MicroRNA-193 Pro-Proliferation Effects for Bone Mesenchymal Stem Cells After Low-Level Laser Irradiation Treatment Through Inhibitor of Growth Family, Member 5

Abstract

The enhanced proliferation of mesenchymal stem cells (MSCs) can be helpful for the clinical translation of cell therapy. Low-level laser irradiation (LLLI) has been demonstrated as regulating MSC proliferation. MicroRNAs (miRNAs) are involved in various pathophysiologic processes in stem cells, but the role of miRNAs in the LLLI-based promotion of MSC proliferation remains unclear. We found that the proliferation level and cell cycle-associated genes in MSCs were increased after LLLI treatment in a time-dependent manner. Microarray assays revealed subsets of miRNAs to be differentially regulated, and these dynamic changes were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) after LLLI. miR-193 was the most highly up-regulated miRNA, and the change in it was related with the proliferation level. Gain-loss function experiments demonstrated that miR-193 could regulate the proliferation of MSCs, including human's and rat's, but could not affect the apoptosis and differentiation level. Blockade of miR-193 repressed the MSC proliferation induced by LLLI. By qRT-PCR, we found that miR-193, in particular, regulated cyclin-dependent kinase 2 (CDK2) expression. Bioinformatic analyses and luciferase reporter assays revealed that inhibitor of growth family, member 5 (ING5) could be the best target of miR-193 to functionally regulate proliferation and CDK2 activity, and the mRNA and protein level of ING5 was regulated by miR-193. Furthermore, the ING5 inhibited by small interfering RNA (siRNA) could up-regulate the proliferation of MSCs and the expression of CDK2. Taken together, these results strongly suggest that miR-193 plays a critical part in MSC proliferation in response to LLLI stimulation, which is potentially amenable to therapeutic manipulation for clinical application.

Effect of low-level laser therapy on mesenchymal stem cell proliferation: a systematic review

“…Table 2 summarizes the main parameters of the lasers used in the articles. There was a preference for using lower doses, being the dose of 0.05 J/cm 2 as the lowest applied [27], 0.5 J/cm 2 the most used [1,9,25,28], and 42 J/cm 2 the highest dose [27]. Within the visible light spectrum, the lower power used in the experiments was 0.02 mW [27] and the highest 119 mW [8], while in the invisible light spectrum, the lower power was 50 mW [10] and the highest 800 mW [30].…”

“…With respect to the intervals of analysis of cell proliferation, the minimum time point used was 0 h [1,8,9,25,26,28] after irradiation and the maximum was 4 weeks after irradiation [4].…”

MicroRNA delivery for regenerative medicine

“…In comparison to viral transduction, nonviral delivery systems such as lipid-based (lipoplex) and polymer-based (polyplex) systems impose a lower degree of genetic perturbation and may be more beneficial to the clinical application of stem cell differentiation. Many cationic lipid-based systems can be selected from a number of commercially available products (Table 3), including Lipofectamine® [139, 141, 142], siPORT ™ [143], Oligofectamine ™ [144], MaxSuppressor ™ In Vivo RNA-LANCEr II [76], HiPerFect [77], Lullaby [145], DharmaFECT® [146], SilentFect ™ [101] and NeuroPorter ™ [147]. Many investigations have validated the possible delivery of miRNA by lipoplexes.…”

The influence of continuous low-intensity laser radiation at the red (635 nm) and green (525 nm) wavelengths on the human mesenchymal stem cells in vitro: a review of the literature and original investigations

“…Исходя из этого рассчитывалась плотность мощности (ПМ) и ЭП для каждого вари-анта освечивания. Требование к равномерной за-светке всей поверхности чашки связано с обеспече-нием лучшего эффекта, чем при точечном воздей-ствии на часть культуры клеток [22,45].…”

“…Несмотря на значительный разброс энергетиче-ских параметров лазерного воздействия в исследо-ваниях по изучению влияния НИЛИ на пролифера-цию МСК, данные литературы в отношении опти-мальной ЭП для непрерывного НИЛИ с длиной волны 633-635 нм находятся в достаточно узком диапазоне -0,3-0,5 Дж/см 2 , при этом ПМ варьи-рует в значительно более широких пределах, от 3-6,61 мВт/см 2 [18][19][20][21][22][23] a -фотография клеток в проходящем свете. Ув.…”