Genipin (GN) is a natural molecule extracted from the fruit of Gardenia jasminoides Ellis according to modern microbiological processes. Genipin is considered as a favorable cross-linking agent due to its low cytotoxicity compared to widely used cross-linkers; it cross-links compounds with primary amine groups such as proteins, collagen, and chitosan. Chitosan is a biocompatible polymer that is currently studied in bone tissue engineering for its capacity to promote growth and mineral-rich matrix deposition by osteoblasts in culture. In this work, two genipin cross-linked chitosan scaffolds for bone repair and regeneration were prepared with different GN concentrations, and their chemical, physical, and biological properties were explored. Scanning electron microscopy and mechanical tests revealed that nonremarkable changes in morphology, porosity, and mechanical strength of scaffolds are induced by increasing the cross-linking degree. Also, the degradation rate was shown to decrease while increasing the cross-linking degree, with the high cross-linking density of the scaffold disabling the hydrolysis activity. Finally, basic biocompatibility was investigated in vitro, by evaluating proliferation of two human-derived cell lines, namely, the MG63 (human immortalized osteosarcoma) and the hMSCs (human mesenchymal stem cells), as suitable cell models for bone tissue engineering applications of biomaterials.

Chitosan is a polysaccharide commonly used, together with its derivatives, in the preparation of hydrogel formulations, scaffolds and films for tissue engineering applications. Chitosan can be used as such, but it is commonly stabilized by means of chemical crosslinkers. Genipin is one of the crosslinkers that has been considered that is a crystalline powder extracted from the fruit of Gardenia jasminoides and processed to obtain an aglycon compound. Genipin is gaining interest in biological applications because of its natural origin and anti-inflammatory actions. In this paper, the ability of chitosan-based materials crosslinked with genipin to exert anti-inflammation properties in applications such as bone regeneration was studied. Powders obtained from chitosan–genipin scaffolds have been tested in order to mimic the natural degradation processes occurring during biomaterials implantation in vivo. The results from osteoblast-like cells showed that specific combinations of chitosan and genipin stimulate high permissiveness towards cells, with higher performance than the pure chitosan. In parallel, evidences from monocyte-like cells showed that the crosslinker, genipin, seems to promote slowing of the monocyte-macrophage transition at morphological level. This suggests a sort of modularity of pro-inflammatory versus anti-inflammatory behavior of our chitosan-based biomaterials. Being both the cell types exposed to microscale powders, as an added value our results bring information on the cell–material interactions in the degradative dynamics of chitosan scaffold structures during the physiological resorption processes.

In order to induce bone regeneration several natural and synthetic materials have been proposed. However, single-phase scaffolds present some insurmountable disadvantages such as poor mechanical strength or brittleness and too low or too high degradation rate. In order to overcome these drawbacks, composite systems can be an interesting and promising option. In the present work a novel hybrid porous scaffold for bone tissue engineering is proposed. Chitosan/Forsterite (Ch/FS) composite scaffolds were prepared by freeze-drying method using a chitosan/forsterite ratio of 90/10. The FS nanopowder (Mg₂SiO₄) is synthesized using a simple solgel based method. The FS composition was checked by XRD analysis. The macrostructure of the Ch/FS scaffolds were analyzed by SEM, the FS distribution within the chitosan matrix observed by EDS, the mechanical strength measured by compression test in PBS and the biocompatibility of the composite on human osteosarcoma cell line (MG-63) verified by MTT assay after 48 hours. The porosity appears interconnected and with a pore size ranging from 1 to 100 μm. The FS is overall distributed within the chitosan matrix. The compression strength of composite scaffolds increased with respect to the pure chitosan scaffolds of more than two times (from 0.8 to 1.9 KPa) and the composites did not show any toxicity effect on human osteosarcoma cells.