Research article
Vascular endothelial and smooth muscle cell galvanotactic response and differential migratory behavior

https://doi.org/10.1016/j.yexcr.2020.112447Get rights and content

Abstract

Chronic disease or injury of the vasculature impairs the functionality of vascular wall cells particularly in their ability to migrate and repair vascular surfaces. Under pathologic conditions, vascular endothelial cells (ECs) lose their non-thrombogenic properties and decrease their motility. Alternatively, vascular smooth muscle cells (SMCs) may increase motility and proliferation, leading to blood vessel luminal invasion. Current therapies to prevent subsequent blood vessel occlusion commonly mechanically injure vascular cells leading to endothelial denudation and smooth muscle cell luminal migration. Due to this dichotomous migratory behavior, a need exists for modulating vascular cell growth and migration in a more targeted manner. Here, we examine the efficacy of utilizing small direct current electric fields to influence vascular cell-specific migration (“galvanotaxis”). We designed, fabricated, and implemented an in vitro chamber for tracking vascular cell migration direction, distance, and displacement under galvanotactic influence of varying magnitude. Our results indicate that vascular ECs and SMCs have differing responses to galvanotaxis; ECs exhibit a positive correlation of anodal migration while SMCs exhibit minimal change in directional migration in relation to the electric field direction. SMCs exhibit less motility response (i.e. distance traveled in 4 h) compared to ECs, but SMCs show a significantly higher motility at low electric potentials (80 mV/cm). With further investigation and translation, galvanotaxis may be an effective solution for modulation of vascular cell-specific migration, leading to enhanced endothelialization, with coordinate reduced smooth muscle in-migration.

Section snippets

Credit author statement

Kaitlyn R Ammann, PhD conceptualized and developed methodology, validated methods, curated and analyzed data, and wrote and revised the manuscript. Marvin J Slepian, MD conceptualized project, reviewed and edited manuscript, and supervised the study.

Galvanotaxis platform design and validation

Cell viability was maintained during galvanotaxis experiments through utilization of a controlled-environment microscope chamber (Fig. 1). Temperature, CO2 levels, and relative humidity were all regulated within the microscope chamber, allowing for live-cell imaging of migrating cells (Fig. 1A). The galvanotaxis platform, consisting of multiple reservoirs, additionally allowed for DCEFs to be applied across the cell chamber without the introduction of electrolysis to the media (Fig. 1B).

Discussion

The objective of this study was to examine the response of endothelial and smooth muscle cells to galvanotactic stimuli and determine the ability of galvanotaxis to differentially modulate migration of vascular ECs versus SMCs. Our results indicate that ECs and SMCs indeed have a differential response to DCEFs, with ECs migrating towards the anode with increasing voltage magnitude, while SMCs remain statistically unresponsive over the range of voltage tested. Similarly, the motility, i.e. the

Conclusions

Application of DCEFs to vascular endothelial and smooth muscle cells notably influences their migration. This influence impacts both migratory direction and extent of migration. Compared to migration distance and displacement, directionality is more evidently dominant in the differential galvanotactic response between vascular ECs and SMCs. This behavior offers potential for a desired clinical therapeutic if advanced in translation, as a means of preferentially advancing endothelial cell

Galvanotaxis System Overview

All experiments were performed within a custom-designed microscope chamber (OKO Labs) for a Zeiss Axiovert 135, and regulated at 37 °C and 5% CO2 (Fig. 1A). A sterile water reservoir was included in the chamber, to maintain high relative humidity. The galvanotaxis platform was held in place with a microscope stage with 60-mm dish housing. Cells in the galvanotaxis chamber were visualized through a 10X objective lens. External to the microscope chamber, a DC power supply (BK Precision 9184B) was

Acknowledgements

The authors acknowledge NIH Cardiovascular Biomedical Engineering Training Grant T32 HL007955 and the Arizona Center for Accelerated Biomedical Innovation (ACABI) for funding support.

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