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Related Experiment Videos

Distributed vasculogenesis from modular agarose-hydroxyapatite-fibrinogen microbeads.

Ana Y Rioja1, Ethan L H Daley1, Julia C Habif1

  • 1Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, United States.

Acta Biomaterialia
|April 3, 2017
PubMed
Summary

Researchers developed novel microbeads for critical limb ischemia treatment. These agarose-hydroxyapatite-fibrinogen microbeads promote blood vessel formation, offering a promising therapeutic angiogenesis strategy.

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Area of Science:

  • Biomaterials Science
  • Regenerative Medicine
  • Vascular Biology

Background:

  • Critical limb ischemia (CLI) severely impairs circulation, leading to pain, poor wound healing, and tissue necrosis.
  • Therapeutic angiogenesis aims to restore blood flow by repairing damaged microvasculature.
  • Current treatments for CLI have limitations, necessitating innovative approaches.

Purpose of the Study:

  • To develop modular, micro-scale constructs (microbeads) for therapeutic angiogenesis in CLI.
  • To create constructs with enhanced handling, in vitro pre-culture capabilities, and the ability to promote microvasculature formation.
  • To evaluate the efficacy of different microbead compositions in supporting endothelial cell network formation.

Main Methods:

  • Engineered microbeads using an agarose base with fibrinogen (FGN) and/or hydroxyapatite (HA) for cell adhesion.
Keywords:
AgaroseEndothelial sproutingFibrinogenHydroxyapatiteMicrobeads

Related Experiment Videos

  • Encapsulated co-cultures of human umbilical vein endothelial cells (HUVEC) and fibroblasts within the microbeads.
  • Characterized microbead size, composition, and cell viability.
  • Embedded cell-seeded microbeads in fibrin hydrogels and assessed HUVEC sprouting and network formation over time.
  • Compared network formation in microbeads with varying compositions (AG, AG+HA, AG+FGN, AG+HA+FGN) and pre-culture durations.

    • Main Results:

    • Microbeads ranged from 80-100µm, with size increasing upon addition of FGN and HA.
    • Hydroxyapatite addition improved microbead yield and fibrinogen distribution.
    • High cell viability was observed across all microbead types.
    • Cell-seeded microbeads in fibrin hydrogels exhibited HUVEC sprouting and inosculation.
    • Pre-culture of AG+HA+FGN microbeads significantly enhanced HUVEC network length compared to other compositions.
    • Composite microbeads promoted more even and widespread endothelial network formation than pure fibrin controls.

    Conclusions:

    • The developed agarose-hydroxyapatite-fibrinogen microbeads effectively support HUVEC sprouting within and between adjacent constructs.
    • These modular microtissues promote distributed vascularization of external matrices.
    • The microbead format and tunable matrix composition offer a promising strategy for revascularization in treating ischemic diseases like CLI.