可见光通过光诱导生物调节诱导鼠骨髓间充质干细胞的成脂分化。
Adipogenic differentiation of murine bone marrow mesenchymal stem cells induced by visible light via photo- induced biomodulation.
机构信息
Department of Chemistry, Center of Nanotechnology and Tissue Engineering - Photobiology and Photomedicine Research Group, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, Ribeirão Preto, University of São Paulo, 14040-901, Brazil.
Center for Cell Therapy and Regional Blood Center, National Institute of Science and Technology in Stem Cell and Cell Therapy, Medical School, University of São Paulo, Ribeirão Preto 14051-140, Brazil.
出版信息
Photodiagnosis Photodyn Ther. 2019 Mar;25:119-127. doi: 10.1016/j.pdpdt.2018.11.013. Epub 2018 Nov 17.
BACKGROUND
Bone marrow mesenchymal stem cells (BM-MSCs) are undifferentiated cells that can proliferate and differentiate into specialized cells for tissue self-repair. Low-level laser (LLL) can induce biomodulatory effects such as cellular proliferation, differentiation, and migration. We investigated the biomodulatory effects of the photoactive compound chloroaluminum phthalocyanine nanoemulsion (AlClPc/NE) on the adipogenic differentiation of BM-MSCs, when combined with LLL (AlClPc/NE-LLL).
METHODS
The BM-MSCs used in this work were isolated from green fluorescent protein-positive (GFP) C57BL6 mice. Cells were first treated with AlClPc/NE, a well-designed photoactive nano-drug and were then subjected to in vitro expansion, morphological and immunophenotypic characterization, and cellular cytotoxicity analysis. Subsequently, BM-MSCs were induced to differentiate into adipocytes by photo-induced biomodulation with AlClPc/NE-LLL.
RESULTS
Our results showed that the isolated cell population was consistent with murine BM-MSCs. The cellular cytotoxicity analysis revealed that the optimal nanoemulsion dose to induce BM-MSC biomodulation was 5.0 μmol/L. Twenty-four hours following treatment with AlClPc/NE, BM-MSC were subjected to visible light irradiation of 20 mJ/cm at 670 nm. Six days after photo-induced biomodulation, cells maintained high GFP expression level, and expressed detectable mRNA levels of adipogenic genes (lipoprotein lipase and PPARγ); formation of lipid vacuoles was observed, and the cells did not show any tumorigenic potential in vivo.
CONCLUSIONS
Our results indicated that photo-induced biomodulation via visible light using AlClPc/NE and LLL can induce adipogenic differentiation of murine BM-MSCs. Therefore, cell therapy with BM-MSCs and photo-induced biomodulation may contribute to the development of new therapeutic strategies that are faster and more effective than traditional methods to trigger MSC differentiation.
背景
骨髓间充质干细胞（BM-MSCs）是未分化的细胞，可以增殖并分化为组织自我修复的特化细胞。低水平激光（LLL）可以诱导细胞增殖、分化和迁移等生物调节效应。我们研究了光活性化合物氯铝酞菁纳米乳液（AlClPc/NE）与 LLL（AlClPc/NE-LLL）联合应用对 BM-MSCs 成脂分化的生物调节作用。
方法
本工作中使用的 BM-MSCs 分离自绿色荧光蛋白阳性（GFP）C57BL6 小鼠。细胞先用 AlClPc/NE 处理，这是一种设计良好的光活性纳米药物，然后进行体外扩增、形态和免疫表型特征以及细胞毒性分析。随后，通过 AlClPc/NE-LLL 的光诱导生物调节作用将 BM-MSCs 诱导分化为脂肪细胞。
结果
我们的结果表明，分离的细胞群体与鼠 BM-MSCs 一致。细胞毒性分析显示，诱导 BM-MSC 生物调节的最佳纳米乳液剂量为 5.0 μmol/L。用 AlClPc/NE 处理 24 小时后，用 670nm 的 20mJ/cm 可见光照射 BM-MSC。光诱导生物调节 6 天后，细胞保持高 GFP 表达水平，并检测到脂肪生成基因（脂蛋白脂肪酶和 PPARγ）的可检测 mRNA 水平；观察到脂质空泡的形成，并且细胞在体内没有显示出任何致瘤潜力。
结论
我们的结果表明，使用 AlClPc/NE 和 LLL 的可见光光诱导生物调节可以诱导鼠 BM-MSCs 的成脂分化。因此，BM-MSCs 细胞治疗和光诱导生物调节可能有助于开发比传统方法更快、更有效的新治疗策略，以触发 MSC 分化。
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