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Effects of Er:YAG laser irradiation on proliferation and alteration of gene expression in the MLO-Y4 osteocyte-like cells

ABSTRACT: Objective: The aim of this study was to investigate the effects of Er:YAG laser irradiation on MLO-Y4 osteocyte-like cells. Materials and Methods: MLO-Y4 cells were seeded before 72h laser-irrdiation and irradiated by Er:YAG laser at energy fluences of 3.1, 6.0 and 8.9 J/cm2. To evaluate thermal effects, cell surface temperature and gene expression of Hspa1a were measured. For evaluation of cell proliferation and cytotoxicity, CCK-8 assay, and Lactate dehydrogenase (LDH) assay of the laser-irradiated cells were performed. RNA-sequencing analysis was performed at 6h after 6.0 J/cm2 laser-irradiation and non-irradiated cells. Result: According to surface temperature and Hspa1a expression, no thermal effects were observed at 6.0 J/cm2 irradiation. CCK-8 assay revealed a significant increase in proliferative activity was observed, although LDH levels showed a slight but significant increase at 6.0 J/cm2 laser-irradiation. RNA-seq analysis identified 337 differently expressed genes. Epha, Itga5, and Tcf7, associated with mechanical stimulation, were highly upregulated in the laser-irradiated cells. KEGG pathway analysis showed upregulation of PI3K-Akt signaling pathway in the laser-irradiated cells. Upregulated DEGs belong to GO term of cellular components “euchromatin” in the laser-irradiated cells. Gene set enrichment analysis showed enrichment of TGF-beta signaling gene set and downregulation of mTORC1 signaling gene set in the laser-irradiated cells. Conclusion: Er:YAG laser irradiation promotes the proliferation of MLO-Y4 cells, osteocyte-like cells, via altering cell proliferation and mechanotransduction-related gene expression.

ORGANISM(S): Mus musculus

PROVIDER: GSE295519 | GEO | 2026/05/20

REPOSITORIES: GEO

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Project description:We report that terahertz (THz) irradiation of mouse mesenchymal stem cells with a pulsed broadband (centered at 10 THz) source, or a single-frequency, 2.52 THz, (SF) laser source, both with weak average power (<1mW/cm2), results in specific heterogenic changes in gene expression. The insignificant differential expression of heat shock and stress related genes as well as our temperature measurements imply a non-thermal response. The microarray survey and RT-PCR experiments demonstrate that at different irradiation conditions distinct groups of genes are activated. Stem cells irradiated for 12 hours with the broadband THz source exhibit an accelerated differentiation toward adipose phenotype, while the 2-hour (broadband or SF) irradiation affects genes transcriptionally active in pluripotent stem cells. Phenotypic and gene expression differences suggest that the THz effect depends on irradiation parameters such as duration and type of THz source, and on the level of stem cell differentiation. Computer simulations of the core promoters of two pluripotency markers reveal association between gene upregulation and propensity for DNA breathing. We propose that THz radiation has potential for non-contact control of cellular gene expression. Growing in petri dish mouse mesenchymal stem cell cultures are irradiated for 2 hours with a single-frequency (2.52 THz) CW laser source. Three independent biological experiments were conducted. In each experiment, a control mMSC culture was placed adjacent to the irradiated sample, and the temperature was kept at 26-27M-BM-:C for both the control and the irradiated sample. Immediately after completing the irradiation, total RNA was extracted from the irradiated sample and the control, and microarrays were used to identify differential changes in gene expression between the irradiated sample and its respective control. In each experiment, the mMSC cultures were synchronized to be at the same differentiation time point immediately before the irradiation.

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Effects of Er:YAG laser irradiation on proliferation and alteration of gene expression in the MLO-Y4 osteocyte-like cells

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