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

Abstract

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.

Photo biomodulation of dental derived stem cells to ameliorate regenerative capacity: In vitro study

“…In previous studies ( Hamblin, 2017 , Migliario et al, 2018 ) whose findings coincided with our findings, as increasing NF-KB levels in laser-treated groups. Others used Alkaline phosphatase, Type 1 collagen, Osteocalcin, and Bone sialoprotein genes for mineralization process ( Mirza et al, 2021 , Karkehabadi et al, 2023 ).…”

Investigation on the Oxytocin Effect on Proliferation and Osteogenic/Odontogenic Differentiation of Human Stem Cells from the Apical Papilla: An in vitro study

“…Mineralization assay (alizarin red staining) Alizarin red staining was performed to evaluate hSCAPs potential of mineralized nodule formation under the differentiation culture condition with or without treatment with OT. At day 21, after washing with phosphate buffered saline, the cells were xed with 10% formaldehyde and incubated at 37°C for 30 minutes in a dark environment with 1% ARS (Sigma-Aldrich, St. Louis, MO, USA) solution to stain calcium nodules (Karkehabadi, Rahmati et al 2023). The cells then were washed four times with PBS and visualized under direct microscope.…”

“…The resultant cell pellet was dissolved in the complete cell culture media containing Dulbecco's Modi ed Eagle's Medium (Gibco, GrandIsland, NY, USA), supplemented with 15% FBS and 1% penicillin and streptomycin. The cells were kept at 37°C, 5% CO2, and the medium was exchanged every 2-3 days (Karkehabadi, Rahmati et al 2023).…”

“…al. 2008, Pereira, Longo et al 2012, Kang, Fan et al 2019,Karkehabadi, Rahmati et al 2023). Moreover, substantial evidence indicates that hSCAPs are capable of differentiating into various cell lineages including osteogenic, odontogenic, neurogenic, adipogenic, chondrogenic, and hepatogenic lineages, which may serve as a promising cell source for stem cell-based therapy(Sonoyama, Liu et al.…”