Enhancement of differentiation and mineralization of human dental pulp stem cells via TGF-β signaling in low-level laser therapy using Er:YAG lasers

IF 2.6 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences Pub Date : 2025-01-19 DOI:10.1016/j.job.2025.100617

Ryo Yoshida , Kazuyuki Kobayashi , Kazuo Onuma , Ryuji Yamamoto , Risako Chiba-Ohkuma , Takeo Karakida , Shunjiro Yamakawa , Noriyasu Hosoya , Yasushi Yamazaki , Yasuo Yamakoshi

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引用次数: 0

Abstract

Objectives

Low-level laser therapy (LLLT) using an erbium-doped yttrium aluminum garnet (Er:YAG) laser provides a non-invasive approach applicable to various dental treatments. Here, we investigated the effects of Er:YAG laser irradiation on human dental pulp stem cells (hDPSCs) in an in vitro experiment.

Methods

The hDPSCs were categorized into four groups: laser-irradiated with activators (VLT: activated vitamin D₃, bone morphogenetic protein receptor inhibitor, and transforming growth factor-beta (TGF-β)) (LLLT(+)VLT), laser-irradiated without activators (LLLT(+)-only), non-irradiated with activators (LLLT(−)VLT), and non-irradiated without activators (control). Cell proliferation, hard tissue differentiation, TGF-β signaling pathway activity, mineralization induction, and gene expression levels were assessed using several approaches, including cell proliferation assays, ALP assays, western blotting, Alizarin Red S staining, X-ray diffraction, and quantitative polymerase chain reaction.

Results

Cell proliferation was similar between the LLLT(+)-only and control groups. The ALP activity was significantly higher in LLLT(+)VLT group than in LLLT(−)VLT group (p < 0.05); however, it was suppressed by TGF-β signaling inhibitors. Western blotting showed enhanced SMAD3 phosphorylation in the LLLT(+)VLT group. The mineralization nodules and mRNA levels of matrix vesicle marker genes were significantly higher in LLLT(+)VLT group, and the nodules were partially composed of hydroxyapatite. The hard tissue formation marker gene expression in LLLT(+)VLT group was significantly higher (p < 0.05) than that in the LLLT(+)-only and control groups; however, it was unchanged or suppressed compared with that in LLLT(−)VLT group.

Conclusions

LLLT using an Er:YAG laser, combined with VLT, may promote the differentiation of hDPSCs into hard tissue-forming cells and enhance mineralization.

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TGF-β信号在Er:YAG低水平激光治疗中促进人牙髓干细胞分化和矿化

目的：使用掺铒钇铝石榴石（Er:YAG）激光器进行低水平激光治疗（LLLT），为各种牙科治疗提供了一种无创的方法。在体外实验中，我们研究了Er:YAG激光照射对人牙髓干细胞（hDPSCs）的影响。方法：将hDPSCs分为激光照射激活剂组（VLT：活化维生素D3、骨形态发生蛋白受体抑制剂和转化生长因子β (TGF-β)）（LLLT(+)VLT）、激光不照射激活剂组（LLLT(+)-）、不照射激活剂组（LLLT(-)VLT）、不照射激活剂组（对照）。采用多种方法评估细胞增殖、硬组织分化、TGF-β信号通路活性、矿化诱导和基因表达水平，包括细胞增殖试验、ALP试验、western blotting、茜素红S染色、x射线衍射和定量聚合酶链反应。结果：LLLT（+）组与对照组细胞增殖无明显差异。LLLT(+)VLT组ALP活性显著高于LLLT(-)VLT组（p < 0.05）；而TGF-β信号抑制剂可抑制其表达。Western blotting显示，LLLT(+)VLT组SMAD3磷酸化增强。LLLT(+)VLT组矿化结节及基质囊泡标记基因mRNA水平显著升高，且矿化结节部分由羟基磷灰石组成。硬组织形成标记基因在LLLT(+)VLT组的表达量显著高于LLLT（+）组和对照组（p < 0.05）；但与LLLT(-)VLT组相比，它没有变化或受到抑制。结论：Er:YAG激光联合VLT可促进hDPSCs向硬组织形成细胞分化，并增强矿化作用。

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