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In Vitro Prevascularization of Self-Assembled Human Bone-Like Tissues and Preclinical Assessment Using a Rat

Fabien Kawecki1,2, Todd Galbraith1, William P Clafshenkel1

  • 1Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Division of Regenerative Medicine, CHU de Québec Research Center-Université Laval, Québec, QC G1J 1Z4, Canada.

Materials (Basel, Switzerland)
|April 30, 2021
PubMed
Summary

Engineered bone tissues with added endothelial cells formed blood vessels in vitro. These prevascularized grafts improved cell survival and bone repair in vivo, despite initial decreases in bone markers.

Keywords:
adiposecalvarial bone defectendothelialmesenchymal stem cellsprevascularizationself-assembly

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Area of Science:

  • Biomaterials Engineering
  • Tissue Engineering
  • Regenerative Medicine

Background:

  • Engineered tissue constructs face challenges with core cell viability and graft integration.
  • In vitro prevascularization aims to improve cell survival and vascular anastomosis for tissue grafts.
  • Human adipose-derived stromal/stem cells (hASCs) are used for osteogenic differentiation in biomimetic bone-like tissues.

Purpose of the Study:

  • To investigate the impact of endothelial cells on osteogenesis and biomineralization in self-assembled bone-like tissues.
  • To evaluate the in vivo efficacy of prevascularized bone-like tissue constructs for rat calvarial bone defect repair.
  • To assess the influence of prevascularization on graft survival and bone healing.

Main Methods:

  • Fabrication of self-assembled bone-like tissues using hASCs with and without endothelial cells.
  • In vitro culture and maturation of engineered tissues for 35 days.
  • In vivo implantation of constructs into rat calvarial bone defects for 12 weeks.
  • Assessment of in vitro bone markers (osteocalcin, hydroxyapatite) and in vivo bone healing and cell survival via imaging.

Main Results:

  • A dense capillary network spontaneously formed in vitro within two weeks.
  • Prevascularized tissues showed reduced osteocalcin levels and hydroxyapatite formation in vitro.
  • In vivo, prevascularized constructs demonstrated improved cell survival and supported bone healing in calvarial defects.
  • Bone repair in prevascularized grafts was comparable to stromal-only grafts, without impediment.

Conclusions:

  • In vitro prevascularization of engineered bone-like tissues is achievable through spontaneous capillary network formation.
  • While in vitro osteogenic markers decreased, prevascularization enhanced in vivo cell survival and functional bone repair.
  • These findings support the engineering of prevascularized bone-like tissues with improved functional outcomes for regenerative applications.