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Background: There is tendency for unavoidable sealer extrusion in some clinical cases. This might adversely affect host stem cells and affect healing. This study aimed to investigate the effect of different sealers on the cytocompatibility and osteogenic potential of human periodontal ligament stem cells (hPDLSCs). Methods: The cytotoxic effect of the extracted elutes of VDW.1Seal (VDW.1), Endosequence BC Sealer HiFlow (ES), GuttaFlow-2 (GF), and ADSeal (AD-S) on the hPDLSCs was determined using the MTT assay. Cell proliferation and migration were assessed by the scratch wound healing assay. Osteogenic differentiation potential was assessed. Measurement of pH values and calcium ions release was performed. Results: GF had a significantly higher percentage of viable cells. The cell migration assay showed that GF demonstrated the lowest open wound area percentage. GF and AD-S showed the highest calcium nodule deposition. GF demonstrated higher ALP activity than ES. Expression of RUNX2 and OC genes was similar for all sealers, while OPG gene expression was significantly higher for VDW.1 and GF. ES and AD-S displayed the highest pH values on day 1. Calcium ion release of ES and VDW.1 was significantly the highest. Conclusions: GuttaFlow-2 and VDW.1Seal sealers have favorable behavior toward host stem cells.

Numerous tissue engineering uses for gingival-derived mesenchymal stem cells (GMSCs) have been demonstrated. Recently, low-level laser therapy (LLLT) has been projected as a factor that can improve MSCs’ regeneration capacity. Therefore, the aim of this research was to examine the impact of LLLT at 1.5 J/cm2 and 3 J/cm2 on the viability and osteo/odontogenic potential of GMSCs. An MTT assay was performed to detect viability. Osteo/odontogenic differentiation was evaluated using Alizarin Red S staining and qRT-PCR for the evaluation of the RUNX2, OC, DMP1, and DSPP genes. A two-way ANOVA with Tukey’s post hoc test was used to determine the statistical significance between groups. The results revealed that LLLT of both energy densities had no cytotoxic effect on GMSC viability. LLLT of 1.5 J/cm2 demonstrated better viability than the higher energy density (3 J/cm2). Furthermore, the osteo/odontogenic differentiation potential was promoted following LLLT radiation, where both groups exhibited mineralized nodule formation, with the low-energy laser having a significantly higher Alizarin Red S stain level. A qRT-PCR analysis revealed higher expression levels of osteogenic and odontogenic markers in the LLLT groups compared to the control group. In conclusion, this study showed the potential application of LLLT as a non-toxic and effective strategy to enhance the regenerative capacity of GMSCs for tissue engineering and clinical treatments in the oral and craniofacial fields.

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Background: Surface roughness of dental implants impacts the survival of adult periodontal stem cells and rate of differentiation. This research was conducted to test how human periodontal ligament stem cells behaved on yttria stabilized tetragonal zirconia polycrystals and polyetheretherketone (PEEK) discs with different surface topographies. Methods: Discs roughening was prepared by sandblasting. Stem cells were cultivated on zirconia discs with a polished surface, PEEK discs with a polished surface, sandblasted zirconia discs and sandblasted PEEK discs. Cells viability was assessed after 24, 48, 72 hours. Scanning electron microscopy was used to examine the adherence and attachment of cells. Osteoblastic differentiation capacity was studied by checking the mineralization clusters development through alizarin red S staining and alkaline phosphatase assay. ANOVA and the Tukey post hoc test were used for the statistical analysis. Results: Polished PEEK discs showed lower cell viability, whereas roughened sandblasted zirconia and PEEK discs showed the highest proliferation rates and cell viability percent. The osteogenic differentiation was enhanced for rough surfaces in comparison to polished surfaces. Sandblasted zirconia and PEEK discs showed a markedly increased mineralized nodule development and ALP enzyme activity compared to the polished surface and control. Conclusions: Micro- topographies creation on the PEEK implant surface enhances stem cell attachment, viability, and osteogenic differentiation.