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Impact of type I collagen enrichment of an alginate-based bioink on the quality of cartilage sustitutes produced by 3D bioprinting
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International audience. INTRODUCTIONIn cartilage engineering, the 3D bioextrusion printing permits to produce a stratified construct that resembles native cartilage. It is therefore necessary to optimize the bio-inks adapted to the different layers. The aim of our work was to evaluate the effect of different amounts of type I collagen added to the bioink on chondrogenic differentiation and extracellular matrix synthesis.EXPERIMENTAL METHODS : MSCs were isolated from human bone marrow following total hip arthroplasty. After their proliferation, cells were seeded in an alginate-based bio-ink at a cell density of 106 cells/mL. Then, we added different amounts of type I collagen (0, 0.5, 1 and 5 mg/syringe) to the bio-ink. The bio-printing patterns were generated using Repetier Host Software® in order to generate a cartilage substitute. Chondrogenic differentiation was induced by adding TGF-β1 (10 ng/mL) for 56 days. The chondrogenic properties of MSCs was evaluated by gene expression analysis and the quality of the extracellular matrix by histology and immunohistochemistry. RESULTS: Gene expression analysis showed that the chondrogenic markers are significantly more expressed under the effect of TGF-β1, associated with a significant synthesis of proteoglycans and type II collagen. A small amount of type I collagen (0.5 and 1 mg) induced an increase in chondrogenic markers associated with a synthesized extracellular matrix richer in type II collagen. Conversely, the highest amount of type I collagen (5 mg) causes an increase in hypertrophic markers and a decrease in the synthesis of cartilaginous matrix constituents associated with an increase in the rate of calcifications within the substitutes.CONCLUSIONS: Our innovative bioink allows the generation of TGF-β-inducible engineered cartilage substitutes. Moreover, a low concentration type I collagen as an additive was interesting for the production of the upper layers of cartilage while the higher concentration of 5 mg could be more suitable for the calcified cartilage layer.This work was supported by « ANR-DGA Bloc Print (N°ANR-16-ASTR-0021) », « Fondation de l’Avenir (N°AP-RM-16-042) », « Université de Lorraine – Région Alsace-Champagne-Ardennes-Lorraine 2016 (N°AAP-002-037) », « CPER IT2MP », « FEDER » and « Fondation pour la Recherche Médicale » (N° ECO201906008942).
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