In this study porous gelatin scaffolds were prepared using in-situ gas foaming, and four crosslinking agents were used to determine a biocompatible and effective crosslinker that is suitable for such a method. Crosslinkers used in this study included: hexamethylene diisocyanate (HMDI), poly(ethylene glycol) diglycidyl ether (epoxy), glutaraldehyde (GTA), and genipin. The prepared porous structures were analyzed using Fourier Transform Infrared Spectroscopy (FT-IR), thermal and mechanical analysis as well as water absorption analysis. The microstructures of the prepared samples were analyzed using Scanning Electron Microscopy (SEM). The effects of the crosslinking agents were studied on the cytotoxicity of the porous structure indirectly using MTT analysis. The affinity of L929 mouse fibroblast cells for attachment on the scaffold surfaces was investigated by direct cell seeding and DAPI-staining technique. It was shown that while all of the studied crosslinking agents were capable of stabilizing prepared gelatin scaffolds, there are noticeable differences among physical and mechanical properties of samples based on the crosslinker type. Epoxy-crosslinked scaffolds showed a higher capacity for water absorption and more uniform microstructures than the rest of crosslinked samples, whereas genipin and GTA-crosslinked scaffolds demonstrated higher mechanical strength. Cytotoxicity analysis showed the superior biocompatibility of the naturally occurring genipin in comparison with other synthetic crosslinking agents, in particular relative to GTA-crosslinked samples.

In this study, a nanostructured scaffold was designed for bone repair using hydroxyapatite (HA) and gelatin (GEL) as its main components. Nanopowders of HA were synthesized, and together with GEL, used to engineer a 3-dimensional nanocomposite combining 3 techniques of layer solvent casting, freeze-drying, and lamination. The results show that the scaffold possesses a 3-dimensional interconnected homogenous porous structure with a porosity of 82% and pore sizes ranging from 300 to 500 mum. It has also been shown that mechanical indices are in the range of spongy bones. Cultured osteoblast-like cells (SaOS-2) have shown an excellent level of cell attachment, migration, and penetration into the porosities of the nanocomposite scaffold. Here, we have shown that by a combination of widely available methods with simple experimental operations, nano-HA powders can be synthesized and used to make 3-dimensional HA/GEL nanocomposites in any desired shape, with mechanical properties comparable to spongy bone.

The current study presents an effective and simple strategy to obtain stable porous scaffolds from gelatin via gas foaming method. The technique exploits the intrinsic foaming ability of gelatin in the presence of CO2 to obtain a porous structure stabilised with glutaraldehyde. The produced scaffolds were characterised using physical and mechanical characterisation methods. The results showed that gas foaming may allow the tailoring of the 3-dimensional structure of the scaffolds with an interconnected porous structure. To assess the effectiveness of preparation method in mitigating the potential cytotoxicity risk of using glutaraldehyde as a crosslinker, direct and indirect cytotoxicity assays were performed at different concentrations of glutaraldehyde. The results indicate the potential of the gas foaming method, in the preparation of viable tissue engineering scaffolds.

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