Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Department of General Surgery, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Photobiomodul Photomed Laser Surg. 2022 Aug;40(8):543-553. doi: 10.1089/photob.2022.0003. Epub 2022 Jul 28.
This study aimed to verify the effect of photobiomodulation therapy (PBMT) with a wavelength of 532 nm on the proliferation and differentiation of tendon-derived stem cells (TDSCs) of Sprague-Dawley (SD) rats. The combination of PBMT and stem cell transplantation with TDSCs provides a new treatment strategy for tendon injury. Nevertheless, the effect of PBMT on the biological behavior of TDSCs and its internal mechanisms remain unclear. TDSCs were isolated from Achilles tendons of SD rats and identified by cell morphology and flow cytometric analysis. Energy density gradient experiment was performed to determine the ideal energy. Then, TDSCs were treated with PBMT using a wavelength of 532 nm at a fluence of 15 J/cm in 532 nm laser group, and the TDSC in control group were not treated with 532 nm laser. Cell response after irradiation was observed to ascertain cell morphology and cell proliferation in the 532 nm laser group and the control group. The RNA expression levels of the key genes of TDSC differentiation, including scleraxis (), tenomodulin (), Mohawk homeobox (), Decorin (), peroxisome proliferator-activated receptor gamma (), SRY-box transcription factor 9 (), and RUNX family transcription factor 2 (), were detected by reverse transcription-polymerase chain reaction. Then, gene chip microarray was used to detect the expression of differential genes after 532 nm laser intervention in TDSCs, and the target genes were screened out to verify the role in this process and . When the 532 nm laser energy density was 15 J/cm, the proliferation capacity of TDSCs was improved (2.73 ± 0.24 vs. 1.81 ± 0.71, < 0.05), and the expression of genes related to tenogenic differentiation of TDSCs was significantly increased ( < 0.01). After RNA sequencing and bioinformatics analyses, we speculated that nuclear receptor subfamily 4 group A member 1 () was involved in the tenogenic differentiation process of TDSCs regulated by 532 nm laser treatment. Subsequent experiments confirmed that regulated the expression of the tenogenic differentiation genes and in TDSCs. A 532 nm laser with 15 J/cm regulated the process of TDSC proliferation and upregulated to stimulate tenogenic differentiation.
本研究旨在验证 532nm 波长的光生物调节疗法（PBMT）对 SD 大鼠肌腱衍生干细胞（TDSCs）增殖和分化的影响。PBMT 联合 TDSC 干细胞移植为肌腱损伤提供了一种新的治疗策略。然而，PBMT 对 TDSC 生物学行为的影响及其内在机制尚不清楚。从 SD 大鼠跟腱中分离 TDSC，通过细胞形态学和流式细胞分析进行鉴定。通过能量密度梯度实验确定理想的能量。然后，在 532nm 激光组中，用 532nm 波长、15J/cm 的光辐照 TDSC，对照组 TDSC 不进行 532nm 激光处理。观察照射后细胞的形态和细胞增殖，以确定 532nm 激光组和对照组的细胞反应。采用逆转录聚合酶链反应检测 TDSC 分化的关键基因，包括 Scleraxis ()、Tenomodulin ()、Mohawk Homeobox ()、Decorin ()、过氧化物酶体增殖物激活受体 γ ()、SRY-box 转录因子 9 () 和 RUNX 家族转录因子 2 () 的 RNA 表达水平。然后，采用基因芯片微阵列检测 532nm 激光干预后 TDSC 中差异基因的表达，筛选出靶基因，验证其在该过程中的作用。当 532nm 激光能量密度为 15J/cm 时，TDSCs 的增殖能力提高（2.73±0.24 比 1.81±0.71， <0.05），TDSCs 肌腱分化相关基因的表达显著增加（ <0.01）。经过 RNA 测序和生物信息学分析，我们推测核受体亚家族 4 组 A 成员 1 () 参与了 532nm 激光处理调节的 TDSC 肌腱分化过程。随后的实验证实，在 TDSCs 中，调节肌腱分化基因的表达。15J/cm 的 532nm 激光调节 TDSC 增殖过程，并上调 来刺激肌腱分化。
