In Vitro Mineralisation of Tissue-Engineered Cartilage Reduces Endothelial Cell Migration, Proliferation and Tube Formation

In Vitro Mineralisation of Tissue-Engineered Cartilage Reduces Endothelial Cell Migration, Proliferation and Tube Formation

Ji, Encheng (Erasmus MC)
Leijsten, Lieke (Erasmus MC)
Witte-Bouma, Janneke (Erasmus MC)
Rouchon, Adelin (University of Basel)
Di Maggio, Nunzia (University of Basel)
Banfi, Andrea (University of Basel)
van Osch, G.J.V.M. (TU Delft Biomaterials & Tissue Biomechanics; Erasmus MC)
Farrell, Eric (Erasmus MC)
Lolli, Andrea (Erasmus MC)

2023

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.

angiogenesis
bone tissue engineering
endochondral ossification
mesenchymal stromal cells
mineralisation

http://resolver.tudelft.nl/uuid:6bf58392-5a6f-4a74-b4ee-e0f7ab4da982

https://doi.org/10.3390/cells12081202

2073-4409

Cells, 12 (8)

Institutional Repository

journal article

© 2023 Encheng Ji, Lieke Leijsten, Janneke Witte-Bouma, Adelin Rouchon, Nunzia Di Maggio, Andrea Banfi, G.J.V.M. van Osch, Eric Farrell, Andrea Lolli