Electrical Stimulation Increases the Secretion of Cardioprotective Extracellular Vesicles from Cardiac Mesenchymal Stem Cells

Abstract

Clinical trials have shown that electric stimulation (ELSM) using either cardiac resynchronization therapy (CRT) or cardiac contractility modulation (CCM) approaches is an effective treatment for patients with moderate to severe heart failure, but the mechanisms are incompletely understood. Extracellular vesicles (EV) produced by cardiac mesenchymal stem cells (C-MSC) have been reported to be cardioprotective through cell-to-cell communication. In this study, we investigated the effects of ELSM stimulation on EV secretion from C-MSCs (C-MSCELSM). We observed enhanced EV-dependent cardioprotection conferred by conditioned medium (CM) from C-MSCELSM compared to that from non-stimulated control C-MSC (C-MSCCtrl). To investigate the mechanisms of ELSM-stimulated EV secretion, we examined the protein levels of neutral sphingomyelinase 2 (nSMase2), a key enzyme of the endosomal sorting complex required for EV biosynthesis. We detected a time-dependent increase in nSMase2 protein levels in C-MSCELSM compared to C-MSCCtrl. Knockdown of nSMase2 in C-MSC by siRNA significantly reduced EV secretion in C-MSCELSM and attenuated the cardioprotective effect of CM from C-MSCELSM in HL-1 cells. Taken together, our results suggest that ELSM-mediated increases in EV secretion from C-MSC enhance the cardioprotective effects of C-MSC through an EV-dependent mechanism involving nSMase2.

Extracellular Vesicles: A New Star for Gene Drug Delivery

“… 94 Zhang et al demonstrated that electrical stimulation method significantly increased the release of cardiac MSC-derived EVs, and the EVs obtained through a neutral sphingomyelinase 2 (nSMase2)-dependent mechanism had an enhanced cardioprotective effect in C-MSCs. 95 , 96 Our group has also developed a cellular nanoporation (CNP) technology 33 as shown in Figure 4 . In this work, cells were stimulated by transient electrical pulses to generate nanopores on the cell membrane, which in turn facilitated the loading of pDNA from the buffer into the cells.…”

Effects of Physical Cues on Stem Cell-Derived Extracellular Vesicles toward Neuropathy Applications

“…As a result, Hu et al highlighted the direct correlation between an upregulation in Ca 2+ ion concentration and the increased secretion of EVs [ 288 , 291 ]. Similarly, Zhang et al exposed cardiac-MSCs (C-MSCs) to low-level ES from 2 to 72 h. There was a significant increase in the nSMase2 protein levels, crucial to EV biogenesis and release [ 292 ]. Compared to C-MSCs (control), C-MSCs (ES) produced a 38% increase in EV particles/mL concentration and diameter [ 292 ].…”

“…Similarly, Zhang et al exposed cardiac-MSCs (C-MSCs) to low-level ES from 2 to 72 h. There was a significant increase in the nSMase2 protein levels, crucial to EV biogenesis and release [ 292 ]. Compared to C-MSCs (control), C-MSCs (ES) produced a 38% increase in EV particles/mL concentration and diameter [ 292 ]. Alternatively, with high voltage and exposure time, the hyperactive rate of nerve action potential proved to be damaging to the cell [ 291 ].…”

“…Alternatively, with high voltage and exposure time, the hyperactive rate of nerve action potential proved to be damaging to the cell [ 291 ]. A complete understanding of ES on the cell has yet to be fully understood; however, ES has been confirmed to alter cellular energy metabolism, morphology, phenotype, and Ca 2+ expression [ 291 , 292 ]. The optimization of ES parameters is necessary to ensure cellular proliferation and the secretion of glutamate and EVs without causing cellular damage or the ability to produce quality EVs.…”

“…Cardiac MSC 1.5 V/1.8 cm 2-72 h; 1.5 V/1.8 cm voltage, 0.5 Hz frequency, pulse width at 5 ms Cultured-cell pacer system (IonOptix) [292] The molecular mechanism of cellular differentiation and increased EV production following ES exposure is inconclusive [299]. However, various studies have concluded on the increased mobility of secreted vesicles that transport vesicular cargo, neurotransmitters, neuromodulators, hormones, and peptides [299].…”