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Influence of Neodymium-Doped Yttrium Aluminum Garnet Laser Irradiation on Cell Proliferation and Bone Morphogenetic Protein-2 Concentrations in MG-63 Osteoblast-Like Cells: An in Vitro Study
Objective: The aim of this in vitro study was to assess the influence of neodymium-doped yttrium aluminum garnet (Nd:YAG) laser irradiation on proliferation and bone morphogenetic protein-2 (BMP-2) concentrations in MG-63 osteoblast-like cells. Methods: We cultured MG-63 cells in plastic 96-well plates and exposed the cultures to Nd:YAG laser irradiation (1064 nm). Independent variables included power (0.25 to 5.0 W) and irradiation time (10 to 60 s). In test and control groups, we assessed BMP-2 protein concentrations using two methods: enzyme-linked immunosorbent assay (ELISA) and magnetic microsphere BMP-2 assay. In addition, we assessed cell viability and proliferation using water soluble tetrazolium (WST-1) and cell counting kit-8 (CCK-8) assays. We evaluated outcome variables 24 and 72 h following laser exposure (individual analyses) and used Mann-Whitney U tests and Kruskal-Wallis tests to determine if outcomes differed across the independent variables of interest. Results: Magnetic microsphere analysis failed to identify any statistically significant influence of laser energy on BMP-2 concentration irrespective of power or irradiation time. In the ELISA analysis, BMP-2 concentration was significantly higher for irradiation time of 10 versus 60 seconds (p=0.009) at the 24-hour assessment only. CCK-8 analysis showed that, compared with control cultures, cell proliferation was significantly higher at the 24-hour recording for cells receiving power of 0.25 W (p=0.0048), 1 W (p=0.0048), 3 W (p=0.0048), or 5 W (p=0.0048). We observed comparable effects on cell proliferation regardless of assessment time (24 or 72 h) or cell proliferation assay (WST-1 or CCK-8). Conclusion: Under the conditions described, Nd:YAG laser irradiation consistently increased proliferation of MG-63 cells. However, in almost all experiments, BMP-2 protein concentrations in control and laser-treated cell cultures were not statistically different.
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