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632.8nm 低强度光生物调节体外增加大鼠骨髓间充质干细胞中 tgfβ3、col2a1 和 sox9 基因的表达。

Low-intensity photobiomodulation at 632.8 nm increases tgfβ3, col2a1, and sox9 gene expression in rat bone marrow mesenchymal stem cells in vitro.

机构信息

Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky ave. 4, St. Petersburg, 195427, Russia.

Vreden National Medical Research Center of Traumatology and Orthopedics, Akademika Baikova st. 8, St. Petersburg, 195427, Russia.

出版信息

Lasers Med Sci. 2022 Feb;37(1):435-441. doi: 10.1007/s10103-021-03279-0. Epub 2021 Feb 26.

Abstract

The high incidence of cartilage destructions, as well as the social and economic importance of this pathology attracted great interest to the problem. At the present time, some data are available about the 632.8 nm low-intensity laser photobiomodulation positive effect on the cartilage tissue proliferation. The effect of this wavelength laser irradiation on the mesenchimal stem cell (MSC) differentiation in the chondrogenic direction was studied. The main aim of this work was to assess the low-intensity photobiomodulation effect on chondrogenesis. In this experiment, the cell model was used to compare the photobiomodulation and cytokine Tgfβ3 (transforming growth factor β 3) effects. Bone marrow MSCs were isolated from Wistar rats and cultured for the third passage. Chondrogenic effects of low-intensity He-Ne laser photobiomodulation and cytokine Tgfβ3 (10 ng/μL) were analyzed and compared after 21 days. The radiation source was the standard LGN-208 helium-neon (He-Ne) laser (632.8 nm, 1.7 mWt). Irradiation was performed cyclically for 15 min with 45-min pauses. The increase of the responsible for chondrogenesis (col2a1, tgfβ3, and sox9) main gene expression under the photobiomodulation at 632.8 nm was evaluated in comparison with Tgfβ3 effect. The tgfβ3, col2a1, and sox9 gene expression increase was obtained in two experimental groups: using the laser photobiomodulation and cytokine Tgfβ3 effect. Gene expression levels of tgfβ3, col2a1, and sox9 were measured using real-time polymerase chain reaction (RT-PCR) according to the -ΔΔCt method. It was found that the responsible for chondrogenesis genes expression (tgfβ3, col2a1, sox9) increased under the action of specific laser photobiomodulation during the observation period (from 0 to 21 days). The chondrogenic differentiation effect under the laser irradiation is less significant than Tgfβ3 cytokine effect.

摘要

软骨破坏的高发病率以及这种病理学的社会和经济重要性引起了人们对该问题的极大兴趣。目前,已经有一些关于 632.8nm 低强度激光光生物调节对软骨组织增殖的积极影响的数据。研究了该波长激光照射对间充质干细胞(MSC)向软骨分化的影响。这项工作的主要目的是评估低强度光生物调节对软骨形成的影响。在这项实验中,使用细胞模型比较了光生物调节和细胞因子 TGFβ3(转化生长因子β3)的作用。从 Wistar 大鼠分离骨髓间充质干细胞并培养至第三代。分析并比较了低强度氦氖激光光生物调节和细胞因子 TGFβ3(10ng/μL)在 21 天后对软骨形成的影响。辐射源为标准 LGN-208 氦氖(He-Ne)激光(632.8nm,1.7mWt)。以 15min 为一个周期进行循环照射,每次照射后休息 45min。与 TGFβ3 作用相比,评估了在 632.8nm 光生物调节下负责软骨形成(col2a1、tgfβ3 等)的主要基因表达的增加。在两个实验组中都获得了 tgfβ3、col2a1 和 sox9 基因的表达增加:使用激光光生物调节和细胞因子 TGFβ3 作用。根据 -ΔΔCt 法,使用实时聚合酶链反应(RT-PCR)测量 tgfβ3、col2a1 和 sox9 基因的表达水平。结果发现,在观察期内(从 0 到 21 天),特定激光光生物调节作用下,负责软骨形成的基因表达(tgfβ3、col2a1、sox9)增加。激光照射下的软骨分化效果不如细胞因子 TGFβ3 明显。

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