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

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

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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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.

Original languageEnglish
Article number1202
JournalCells
Volume12
Issue number8
DOIs
Publication statusPublished - 20 Apr 2023

Bibliographical note

Funding Information:
The project was supported by the following funding: the European Union Horizon 2020 Research and Innovation Program under grant agreement 801159; the Dutch Research Council (NWO)-XS Science grant OCENW.XS5.074; Erasmus MC-Health~Holland TKI-LSH grant EMCLSH20022. E.J. is supported by the China Scholarship Council (CSC, NO. 202207720011) and Wenzhou Wangqiao Orthopedic Hospital.

Publisher Copyright:
© 2023 by the authors.

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