Constantin Alina, Dumitrescu Madalina, Mihai Corotchi Maria Cristina, Jianu Dana, Simionescu Maya
Institute of Cellular Biology and Pathology "Nicolae Simionescu", Bucharest, Romania.
PROESTETICA Medical Center, Bucharest, Romania.
Lasers Med Sci. 2017 Jan;32(1):117-127. doi: 10.1007/s10103-016-2093-6. Epub 2016 Oct 20.
CO laser has a beneficial effect on stem cells by mechanisms that are not clearly elucidated. We hypothesize that the effect of fractional CO laser on human adipose-derived stem cells (ADSC) could be due to changes in redox homeostasis and secretion of factors contributing to cellular proliferation and angiogenic potential. ADSC incubated in medium containing 0.5 or 10 % FBS were exposed to a single irradiation of a 10,600-nm fractional CO laser; non-irradiated ADSC were used as control. Viability/proliferation of ADSC was assessed by MTT assay; the intracellular reactive oxygen species (ROS) levels and the mitochondrial membrane potential (∆Ψ) were determined with DCFH-DA and JC-1 fluorescent probes, respectively. Molecules secreted by ADSC in the medium were determined by ELISA assay, and their capacity to support endothelial tube-like formation by the Matrigel assay. The results showed that compared to controls, ADSC kept in low FBS medium and irradiated with CO laser at 9 W exhibited: (a) increased proliferation (∼20 %), (b) transient increase of mitochondrial ROS and the capacity to restore Δψ after rotenone induced depolarization, and (c) augmented secretion in the conditioned medium of MMP-2 (twofold), MMP-9 (eightfold), VEGF (twofold), and adiponectin (∼50 %) that have the capacity to support angiogenesis of endothelial progenitor cells. In conclusion, the mechanisms underlying the benefic effect of CO laser on ADSC are the activation of the redox pathways which increases cell proliferation and enhances secretion of angiogenic molecules. These results explain, in part, the mechanisms involved in the increased regenerative potential of CO laser-exposed ADSC that could be exploited for clinical applications.
CO激光通过尚未完全阐明的机制对干细胞产生有益作用。我们推测,分数CO激光对人脂肪来源干细胞(ADSC)的作用可能归因于氧化还原稳态的变化以及有助于细胞增殖和血管生成潜能的因子分泌。将在含有0.5%或10%胎牛血清(FBS)的培养基中培养的ADSC暴露于波长为10600nm的分数CO激光单次照射下;未照射的ADSC用作对照。通过MTT法评估ADSC的活力/增殖;分别用DCFH-DA和JC-1荧光探针测定细胞内活性氧(ROS)水平和线粒体膜电位(∆Ψ)。通过ELISA测定法测定ADSC在培养基中分泌的分子,并通过基质胶测定法测定它们支持内皮管样形成的能力。结果表明,与对照组相比,在低FBS培养基中培养并用9W的CO激光照射的ADSC表现出:(a)增殖增加(约20%),(b)线粒体ROS短暂增加以及在鱼藤酮诱导的去极化后恢复∆Ψ的能力,以及(c)条件培养基中MMP-2(两倍)、MMP-9(八倍)、VEGF(两倍)和脂联素(约50%)的分泌增加,这些分子具有支持内皮祖细胞血管生成的能力。总之,CO激光对ADSC有益作用的潜在机制是氧化还原途径的激活,这增加了细胞增殖并增强了血管生成分子的分泌。这些结果部分解释了CO激光照射的ADSC再生潜能增加所涉及的机制,这些机制可用于临床应用。