Induction of mesenchymal stem cell differentiation and cartilage formation by cross-linker-free collagen microspheres

Marc Mathieu (1) , S. Vigier (1, 2, 3) , M-N Labour (2, 3) , C. Jorgensen (1, 4) , E. Belamie (2, 3) , D. Noël (1)

1 Cellules souches mésenchymateuses, environnement articulaire et immunothérapies de la polyarthrite rhumatoide
2 EPHE - École Pratique des Hautes Études
3 ICGM ICMMM - Institut Charles Gerhardt Montpellier - Institut de Chimie Moléculaire et des Matériaux de Montpellier
4 CHU - Hôpital Lapeyronie [Montpellier]

Because of poor self-healing ability, joint cartilage can undergo irreversible degradation in the course of various diseases or after injury. A promising approach for cartilage engineering consists of using of mesenchymal stem cells (MSC) and a differentiation factor combined with an injectable carrier biomaterial. We describe here a novel synthesis route for native collagen microspheres that does not involve the use of potentially toxic crosslinking agents. An emulsion was formed between a type I collagen solution and perfluorinated oil, stabilised by a biocompatible triblock perfluorinated copolymer surfactant. Spherical microparticles of fibrillar collagen were formed through a sol-gel transition induced by ammonia vapours. Electron microscopy observations showed that these self-cross-linked microspheres were constituted by a gel of striated collagen fibrils. Microspheres that were loaded with transforming growth factor beta (TGF-β)3 progressively released this differentiation factor over a four weeks period. Human MSC rapidly adhered to TGF-β3-loaded microspheres and, after 21 d of culture, exhibited typical chondrocyte morphology and produced an uncalcified matrix made of the predominant cartilage components, aggrecan and type II collagen, but devoid of the hypertrophic marker type X collagen. Subcutaneous co-injection of MSC and TGF-β3-loaded microspheres in mice consistently led to the formation of a cartilage-like tissue, which was however hypertrophic, calcified and vascularised. In conclusion, we developed cross-linker free collagen microspheres that allowed chondrogenic differentiation of MSC in vitro and in vivo.

Mots clés: Biomaterial; cartilage engineering; chondrogenesis; collagen; injectable; mesenchymal stem cell; microspheres; safety; self-assembly; transforming growth factor-β.; cartilage engineering; transforming growth factor-β

Domaines: Sciences du Vivant [q-bio]

Marc Mathieu, S. Vigier, M-N Labour, C. Jorgensen, E. Belamie, et al.. Induction of mesenchymal stem cell differentiation and cartilage formation by cross-linker-free collagen microspheres. eCells and Materials Journal, 2014, 28, pp.82-97. ⟨10.22203/eCM.v028a07⟩. ⟨hal-03116260⟩