. 2022 Jan 26;10:813885. doi: 10.3389/fcell.2022.813885

TABLE 1.

Summary of studies showing a correlation between the levels of total or specific cell type EVs subpopulations, measured by flow cytometry, in circulation (plasma) and cardiovascular diseases.

Disease/patient group (n = number)	EVs type	Outcome	References
844 individuals of the Framingham Offspring cohort (mean age 66 + 9 years, 57% women) without cardiovascular disease	Endothelial (e)EVs subsets	Association of eEVs with hypertension, triglycerides and metabolic syndrome and the Framingham risk score	Amabile et al. (2014)
	CD144+
	CD31⁺CD41⁻
Systemic review and meta-analysis of 48 studies involving 2,460 patients with type 2 DM and 1,880 non-diabetic controls	Specific surface markers	Total EVs, pEVs, mEVs and eEVs were higher in type 2 DM vs. controls	(Li et al., 2016)
Three groups: 1) untreated patients with severe uncontrolled hypertension, n = 24; 2) untreated patients with established mild hypertension, n = 19; and 3) normotensive volunteer subjects n = 16	eEVs: CD31⁺CD42⁻	eEVs and pEVs were significantly increased in the severely hypertensive group	Preston et al. (2003)
	Platelet (p)EVs: CD41⁺		Preston et al. (2003)
Two groups: 1) High cardiovascular risk patients (n = 37); 2) age, gender, and treatment-matched controls (n = 37)	Total EVs (Annexin V+)	Total EVs, pEVs, mEVs, and TF + EVs were significantly elevated in high risk patients vs. controls	Suades et al. (2015)
	pEVs
	TSP1+
	PAC+
	PAC + CD62P+	Levels of TF + mEVs and pEVs were associated to atherosclerotic burden
	Tissue factor (TF)+ pEVs: CD142 + TSP1+
	TF + monocyte (m)EVs: CD142 + CD14⁺
Four groups: 40 patients with myocardial infarction (MI), 30 unstable angina (UA), 20 stable angina (SA), and 20 healthy individuals	pEVs: CD41⁺	eEVs and pEVs were significantly elevated in MI and UA vs SA and control	Cui et al. (2013)
	mEVs: CD14⁺	No differences were observed in mEVs and lEVs among the groups
	Lymphocytes (l)EVs: CD4⁺
	eEVs: CD144+	TF + EVs were higher in MI and UA
	TF + EVs: CD142+	eEVs and pEVs levels correlated with IL-6 or CRP in coronary heart disease patients
Patients with newly diagnosed acute coronary syndrome (ACS) were divided into 3 groups: 1) UA (n = 36), 2) NSTEMI (n = 22), and 3) STEMI (n = 42); and an additional group of n = 10 age and sex matched controls	Total EVs: Lactadherin+	Total EVs, pEVs and eEVs were higher in ACS groups vs controls	Liu et al. (2016)
	pEVs: CD41a+
	eEVs: CD31⁺	Leu- and eryEVs were higher in the STEMI group vs UA and NSTEMI groups (both p < 0.05)
	mEVs: CD14⁺
	B-cells EVs: CD19⁺	In vitro, EVs form ACS patients displayed procoagulant activity
	T cells EVs: CD3⁺
	Erythrocyte (ery)EVs: CD235a + TF + EVs: CD142+
CAD patients undergoing endarterectomy (n = 42), and age- and sex-matched controls (n = 73)	Ann V + EVs	Annexin V + EVs and pEVs subsets, were higher in cases vs. controls	Wekesa et al. (2014)
	pEVs subsets
	CD41⁺
	Ann V + CD41⁺	eEVs subsets were higher in patients with unstable vs. stable plaques
	eEVs subsets
	CD31⁺CD41⁻
	CD144+	eEVs and pEVs were significantly higher in patients with carotid stenosis vs. controls
	CD146+
	CD105+
Patients undergoing carotid endarterectomy: n = 19 asymptomatic and n = 51 symptomatic patients (n = 51); and n = 20 healthy age-matched controls	eEVs: Ann V + CD31⁺CD42b−		Schiro et al. (2015)
	pEVs: Ann V + CD31⁺CD42b+	No differences were observed between asymptomatic vs. symptomatic patients
STEMI patients (n = 40) treated by percutaneous coronary intervention (PCI); age, gender, risk factors and pharmacological treatments matched control group (n = 20); and patients recovering from STEMI (n = 20)	Total EVs (Ann V+)	STEMI patients present increased levels of total EVs, LeuEVs subsets, eEVs subsets and PEVs	Suades et al. (2016)
	eEVs
	CD31⁺
	CD146+
	CD62E+
	pEVs: CD61⁺
	Leukocytes (leu)EVs: CD45⁺
	lEVs: CD45+/CD3+
	mEVs: CD14⁺
	neutrophil EVs: CD66b+
	TF + EVs: CD142+
17 healthy volunteers and 13 ACS.	Magnetic nanoparticles conjugated with anti-CD63/CD31 or anti-CD31 for eEVs, or with anti- CD63/CD41a or anti-CD41a antibodies for pEVs	ACs patients presented increased levels of EVs, mainly of platelet origin	Vagida et al. (2016)
44 end-stage renal failure patients (ESRF), and 32 healthy subjects	Ann V + EVs	Annexin V + EVs, eEVs, pEVs and eryEVs were increased in ESRF patients vs. controls	Amabile et al. (2005)
	eEVs subsets
	CD31⁺
	CD144+
	pEVs: CD41⁺
	eryEVs: CD235a+	Only eEVs correlated with arterial dysfunction
	Lymphocyte EVs: CD3⁺
	myeloid EVs: CD11b+
	LeuEVs: CD45⁺
	Neutrophil EVs: CD66b+
232 patients with DM and 102 controls	eEVs: CD144 + CD42b−	eEVs levels were increased in DM vs. control	Koga et al. (2005)
232 patients with DM and 102 controls	In DM patients, eEVs were associated to a higher risk for CAD		Koga et al. (2005)
CAD patients (n = 50)	eEVs: CD31 + Ann V+	Increased eEVs correlated with worse endothelial-dependent vasodilatation and independently predicted severe endothelial dysfunction	Werner et al. (2006)
CAD patients (n = 200)	eEVs: CD31 + Ann V+	eEVs were increased in patients with a first major adverse cardiovascular and cerebral events (MACCE)	Sinning et al. (2011)
CAD patients (n = 200)		In the follow up eEVs were independently associated to higher risk of CV death, need for revascularization or MACCE.	Sinning et al. (2011)
Healthy controls (n = 80), chest paint patients: non-CAD (n = 94), SA (n = 111), and ACS (n = 145)	eEVs: CD146+	The levels of eEVs were increased in ACS > SA > non-CAD > controls	Fan et al. (2014)
		eEVs levels were associated to higher risk of MACE in ACS group	Fan et al. (2014)
STEMI patients (n = 51) and age-matched controls (n = 50)	Ann V + EVs	eryEVs were increased in STEMI patients vs. controls	Giannopoulos et al. (2014)
	pEVs: CD41⁺	No differences were found in pEvs
	eryEVs: CD235a+	eryEVs levels were independently associated to a higher risk of MACE during the follow-up
Stroke patients: 1) mild stroke, n = 20; 2) moderate–severe stroke, n = 21; 3) age-matched controls, n = 23	eEVs subsets	PS + eEVs were increased in stroke patients vs. controls	Simak et al. (2006)
	CD105 + CD41a-CD45− (E + eEVs)	All eEVs subsets were elevated in moderate–severe stroke patients vs. controls
	CD105 + CD144+ (C + eEVs)
	CD105 + PS + CD41a− (PS + eEVs)	Brain lesion volume was correlated E + eEVs, PS + eEVs and I+ eEVs levels
	CD105 + CD54⁺CD45− (I + eEVs)
Patients with acute stroke (n = 73), and patients with vascular risk factors but no stroke events (n = 275)	eEVs subsets	Levels of CD31+/AnnV+ and CD62E + eEVs subsets were greater in acute stroke patients vs. controls	Jung et al. (2009)
	CD31+/CD42b-	CD62E + eEVs were strongly associated with stroke severity and infarct volume
	CD31+/AV+
		CD62E+
Patients with acute ischemic stroke (n = 68), and age- and sex-matched controls (n = 61)	eEVs subsets	CD144+/CD41a−, CD31⁺CD41a−, CD62E+, and Annexin V + CD62E + eEVs, were significantly increased in acute ischemic stroke patients vs. controls	Li and Qin (2015)
	CD144 + CD41a−
	CD31⁺CD41a−
	CD62E+	CD144+/CD41a− eEVs were correlated with stroke severity
	Ann V + CD62E+
		pEVs: CD41a + CD144−
18 PAD patients and 12 asymptomatic controls	Total EVs: Lactadherin+	PAD patients presented increased levels of eEVs carrying the monomeric form of C-reactive protein (mCRP)	Crawford et al. (2016)
	pEVs: CD41a+
	eEVs subsets
	CD31⁺
	CD144 +
	LeuEVs: CD45⁺	Control subjects on statins presented a reduction in mCRP + eEVs
	mEVs: CD14⁺
	B-cells EVs: CD19⁺
	T cells EVs: CD3⁺
	Neutrophil EVs: CD66b+
		eryEVs: CD235a+
		Monomeric (m) or pentameric (p) CRP + EVs
PAD patients (n = 50) and controls (n = 50)	eEVs: CD144+	PAD patients present increased levels of shh+ in all EVs subpopulations	Giarretta et al. (2018)
	pEVs: CD42b+
	LeuEVs: CD45⁺	Shh + eEVs levels correlated with the number of collateral vessels in ischemic thighs of PAD patients (n = 18)
	eryEVs: CD235+
	Sonic Hedgedog (Shh)+EVs
PAD patients (n = 50) and controls (n = 50)	pEVs: CD41⁺	Increased levels of pEVs in PAD patients vs controls	Zeiger et al. (2000)
PAD patients, n = 23 with severe disease (critical limb ischemia, CLI), 36 with moderate disease (intermittent claudication, IC), and n = 30 healthy controls	pEVs: CD61⁺CD42b+	Gradual increased in pEVs levels according ro severity (CLI > IC > controls)	Tan et al. (2005)
Patients presenting stable angina (n = 10), peripheral arterial disease (n = 10), NSTEMI (n = 11) and STEMI myocardial infarction (n = 10), age- and sex matched older controls n = 10 and young healthy individuals (n = 10)	pEVs subsets	96% of the detected EVs were from platelet origin	van der Zee et al. (2006)
	Ann V + CD61⁺CD62P+	96% of the detected EVs were from platelet origin
	Ann V + CD61⁺CD63⁺	CD62P + pEVs increased in patients with NSTEMI and STEMI vs. older controls
	eEVs: CD62E
	EryEVs: CD235a
	T- cells EVs	CD63+pEVs- were increased in patients with PAD, NSTEMI, and STEMI vs. older controls
	CD4 +
	CD8⁺
	mEVs: CD14⁺
	B cells EVs: CD20⁺
	Neutrophil EVs: CD66e+
PAD patients (n = 45)	AnnexinV eEVs: CD62E+ pEVs: CD41/61+ LeuEVs: CD11b+ eryEVs: CD235a+	In plasma of PAD patients pEVs were the most abundant subpopulation, followed by eryEVs, eEVs and LeuEVs	Saenz-Pipaon et al. (2020)
	More than 85% of pEVs and eryEVs were Ann V+, while the percentage was lower for eEVs (70%) and LeuEVs (40%)
	The number pEVs were inversely correlated with procoagulant activity of plasma
14 PAD patients and 15 normal controls PAD patients were treated with cilostazol (2 weeks) or cilostazol with dipyridamole (14 weeks)	pEVs: CD42⁺	PAD patients presented increased levels of pEVs	Nomura et al. (2004)
		Cilostazol, and further, cilostazol with dipyridamole decreased pEVs levels in PAD patients	Nomura et al. (2004)
PAD patients (n = 19) randomly assigned to Atorvastatin or placebo treatment for 8 weeks	Total EVs: lactadherin	Atorvastatin treatment reduced the number of CD142+, CD62P+ and CD61⁺ pEVs vs placebo treated PAD patients	Mobarrez et al. (2011)
	pEVs
	CD42a + CD142+
	CD42a + CD62P+
	CD42a + CD61⁺
PAD patients (n = 19) randomly assigned to Atorvastatin or placebo treatment for 8 weeks	Total EVs	Both CD144 + eEVs and CD144 + CD142+ eEVs were increased in patients on atorvastatin vs. placebo	Mobarrez et al. (2012)
	eEVs
	Lactdherin + CD144+
	Lactadherin + CD144 + CD142+
22 patients with severe aortic stenosis (AS) and 18 controls	eEVs: CD62E+	pEVs, LeuEVs and eEVs were increased in AVS patients vs. control	Diehl et al. (2008)
	pEVs
	CD31⁺ CD61⁺	pEVs levels were correlated with shear stress and eEVs with the number of blood monocytes
	CD62P+
	LeuEVs: CD11b+
Patients with severe AS. n = 28 with low coronary calcification (CAC) score, and n = 27 with high CAC score	eEVs	The levels of pEVs and CD62E + eEVs were increased in high CAC score patients vs. low CAC score group, and correlated to the calcium score	Horn et al. (2016)
	CD144+
	CD62E+
	CD31⁺CD41⁻	EVs thrombin generation activity was higher in patients with high CAC score
	pEVs: CD41⁺
	EVs trombin generation activity
56 severe AS patients undergoing transcatheter aortic valve implantation (TAVI)	eEVs	All eEVs subpopulations decreased 3 months after TAVI, along with an increase in the endothelial function	Horn et al. (2015)
	CD144+
	CD62E+
	CD31⁺CD41⁻
	pEVs: CD41⁺
92 severe AS patients undergoing TAVI	eEVs	The levels of CD62E + eEVs decreased gradually from pre-TAVI to post-TAVI (1 week, 1, 3 and 6 months) determination	Jung et al. (2017)
	CD31 + Annexin+
	CD31 + Annexin−
	CD31⁺CD42b−	In contrast, circulating PEVs increased gradually after TAVI
	CD62E+
	pEVs: CD31⁺CD42b+
Patients with severe AS selected for percutaneous replacement of the aortic valve (n = 9)	eEVS: CD31 + Ann V+	No differences were observed between pre- and post-operative (5 days) levels of eEVs, pEVs or LeuEVs	Marchini et al. (2016)
	pEVs: CD41 + Ann V+
	LeuEVs: CD45 + Ann V+
135 patients undergoing surgical aortic valve replacement	small (s)EVs were quantified by nanoparticle tracking analysis (NTA)	sEVs decreased 24 h post-surgery, and recovered to pre-operative levels 7 days and 3 months post-procedure	Weber et al. (2020)
135 patients undergoing surgical aortic valve replacement		No association between sEVs and echocardiographic parameters before or after surgery (7 days and 3 months) were observed sEVs levels were correlated to prosthesis patients mismatch parameters at month 3 post-surgery	Weber et al. (2020)
AAA patients (blood samples and mural thrombi, n = 20), and sex and age-matched healthy individuals (blood samples, n = 20)	Annexin V + EVs	Circulating total EVs were significantly increased in AAA patients vs. controls	Touat et al. (2006)
	pEVs: CD41
	neutrophil EVs CD15
	mEVs: CD14	Locally, luminal thrombus layers released larger quantities of annexin V-positive EV, mainly of platelet and neutrophil origin, compared to the intermediate and abluminal layers
	eEVs: CD106
	eryEVs: CD235
Controls (n = 66) and thoracic AA (TAA) patients associated to bicuspid aortic valves (BAV) (n = 15), or other origins (degenerative, n = 23)	Ann V + EVs	The levels of EVs and pEVs were higher in TAA groups vs. control	Touat et al. (2008)
	pEVs: Ann V + CD41⁺		Touat et al. (2008)

AAA: abdominal aortic aneurysm; ACS: acute coronary syndrome; AS: aortic stenosis; TAA: Thoracic aortic aneurysm; TAVI: transcatheter aortic valve implantation; CAD: coronary artery disease; CAC: coronary calcification score; DM: diabetes mellitus; ESRF: end-stage renal failure; CLI: critical limb ischemia; IC: Intermittent claudication; MI: myocardial infarction; PAD: peripheral arterial disease; SA: stable angina; UA: unstable angina; Ann V: Annexin V; EVs: Extracellular vesicles; eEVs: endothelial EVs; eryEVs: erythrocyte EVs; LeuEVs: leukocyte EVs; lEVs: Lymphocyte EVs; mEVs: monocyte EVs; pEVs: platelet EVs; (N)STMI: (non) ST Segment Elevation Myocardial Infarction; TF: Tissue factor; TSP-1: Thrombospondin-1; T; PAC: activated αIIbβ3-integrin.