Table 1.

Summary of the EC–VSMC crosstalk, define by different in vitro models.

EC–VSMC Communication		Methodology	Molecule/Signal Pathway	Results	References
Paracrine	Soluble Factors	EC–VSMC coculture in opposite sides of transwell	↑VEGF, PDGF-AA, PDGF-BB, and TGF-β in VSMCs ↓bFGF	Coculture affect gene and protein expression of angiogenic factors	[26,27,28]
		Conditioned culture media	↑TF	EC suppress the proliferation of co-existing VSMCs	[29]
		Coculture flow chamber system	↑ICAM-1, VCAM-1 and E-selectin gene expression	Under static conditions, coculture with VSMCs induces adhesion proteins expression in ECs	[30]
		Coculture flow chamber system	↑GRO-α, MCP-1	Under static conditions, coculture with VSMCs induces GRO-α, MCP-1 in ECs	[31]
		Microcarrier coculture system	LDL	EC influenced VSMC’s LDL metabolism	[33,34]
		Conditioned culture media/Ex vivo aortic ring	eNOS, cGMP, endothelin, AngII	Regulation of the vascular tone	[33,39,40,46]
		Ex vivo aortic ring	Perlecan	Mechanotransduction in EC controls VSMCs proliferation	[48]
		Coculture flow chamber system	PDGF-BB	EC triggers proliferation and migration of VSMCs Synthetic VSMCs modulate anti-angiogenic effect over EC	[53,54,56]
		Coculture flow chamber system	TGF-β1	EC modulates VSMCs phenotypic switching and extracellular matrix synthesis	[55]
		Spheroids coculture	Ang-1/Ang-2	Desestabilization of the quiescent endothelium	[61]
		In vitro model of a vessel-like construct	mTOR	VSMCs regulates EC response to flow and injury	[68]
	Extracellular vesicles	Conditioned culture media/Boyden chamber assay	miR143/145 miR-206 miR-126	Endothelial EVs regulate VSMCs phenotypic changes	[79,80,81,82,83,84,85,86]
		Conditioned culture media/Boyden chamber assay	miR-221/miR-222 miR-155 miR-1246, miR-182, miR-486	VSMCs EVs regulate endothelial permeability, migration and vascular calcification	[90,91,92,93]
Parenchymal players		3D Bioprinted gelatin hydrogel platform	Collagen I, IV, fibronectin, heparan sulfate chains	Extracellular matrix presentation modulates VSMCs mechanostransduction	[103,104,105]
Direct contact	Myoendothelial gap junctions (connexins)	EC–VSMCs coculture in opposite sides of small pore transwell	Second messengers (Ca2+, IP3, camp)	Vascular constriction-relaxation. Phenotypic changes	[113,114,115,116,117,118]
	Notch signaling	EC–VSMC coculture in opposite sides of small pore transwell Human-derived blood vessels organoids	Notch3 receptor BMPR2-Notch1 DII4 and Notch3	VSMCs phenotypic switching, EC regeneration and maintainer of EC monolayer integrity Regulators of diabetic vasculopathy	[125,126,127,128,147,148,149]
		Spheroids coculture	Ephrin-B2	VSCMs migration and EC adhesion	[129,130,131]
		Spheroids coculture	24-dehydrocholesterol reductase	EC control VSMCs cholesterol levels	[144]
		3D tubular artery-like constructs	Glucose metabolism	Investigation of late atherosclerosis lesion	[145,146]