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- Title
In Vitro Mineralisation of Tissue-Engineered Cartilage Reduces Endothelial Cell Migration, Proliferation and Tube Formation.
- Authors
Ji, Encheng; Leijsten, Lieke; Witte-Bouma, Janneke; Rouchon, Adelin; Di Maggio, Nunzia; Banfi, Andrea; van Osch, Gerjo J. V. M.; Farrell, Eric; Lolli, Andrea
- Abstract
Tissue engineering bone via endochondral ossification requires the generation of a cartilage template which undergoes vascularisation and remodelling. While this is a promising route for bone repair, achieving effective cartilage vascularisation remains a challenge. Here, we investigated how mineralisation of tissue-engineered cartilage affects its pro-angiogenic potential. To generate in vitro mineralised cartilage, human mesenchymal stromal cell (hMSC)-derived chondrogenic pellets were treated with β-glycerophosphate (BGP). After optimising this approach, we characterised the changes in matrix components and pro-angiogenic factors by gene expression analysis, histology and ELISA. Human umbilical vein endothelial cells (HUVECs) were exposed to pellet-derived conditioned media, and migration, proliferation and tube formation were assessed. We established a reliable strategy to induce in vitro cartilage mineralisation, whereby hMSC pellets are chondrogenically primed with TGF-β for 2 weeks and BGP is added from week 2 of culture. Cartilage mineralisation determines loss of glycosaminoglycans, reduced expression but not protein abundance of collagen II and X, and decreased VEGFA production. Finally, the conditioned medium from mineralised pellets showed a reduced ability to stimulate endothelial cell migration, proliferation and tube formation. The pro-angiogenic potential of transient cartilage is thus stage-dependent, and this aspect must be carefully considered in the design of bone tissue engineering strategies.
- Subjects
CELL migration; ENDOCHONDRAL ossification; ENDOTHELIAL cells; CARTILAGE; UMBILICAL veins; TISSUE engineering; STROMAL cells
- Publication
Cells (2073-4409), 2023, Vol 12, Issue 8, p1202
- ISSN
2073-4409
- Publication type
Article
- DOI
10.3390/cells12081202