Micro/nano materials regulate cell morphology and intercellular communication by extracellular vesicles.

Extracellular vesicles (EVs) have emerged as important nano-cargo carriers for cell-cell communication, yet how biophysical factors regulate EV-mediated signaling is not well understood. Here we show that microgrooves can modulate the morphology of endothelial cells (ECs), and regulate the phenotype of smooth muscle cells (SMCs) through EVs in co-culture. Elongated ECs, in comparison with polygonal ECs, increased the expression of contractile markers in SMCs. Depletion of EVs in the culture medium abolished this effect. Further analysis demonstrated that elongated ECs significantly upregulated miR-143/miR-145, leading to the increase of these microRNAs in EC-secreted EVs that were transferred to SMCs under a co-culture condition. Inhibition of EV secretion from ECs abolished the EC-SMC communication and the increased expression of SMC contractile markers. Moreover, electrospun nano-fibrous scaffolds with aligned fibers had the same effects as microgrooves to induce EC secretion of EVs to regulate SMC phenotypic marker expression. These results demonstrate that micro and nano materials can be used to engineer cell morphology and regulate EV secretion for cell-cell communication, which will have significant implications in the engineering of blood vessels and other tissues. STATEMENT OF SIGNIFICANCE: By manipulating EC morphology with micro/nano materials, we show that EV-mediated signaling can regulate SMC phenotypic marker expression. This is a very thorough and unique study to demonstrate the function of extracellular vesicles (EVs) as important nano-carriers in cell-cell communication. The originality of this study is to demonstrate that EC morphology modulates the phenotype of smooth muscle cells via extracellular vesicles enclosing miR143/miR145. These findings underscore the important role of biophysical changes in cell-cell communications, and provide a rational basis for engineering micro/nano materials to control cell-cell communications for cell and tissue engineering.

Endothelial Cell Morphology Regulates Inflammatory Cells Through MicroRNA Transferred by Extracellular Vesicles.

Vascular inflammation plays an important role in the pathogenesis and the development of cardiovascular diseases such as arteriosclerosis and restenosis, and the dysfunction of endothelial cells (ECs) may result in the activation of monocytes and other inflammatory cells. ECs exhibit an elongated morphology in the straight part of arteries but a cobblestone shape near the pro-atherogenic region such as branch bifurcation. Although the effects of hemodynamic forces on ECs have been widely studied, it is not clear whether the EC morphology affects its own function and thus the inflammatory response of monocytes. Here we showed that elongated ECs cultured on poly-(dimethyl siloxane) membrane surface with microgrooves significantly suppressed the activation of the monocytes in co-culture, in comparison to ECs with a cobblestone shape. The transfer of EC-conditioned medium to monocytes had the same effect, suggesting that soluble factors were involved in EC-monocyte communication. Further investigation demonstrated that elongated ECs upregulated the expression of anti-inflammatory microRNAs, especially miR-10a. Moreover, miR-10a was found in the extracellular vesicles (EVs) released by ECs and transferred to monocytes, and the inhibition of EV secretion from ECs repressed the upregulation of miR-10a. Consistently, the inhibition of miR-10a expression in ECs reduced their anti-inflammatory effect on monocytes. These results reveal that the EC morphology can regulate inflammatory response through EVs, which provides a basis for the design and the optimization of biomaterials for vascular tissue engineering.