Collagen-based hydrogels are an attractive option in the field of cartilage regeneration with features of high biocompatibility and low immunogenic response. Crosslinking treatments are often employed to create stable 3D gels that can support and facilitate cell embodiment. In this study, we explored the properties of JellaGel™, a novel jellyfish material extracted from Rhizostoma pulmo. In particular, we analyzed the influence of genipin, a natural crosslinker, on the formation of 3D stable JellaGel™ hydrogels embedding human chondrocytes. Three concentrations of genipin were used for this purpose (1 mM, 2.5 mM, and 5 mM). Morphological, thermal, and mechanical properties were investigated for the crosslinked materials. The metabolic activity of embedded chondrocytes was also evaluated at different time points (3, 7, and 14 days). Non-crosslinked hydrogels resulted in an unstable matrix, while genipin-crosslinked hydrogels resulted in a stable matrix, without significant changes in their properties; their collagen network revealed characteristic dimensions in the order of 20 µm, while their denaturation temperature was 57 °C. After 7 and 14 days of culture, chondrocytes showed a significantly higher metabolic activity within the hydrogels crosslinked with 1 mM genipin, compared to those crosslinked with 5 mM genipin.

A stable adhesion to the cartilage is a crucial requisite for hydrogels used for cartilage regeneration. Indeed, a weak interface between the tissue and the implanted material may produce a premature detachment and thus the failure of the regeneration processes. Fibrin glue, cellulose nanofibers and catecholamines have been proposed in the state-of-the-art as primers to improve the adhesion. However, no studies focused on a systematic comparison of their performance. This work aims to evaluate the adhesion strength between ex vivo cartilage specimens and polysaccharide hydrogels (gellan gum and methacrylated gellan gum), by applying the mentioned primers as intermediate layer. Results show that the fibrin glue and the cellulose nanofibers improve the adhesion strength, while catecholamines do not guarantee reaching a clinically acceptable value. Stem cells embedded in gellan gum hydrogels reduce the adhesion strength when fibrin glue is used as a primer, being anyhow still sufficient for in vivo applications.

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