. 2021 Feb 25;11(3):570. doi: 10.3390/nano11030570

Table 2.

Representative preclinical efficacy studies from the last five years, using extracellular vesicles as therapeutic agents for myocardial infarction.

Cell Source	Isolation Method	Animal Model	Dose	Administration Route and Time Post-MI	Reparative Effect	Molecule/Mechanism Involved	Ref
MSCs
Rat BM-MSCs	Total Exosome Isolation Kit (Invitrogen)	Rat, permanent	20 µg	IM; immediate	Improved cardiac function Reduced fibrosis Reduced inflammation	-	[62]
Mouse BM-MSCs	Density- gradient UC	Mouse, I/R	50 µg	IM; immediate after reperfusion	Reduced infarct size Alleviated inflammation (polarization of macrophages to M2 phenotype)	Inhibition of TLR4 by miR-182	[63]
Proinflammatory rat BM-MSCs	Density-gradient UC	Mouse, permanent	50 µg	IM; immediate	Reduced inflammation Anti-inflammatory macrophage polarization Reduced cardiomyocyte apoptosis	Suppression of NF-κB and regulation of AKT1/AKT2	[64]
BM-MSCs	UC	Rat, permanent	10 µg EVs (and 2×10⁶ BM-MSCs)	IM; at 30 min	Improved cardiac function Reduced infarct size and fibrosis Increased vascularization Reduced inflammation Enhanced recruitment of IV-infused MSCs	-	[65]
ATV-pre-treated rat BM-MSCs	UC	Rat, permanent	10 µg	IM; immediate	Improved cardiac function Reduced infarct size Decreased cardiomyocyte apoptosis Increased angiogenesis	lncRNA H19 and miR-675	[66]
Mouse BM-MSCs	UC	Mouse, permanent	-	IV; immediate and day 6	Improved cardiac function Angiogenesis Reduced fibrosis	miR-210 and Efna3 gene suppression	[67]
Mouse BM-MSCs	UC	Mouse, permanent	EVs derived from 2×10⁷ cells	IM; immediate	Improved cardiac function Angiogenesis Decreased scar size Reduced cardiomyocyte survival Activation of resident CPCs	miR-210	[68]
Rat BM-MSCs	Total Exosome Isolation Kit (Invitrogen)	Rat, I/R	5 µg	IM; prior to reperfusion	Decreased cardiomyocyte apoptosis Reduced infarct size Improved heart function by an enhanced autophagy	AMPK and AKT pathways	[69]
Mouse BM-MSCs	UC	Mouse, I/R	12.5 µg/ 5.62×10⁵ EVs	IM; 24h prior to ischemia	• Decreased infarct size	Reduced expression of pro-apoptotic genes PDCD4, PTEN, Peli1 and FasL via miR-21a-5p	[70]
Mouse BM-MSCs	UC	Mouse, permanent	200 µg	IM; immediate	Improved cardiac function Reduced infarct size	miR-125b	[71]
BM-MSCs	ExoQuick	Rat, permanent	-	IM; immediate	Reduced infarct size Alleviated cardiomyocyte apoptosis Improved cardiac function	miR-24	[72]
Rat ADSCs	UC	Rat, permanent	2.5×10¹² particles	IV; at 1h	Decreased fibrosis Decreased cell apoptosis Attenuated inflammation via anti-inflammatory macrophage polarization Improved cardiac function	S1P/SK1/S1PR1 activation	[73]
Rat ADSCs	Ultrafiltration and UC	Rat, I/R	400 µg	IV; at reperfusion	Reduced infarct area Attenuated apoptosis Reduced serum levels of cardiac damage markers	Wnt/β-catenin activation	[74]
Human umbilical cord MSCs	Density-gradient UC	Rat, permanent	400 µg and 800 µg	IV; once daily for 7 days	• Safety: no effect on hemolysis, no vascular and muscle stimulation, no side effects on hematology indexes, liver and renal function, and protective effect on weight loss	-	[75]
Human umbilical cord MSCs	ExoQuick-TC (System Biosciences)	Rat, permanent	400 µg	IM; immediate	Increased density of myofibroblasts Attenuated inflammation Reduced cardiomyocyte apoptosis	-	[76]
Human umbilical cord MSCs	Density-gradient UC	Rat, permanent	400 µg	IV; immediate	Improved cardiac function Increased cardiomyocyte survival	Upregulation of Smad7 by inhibition of miR-125b-5p	[77]
Cardiac MSCs	Precipitacion with PEG	Mouse, permanent	50 µg	IM; immediate	Improved cardiac function Increased scar thickness Angiogenesis Cardiomyocyte proliferation	-	[78]
CDCs
Human CDCs	Ultrafiltration and precipitation with PEG	Pig, I/R	7.5 mg	IC; 30 min after reperfusion IM; 30 min after reperfusion	Decreased infarct size and preserved LV function Reduced leukocyte infiltration Reduced fibrotic mass Higher arteriolar density	-	[79]
Porcine CDCs	Ultrafiltration followed by Field-Flow Fractionation	Pig, I/R	9.16 mg	IM; at 72h after reperfusion	Inconclusive; tendency to reduce infarct size and increase cardiac function Increased M2 macrophages	-	[80]
Human CDCs	Ultrafiltration and PEG precipitation	Pig, I/R	7.5 mg	IM; at 20 min after reperfusion	Preserved cardiac function Reduced microvascular occlusion Attenuated infarct size Reduced CD68⁺ macrophages infiltration	Regulation of gene expression by miRNA	[81]
Human CDCs	Ultrafiltration and precipitation with PEG	Rat, I/R	350 µg	IM; at 30 min after reperfusion	Preserved cardiac function Reduced infarcted area	-	[81]
Human CDCs	ExoQuick (precipitation)	Rat, permanent	250 µg	IM; at 4 weeks	Improved cardiac function Reduced scar mass Increased wall thickness Increased capillary and microvessel density	Regulation of gene expression by miRNA	[82]
CPCs
Human CPCs	Density-gradient UC	Mice, permanent	8 µg	IM; at 15 min	Reduced infarct size Increased proliferation of cardiomyocytes and endothelial cells	Activation of endoglin in endothelial cells	[83]
Rat CPCs	UC	Rat, I/R	5 µg/kg	IM; during reperfusion	Reduced infarct size Increased cardiac contractility	Decreased levels of collagen I, collagen III, vimentin and CTGF Regulation of gene expression via miRNA	[84]
Human CPCs	UC	Rat, permanent and I/R	10¹¹ particles	IM; at 1h after permanent ligation or at reperfusion	Increased cardiac function Reduced scar size Increased blood vessel density Decreased CD68⁺ macrophages	miR-146a-3p, miR-132, and miR-181a PAPP-A IGF-1	[85]
iPS
Human iPS	UC	Mouse, permanent	3×10¹⁰ particles	Transcutaneous echo-guided IM; at 3 weeks	• Increased cardiac function	Regulation of gene expression via miRNA	[86]
Human iPS	UC	Mouse, permanent	100 µg (10¹⁰ particles)	IM; at 2 days or 3 weeks	No detectable humoral or immune response Decreased pro-inflammatory monocytes and cytokines	-	[87]
Mouse iPS	UC	Mouse, I/R	100 µg	IM; at 48h after reperfusion	Preserved cardiac function Improved systolic infarct wall thickness Smaller LV end-systolic volume Reduced apoptosis in myocytes Increased capillary density No tumor formation	Regulation of gene expression via miRNA and metabolic regulation via protein delivery (in silico analysis)	[88]
ESC
Human ESC	UC	Mouse, permanent	20 µg	IM; immediate	Improved cardiac function and LV systolic dimension Reduced scar size Decreased cardiomyocyte apoptosis Higher number of endothelial cells	Targeting miR-497 by lncRNA MALAT1	[89]
Human ESC	UC	Mouse, permanent	-	Transcutaneous echo-guided IM; at 2-3 weeks	Decreased LV end-systolic and diastolic volume Reduced fibrosis Smaller cardiomyocytes	Gene regulation of DNA repair, cell survival, cell cycle progression and cardiomyocyte contractility (in silico)	[90]
Mouse ESC	UC	Mouse, permanent	-	IM; immediate	Enhanced contractility and decreased LV end-systolic diameter Increased capillary density Reduced apoptosis Elevated cardiomyocyte proliferation	Regulation of CPC cell cycle and association with proliferation and survival mediated by miR-294	[91]

ADSCs: adipose tissue-derived mesenchymal stem cells; ATV: atorvastatin; BM-MSCs: bone marrow-derived mesenchymal stromal cells; CDCs: cardiosphere-derived cells; CPCs: cardiac progenitor cells; CTGF: connective tissue growth factor; ESC: embryonic stem cells; EVs: extracellular vesicles; I/R: ischemia/reperfusion; IC: intracoronary; IGF-1: insulin-like growth factor-1; IM: intramyocardially; iPS: induced pluripotent stem cells; IV: intravenously; lncRNA: long non-coding RNA; LV: left ventricle; MALAT1: metastasis-associated lung adenocarcinoma transcript 1; MI: myocardial infarction; miRNA: microRNA; MSCs: mesenchymal stromal cells; NF-κB: nuclear factor kappa B; PAPP-A: pregnancy-associated plasma protein A; PEG: polyethylene glycol; S1P: sphingosine 1-phosphate; S1PR1: sphingosine-1-phosphate receptor 1; SK1: sphingosine kinase 1; Smad7: mothers against decapentaplegic homolog 7; TLR4: toll-like receptor 4; UC: ultracentrifugation.