performed using a one-way analysis of variance and the Bonferroni post hoc test (p < 0.05). The cytotoxicities of BC and BR were less than that of AH Plus. In the presence of 1:2 BR, the cell proliferation was significantly lower than the control. At 1:8 and 1:32 concentrations, both the tricalcium silicate sealers led to similar cellular proliferation. Cells in BC and BR sealers' extracts spread better than those in AH Plus extract.

Cytotoxicity and proliferative effects of Iodoform-containing root canal-filling material on RAW 264.7 macrophage and RKO epithelial cell lines.

OBJECTIVE: The present study investigated the effect of the Iodoform-containing root canal filling material on the viability of cultured macrophages and epithelial cells, and on cytokine secretion. DESIGN: The effect of Endoflas F.S. on the proliferation of a RAW 264.7 macrophage cell line and on a RKO epithelial cell line, and on the production of tumour necrosis factor alpha (TNFα) from macrophages was examined. Cell vitality was evaluated using a colourimetric XTT (sodium 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)-carbonyl]-2H-tetrazolium inner salt) assay. The presence of cytokines was determined by two-site enzyme-linked immunosorbent assay (ELISA). RESULTS: Direct exposure of Endoflas F.S. and its media, up to a dilution of 1/8, decreased the viability of macrophages and epithelial cells by ∼70% compared to control media (P<0.05). Media dilution from 1/16 to 1/1024 demonstrated a proliferative effect, increasing cell viability by about 60% compared to media without Iodoform-containing root canal filling material. CONCLUSIONS: Direct and indirect exposure to high concentrations of iodoform-containing root canal filling material showed a cytotoxic effect on macrophages and epithelial cells, while low concentrations induced cell proliferation.

Rat subcutaneous tissue response to MTA Fillapex® and Portland cement

The aim of this study was to evaluate the response of rat subcutaneous tissue to MTA Fillapex® (Angelus), an experimental root canal filling material based on Portland cement and propylene glycol (PCPG), and a zinc oxide, eugenol and iodoform (ZOEI) paste. These materials were placed in polyethylene tubes and implanted into the dorsal connective tissue of Wistar rats for 7 and 15 days. The specimens were stained with hematoxylin and eosin, and evaluated regarding inflammatory reaction parameters by optical microscopy. The intensity of inflammatory response against the sealers was analyzed by two blinded and previously calibrated examiners for all experimental periods (kappa=0.96). The histological evaluation showed that all materials caused a moderate inflammatory reaction at 7 days, which subsided with time. A greater inflammatory reaction was observed at 7 days in the tubes filled with ZOEI paste. Tubes filled with MTA Fillapex presented some giant cells, macrophages and lymphocytes after 7 days. At 15 days, the presence of fibroblasts and collagen fibers was observed indicating normal tissue healing. The tubes filled with PCPG showed similar results to those observed in MTA Fillapex. At 15 days, the inflammatory reaction was almost absent at the tissue, with several collagen fibers indicating normal tissue healing. Data were analyzed by the nonparametric Kruskal-Wallis test (α=0.05). Statistically significant difference (p<0.05) was found only between PCPG at 15 days and ZOEI at 7 days groups. No significant differences were observed among the other groups/periods (p>0.05). MTA Fillapex and Portland cement added with propylene glycol had greater tissue compatibility than the PCPG paste.

O objetivo deste estudo foi avaliar a resposta do tecido subcutâneo de rato ao MTA Fillapex® (Angelus), a um cimento endodôntico experimental à base de cimento Portland e propilenoglicol, e à pasta de óxido de zinco e eugenol com iodofórmio. Estes materiais foram colocados em tubos de polietileno e implantados no tecido conjuntivo do dorso de ratos Wistar, por 7 e 15 dias. Os espécimes foram corados com hematoxilina e eosina e os parâmetros de reação inflamatória foram avaliados em microscópio óptico. A intensidade da resposta inflamatória provocada pelos cimentos foi analisada em todos os períodos por dois observadores previamente calibrados (kappa 0,96) e sem conhecimento dos grupos experimentais. O exame histológico mostrou que todos os materiais provocaram reação inflamatória moderada aos 7 dias que regrediu com o tempo. A maior resposta inflamatória do tecido foi observada aos 7 dias, nos tubos preenchidos com pasta de Óxido de Zinco e Eugenol com Iodofórmio. Os tubos com MTA Fillapex apresentaram algumas células gigantes, macrófagos e linfócitos após 7 dias. Aos 15 dias, a presença de fibroblastos e fibras de colágenas foi observada, indicando processo de cicatrização do tecido. Os tubos com o cimento Portland mostraram resultados semelhantes aos observados no grupo MTA Fillapex. Aos 15 dias, a reação inflamatória apresentada foi praticamente ausente, com muitas fibras colágenas, indicando cicatrização normal do tecido. A análise estatística mostrou diferença estatisticamente significante entre o grupo de cimento Portland (15 dias) e óxido de zinco eugenol com Iodofórmio (7 dias) (p<0,05). Nos outros grupos não houve diferença estatística significante. MTA Fillapex e cimento Portland são mais biocompatíveis do que os outros cimentos testados.

Cytotoxicity of newly developed ortho MTA root-end filling materials.

INTRODUCTION: Various materials have been advocated for use as root-end filling materials. The purpose of the present in vitro study was to compare the cytotoxicity of 4 root-end filling materials: glass ionomer cement (GIC; Fuji II, GC Corp, Tokyo, Japan), reinforced zinc oxide-eugenol cement (IRM; Dentsply Tulsa Dental, Tulsa, OK), and 2 types of mineral trioxide aggregate. METHODS: This study used MG-63 cells derived from a human osteosarcoma. To quantitatively evaluate the cytotoxicity of test materials, the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Each specimen was examined by scanning electron microscopy for the observation of cell morphology. RESULTS: The XTT assay showed that the cell viability of ProRoot MTA (Dentsply Tulsa Dental) was higher than that of GIC and Ortho MTA (BioMTA, Seoul, Republic of Korea) at all time points. IRM showed significantly lower cell viability than the other groups. The scanning electron microscopic analysis revealed that elongated, dense, and almost confluent cells were observed in the cultures of GIC, Ortho MTA, and ProRoot MTA specimens. In contrast, cells on the surface of IRM were rounded in shape, and the numbers and the density of the cells were smaller than that in the other groups. CONCLUSIONS: ProRoot MTA and GIC showed good biocompatibility in this study. However, Ortho MTA showed lower biocompatibility compared with ProRoot MTA and GIC.

Reaction of rat subcutaneous connective tissue to a mineral trioxide aggregate-based and a zinc oxide and eugenol sealer.

INTRODUCTION: The purpose of this study was to evaluate the subcutaneous connective tissue reaction in rats to a mineral trioxide aggregate (MTA)-based endodontic sealer Fillapex (Angelus, Londrina, PR, Brazil) and compare it with Grossman sealer (Farmadental, Buenos Aires, Argentina). METHODS: Sterile medical-grade silicone tubes containing the test materials were implanted in 24 Wistar rats. After 10, 30, and 90 days, the animals (n = 8 per period) were euthanized, and the implants along with their surrounding tissues were dissected, fixed, and processed for histologic evaluation. A 4-category evaluation system was used to evaluate the microscopic observations. The tissue response on the lateral walls of the silicone tubes was used as the negative control. The data were analyzed for statistical significance using the Wilcoxon signed rank, Kruskal-Wallis, and Dunn tests. RESULTS: Fillapex showed a severe tissue reaction for all 3 observation periods. Grossman sealer showed similar features after 10 and 30 days, but the reaction decreased slightly after 90 days. In contrast, the negative controls did not show adverse reactions in any sample of the 3 time periods. After 10 and 30 days, no statistically significant differences were found between Fillapex and Grossman sealer (P > .05); however, the difference was significant after 90 days (P < .05). For all experimental periods, there were statistically significant differences between both Fillapex and Grossman sealer and the negative controls (P < .05). CONCLUSIONS: It was concluded that both MTA-Fillapex and Grossman sealer remained toxic to subcutaneous tissues in rats after 90 days.

Cytotoxicity evaluation of Gutta Flow and Endo Sequence BC sealers.

OBJECTIVE: This study evaluated the cytotoxicity of GuttaFlow and EndoSequence BC sealers and compared them with AH Plus and Tubli-Seal sealers. STUDY DESIGN: Samples (0.5 mg) of freshly mixed or set BC, GuttaFlow, AH Plus, and Tubli-Seal sealers were eluted with 300, 600, and 1,000 µL cell culture medium for 24 and 72 hours. L929 cells were seeded into 96-well plates at 3 × 10(4) cells/well and cultured with 100 µL eluate from each eluate group. Cells cultured only with culture medium served as control. After 24 hours' incubation, the cytotoxicity was evaluated by MTT assay. Cell viability was calculated as the percentage of the control group, and the results were analyzed with a one-way analysis of variance. RESULTS: For the freshly mixed sealer, cell viability in the AH Plus group was less than in all of the other 3 sealer groups. The Tubli-Seal sealer group had less cell viability than the EndoSequence BC and GuttaFlow sealer groups. For the set sealer, the Tubli-Seal and AH Plus groups had less cell viability than the EndoSequence BC and GuttaFlow sealer groups. There was no cell viability difference between the EndoSequence BC and GuttaFlow sealer groups in the either freshly mixed or set sealer group. CONCLUSIONS: The GuttaFlow and EndoSequence BC sealers have lower cytotoxicity than the AH Plus and Tubli-Seal sealers.

Human periodontal ligament cell response to a newly developed calcium phosphate-based root canal sealer.

OBJECTIVES: The aim of this study was to investigate the cellular effects of newly developed calcium phosphate-based sealers (CAPSEAL I and II) using cultured human periodontal ligament cells (HPDLCs) compared with epoxy resin sealer (AH26; Dentsply, DeTrey, Konstanz, Germany), zinc oxide eugenol [ZOE] sealer (extended working time [EWT]; Kerr Corporation, Orange, CA), and CPC sealer (Sankin apatite sealer; Sankin-kogyo, Tokyo, Japan). METHODS: Cell viability by -(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazoliumbromide assay, cell attachment by scanning electron microscopy, osteoblastic differentiation and inflammatory mediators by reverse-transcriptase polymerase chain reaction, Western blotting, and alizarin red staining were evaluated. RESULTS: The cytotoxicities of CAPSEAL I and II were less than those of AH 26 and EWT after 1 and 14 days. Cells on CAPSEAL I and II were spread better as compared with those on other sealers. Mineralization after 14 days and the expression of osteoblastic differentiation markers such as alkaline phosphate and osteonectin messenger RNA increased in CAPSEAL I- and II-exposed HPDLCs after 1 and 3 days, whereas the production of inflammatory mediators, including cyclooxygenase-2, inducible nitric oxide synthetase, and reactive oxygen species (ROS), were lower than in other sealers. CONCLUSIONS: These results suggest that both CAPSEAL I and II show less cytotoxicity and inflammatory mediators compared with other sealers and have the potential to promote bone regeneration as root canal sealers.

Comparision of biocompatibility and cytotoxicity of two new root canal sealers.

The aim of this study was to investigate the remote organ toxicity and connective tissue reaction of two new root canal sealers ("GuttaFlow(®)" and "EndoREZ(®)") and to compare them with zinc oxide eugenol sealer using biochemical and histopathological parameters. A total of 60 white albino Wistar rats were used in the study. 0.1ml of GuttaFlow(®), EndoREZ(®) or Kerr Pulp Canal Sealer(®) were administered subcutaneously into the mid-dorsal thoracic region of rats (15 in each group). Control rats were given saline only. Rats were decapitated after 24h, on day 7 and on day 30 of the experiment and tissue samples from lung, liver, kidney and skin were removed for the determination of malondialdehyde (MDA) and glutathione (GSH) levels. In parallel, tissues were also examined histologically. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) levels, and creatinine and blood urea nitrogen (BUN), concentrations (BUN) were measured to assess liver and kidney functions, respectively. Tumor necrosis factor (TNF) and lactate dehydrogenase (LDH) were also assayed in serum samples. No statistical differences were found among the control and EndoREZ(®), GuttaFlow(®) and Kerr Pulp Canal sealers regarding tissue MDA, GSH levels or serum parameters (p>0.05) at all time points examined. Both of the new root canal sealers showed good compatibility and acceptable tissue toxicity.

Cytotoxicity evaluation of two root canal sealers and a commercial calcium hydroxide paste on THP1 cell line by Trypan Blue assay.

OBJECTIVE: The aim of this investigation was to evaluate the cytotoxicity of two brands of root canal sealers, epoxy-resin based and zinc oxide-eugenol based, and one commercial calcium hydroxide paste on a monocyte cell line THP-1. MATERIAL AND METHODS: Undiluted (crude extract) and diluted extracts to 10%, 1%, 0.1%, 0.01%, 0.001% and 0.0001% of the sealers were tested for cytotoxicity to THP-1 cells using the trypan blue assay. Extracts were obtained according to ISO standard. Data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests at 5% significance level. RESULTS: Crude extract of AH Plus and Fill Canal killed approximately 90% of THP-1 cells versus 36% of THP-1 cells killed by L&C crude extract (p<0.05). Ten-fold dilutions of L&C, Fill Canal and AH Plus killed 24, 35 and 61% of THP-1 cells (p<0.05), respectively. Dilutions lesser than 1% caused minimal cell death as compared to the control groups (p>0.05), except for L&C 1% extract. CONCLUSIONS: The results revealed that the L&C paste crude extract was less cytotoxic to THP-1 cells than AH Plus or Fill Canal crude extracts.

Cytotoxicity evaluation of two root canal sealers and a commercial calcium hydroxide paste on THP1 cell line by Trypan Blue assay

OBJECTIVE: The aim of this investigation was to evaluate the cytotoxicity of two brands of root canal sealers, epoxy-resin based and zinc oxide-eugenol based, and one commercial calcium hydroxide paste on a monocyte cell line THP-1. MATERIAL AND METHODS: Undiluted (crude extract) and diluted extracts to 10 percent, 1 percent, 0.1 percent, 0.01 percent, 0.001 percent and 0.0001 percent of the sealers were tested for cytotoxicity to THP-1 cells using the trypan blue assay. Extracts were obtained according to ISO standard. Data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests at 5 percent significance level. RESULTS: Crude extract of AH Plus and Fill Canal killed approximately 90 percent of THP-1 cells versus 36 percent of THP-1 cells killed by L&C crude extract (p<0.05). Ten-fold dilutions of L&C, Fill Canal and AH Plus killed 24, 35 and 61 percent of THP-1 cells (p<0.05), respectively. Dilutions lesser than 1 percent caused minimal cell death as compared to the control groups (p>0.05), except for L&C 1 percent extract. CONCLUSIONS: The results revealed that the L&C paste crude extract was less cytotoxic to THP-1 cells than AH Plus or Fill Canal crude extracts.

Cytotoxicity evaluation of Activ GP and Resilon sealers in vitro.

OBJECTIVE: This study is to evaluate the cytotoxicity of Activ GP and RealSeal sealers in a cell culture system in vitro, and to compare them with traditional AH 26 and Kerr sealers. STUDY DESIGN: Samples of 0.5 mg freshly mixed or set RealSeal, Activ GP, AH 26, and Kerr sealers were eluted with 200, 400, 800, and 1,200 microL cell culture medium for 24 and 72 hours. L929 cells were seeded into 96-well plates at 3 x 10(4) cells/well and cultured with 100 microL eluate from each eluate group. Cells cultured with culture medium only served as a control. After 24 hours' incubation the cytotoxicity was evaluated by MTT assay. Cell viability was calculated as the percentage of the control group, and the results were analyzed with 1-way analysis of variance. RESULTS: For the freshly mixed sealer, cell viability in the AH 26 group was less than in all of the other 3 sealer groups. The Kerr sealer group had greater cell viability than RealSeal and Activ GP groups. For the set sealer, cell viability in the AH 26 group was greater than in all of the other 3 groups. Cell viability in the RealSeal group was less than in the Kerr and Activ GP groups. CONCLUSION: Freshly mixed RealSeal and Activ GP sealers have lower cytotoxicity than AH 26 sealer and more cytotoxicity than Kerr sealer. When sealers are set, RealSeal sealer has more cytotoxicity than AH 26 and Kerr sealer. Activ GP sealer has more cytotoxicity than AH 26 and is similar to Kerr sealer.

Up-regulation of receptor activator nuclear factor-kappaB ligand expression by root canal sealers in human osteoblastic cells.

Histologic investigations have demonstrated that root canal sealers can induce mild to severe inflammatory alternations. A recently identified tumor necrosis factor family molecule, receptor activator of nuclear factor-kappaB ligand (RANKL), plays a critical role in the development of osteoclasts that result in bone resorption. The aim of this study was to investigate the effects of root canal sealers AH26, Canals, and N2 on the expression of RANKL in human osteoblast cell line U2OS cells. Freshly mixed materials were filled in glass rings (4 mm height and 10 mm in diameter) and eluted in 10 mL of culture medium for 1 day. Subsequently, various dilutions (final dilution: 1/2, 1/4, and 1/8) of these extraction media were prepared for this study. Cytotoxicity was measured by the propidium iodide fluorescence assay. The expression of RANKL was measured by reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay. The results showed that AH26, Canals, and N2 were cytotoxic to U2OS cells in a concentration-dependent manner (P < .05). The exposure of U2OS cells to root canal sealers resulted in the up-regulation of RANKL mRNA and protein expression (P < .05). The expression of RANKL was up-regulated by root canal sealers in the following order: N2 > AH26 > Canals. Taken together, the activation of RANKL might play an important role in the pathogenesis of periapical bone destruction induced by root canal sealers.

Analysis of tissue reactions to methacrylate resin-based, epoxy resin-based, and zinc oxide-eugenol endodontic sealers.

The purpose of this study was to investigate the reaction of the subcutaneous connective tissue of rats to methacrylate resin-based sealer (EndoREZ), epoxy resin-based sealer (AH Plus), and zinc oxide-eugenol sealer (EndoFill). Polyethylene tubes containing the test materials were implanted in 18 rats. After 7, 30, and 60 days, tissues were collected for biopsy and fixed and processed for histologic evaluation. Observations were made of the cellular inflammatory component, the fibrous condensation, and the abscess formation. Comparisons between groups and times were made with the Friedman and Kruskall-Wallis tests. EndoREZ and EndoFill sealers showed a more intense and longer-lasting inflammation. With AH Plus, the inflammatory reaction showed a tendency to diminish over time. The only group to show a statistically significant reduction in inflammation during the 60-day period was the control group. None of the materials tested proved to have ideal characteristics for biocompatibility.

In vitro cytotoxicity evaluation of a self-adhesive, methacrylate resin-based root canal sealer.

This study compared the cytotoxicity of MetaSEAL (Parkell Inc, Farmington, NY), a methacrylate resin-based sealer with an epoxy resin-based (AH Plus Jet; Dentsply Caulk, Milford, DE) and a zinc oxide-eugenol-based sealer (Pulp Canal Sealer; SybronEndo, Orange, CA). Five-millimeter diameter disks prepared from the respective sealer and disks prepared from Teflon (negative control) and polymethyl methacrylate (positive control) were placed in direct contact with a rat osteosarcoma (ROS) 17/2.8 rat osteoblast-like cell line at six intervals after setting completely at 72 hours and for 5 succeeding weeks after the disks were immersed in simulated body fluid. Succinate dehydrogenase activity was evaluated by using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. All sealers exhibited severe toxicity at 72 hours, after which toxicity decreased gradually over the experimental period except for Pulp Canal Sealer, which remained severely toxic. MetaSEAL was more toxic than AH Plus Jet during the first week. Both were similar to the toxicity profile of the positive control after the first week, which was probably diffusion controlled.

Cytotoxicity evaluation of four endodontic sealers

This study evaluated in vitro the cytotoxicity of four root canal sealers (Topseal, EndoRez, TubliSeal and Kerr Pulp Canal Sealer E.W.T.) and their effects on reactive oxygen/nitrogen intermediate induction by mouse peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 mL 10 mM phosphate-buffered saline, washed twice and resuspended (106 cells/mL) in appropriate medium for each test. Cytotoxicity was determined by the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. Sealer suspensions were obtained in two different concentrations from each material: 18 mg/mL and 9 mg/mL, established according to compatibility parameters following MTT assay. Comparing the sealers, H2O2 release at concentrations of 9 mg/mL and 18 mg/mL was similar: Topseal > positive control (medium + cells + 5 mg/mL zimozan solution) > EndoRez > TubliSeal > Kerr Pulp E.W.T. > negative control (medium + cells). NO release at concentration of 9 mg/mL was: positive control (medium + cells + 10 µg/mL LPS solution) > Topseal > Kerr Pulp E.W.T. > TubliSeal = EndoRez > negative control (medium + cells); at concentration of 18 mg/mL was: positive control > Topseal > Kerr Pulp E.W.T > TubliSeal > EndoRez > negative control. Based on the results, it may be concluded that Topseal presented the highest cytotoxicity among the tested sealers, releasing higher concentrations of NO and H2O2 in macrophage culture.

Este estudo avaliou in vitro a citotoxicidade de quatro cimentos obturadores (Topseal, EndoRez, TubliSeal e Kerr Pulp Canal Sealer E.W.T) e seus efeitos na liberação de reativos intermediários do oxigênio e do nitrogênio em cultura de macrófagos peritoniais de ratos.Tioglicolato foi utlizado para se obter células peritoneias de camundongos. A cavidade peritoneal foi irrigada com 5 mL de solução salina 10 mM. As células foram lavadas duas vezes e foi feita uma suspensão (106 células/mL) em meio apropriado para cada um dos testes. A citotoxicidade dos cimentos foi determinada pela presença de peróxido de hidrogênio (H2O2) e óxido nítrico (NO) pela oxidação peroxidase-dependente do vermelho fenol e pela reação de Griess, respectivamente. Suspensões de cimento foram obtidas em duas diferentes concentrações para cada material: 18 mg/mL e 9 mg/mL, estabelecidas previamente pelo teste de viabilidade celular MTT. Comparando os cimentos, a liberação de H2O2 foi similar nas duas concentrações: Topseal > controle positivo (meio + células + Zimozan a 5mg/mL ) > EndoRez > TubliSeal > Kerr Pulp E.W.T. > controle negativo (meio + células). A liberação de NO na concentração de 9 mg/mL foi: de 9 mg/mL foi: controle positivo (meio + células + solução de LPS a 10 »g/mL) > Topseal > Kerr Pulp E.W.T. > TubliSeal = EndoRez > controle negativo (meio + células); e na concentração de 18 mg/mL; e na concentração de 18 mg/mL: controle positivo > Topseal > Kerr Pulp E.W.T > TubliSeal > EndoRez > controle negativo. Baseado nos resultados, pode-se concluir que o Topseal apresentou a maior citotoxicidade dentre os cimentos avaliados, liberando as mais altas concentrações de NO e H2O2 em cultura de macrófagos.

Behaviour of bone marrow osteoblast-like cells on mineral trioxide aggregate: morphology and expression of type I collagen and bone-related protein mRNAs.

AIM: To investigate the in vitro behaviour of rat bone marrow cells (RBM) on mineral trioxide aggregate (MTA) (ProRoot, MTA Root Canal Repair Material; Dentsply Tulsa, Tulsa, OK, USA) compared with intermediate restorative materials (IRM) (Dentsply Caulk, Milford, DE, USA). METHODOLOGY: RBM were obtained from rat femur and were primary cultured and then subcultured. Cells were then seeded on three dishes of each material, and cultured for 3 days, after which they were evaluated morphologically using scanning (SEM) and transmission (TEM) electron microscopy. Furthermore, the calcium released from hydrated material, the cell proliferation ratio and alkaline phosphatase (ALP) activity were analysed, and the expression of type I collagen and bone-related protein mRNAs were evaluated. The data were averaged and analysed via one-way analysis of variance (anova) and were then compared by the Scheffe's test. RESULTS: SEM showed that RBM attached to MTA and had a flattened appearance without nuclear protrusions and microspikes. TEM showed that the cells attached in the same manner as the control group, but gaps larger than 2 microm were frequently seen. The calcium released from hydrated MTA was about 130 ppm after 3 days of immersion in saline. The ALP activity was similar to the control group. Cell proliferation and expression of type I collagen mRNA was significantly lower, while the expression of osteopontin mRNA was significantly higher than the control group at the third day of culture. In IRM groups, a few rounded cells were observed on the material but no living cells were seen. CONCLUSIONS: MTA is a material of low toxicity which does not inhibit cell growth, but does suppress the differentiation of osteoblast-like cells.

Citotoxidade de três cimentos obturadores do sistema de canais radiculares sobre cultura de células L929 / Citotoxicity of three filling cements of radicular canals system over L929 culture cells

O objetivo desse estudo in vitro foi avaliar a citotoxicidade dos cimentos Intrafill e Pulp Canal Sealer, ambos à base de óxido de zinco eugenol e um outro cimento, o Sealapex, à base de hidróxido de cálcio. Esse experimento foi realizado usando linhagem de células L 929 (de fibroblastos de camundongos). A citotoxicidade foi avaliada in vitro, usando-se o método de difusão em Agar, depois de 48 horas do endurecimento do cimento, o corante utilizado foi o vermelho neutro. Os resultados mostraram diferenças nas médias dos valores da viabilidade celular pelo teste ANOVA. Após o período determinado pela amostra, o Intrafill, e o Sealapex apresentaram alterações morfológicas celulares com maior citotoxicidade quando comparados ao Pulp Canal Sealer.

A comparative histological evaluation of the biocompatibility of materials used in apical surgery.

AIM: To evaluate the biological properties of a variety of materials that could be used in apical surgery. METHODOLOGY: The intraosseous implant technique recommended by the FDI (1980) and ADA (1982) was used to test the following materials: zinc oxide-eugenol (ZOE), mineral trioxide aggregate (MTA), and Z-100 light-cured composite resin. Thirty guinea-pigs, 10 for each material, divided into experimental periods of 4 and 12 weeks, received one implant on each side of the lower jaw symphysis. The connective tissue response alongside the lateral wall outside the cup served as a negative control for the technique. At the end of the observation periods, the animals were killed and the specimens prepared for routine histological examination to evaluate their biocompatibility. RESULTS: The reaction of the tissue to the materials diminished with time. The ZOE cement was highly toxic during the 4-week experimental period, but this profile changed significantly after 12 weeks, when it showed biocompatible characteristics. MTA and Z-100 showed biocompatibility in this test model at both time periods. CONCLUSIONS: MTA and Z-100 composite were biocompatible at 4 and 12 weeks in this experimental model.

Biocompatibility of an experimental glass-ionomer cement sealer in rat mandibular bone.

OBJECTIVE: The purpose of this study was to compare an experimental glass-ionomer cement sealer, KT-308, with a conventional zinc oxide-eugenol sealer, Canals, in terms of tissue compatibility and solubility. STUDY DESIGN: Tissue reactions were examined under light and electron microscopes at 3 and 20 days after the implantation of either freshly mixed KT-308 sealer or Canals sealer into prepared cavities in rat mandibles. RESULTS: At 3 days after implantation, no inflammatory reaction was seen around KT-308 sealer, which was in direct contact with the bone surface. In contrast, Canals sealer elicited an initially severe inflammation in the surrounding tissue. At 20 days, the majority of KT-308 sealer remained in the bone cavity. Canals sealer was largely absorbed and surrounded by fibrous tissue with many macrophages. An ultrastructural examination also revealed that no intervening tissue was present between the cut bone surface and the glass-ionomer cement sealer and that disintegrated zinc oxide-eugenol particles were engulfed by macrophages. CONCLUSIONS: KT-308 sealer possesses better tissue compatibility and lower solubility compared with a conventional zinc oxide-eugenol sealer, suggesting its potential for use as a root canal sealant.

Tissue reaction initiated by different sealers.

AIMS: To analyse the type and degree of inflammatory reaction initiated by four sealers. METHODOLOGY: Twenty-four root canals of Macaca mulatta monkeys were filled within the canal and eight were overfilled with AH26, Apexit, Endomethasone or Grossman's sealers. The result of the treatment was evaluated after 6 months by histological assessment of the periapical tissues. RESULTS: In the group of root canals filled within the root, no inflammatory reaction was detected in specimens of Apexit and Grossman's sealers, but the other two sealers initiated different degrees of lymphocytic/plasmocytic tissue reactions. Endomethasone initiated a mild lymphocytic/plasmocytic reaction in three of the nine cases and AH26 caused mild lymphocytic/plasmocytic infiltration in two of the seven cases. In the group of overfilled root canals, all four sealers initiated inflammatory reactions. The periapical tissue reactions of overfilled root canals were similar to reactions detected in cases filled within the canal. However, additional histological features developed in specimens of Endomethasone and AH26: Endomethasone initiated a foreign body-type granulomatous reaction around the sealer particles and AH26 particles were engulfed by macrophages. The overfilled root canals of Apexit and Grossman's sealers initiated only lymphocytic/plasmocytic reactions. CONCLUSIONS: This study suggests that sealers with different chemical compositions initiate different histological reactions. It also emphasizes the importance of confining root filling to the canal system because all sealers initiate inflammatory reactions when they are present in the apical tissues