Biological effects of acid-eroded MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells.

OBJECTIVE: The aim of this study was to analyze the biological effects of MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells (hPDLSCs) after exposure to acidic and neutral environments. MATERIALS AND METHODS: Discs of each material (n = 30) were exposed to phosphate buffered saline (pH = 7.4) or butyric acid (pH = 5.2) for 7 days, and biological testing was carried out in vitro on hPDLSCs. Cell viability and apoptosis assays were performed using eluates of each root-end filling material. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the root-end filling materials was determined by energy-dispersive x-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by ANOVA and Tukey test (p < 0.05). RESULTS: Under an acidic environment, both materials displayed similar ion release abilities, with the increased release of Si and Ca ions. Substantial changes in microstructure were observed for both materials after exposure to acidic pH. In addition, material exposure to an acidic environment showed a similar degree of cell adherence, and, surprisingly, MTA Repair HP exhibited higher cell viability rates at pH 5.2 than ProRoot MTA. CONCLUSIONS: Exposure to an acidic environment promoted Si and Ca ion release from ProRoot MTA and MTA Repair HP. Moreover, we observed optimal biological properties of ProRoot MTA and MTA Repair HP in terms of cell viability, cell death, and cell attachment in both environments. CLINICAL RELEVANCE: These results may suggest that MTA Repair HP and ProRoot exhibited optimal biological properties in terms of cell viability, cell death and cell attachment in acidic environment, being considered as materials for root-end filling and perforations.

Physicochemical, cytotoxicity and in vivo biocompatibility of a high-plasticity calcium-silicate based material.

The purpose of this work was to evaluate the physicochemical properties, the cytotoxicity and in vivo biocompatibility of MTA Repair HP (MTA HP) and White MTA (WMTA). The setting time, flow, radiopacity and water solubility were assessed. To the cytotoxicity assay, primary human osteoblast cells were exposed to several dilutions of both materials eluates. MTT assay, apoptosis assay and cell adhesion assay were performed. The in vivo biocompatibility was evaluated through histological analysis using different staining techniques. No differences were observed between MTA HP and WMTA for setting time, radiopacity, solubility and water absorption (P > 0.05). However, MTA HP showed a significantly higher flow when compared to WMTA (P < 0.05). Cell viability results revealed that the extracts of WMTA and MTA HP promoted the viability of osteoblasts. After incubation of cells with the endodontic cement extracts, the percentage of apoptotic or necrotic cells was very low (<3%). Furthermore, SEM results showed a high degree of cell proliferation and adhesion on both groups. MTA HP showed similar in vivo biocompatibility to the WMTA and the control group in all time-points. The MTA HP presented adequate physicochemical and biological properties with improved flow ability when compared to WMTA. Such improved flow ability may be a result of the addition of a plasticizing agent and should be related to an improvement in the handling of MTA HP.

Biocompatibility of New Pulp-capping Materials NeoMTA Plus, MTA Repair HP, and Biodentine on Human Dental Pulp Stem Cells.

INTRODUCTION: The aim of the present study was to evaluate the in vitro cytotoxicity of MTA Repair HP, NeoMTA Plus, and Biodentine, new bioactive materials used for dental pulp capping, on human dental pulp stem cells (hDPSCs). METHODS: Biological testing was carried out in vitro on hDPSCs. Cell viability and cell migration assays were performed using eluates of each capping material. To evaluate cell morphology and cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the pulp-capping materials was determined by energy-dispersive X-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by analysis of variance and Tukey test (P < .05). RESULTS: Cell viability was moderate after 24 and 48 hours in the presence of MTA Repair HP and NeoMTA Plus, whereas at 48 and 72 hours, Biodentine showed higher rates of cell viability than MTA Repair HP and NeoMTA Plus (P < .001). A cell migration assay revealed adequate cell migration rates for MTA Repair HP and NeoMTA Plus, both similar to the control group rates, meanwhile the highest cell migration rate was observed in the presence of Biodentine (P < .001). Scanning electron microscope studies showed a high degree of cell proliferation and adhesion on Biodentine disks but moderate rates on MTA Repair HP and NeoMTA Plus disks. Energy-dispersive X-ray pointed to similar weight percentages of C, O, and Ca in all 3 materials, whereas other elements such as Al, Si, and S were also found. CONCLUSIONS: The new pulp-capping materials MTA Repair HP, NeoMTA Plus, and Biodentine showed a suitable degree of cytocompatibility with hDPSCs, and good cell migration rates, although Biodentine showed higher rates of proliferation time-dependent.

Thermo-setting glass ionomer cements promote variable biological responses of human dental pulp stem cells.

OBJECTIVE: To evaluate the in vitro cytotoxicity of Equia Forte (GC, Tokyo, Japan) and Ionostar Molar (Voco, Cuxhaven, Germany) on human dental pulp stem cells (hDPSCs). METHODS: hDPSCs isolated from third molars were exposed to several dilutions of Equia Forte and Ionostar Molar eluates (1/1, 1/2 and 1/4). These eluates were obtained by storing material samples in respective cell culture medium for 24h (n=40). hDPSCs in basal growth culture medium were the control. Cell viability and cell migration assays were performed using the MTT and wound-healing assays, respectively. Also, induction of apoptosis and changes in cell phenotype were evaluated by flow cytometry. Changes in cell morphology were analysed by immunocytofluorescence staining. To evaluate cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy (SEM). The chemical composition of the materials was determined by energy dispersive X-ray (EDX) and eluates were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS). Statistical analysis was performed with analysis of variance (ANOVA) and Student's t-test (α<0.05). RESULTS: Undiluted Equia Forte extracts led to a similar cell proliferation rates than the control group from 72h onwards. There were no significance differences between Equia Forte and Ionostar Molar in terms of cell apoptosis and phenotype. However, in presence of Equia extracts the migration capacity of hDPSCs was higher than in presence of Ionostar Molar (p<0.05). Also, SEM studies showed a higher degree of cell attachment when Equia Forte extracts were used. Finally, EDX analysis pointed to different weight percentages of C, O and Ca ions in glass ionomer cements, while other elements such as La, Al, Si, W, Mo and F were also detected. SIGNIFICANCE: In summary, Equia Forte promoted better biological responses in hDPSCs than Ionostar Molar.

Cytotoxicity of GuttaFlow Bioseal, GuttaFlow2, MTA Fillapex, and AH Plus on Human Periodontal Ligament Stem Cells.

INTRODUCTION: The aim of the present study was to evaluate the in vitro cytotoxicity of endodontic sealers (GuttaFlow Bioseal, GuttaFlow2, and MTA Fillapex) on human periodontal ligament stem cells (hPDLSCs). As a reference, AH Plus was compared with the more recent endodontic sealers regarding cell viability and cell attachment. METHODS: Biological testing was carried out in vitro on hPDLSCs. Cell viability assay was performed by using eluates from each endodontic sealer. To assess cell morphology and attachment to the different sealers, the hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. Chemical composition of the sealers was determined by energy-dispersive x-ray, and eluates were analyzed by inductively coupled plasma mass spectrometry. Statistical differences were assessed by analysis of variance and Tukey test (P < .05). RESULTS: Cell viability was evident after 24 hours in the presence of GuttaFlow Bioseal and GuttaFlow 2 but not in the case of AH Plus or MTA Fillapex. At 168 hours, GuttaFlow Bioseal and GuttaFlow 2 exhibited high and moderate cell viability, respectively, whereas AH Plus and MTA Fillapex revealed low rates of cell cell viability (P < .001). Finally, scanning electron microscopy studies revealed a high degree of proliferation, cell spreading, and attachment, especially when using GuttaFlow Bioseal disks. CONCLUSIONS: GuttaFlow Bioseal and GuttaFlow2 showed lower cytotoxicity than MTA Fillapex and AH plus. Further in vitro and in vivo investigations are required to confirm the suitability of GuttaFlow Bioseal for clinical application.

Efectos biológicos de los cementos MTA Repair HP y ProRoot MTA tras ser expuestos a diferentes pH sobre células madre de ligamento periodontal / Biological effects of MTA Repair HP and ProRoot MTA exposed to different pH on human periodontal ligament stem cells

Objetivos. El propósito de este trabajo fue evaluar los efectos biológicos de MTA Repair HP y ProRoot MTA en células madre procedentes de ligamento periodontal (hPDLSCs) tras ser ex-puestos los cementos a ambiente ácido y neutro. Material y Métodos: Discos de cada material (n=30) fueron ex-puestos a un tampón fosfato-salino (pH 7.4) o a ácido butírico (pH 5.2) durante 7 días, posteriormente se realizaron pruebas biológicas in vitro usando hPDLSCs. A partir de eluatos de los diferentes materiales de obturación a retro, se analizaron pruebas de viabilidad celular y apoptosis. Para evaluar la adhesión celular, hPDLSCs se sembraron directamente sobre la superficie de los materiales y se observaron bajo microscopio electrónico de barrido. Para analizar estadísticamente los resultados se usaron los test ANOVA y Tukey test (p < 0.05). Resultados: Los cementos endodónticos expuestos a ambiente ácido mostraron un similar grado de adhesión celular, y sorprendentemente, MTA Repair HP a pH 5.2 exhibió una mayor viabilidad celular que ProRootMTA (p<0.05). A pH 7.4, ProRooT MTA obtuvo una tasa mayor de viabilidad celular que con MTA Repair HP. Conclusiones: Los materiales ProRoot MTA y MTA Repair HP presentaron adecuadas propiedades biológicas en los diferentes ambientes, en términos de viabilidad celular, apoptosis y adhesión

Objectives: The purpose of this work was to evaluate the biolo-gical effects of MTA Repair HP and ProRoot MTA on stem cells from periodontal ligament (hPDLSCs) after exposure to acidic and neutral environments. Material and Methods: Discs of each material (n=30) were ex-posed to phosphate buffered saline (pH = 7.4) or butyric acid (pH = 5.2) for 7 days, and biological testing was carried out in vitro on hPDLSCs. Cell viability and apoptosis assays were performed using eluates of each root-end filling material. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. Statistical differences were assessed by ANOVA and Tukey test (p < 0.05).Results: Endodontic cements exposure to an acidic environment showed a similar degree of cell adherence, and, surprisingly, MTA Repair HP exhibited higher cell viability rates at pH 5.2 than Pro-Root MTA, whereas ProRoot MTA 7.4 showed higher cell viability rates than MTA Repair HP. Conclusions: Adequate biological properties of ProRoot MTA and MTA Repair HP in terms of cell viability, cell death and cell attach-ment were observed in both environments