Ming-You Shie

The Effect of Setting Accelerator on Properties of Mineral Trioxide Aggregate

The purpose of this study was to investigate the effect of a sodium phosphate dibasic (Na2HPO4) setting accelerator on the properties of white-colored mineral trioxide aggregate (WMTA). Setting times were measured by using a Gilmore needle. Changes in the pH value, diametral tensile strength, and phase composition of WMTA were evaluated. By using a 15% Na2HPO4 solution as a liquid phase mixed with WMTA, the setting time was significantly reduced to 26 minutes rather than the usual 3 hours. The 15% Na2HPO4 promoted WMTA to achieve a maximum diametral tensile strength of 4.9 MPa at an initial 6-hour aging time and 1 MPa for the cement mixed with water. The pH value of the 15% Na2HPO4-mixed cement was changed from an initial 11.0 to a high of 13.2, which was similar to the results using water. The results suggest that the Na2HPO4 solution may be an effective setting accelerator for WMTA.

Comparison of Calcium and Silicate Cement and Mineral Trioxide Aggregate Biologic Effects and Bone Markers Expression in MG63 Cells

Bone cell (MG63) biocompatibility and bone marker expression were compared after calcium and silicate base cement (CS) and mineral trioxide aggregate (MTA) treatment. X-ray diffraction was used to identify material surface structure, and tetrazolium bromide colorimetric assay was used to evaluate the cell viability. The relative mitogen activation protein kinase expression was compared with Western blot, and bone marker expression was evaluated with reverse transcriptase polymerization chain reaction. The results showed that CS and MTA are similar chemical structures and biocompatible with MG63 cells. CS and MTA cements showed good MG63 cell proliferation by high phosphor extracellular signal-regulated kinase expression levels. CS and MTA cements showed the evident type I collagen, osteocalcin, alkaline phosphatase, bone sialoprotein, and osteopontin expression. Both MTA and CS cements are biocompatible and appear to have osetoconduction effects on bone cells.

The Physical and Cytological Properties of White MTA Mixed with Na2HPO4 as an Accelerant

The purpose of the present study was to evaluate the physicochemical and cytologic properties of mineral trioxide aggregate (MTA) mixed with distilled water and Na(2)HPO(4) buffer solution. The MTA setting time and pH value were evaluated. An MTA micrograph on scanning electron microscopy (SEM) was observed. Mouse fibroblasts (an L929 cell line) were used to test the toxicity of MTA after the first and seventh day of treatment by mitochondrial colorimetric assay. The results show the Na(2)HPO(4) buffer group reduced the MTA setting time, and the pH value as in the distilled water group is similar with the Na(2)HPO(4) buffer group. The mixed MTAs XRD produced similar peaks of the distilled water and Na(2)HPO(4) buffer solution groups. The L929 cell survival rate of distilled water and H(2)PO(4) buffer solution groups did not exhibit any significant difference (p < 0.05). There are differences in SEM observations both of the MTA surface and of the cells in culture on the surface of the MTA with Na(2)HPO(4) versus distilled water. The results suggest that 15% Na(2)HPO(4) buffer can be successfully used as an accelerator of MTA.

The Effect of a Physiologic Solution pH on Properties of White Mineral Trioxide Aggregate

The purpose of this study was to investigate the effect of two solutions differing by pH (6.4 and 4.0) and the use of a setting accelerator (15% Na(2)HPO(4) solution) on the properties of white-colored mineral trioxide aggregate (WMTA). These studies indicated that pH 4.0 had a deleterious effect on the morphology of WMTA mixed with water, however, not for WMTA mixed with Na(2)HPO(4). When immersed in a pH 4.0 solution for 7 days, WMTA mixed with water or Na(2)HPO(4) achieved a diametral tensile strength of 7.9 and 9.0 MPa, respectively, which was significantly lower (p < 0.05) than those obtained at pH 6.4 (11.2 and 12.0 MPa) but significantly higher (p < 0.05) than day 0 samples (4.4 and 4.8 MPa). Mixing WMTA with the accelerator did not significantly affect the microstructure, solubility, or strength in an acidic environment.

The synergistic effects of fibroblast growth factor-2 and mineral trioxide aggregate on an osteogenic accelerator in vitro.

AIM To examine the effects of mineral trioxide aggregate (MTA)/fibroblast growth factor-2 (FGF-2) on material properties and in vitro human dental pulp cell (hDPCs) behaviour. METHODOLOGY The setting time and diametral tensile strength (DTS) of MTA and MTA/FGF-2 were measured. The structure of specimens before and after soaking in DMEM was examined under a scanning electron microscope. Alamar Blue was used for evaluating hDPCs proliferation. An enzyme-linked immunosorbent assay was employed to determine ALP and osteocalcin (OC) expression in hDPCs cultured on cements. The effect of small interfering RNA (siRNA) transfection targeting fibroblast growth factor receptor (FGFR) was also evaluated. One-way analysis of variance was used to evaluate the significance of the differences between the mean values. RESULTS Setting time and DTS data were not found to be significant (P > 0.05) between MTA with and without FGF-2. Cell proliferation and differentiation increased significantly (P < 0.05) with FGF-2 mixed MTA. After siRNA transfection with FGFR, the proliferation and differentiation behaviour of the hDPCs appreciably decreased when cultured on an MTA/FGF-2 composite. In contrast, no significant amounts (P > 0.05) of ALP and OC were secreted by hDPCs seeded on MTA. CONCLUSIONS Mineral trioxide aggregate with FGF-2 content enhanced the higher expression of hDPCs proliferation and osteogenic differentiation as compared to pure MTA cement.

Using calcium silicate to regulate the physicochemical and biological properties when using β-tricalcium phosphate as bone cement

β-Tricalcium phosphate (β-TCP) is an osteoconductive material. For this research we have combined it with a low degradation calcium silicate (CS) to enhance its bioactive and osteostimulative properties. To check its effectiveness, a series of β-TCP/CS composites with different ratios were prepared to make new bioactive and biodegradable biocomposites for bone repair. Regarding the formation of bone-like apatite, the diametral tensile strength as well as the ion release and weight loss of composites were compared both before and after immersions in simulated body fluid (SBF). In addition, we also examined the behavior of human dental pulp cells (hDPCs) cultured on β-TCP/CS composites. The results show that the apatite deposition ability of the β-TCP/CS composites improves as the CS content is increased. For composites with more than a 60% CS content, the samples become completely covered by a dense bone-like apatite layer. At the end of the immersion period, weight losses of 24%, 32%, 34%, 38%, 41%, and 45% were observed for the composites containing 0%, 20%, 40%, 80%, 80% and 100% β-TCP cements, respectively. In addition, the antibacterial activity of CS/β-TCP composite improves as the CS-content is increased. In vitro cell experiments show that the CS-rich composites promote human dental pulp cell (hDPC) proliferation and differentiation. However, when the CS quantity in the composite is less than 60%, the quantity of cells and osteogenesis protein of hDPCs is stimulated by Si released from the β-TCP/CS composites. The degradation of β-TCP and the osteogenesis of CS give strong reason to believe that these calcium-based composite cements will prove to be effective bone repair materials.

Regulation of physicochemical properties, osteogenesis activity, and fibroblast growth factor-2 release ability of β-tricalcium phosphate for bone cement by calcium silicate.

β-Tricalcium phosphate (β-TCP) is an osteoconductive material. For this research we have combined it with a low degradation calcium silicate (CS) to enhance its bioactive and osteostimulative properties. To check its effectiveness, a series of β-TCP/CS composites with different ratios were prepared to make new bioactive and biodegradable biocomposites for bone repair. Formation of bone-like apatite, the diametral tensile strength, and weight loss of composites were considered before and after immersion in simulated body fluid (SBF). In addition, we also examined the effects of fibroblast growth factor-2 (FGF-2) released from β-TCP/CS composites and in vitro human dental pulp cell (hDPC) and studied its behavior. The results showed that the apatite deposition ability of the β-TCP/CS composites was enhanced as the CS content was increased. For composites with more than 50% CS contents, the samples were completely covered by a dense bone-like apatite layer. At the end of the immersion point, weight losses of 19%, 24%, 33%, 42%, and 51% were observed for the composites containing 0%, 30%, 50%, 70% and 100% β-TCP cements, respectively. In vitro cell experiments show that the CS-rich composites promote human dental pulp cell (hDPC) proliferation and differentiation. However, when the CS quantity in the composite is less than 70%, the amount of cells and osteogenesis protein of hDPCs was stimulated by FGF-2 released from β-TCP/CS composites. The combination of FGF-2 in degradation of β-TCP and osteogenesis of CS gives a strong reason to believe that these calcium-based composite cements may prove to be promising bone repair materials.

Expression of the Inflammatory Marker Cyclooxygenase-2 in Dental Pulp Cells Cultured with Mineral Trioxide Aggregate or Calcium Silicate Cements

INTRODUCTION Mineral trioxide aggregate (MTA) and calcium silicate (CS) cements exhibit acceptable physical and chemical properties. The aim of the present study was to evaluate the effects of MTA and CS cements on inflammatory reactions in primary cultured human dental pulp cells. METHODS The mitochondrial colorimetric assay was used to evaluate pulp cell survival rates. Fluorescent immunohistochemistry was used to observe focal adhesion kinase (FAK) and cyclooxygenase-2 (COX-2) distributions in the cells. Reverse transcription-polymerase chain reaction was used to assess COX-2 expression. RESULTS The results showed that MTA and CS are biocompatible with pulp cells (P>.05). FAK was well-distributed in pulp cells in contact with both cements. Both MTA and CS cements induced pulp cell inflammation as evidenced by increased COX-2 expression. CONCLUSIONS The present study demonstrated that MTA and CS cements are biocompatible with primary cultured pulp cells. Both cements can induce inflammatory COX-2 expression in the pulp cells.

The Role of Integrin αv in Proliferation and Differentiation of Human Dental Pulp Cell Response to Calcium Silicate Cement

INTRODUCTION It has been proved that integrin αv activity is related to cell proliferation, differentiation, migration, and organ development. However, the biological functions of integrin αv in human dental pulp cells (hDPCs) cultured on silicate-based materials have not been explored. The aim of this study was to investigate the role of integrin αv in the proliferation and odontogenic differentiation of hDPCs cultured with the effect of calcium silicate (CS) cement and β-tricalcium phosphate (TCP) cement. METHODS In this study, hDPCs were cultured on CS and TCP materials, and we evaluated fibronectin (FN) secretion and integrin αv expression during the cell attachment stage. After small interfering RNA transfection targeting integrin αv, the proliferation and odontogenesis differentiation behavior of hDPCs were analyzed. RESULTS The results indicate that CS releases Si ion-increased FN secretion and adsorption, which promote cell attachment more effectively than TCP. The CS cement facilitates FN and αv subintegrin expression. However, the FN adsorption and integrin expression of TCP are similar to that observed in the control dish. Integrin αv small interfering RNA inhibited odontogenic differentiation of hDPCs with the decreased formation of mineralized nodules on CS. It also down-regulated the protein expression of multiple markers of odontogenesis and the expression of dentin sialophosphoprotein protein. CONCLUSIONS These results establish composition-dependent differences in integrin binding and its effectiveness as a mechanism regulating cellular responses to biomaterial surface.

Enhanced adhesion and differentiation of human mesenchymal stem cell inside apatite-mineralized/poly(dopamine)-coated poly(ε-caprolactone) scaffolds by stereolithography

The purpose of this study is to develop PCL scaffolds using stereolithography technology and induced modifications using a poly dopamine (PDA)-coated/HA precipitate to stimulate human mesenchymal stem cells (hMSCs). The chemical composition and surface properties of HA/PDA/PLA were characterized by XPS. HA/PDA/PLA modulated hMSCs responses in several ways. The extracellular matrix (collagen and fibronectin) adsorption was significantly higher on the substrates with the highest amount of PDA coating than on pure PCL. Increased focal adhesion kinase (FAK) levels and enhanced cell attachment were observed upon an increase in PDA content. The proliferation, alkaline phosphatase, osteogenesis-related proteins (OPN and BSP), and angiogenesis-related protein (vWF and ang-1) secretion of hMSCs were significantly stimulated when the PDA-coated concentration was increased. The PDA-coated/HA precipitate increases osteogenesis and angiogenesis of hMSCs cultured with a PCL scaffold. Our results demonstrate that this simple, bio-inspired surface modification of the organic PCL scaffold using PDA is a very promising tool for regulating cell behaviour, and may serve as an effective stem cell delivery carrier for bone tissue engineering.