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Label-free relative quantification of secreted proteins as a non-invasive method for the quality control of chondrogenesis in bioengineered substitutes for cartilage repair.

Abstract：

: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 are 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 with the aim of characterizing the implants. The limits and advantages of analysing, by label-free liquid chromatography-coupled matrix-assisted laser desorption and ionization (LC-MALDI) mass spectrometry, the secreted proteome released into culture medium by engineered cartilage tissues were investigated and compared with more classically used methods for biomaterial characterization. This method did not require sacrificing the biomaterials and robustly evidenced their chondrogenic statuses. In more detail, the method highlighted differences between batches prepared from distinct donors. It was adapted to distinct scaffolds and allowed a comparison of 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 the 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.

journal_name

J Tissue Eng Regen Med

journal_title

Journal of tissue engineering and regenerative medicine

authors

Henrionnet C,Gillet P,Mainard D,Vincourt JB,Pinzano A

doi

10.1002/term.2454

subject

Has Abstract

pub_date

2018-03-01 00:00:00

pages

e1757-e1766

issue

eissn

1932-6254

issn

1932-7005

journal_volume

pub_type

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Label-free relative quantification of secreted proteins as a non-invasive method for the quality control of chondrogenesis in bioengineered substitutes for cartilage repair.