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Strategies for improved endothelial cell adhesion in microphysiological vascular model systems.

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  • 1Department of Biomedical Engineering, Duke University, Durham, North Carolina, United States of America.

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|May 19, 2025
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Summary

This protocol optimizes endothelial cell seeding in tissue-engineered blood vessels (TEBVs) for disease modeling. It ensures sustained endothelial coverage and functionality under flow, improving TEBV utility for research.

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Area of Science:

  • Biomedical Engineering
  • Vascular Biology
  • Tissue Engineering

Background:

  • Tissue-engineered blood vessels (TEBVs) are crucial in vitro models for studying vascular diseases like atherosclerosis.
  • Maintaining a functional, confluent endothelial monolayer in TEBVs under shear stress is a significant challenge.
  • Current methods often struggle with consistent endothelial cell attachment and coverage.

Purpose of the Study:

  • To develop and optimize a protocol for robust endothelialization of human TEBVs.
  • To identify key factors influencing endothelial cell adherence and survival within engineered vessels.
  • To enhance the reliability of TEBVs as disease models by ensuring endothelial integrity.

Main Methods:

  • Optimization of endothelial cell (EC) seeding density, rotation time, and perfusion parameters.
  • Redesign of TEBV chambers with integrated viewing windows for real-time monitoring.
  • Assessment of EC coverage, alignment, and responsiveness to inflammatory stimuli using fluorescence microscopy.

Main Results:

  • Achieved sustained and confluent endothelial cell luminal coverage in TEBVs.
  • Demonstrated EC alignment with the direction of applied physiological shear stress.
  • Confirmed EC responsiveness to inflammatory stimuli, indicating functional endothelium.
  • Developed a streamlined monitoring system for efficient assessment of endothelialization.

Conclusions:

  • The optimized protocol significantly improves endothelialization success in TEBVs.
  • This method enhances the utility of TEBVs for in vitro vascular disease research.
  • The redesigned chambers facilitate rapid assessment and fabrication of functional engineered vessels.