Label-free relative quantification of secreted proteins as a non-invasive method for the quality control of chondrogenesis in bioengineered substitutes for cartilage repair.
Fiche publication
Date publication
mai 2017
Journal
Journal of tissue engineering and regenerative medicine
Auteurs
Membres identifiés du Cancéropôle Est :
Pr GILLET Pierre, Pr MAINARD Didier, Dr VINCOURT Jean-Baptiste
Tous les auteurs :
Henrionnet C, Gillet P, Mainard D, Vincourt JB, Pinzano A
Lien Pubmed
Résumé
Cartilage tissue engineering is making progress but the competing available strategies still leave room for improvement and consensual overviews regarding the best combinations of scaffolds and cell sources are limited by the capacity to compare them directly. In addition, because most strategies involve autologous cell transfer, once these will be optimized, the resulting implants require individual quality control prior to grafting in order to emphasize patient-to-patient differential responsiveness to engineering processes. Here, cartilage substitutes prepared from human mesenchymal stem cells undergoing chondrogenic differentiation within distinct scaffolds were used as pilot samples to investigate the pertinence of a novel method aiming at characterizing the implants. The limits and advantages of analyzing, by label-free liquid-chromatography-coupled MALDI mass spectrometry (LC-MALDI), the secreted proteome released into culture medium by engineered cartilage tissues were investigated and compared to more classically used methods for biomaterial characterization. This method did not require sacrificing the biomaterials and robustly evidenced their chondrogenic statuses. In more details, the method highlighted differences between batches prepared from distinct donors. It was adapted to distinct scaffolds and allowed comparing the influence of individual engineering steps, such as growth factor combinations and oxygen tension. Finally, it evidenced subtle changes between replicate substitutes within a series, thereby distinguishing least and most accomplished ones. We conclude that relative quantification of secreted proteins through label-free LC-MALDI will be useful not only to orientate engineering methodologies but also to ultimately provide non-invasive quality control of engineered tissue substitutes for the repair of cartilage and possibly other connective tissues.
Mots clés
LC-MALDI, chondrocyte, collagen, mass spectrometry, mesenchymal stem cell, secretome
Référence
J Tissue Eng Regen Med. 2017 May;: