Microcomputed Tomography-Based Analysis of Neovascularization within Bioengineered Vascularized Tissues

Idan Redenski, Shaowei Guo, Majd MacHour, Ariel Szklanny, Shira Landau, Dana Egozi, Yankel Gabet, Shulamit Levenberg

Research output: Contribution to journalArticlepeer-review

Abstract

In the field of tissue engineering, evaluating newly formed vascular networks is considered a fundamental step in deciphering the processes underlying tissue development. Several common modalities exist to study vessel network formation and function. However, a proper methodology that allows through three-dimensional visualization of neovessels in a reproducible manner is required. Here, we describe in-depth exploration, visualization, and analysis of vessels within newly formed tissues by utilizing a contrast agent perfusion protocol and high-resolution microcomputed tomography. Bioengineered constructs consisting of porous, biocompatible, and biodegradable scaffolds are loaded with cocultures of adipose-derived microvascular endothelial cells (HAMECs) and dental pulp stem cells (DPSCs) and implanted in a rat femoral bundle model. After 14 days of in vivo maturation, we performed the optimized perfusion protocol to allow host penetrating vascular visualization and assessment within neotissues. Following high-resolution microCT scanning of DPSC:HAMEC explants, we performed the volumetric and spatial analysis of neovasculature. Eventually, the process was repeated with a previously published coculture system for prevascularization based on adipose-derived mesenchymal stromal cells (MSCs) and HAMECs. Overall, our approach allows a comprehensive understanding of vessel organization during engraftment and development of neotissues.

Original languageEnglish
Pages (from-to)232-241
Number of pages10
JournalACS Biomaterials Science and Engineering
Volume8
Issue number1
DOIs
StatePublished - 10 Jan 2022

Keywords

  • angiogenesis
  • bioengineered constructs
  • microcomputed tomography
  • microfil
  • perfusion protocol
  • prevascularization

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