This study aimed to determine the most suitable recombinant fortilin and evaluate the biological activities of glass ionomer cement (GIC) incorporated with fortilin on human dental pulp stem cells (hDPSCs). Full-length and three fragments of Penaeus merguiensis fortilin were cloned and examined for their proliferative and cytoprotective effects on hDPSCs by MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay. Human DPSCs were cultured with GIC supplemented with fortilin, tricalcium phosphate, or a combination of tricalcium phosphate and fortilin, designated as GIC + FL, GIC + TCP, and GIC + TCP + FL, respectively (n = 4 for each group). At given time points, hDPSCs were harvested and analyzed by MTT, quantitative reverse transcription polymerase chain reaction, alkaline phosphatase activity, and Alizarin Red assays. The full-length fortilin promoted cell proliferation and significantly increased cell survival. This protein was subsequently added into the GIC along with tricalcium phosphate to investigate the biological activities. All experimental groups showed reduced cell viability after treatment with modified GICs on days 1 and 3. The GIC + TCP + FL group significantly promoted odontoblastic differentiation at particular time points. In addition, alkaline phosphatase activity and calcium phosphate deposit were markedly increased in the GIC + TCP + FL group. Among all experimental groups, the GIC incorporated with fortilin and tricalcium phosphate demonstrated the best results on odontogenic differentiation and mineral deposition in hDPSCs.

Fortilin is a multi-functional protein involved in several cellular processes. It has been shown promising potential to be a bioactive molecule that can be incorporated in the dental materials. This study aimed to compare the biocompatibility and mineralization activities of modified glass ionomer cement (Bio-GIC) and Biodentine by direct and indirect method on human dental pulp stem cells (hDPSCs). Conventional glass ionomer cement (GIC), Bio-GIC (GIC supplemented with chitosan, tricalcium phosphate, and recombinant fortilin from Fenneropenaeus merguiensis), and Biodentine were examined in this study. Recombinant fortilin was purified and tested for its cytotoxicity by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. Human DPSCs were treated with different material eluate for particular time intervals. At given time points, viability of hDPSCs was examined using MTT assay and calcium deposition was assessed by Alizarin red staining assay. Comparisons of the data among groups were analyzed by analysis of variance and Tukey's multiple comparisons. All test materials demonstrated no cytotoxicity. In addition, Bio-GIC promoted cell proliferation at 72 h. For direct and indirect method, cells treated with Bio-GIC demonstrated significantly higher calcium deposition than other groups (P < 0.05). Bio-GIC and Biodentine are not cytotoxic to hDPSCs. Bio-GIC demonstrates enhanced calcium deposition comparable to Biodentine. Bio-GIC may be further developed as a bioactive material for dentin regeneration.

This study modified glass ionomer cement (GIC) by adding mimicked biological molecules to reduce cell death. GIC was modified to BIOGIC by adding chitosan and bovine serum albumin for enhancing protein release. The BIOGIC was supplemented with tricalcium phosphate (TCP) and recombinant translationally controlled tumor protein (TCTP) to improve its biological properties. Four groups of materials, GIC, BIOGIC, BIOGIC+TCP, and BIOGIC + TCP + TCTP, were examined by XRD and SEM-EDX. TCTP released from the specimens was determined by an ELISA method. Human dental pulp stem cells (hDPSCs) were harvested and analyzed by MTT assay, apoptosis, gene expression, and cell differentiation. All groups had the same crystallization characteristic peaks of La₂O₃. The elemental compositions composed of La, Si, and Al are the main inorganic components. The results show that BIOGIC + TCP + TCTP presented significantly higher percentages of cell viability than other groups on day 1 to day 23 (p < 0.05), but were not different after day 24 to day 41 and had reduced cell apoptosis including BAX, TPT1, BCL-2, and Caspase-3. The BIOGIC + TCP + TCTP demonstrated higher odontoblast mineralization and differentiation markers including ALP activity, DSPP, DMP-1, ALP, BMP-2, and OPN. It enhanced cell proliferation and differentiation as well as mineralization with down-regulation of genes related to apoptosis compared with other groups.

Purpose: This study aimed to investigate the minimum inhibitory concentration (MIC) and biological properties of modified glass-ionomer cement (BIO-GIC) supplemented with chlorhexidine diacetate (CHX).Methods: The BIO-GIC was originally a combining formulation of conventional glass-ionomer cement, chitosan, albumin and tricalcium phosphate. CHX was incorporated at various concentrations of 1.25%, 2%, 2.5%, 5%, 7.5% and 10%. Antimicrobial activity against certain microorganisms related to cariogenesis Streptococcus mutans (S. mutans), Streptococcus sanguinis (S. sanguinis), and Enterococcus faecalis (E. faecalis), was evaluated by microdilution test. One effective concentration of CHX was mixed with Penaeus merguiensis-Translationally Controlled Tumor Protein (Pmer-TCTP) and further conducted by biological tests including cell viability and mineralization in dental pulp.Results: S. mutans were inhibited at 7.5% CHX-modified BIO-GIC, while S. sanguinis and E. faecalis showed no inhibition at any concentration. Cell viability significantly decreased with 7.5% CHX-modified BIO-GIC after 72 h. Dental pulp exposed to this group formed smaller and fewer calcium nodules but adding Pmer-TCTP reduced cell death and slightly enhanced mineralization.Conclusion: 7.5% CHX-modified BIO-GIC shows potential for inhibiting S. mutans and having potential cell viability and mineralization in dental pulp cells.

Purpose: This study aimed to investigate the antibacterial activities of the ethanolic leaf extract of Rhodomyrtus tomentosa against Enterococcus faecalis (E. faecalis) and its cytocompatibility to human periodontal ligament (hPDL) cells.Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the ethanolic extract of Rhodomyrtus tomentosa leaf (RTE) against E. faecalis were determined using the broth microdilution method. Biofilm inhibition was assessed using the crystal violet assay. Ultrastructural changes were observed via transmission electron microscopy (TEM), whereas biofilm and dentin surface change of root specimens were examined using scanning electron microscopy (SEM). Cytotoxicity was evaluated on hPDL cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Results: RTE exhibited potent antibacterial activity with MIC and MBC of 1 and 32 µg/mL, respectively. RTE significantly inhibited biofilm formation in a concentration-dependent manner and outperformed sodium hypochlorite (NaOCl) at sub-MIC levels. TEM revealed morphological disruption of E. faecalis cells following RTE treatment. SEM confirmed effective bacterial reduction without causing erosion of radicular dentin. RTE at sub-MICs demonstrated relatively low cytotoxicity compared with NaOCl.Conclusion: RTE exhibited strong antibacterial and antibiofilm properties while preserving dentin structure and demonstrating cytocompatibility.