Effect of magnesium oxide nanoparticles and LED irradiation on the viability and differentiation of human stem cells of the apical papilla.

PURPOSE: Currently, regenerative endodontic treatments are gaining more and more attention, and stem cells play a significant role in these treatments. In order to enhance stem cell proliferation and differentiation, a variety of methods and materials have been used. The purpose of this study was to determine the effects of magnesium oxide nanoparticles and LED irradiation on the survival and differentiation of human stem cells from apical papilla. METHODS: The MTT test was used to measure the cell survival of SCAPs that had been exposed to different concentrations of magnesium oxide nanoparticles after 24 and 48 h, and the concentration with the highest cell survival rate was picked for further studies. The cells were classified into four distinct groups based on their treatment: (1) control, which received no exposure, (2) exposure to magnesium oxide nanoparticles, (3) exposure to light emitting diode (LED) irradiation (635 nm, 200 mW/cm2) for 30 s, (4) exposure simultaneously with magnesium oxide nanoparticles and LED irradiation. A green approach was employed to synthesize magnesium oxide nanoparticles. Quantitative real time PCR was used to measure the gene expression of osteo/odontogenic markers such as BSP, DSPP, ALP and DMP1 in all four groups after treatment, and Alizarin red S staining (ARS) was used to determine the osteogenic differentiation of SCAPs by demonstrating the Matrix mineralization. RESULTS: The highest viability of SCAPs was observed after 24 h in concentration 1 and 10 µg/mL and after 48 h in concentration 1 µg/mL, which were not significantly different from the control group. In both times, the survival of SCAPs decreased with increasing concentration of magnesium oxide nanoparticles (MgONPs). According to the results of Real-time PCR, after 24 and 48 h, the highest differentiation of BSP, DMP1, ALP and DSPP genes was observed in the LED + MgONPs group, followed by MgONPs and then LED, and in all 3 experimental groups, it was significantly higher than control group (P < 0.05). Also, after 24 and 48 h, the density of ARS increased in all groups compared to the control group, and the highest density was observed in the MgONPs + LED and MgONPs groups. CONCLUSION: This research concluded that exposure to SCAPs, MgONPs, and LED irradiation has a significant effect on enhancing gene expression of odontogenic/osteogenic markers and increasing matrix mineralization.

Effect of copper oxide nanoparticles and light-emitting diode irradiation on the cell viability and osteogenic/odontogenic differentiation of human stem cells from the apical papilla.

OBJECTIVES: This experimental study aimed to assess the effect of copper oxide nanoparticles (CuONPs) and light-emitting diode (LED) irradiation on the cell viability and osteogenic/odontogenic differentiation of human SCAPs. METHODS: After the culture of SCAPs, the effects of different concentrations of CuONPs on cell viability were evaluated by the methyl thiazolyl tetrazolium (MTT) assay after 24 and 48 h, and the optimal concentration was determined (n = 12). SCAPs were then divided into four groups based on the type of treatment: (I) no-treatment control group, (II) exposure to CuONPs, (III) LED irradiation (635 nm, 200 mW/cm2) for 30 s, and (IV) exposure to CuONPs combined with LED irradiation. CuONPs were synthesized by a green technique, which was based on reduction and simultaneous stability of copper ions by using the pomegranate peel extract. After treatments, the expression of osteogenic/odontogenic markers including dentin sialophosphoprotein (DSPP), bone sialoprotein (BSP), alkaline phosphatase (ALP), and dentin matrix acidic phosphoprotein 1 (DMP1) was evaluated in all four groups using quantitative real-time polymerase chain reaction (PCR) (n = 16). Also, osteogenic differentiation of SCAPs was evaluated qualitatively by alizarin red staining (ARS) to assess the matrix mineralization (n = 4). SPSS version 18 was used for data evaluation. The Kruskal-Wallis and Mann-Whitney tests were used to compare the groups. RESULTS: Exposure to 1 µg/mL CuONPs resulted in maximum viability of SCAPs. Concentrations of CuONPs over 10 µg/mL significantly decreased the viability of SCAPs. Real-time PCR showed that the expression of DMP1, BSP, ALP, and DSPP in CuONPs + LED and LED groups was significantly higher than that in CuONPs and control groups at both 24 and 48 h (P < 0.05). The density of ARS increased in all experimental groups after 24 h, and in CuONPs + LED and CuONPs groups after 48 h, compared to the control group. CONCLUSION: Addition of CuONPs and LED irradiation of SCAPs in the culture medium significantly enhanced their osteogenic/odontogenic differentiation.