Alginate-based hydrogels functionalised at the nanoscale using layer-by-layer assembly for potential cartilage repair

Injuries to articular cartilage are frequently difficult to repair, in part because of the poor regenerative capacity of this tissue. To date, no successful system for complete regeneration of the most challenging cartilage defects has been demonstrated. The aim of this work was to develop functionalised hydrogels at the nanoscale by Layer-by-Layer (LbL) assembly to promote cartilage healing. Hydrogels, based on sodium alginate (NaAlg) and gelatin (G), were prepared by an external gelation method consisting of CaCl₂ diffusion and genipin addition for G crosslinking. Successively, hydrogels were coated with G to obtain a positive charge on the surface, then functionalised by LbL assembly to create 16 nanolayers, based on poly(styrene sulfonate)/poly(allyl amine) (PSS/PAH), including a specific peptide sequence (CTATVHL) and transforming growth factors β1 (TGF-β1). Physico-chemical properties were evaluated by XPS, ATR-FTIR and rheological analyses while in vitro cytocompatibility was studied using bovine articular chondrocytes (BAC). XPS spectra showed N_1s and S_2p peaks, indicating that PAH and PSS have been introduced with success. ATR-FTIR indicated the specific PAH and PSS absorption peaks. Finally, the biomolecule incorporation influenced positively the processes of BAC adhesion and proliferation, and glycosamynoglycan secretion. The functionalised alginate-based hydrogels described here are ideally suited to chondral regeneration in terms of their integrity, stability, and cytocompatibility.