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Supplement of nitric oxide through calcium carbonate-based nanoparticles contributes osteogenic differentiation of mouse embryonic stem cells.
Tissue & Cell ( IF 2.6 ) Pub Date : 2020-05-24 , DOI: 10.1016/j.tice.2020.101390
Seong Yeong An 1 , Hong Jae Lee 1 , Sang Cheon Lee 1 , Jung Sun Heo 1
Affiliation  

This study investigated the delivery of S-nitrosothiol (GSNO) as a nitric oxide (NO) donor loaded into calcium carbonate-based mineralized nanoparticles (GSNO-MNPs) to regulate cell signaling pathways for the osteogenic differentiation of mouse embryonic stem cells (ESCs). GSNO-MNPs were prepared by an anionic block copolymer template-mediated calcium carbonate (CaCO3) mineralization process in the presence of GSNO. GSNO-MNPs were spherical and had a narrow size distribution. GSNO was stably loaded within the MNPs without denaturation. TEM analysis also demonstrated the localization of GSNO-MNPs within membrane-bound structures in the cell, indicating the successful introduction of GSNO-MNPs into the cytosol of ESCs. Intracellular levels of NO and cGMP were significantly increased upon treatment with GSNO-MNPs, compared with the control group. When cells were exposed to GSNO-MNPs, the effects of nanoparticles on cell viability were not statistically significant. GSNO-MNPs treatment increased ALP activity assay and intracellular calcium levels. Real-time RT-PCR also revealed highly increased expression levels of the osteogenic target genes ALP, osteocalcin (OCN), and osterix (OSX) in GSNO-MNP-treated ESCs. The protein levels of OSX and Runt-related transcription factor 2 (RUNX2) showed similar patterns of expression based on real-time RT-PCR. These results indicate that GSNO-MNPs influenced the osteogenic differentiation of ESCs. Transcriptome profiling identified several significantly enriched and involved biological networks, such as RAP1, RAS, PI3K-AKT, and MAPK signaling pathways. These findings suggest that GSNO-MNPs can modulate osteogenic differentiation in ESCs via complex molecular pathways.



中文翻译:

通过基于碳酸钙的纳米颗粒补充一氧化氮有助于小鼠胚胎干细胞的成骨分化。

这项研究调查了将S-亚硝基硫醇(GSNO)作为一氧化氮(NO)供体的传递方式,该供体被装载到基于碳酸钙的矿化纳米颗粒(GSNO-MNPs)中,以调节细胞信号传导途径,用于小鼠胚胎干细胞(ESCs)的成骨分化。 。GSNO-MNPs是通过阴离子嵌段共聚物模板介导的碳酸钙(CaCO 3)在GSNO存在下的矿化过程。GSNO-MNP为球形,尺寸分布较窄。GSNO被稳定地装载在MNP内而没有变性。TEM分析还证实了GSNO-MNP在细胞膜结合结构内的定位,表明将GSNO-MNP成功地引入了ESC的细胞质中。与对照组相比,用GSNO-MNPs治疗后,细胞内NO和cGMP水平显着增加。当细胞暴露于GSNO-MNPs时,纳米颗粒对细胞生存力的影响没有统计学意义。GSNO-MNPs处理增加了ALP活性测定和细胞内钙水平。实时RT-PCR还显示了在GSNO-MNP处理的ESC中成骨靶基因ALP,骨钙蛋白(OCN)和osterix(OSX)的表达水平大大提高。OSX和Runt相关转录因子2(RUNX2)的蛋白质水平显示基于实时RT-PCR的相似表达模式。这些结果表明,GSNO-MNPs影响ESCs的成骨分化。转录组谱分析确定了几个显着丰富和参与的生物网络,例如RAP1,RAS,PI3K-AKT和MAPK信号通路。这些发现表明,GSNO-MNP可以通过复杂的分子途径调节ESC中的成骨分化。和MAPK信号通路。这些发现表明,GSNO-MNP可以通过复杂的分子途径调节ESC中的成骨分化。和MAPK信号通路。这些发现表明,GSNO-MNP可以通过复杂的分子途径调节ESC中的成骨分化。

更新日期:2020-05-24
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