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Evaluation of Redox Activity of Human Myocardium‐derived Mesenchymal Stem Cells by Scanning Electrochemical Microscopy
Electroanalysis ( IF 2.7 ) Pub Date : 2020-02-14 , DOI: 10.1002/elan.201900723
Jurate Petroniene 1 , Inga Morkvenaite‐Vilkonciene 2, 3 , Rokas Miksiunas 4 , Daiva Bironaite 4 , Almira Ramanaviciene 5 , Lina Mikoliunaite 1 , Aura Kisieliute 1 , Kestutis Rucinskas 6 , Vilius Janusauskas 6 , Ieva Plikusiene 1 , Siegfried Labeit 7 , Arunas Ramanavicius 1, 8
Affiliation  

In this study the redox activity of human myocardium‐derived mesenchymal stem cells (hmMSC) were investigated by redox‐competition (RC‐SECM) and generation‐collection (GC‐SECM) modes of scanning electrochemical microscopy (SECM), using 2‐methylnaphthalene‐1,4‐dione (menadione, MD) as a redox mediator. The redox activity of human healthy and dilated hmMSCs was evaluated by measuring reduction of MD. Measurements were performed by approaching and retracting the UME from the surface of growing hmMSC cells. The current study shows that the RC‐SECM mode can be applied to investigate integrity of cell membranes, whereas the most promising results were observed by using the GC‐SECM mode and applying the Hill's equation for the calculation/fitting of dependencies of electrical current vs menadione concentration. The calculated apparent Michaelis constant (K M) for the production of menadiol (MDH2) in the pathological hmMSC cells was 14.4 folds higher compared to that of the healthy hmMSC revealing the lover redox activity of pathological cells. Moreover, the calculated Hill's coefficient n shows a negative cooperative binding between MD and healthy hmMSC and positive cooperative binding between MD and pathological hmMSC. It means that healthy hmMSC is of lower affinity to MD, which is also related to the better membrane integrity of healthy cells. Data of this study demonstrate that SECM can be applied to investigate intracellular redox and membrane changes ongoing in human dilated myocardium‐derived hmMSC in order to improve their functioning and further regenerative potential.

中文翻译:

扫描电化学显微镜评价人心肌间充质干细胞的氧化还原活性

在这项研究中,使用2-甲基萘通过扫描电化学显微镜(SECM)的氧化还原竞争(RC-SECM)和世代收集(GC-SECM)模式研究了人心肌来源的间充质干细胞(hmMSC)的氧化还原活性。 -1,4-二酮(menadione,MD)作为氧化还原介体。通过测量MD​​的降低来评估人类健康和扩张的hmMSC的氧化还原活性。通过从生长中的hmMSC细胞的表面接近和缩回UME进行测量。当前的研究表明RC‐SECM模式可用于研究细胞膜的完整性,而使用GC‐SECM模式并使用Hill方程计算/拟合电流甲萘醌浓度。计算出的生产薄荷醇(MDH 2)的表观米氏常数(K M)在病理hmMSC细胞中比健康hmMSC高14.4倍,揭示了病理细胞的爱好者氧化还原活性。此外,计算的希尔氏系数n表示MD与健康hmMSC之间为负协同结合,而MD与病理hmMSC之间为正协同结合。这意味着健康的hmMSC对MD的亲和力较低,这也与健康细胞的更好的膜完整性有关。这项研究的数据表明,SECM可用于研究人扩张型心肌源hmMSC中正在进行的细胞内氧化还原和膜变化,以改善其功能并进一步再生。
更新日期:2020-02-14
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