The effects of Biodentine/polycaprolactone three-dimensional-scaffold with odontogenesis properties on human dental pulp cells.

AIM: To determine the feasibility of using three-dimensional printed Biodentine/polycaprolactone composite scaffolds for orthopaedic and dental applications. The physicochemical properties and the odontogenic differentiation of human dental pulp cells (hDPCs) were investigated. METHODOLOGY: Biodentine was well-suspended in ethanol and dropped slowly into molten polycaprolactone with vigorous stirring. The Biodentine/polycaprolactone composite scaffolds were then fabricated into controlled macropore sizes and structures using an extrusion-based three-dimensional (3D) printer. The mechanical properties, bioactivity, and the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs) cultured on the scaffolds were evaluated. RESULTS: Biodentine/polycaprolactone scaffolds had uniform macropores 550 µm in size with established interconnections and a compressive strength of 6.5 MPa. In addition, the composite scaffolds exhibited a good apatite-forming ability and were capable of supporting the proliferation and differentiation of hDPCs. CONCLUSION: The composite scaffolds fabricated by an extrusion-based 3D printing technique had similar characteristics to Biodentine cement, including bioactivity and the ability to promote the differentiation of hDPCs. These results indicate that the composite scaffold would be a candidate for dental and bone regeneration.

Role of the p38 pathway in mineral trioxide aggregate-induced cell viability and angiogenesis-related proteins of dental pulp cell in vitro.

AIM: To investigate the influence of mineral trioxide aggregate (MTA) on angiogenesis of primary human dental pulp cells (hDPCs) via the MAPK pathway, in particular p38. METHODOLOGY: Human dental pulp cells were cultured with MTA to angiogenesis, after which cell viability, ion concentration, osmolality, NO secretion, the von Willebrand factor (vWF) and angiopoietin-1 (Ang-1) protein expression were examined. PrestoBlue(®) was used for evaluating the proliferation of hDPCs. An enzyme-linked immunosorbent assay was employed to determine vWF and Ang-1 protein secretion in hDPCs cultured on MTA and the control. Cells cultured on the tissue culture plate without the cement were used as the control. The t-test was used to evaluate the significance of the differences between the mean values. RESULTS: Mineral trioxide aggregate elicited a significant (P < 0.05) increased viability compared with the control (15%, 16% and 13% on days 1, 3 and 5 of cell seeding, respectively). MTA consumed calcium and phosphate ions, and released more Si ions in the medium. MTA significantly (P < 0.05) increased the osmolality of the medium to 313, 328 and 341 mOsm kg(-1) after 1, 3 and 5 days, respectively. P38 was activated through phosphorylation, and the phosphorylation kinase was investigated in the cell system after being cultured with MTA. Expression levels for Ang-1 and vWF in hDPCs on MTA were higher than those of the MTA + p38 inhibitor (SB203580) group (P < 0.05) at all of the time-points. CONCLUSIONS: Mineral trioxide aggregate was able to activate the p38 pathway in hDPCs cultured in vitro. Moreover, Si increased the osmolality required to facilitate the angiogenic differentiation of hDPCs via the p38 signalling pathway. When the p38 pathway was blocked by SB203580, the angiogenic-dependent protein secretion decreased. These findings verify that the p38 pathway plays a key role in regulating the angiogenic behaviour of hDPCs cultured on MTA.

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.

Effects of altering the Si/Ca molar ratio of a calcium silicate cement on in vitro cell attachment.

AIM: To examine the effects of altering the Si/Ca molar ratio (6 : 4, 5 : 5, and 4 : 6) of a quick-setting calcium silicate cement on in vitro cell attachment. METHODOLOGY: Working time and setting time of three different calcium silicate cements were measured. Alamar Blue was used for real-time and repeated monitoring of cell attachment and proliferation. The Si and Ca ion concentrations of the cell culture medium in the presence of three different calcium silicate cements seeded with MG63 cells were measured. Kinetic immunofluorescent staining of MG63 cells was performed during cell attachment and spreading. Reverse transcription-polymerase chain reaction was employed to determine gene expression in MG63 cells cultured on the cements. One-way analysis of variance was used to evaluate the significance of the differences between the mean values. RESULTS: The working time (4-7 min) and setting time (17-24 min) of the cements were shortened with an increase in the Ca content of the calcium silicate powders after mixing the powder with water. In contrast, the higher the Si content in the cement, the more the MG63 cells attached to the cement at all culture time-points, accompanying by the formation of more obvious actin stress fibres. Cell proliferation and differentiation increased significantly (P < 0.05) with an increase in the Si content of the calcium silicate cements. Si ion concentration of the culture medium increased significantly (P < 0.05) with increasing cement Si content and culture time-points. CONCLUSIONS: The higher Si content cement enhanced the higher expression of cell attachment, proliferation and differentiation as compared to the lower Si content cement.

Human dental pulp cell responses to new calcium silicate-based endodontic materials.

AIM: To evaluate human dental pulp cell responses to radiopaque dicalcium silicate cement and white-coloured mineral trioxide aggregate (WMTA). METHODOLOGY: Flow cytometry was employed to quantify the phase percentage of pulp cell cycle. Alamar Blue was used for real-time and repeated monitoring of cell proliferation. Reverse transcription-polymerase chain reaction was performed to determine gene expression in pulp cells cultured on the cements. RESULTS: The cells cultured on the radiopaque dicalcium silicate cement had similar S and G2 phases in the cell cycle and proliferation to WMTA at all culture times. In addition, the two materials presented the same evolution with similar values in interleukin-1, inducible nitric oxide synthase, alkaline phosphatase, osteocalcin and bone sialoprotein gene expression at all culture times. CONCLUSIONS: The dental pulp cell responses to radiopaque dicalcium silicate cement were similar to those reported for WMTA in terms of cell cycle, proliferation, immunocompatibility and osteogenic differentiation.