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软骨细胞样细胞经蓝光照射后的软骨形成相关 mRNA 表达。

Chondrogenic mRNA expression in prechondrogenic cells after blue laser irradiation.

机构信息

Frontier Research Center, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.

出版信息

J Photochem Photobiol B. 2010 Mar 8;98(3):211-5. doi: 10.1016/j.jphotobiol.2010.01.008. Epub 2010 Jan 25.

Abstract

Low-level laser therapy (LLLT) has been used as a method for biostimulation. Cartilage develops through the differentiation of mesenchymal cells into chondrocytes, and differentiated chondrocytes in articular cartilage maintain cartilage homeostasis by synthesizing cartilage-specific extracellular matrix. The aim of this study is to evaluate the enhancement of chondrocyte differentiation and the expression levels of chondrogenic mRNA in prechondrogenic ATDC5 cells after laser irradiation. For chondrogenic induction, ATDC5 cells were irradiated with a blue laser (405 nm, continuous wave) at 100 mW/cm(2) for 180 s following incubation in chondrogenic differentiation medium. Differentiation after laser irradiation was quantitatively evaluated by the measurement of total collagen contents and chondrogenesis-related mRNAs. The total amount of collagen and mRNA levels of aggrecan, collagen type II, SOX-9, and DEC-1 were increased relative to those of a non-laser irradiated group after 14 days of laser irradiation. On the other hand, Ap-2alpha mRNA, a negative transcription factor of chondrogenesis, was dramatically decreased after laser irradiation. In addition, intracellular reactive oxygen species (ROS) were generated after laser irradiation. These results, for the first time, provide functional evidence that mRNA expression relating to chondrogenesis is increased, and Ap-2alpha is decreased immediately after laser irradiation. As this technique could readily be applied in situ to control the differentiation of cells at an implanted site within the body, this approach may have therapeutic potential for the restoration of damaged or diseased tissue.

摘要

低水平激光疗法(LLLT)已被用作生物刺激的一种方法。软骨通过间充质细胞分化为软骨细胞而发育,关节软骨中的分化软骨细胞通过合成软骨特异性细胞外基质来维持软骨的稳态。本研究的目的是评估激光照射后对前软骨细胞 ATDC5 细胞的软骨细胞分化和软骨形成相关 mRNA 表达水平的增强作用。为了进行软骨诱导,在软骨分化培养基中孵育后,用蓝色激光(405nm,连续波)以 100mW/cm2 照射 ATDC5 细胞 180s。通过测量总胶原蛋白含量和与软骨形成相关的 mRNAs 来定量评估激光照射后的分化。与未激光照射组相比,激光照射 14 天后,胶原蛋白总量和聚集蛋白聚糖、II 型胶原、SOX-9 和 DEC-1 的 mRNA 水平增加。另一方面,软骨形成的负转录因子 Ap-2alpha 的 mRNA 水平在激光照射后显著降低。此外,激光照射后会产生细胞内活性氧(ROS)。这些结果首次提供了功能证据,表明与软骨形成相关的 mRNA 表达增加,并且 Ap-2alpha 在激光照射后立即减少。由于该技术可以很容易地应用于体内植入部位的细胞分化的原位控制,因此该方法可能具有恢复受损或患病组织的治疗潜力。

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