Lee Sung-Bin, Lee Hyunjin, Park Jun-Beom
Department of Medicine, Graduate School, The Catholic University of Korea, Seoul, 06591, Republic of Korea.
Department of Periodontics, Seoul St Mary's Hospital, College of Medicine, The Catholic University of Korea, 222, Banpo-daero, Seocho-gu, Seoul, 06591, Republic of Korea.
Sci Rep. 2025 Jul 2;15(1):23326. doi: 10.1038/s41598-025-05889-y.
Modulation of cellular activity by low-level laser therapy (LLLT) has been widely studied, particularly in regenerative medicine. This study examined the effects of LLLT on the viability, osteogenic differentiation, and mineralization of gingiva-derived mesenchymal stem cells (GMSCs) in two-(2D) and three-dimensional (3D) cultures. GMSCs were treated with LLLT at 980 nm and 808 nm using different energy densities and irradiation frequencies. Cell viability was assessed using a colorimetric assay and live/dead staining. Osteogenic differentiation was assessed through alkaline phosphatase activity and real-time polymerase chain reaction (RT-PCR) analysis of RUNX2 and COL1A1 mRNA expression. Mineralization was analyzed using Alizarin Red S staining. LLLT enhanced cell viability without inducing significant morphological changes, with more pronounced effects in 3D spheroids than in 2D monolayers. Osteogenic differentiation and mineralization were significantly increased in LLLT-treated groups. Gene expression analyses confirmed the upregulation of key osteogenic markers, reinforcing LLLT's role in promoting osteogenesis. These findings indicate that LLLT is a non-invasive and effective approach to promoting osteogenic differentiation and mineralization. Notably, its effects were more evident in 3D culture systems, which better mimic in vivo conditions. This study highlights the therapeutic potential of LLLT in tissue engineering and regenerative medicine, emphasizing its applications in bone regeneration.
低强度激光疗法（LLLT）对细胞活性的调节作用已得到广泛研究，尤其是在再生医学领域。本研究考察了LLLT在二维（2D）和三维（3D）培养体系中对牙龈间充质干细胞（GMSCs）的活力、成骨分化及矿化的影响。使用不同能量密度和照射频率，在980nm和808nm波长下对GMSCs进行LLLT处理。采用比色法和活/死细胞染色评估细胞活力。通过碱性磷酸酶活性以及对RUNX2和COL1A1 mRNA表达进行实时聚合酶链反应（RT-PCR）分析来评估成骨分化。使用茜素红S染色分析矿化情况。LLLT提高了细胞活力，且未诱导明显的形态学变化，在3D球体中的作用比在2D单层细胞中更显著。LLLT处理组的成骨分化和矿化显著增加。基因表达分析证实关键成骨标志物上调，进一步证明了LLLT在促进成骨过程中的作用。这些发现表明，LLLT是促进成骨分化和矿化的一种非侵入性有效方法。值得注意地是，其作用在更好模拟体内条件的3D培养体系中更为明显。本研究突出了LLLT在组织工程和再生医学中的治疗潜力，强调了其在骨再生中的应用。
