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Intracanal medicaments with maximal antimicrobial efficacy and minimal damage to resident stem cells are essential for successful regenerative endodontic procedures. 2-Hydroxyisocaproic acid (HICA) could have the attributes of a potential intracanal medicament. This study evaluates its cytotoxicity, genotoxicity, and effects on the odontogenic and osteogenic differentiation of the stem cells of the apical papilla (SCAP). Cytotoxicity and cell viability assays were performed on cells treated for 24, 48, and 72 h with varying concentrations of HICA and compared to the standard intracanal medicament, calcium hydroxide. The genotoxicity was assessed via immunofluorescence for two markers of DNA double-strand breaks: phosphorylated γH2AX and 53BP1. The SCAP differentiation was evaluated based on the alkaline phosphatase activity, Alizarin Red staining, and expression of odontogenic and osteogenic genes (DSPP1, BSP1, OCN, RUNX2) in the presence of selected HICA concentrations. HICA was not cytotoxic at concentrations up to 10 mg/mL, regardless of the exposure time, although it was cytostatic at all tested concentrations. HICA was not genotoxic at concentrations below 5 mg/mL. No difference in cytotoxicity or genotoxicity was found between HICA and calcium hydroxide at 1 mg/mL. HICA retained about 70% of the osteogenic differentiation potential at 1 mg/mL. Within the limitations of this in vitro study, we show that HICA at 1 mg/mL could be a potential intracanal medicament for REPs.
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Advanced platelet-rich fibrin (A-PRF) provides the scaffold and growth factors necessary for stem cells to proliferate and differentiate in successful regenerative endodontic procedures. This study investigates the release of transforming growth factor-ß1 (TGF-ß1), platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF) from A-PRF in cell culture media in the presence and absence of mineral trioxide aggregate (MTA) or Biodentine. Additionally, this research assesses the viability and migration of stem cells of the apical papilla (SCAP) in previously conditioned media. A-PRF obtained from 14 participants were incubated for 7 days in cell culture media alone or via layering with MTA or Biodentine discs and the release of selected growth factors in the media was evaluated using ELISA. The viability of SCAP grown in conditioned media was measured using the CCK8 assay, while SCAP migration was assessed via a transwell assay by counting migrated cells. The release of TGF-ß1, PDGF, and VEGF was significantly higher in media with A-PRF alone than in the presence of either calcium-based silicate material (p < 0.05), which showed no difference from the no-A-PRF control (p < 0.05). None of the tested growth factors released in the A-PRF-conditioned media correlated with clot weight. A-PRF-conditioned media, both with and without calcium-based silicate materials, did not impact SCAP viability and migration (p > 0.05). This study shows that SCAP behavior is not impacted by the decrease in growth factor released in the presence of calcium-based silicate materials and that their role in REPs warrants further investigation.
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INTRODUCTION: In 1890, Breuer applied an electrical field concept to root canals in an attempt at sterilization. Recently, Balson and Dovgan have renewed interest in the use of electricity to aid in canal disinfection. The purpose of this investigation is to determine if the use of the apex locator or apex locator in conjunction with low speed handpiece causes a previously unreported electrophoretic effect. MATERIALS: Forty low-fat, dried sirloin steak cubes separated into four groups of 10 and immersed in 5.25% sodium hypochlorite for two minutes while connected to 1) an apex locator with no batteries, 2) one with batteries, or an apex locator and low speed handpiece set at either 3) zero or 4) 800 rotations per minute. Samples were dried and percentage of weight loss was calculated. ANOVA and Tukey Multicomparison tests were performed for statistical significance of α ≤ 0.05. RESULTS: ANOVA test and Tukey Multicomparison tests showed that there was a significant difference (α ≤ 0.05) between the groups. They revealed statistically significant differences between 800 rotations per minute and all other groups. This study showed that there were no significant differences between an apex locator with no batteries, one used with batteries and one used in conjunction with the low speed handpiece at zero rotations per minute. However, it was shown that there was significantly more tissue dissolution with an apex locator with low speed handpiece at 800 rotations per minute compared to the first three groups. CONCLUSION: These results did not support the original hypothesis that the apex locator with or without the low speed handpiece has an electrophoretic effect. However, the results do support the use of sodium hypochlorite agitation for increased tissue dissolution.
