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增强成骨细胞分化和骨修复:光生物调节对脂肪基质细胞的启动效应。

Enhancing osteoblast differentiation and bone repair: The priming effect of photobiomodulation on adipose stromal cells.

机构信息

School of Dentistry, University of Sao Paulo, Sao Paulo, Brazil.

Bone Research Lab, School of Dentistry of Ribeirao Preto, University of Sao Paulo, Sao Paulo, Brazil.

出版信息

J Photochem Photobiol B. 2024 Nov;260:113040. doi: 10.1016/j.jphotobiol.2024.113040. Epub 2024 Oct 2.

Abstract

Cellular therapy using adipose tissue-derived mesenchymal stromal cells (at-MSCs) has garnered attention for the treatment of bone defects. Therefore, preconditioning strategies to enhance the osteogenic potential of at-MSCs could optimize cell therapy outcomes, and photobiomodulation (PBM) therapy has emerged as an effective, noninvasive, and low-cost alternative. This study explored the impacts of PBM on at-MSCs differentiation and the subsequent repair of bone defects treated with cell injection. Rat at-MSCs were cultured and irradiated (at-MSCs) following the PBM protocol (660 nm; 20 mW; 0.714 W/cm; 0.14 J; 5 J/cm). Cellular differentiation was assessed based on the expression of gene and protein markers. Reactive oxygen species (ROS) were detected using fluorescence. At-MSCs were injected into 5-mm calvarial lesions, and bone formation was analyzed using micro-CT and histological evaluations. At-MSCs were used as control. Data were analyzed using the ANOVA or t-test. At-MSCs exhibited high levels of gene and protein runt-related transcription factor-2 (Runx2) and alkaline phosphatase (Alp) expression. PBM increased ALP activity and significantly reduced ROS levels. In addition, PBM increased the expression of Wnt pathway-associated genes. In vivo, there was an increase in the morphometric parameters, including bone volume, percentage of bone volume, bone surface area, and trabecular number, in at-MSCs-treated defects compared with those in the control. These findings suggest that PBM enhances the osteogenic potential of at-MSCs, thereby supporting the advancement of improved cellular therapies for bone regeneration.

摘要

利用脂肪组织来源的间充质基质细胞(at-MSCs)进行细胞治疗已引起人们对治疗骨缺损的关注。因此,预处理策略可增强 at-MSCs 的成骨潜力,从而优化细胞治疗效果,光生物调节(PBM)治疗已成为一种有效、无创且低成本的替代方法。本研究探讨了 PBM 对 at-MSCs 分化的影响,以及随后通过细胞注射治疗骨缺损的修复。大鼠 at-MSCs 经培养后,按照 PBM 方案进行照射(660nm;20mW;0.714W/cm;0.14J;5J/cm)。通过基因和蛋白标志物的表达评估细胞分化。使用荧光检测活性氧(ROS)。将 at-MSCs 注射到 5mm 颅骨缺损处,并用 micro-CT 和组织学评估分析骨形成。at-MSCs 作为对照。使用 ANOVA 或 t 检验分析数据。at-MSCs 表现出高水平的基因和蛋白 runt 相关转录因子 2(Runx2)和碱性磷酸酶(Alp)表达。PBM 增加了 ALP 活性,并显著降低了 ROS 水平。此外,PBM 增加了 Wnt 通路相关基因的表达。在体内,与对照组相比,at-MSCs 治疗的缺陷中,形态计量学参数(包括骨体积、骨体积百分比、骨表面面积和小梁数量)增加。这些发现表明,PBM 增强了 at-MSCs 的成骨潜力,从而支持改进的细胞疗法在骨再生方面的进展。

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