Comparative biological properties of resin-free and resin-based calcium silicate-based endodontic repair materials on human periodontal ligament stem cells.

OBJECTIVES: To investigate the effect of three different calcium silicate-based materials (CSBM) on the biological behavior of human periodontal ligament stem cells (hPDLSCs). METHODS: Eluates of Biodentine, NeoPutty and TheraCal PT prepared at 1:1, 1:2, and 1:4 ratios were extracted under sterile conditions. The cytotoxicity of the extracts to the hPDLSCs was assessed using the MTT assay. Scratch wound healing assay was utilized for assessing cell migration. Scanning electron microscopy was used to detect cell attachment and morphology. Calcium ion release was measured using inductively coupled plasma-optical emission spectrometry; the pH-value was evaluated with a pH-meter. ANOVA with post hoc Tukey test was used for statistical analysis. RESULTS: Cell viability was significantly higher for Biodentine and NeoPutty at day 1 with all dilutions (p < 0.05), while at day 3 and day 7 with dilutions 1:2 and 1:4; all materials showed similar behavior (p > 0.05). Biodentine had the highest percentage of cell migration into the scratched area at day 1 for all dilutions (p < 0.05). Stem cells were attached favorably on Biodentine and NeoPutty with evident spreading, and intercellular communications; however, this was not shown for TheraCal PT. Biodentine showed the highest pH values and calcium ion release (p < 0.05). CONCLUSIONS: The resin-free CSBM showed better performance and favorable biological effects on hPDLSCs and were therefore considered promising for usage as endodontic repair materials. CLINICAL SIGNIFICANCE: Proper selection of materials with favorable impact on the host stem cells is crucial to ensure outcome in different clinical scenarios.

Effect of rapamycin on human periodontal ligament stem cells that have been exposed to sodium hypochlorite.

AIMS: The present study investigated the effect of rapamycin on the viability and osteogenic differentiation potential of human periodontal ligament stem cells (hPDLSCs) in the presence of sodium hypochlorite (NaOCl). MAIN METHODS: After determining the minimum inhibitory concentration of NaOCl and optimum concentration of rapamycin, the viability of hPDLSCs was evaluated using the MTT assay subsequent to their exposure to NaOCl, rapamycin, or a combination of both. Osteogenic differentiation was evaluated by the cell mineralization assay performed by alizarin red S staining, alkaline phosphatase activity, and monitoring the expression of osteogenic genes markers Runt-related transcription factor 2, osteocalcin, and osteoprotegerin, using real-time quantitative polymerase chain reaction (RT-qPCR). The expression of autophagy-related genes PI3K, Akt, and mTOR, was also analyzed with RT-qPCR. KEY FINDINGS: Stem cells treated with rapamycin showed the highest percentage of viable cells in the presence of NaOCl. The same trend was observed for all osteogenic differentiation assays. The hPDLSCs treated with rapamycin demonstrated the highest calcium nodule deposition, alkaline phosphatase activity, and the expression of osteogenic gene markers. These effects were not adversely affected by the presence of NaOCl. Rapamycin significantly inhibited mTOR gene expression, while there were no differences in the gene expression of PI3K and Akt. SIGNIFICANCE: Rapamycin counteracts the cytotoxic effect of NaOCl by enhancing the viability and osteogenic differentiation potential of hPDLSCs. Rapamycin appears to accomplish these processes via autophagy activation, by inhibiting mTOR gene expression. The incorporation of rapamycin in regenerative endodontic therapy may encourage a higher success rate.

Fracture resistance of root canal-treated molars restored with ceramic overlays with/without different resin composite base materials: an in vitro study.

The objective of the study was to evaluate the effect of different restorative protocols on fracture resistance of root canal-treated molars. 48 mandibular first molars were used and divided into six groups (n = 8); G1 (negative control): teeth kept intact. G2 (positive control): teeth had root canal treatment and standard MOD cavity preparations but kept unrestored. G3: prepared as G2 and directly restored with VitaEnamic ceramic overlays (CO). G4: as G3, but the pulp chamber was restored first with smart dental restorative (SureFil SDR flow = SDR) bulk-fill flowable composite base. G5: as G3, but the pulp chamber was restored first with SonicFill (SF) bulk-fill composite base. G6: as G3, but the pulp chamber was restored first with a fiber-reinforced composite (FRC) base. All samples were subjected to thermocycling between 5 °C and 55 °C in a water bath for a total of 2000 cycles with 10 s dwell time. Then specimens were individually mounted on a computer-controlled testing machine with a load cell of 5 kN, and the maximum load to produce fracture (N) was recorded. Data were analyzed using one-way ANOVA followed by Tukey's post hoc test (P = 0.05). There was a significant difference between the groups (P < 0.001). Teeth restored with FRC and ceramic overlays had the highest load-bearing capacity. Pulp chamber restoration with either FRC or SDR before ceramic overlay fabrication provided significantly better tooth reinforcement than ceramic overlay alone (P < 0.001). Fracture modes were analyzed to determine the type of fracture as repairable or catastrophic, where FRC + CO and SDR + CO groups had favorable fracture modes that were mostly repairable. When restoring root canal-treated molars with overlays, the pulp chamber should be sealed with either FRC or SDR to ensure the best possible fracture resistance. The clinical relevance of the study is that a new simple restorative protocol is presented to enhance the survival of root canal-treated molars using ceramic overlays.