Artificial bone composites exhibit distinctive features by comparison to natural tissues, due to a lack of self-organization and intimate interaction apatite-matrix. This explains the need of "bio-inspired materials", in which hydroxyapatite grows in contact with self-assembling natural polymers. The present work investigates the function of a rational design in the hydroxyapatite-forming potential of a common biopolymer. Gelatin modified through intrinsic interactions with calcium alginate led through freeze-drying to porous hydrogels, whose architecture, constitutive features and chemistry were investigated with respect to their role on biomineralization. The apatite-forming ability was enhanced by the porosity of the materials, while the presence of alginate-reinforced Gel elastic chains, definitely favored this phenomenon. Depending on the concentration, polysaccharide chains act as "ionic pumps" enhancing the biomineralization. The mineralization-promoting effect of the peptide-polysaccharide network strictly depends on the hydrogels structural, compositional and morphological features derived from the interaction between the above mentioned two components.
Non-biodegradable porous polystyrene (PS) scaffolds, composed of microfibers, have been prepared by electrospinning for the reconstruction of large bone defects. PS microfibers were prepared by incorporating β-TCP grains inside the polymer or grafting gold nanoparticles surface functionalized with mercaptosuccinic acid. Cytocompatibility of the three types of scaffolds (PS, β-TCP-PS and Au-PS) was studied by seeding human mesenchymal stem cells. Biocompatibility was evaluated by implanting β-TCP-PS and Au-PS scaffolds into a critical size (4mm) calvarial defect in mice. Calvaria were taken 6, 9, and 12 weeks after implantation; newly formed bone and cellular response was analyzed by microcomputed tomography (microCT) and histology. β-TCP-PS scaffolds showed a significantly higher cell proliferation in vitro than on PS or Au-PS alone; clearly, the presence of β-TCP grains improved cytocompatibility. Biocompatibility study in the mouse calvaria model showed that β-TCP-PS scaffolds were significantly associated with more newly-formed bone than Au-PS. Bone developed by osteoconduction from the defect margins to the center. A dense fibrous connective tissue containing blood vessels was identified histologically in both types of scaffolds. There was no inflammatory foci nor giant cell in these areas. AuNPs aggregates were identified histologically in the fibrosis and also incorporated in the newly-formed bone matrix. Although the different types of PS microfibers appeared cytocompatible during the in vitro experiment, they appeared biotolerated in vivo since they induced a fibrotic reaction associated with newly formed bone.
Bioink-formulations based on gelatin methacrylate combined with oxidized cellulose nanofibrils are employed in the present study. The parallel investigation of the printing performance, morphological, swelling, and biological properties of the newly developed hydrogels was performed, with inks prepared using methacrylamide-modified gelatins of fish or bovine origin. Scaffolds with versatile and well-defined internal structure and high shape fidelity were successfully printed due to the high viscosity and shear-thinning behavior of formulated inks and then photo-crosslinked. The biocompatibility of 3D-scaffolds was surveyed using human adipose stem cells (hASCs) and high viability and proliferation rates were obtained when in contact with the biomaterial. Furthermore, bioprinting tests were performed with hASCs embedded in the developed formulations. The results demonstrated that the designed inks are a versatile toolkit for 3D bioprinting and further show the benefits of using fish-derived gelatin for biofabrication.
In this work we report the potential of nanostructured fibers consisting of nanodiamond particles (NDPs) and fish gelatin (FG) to modulate the mechanical properties of fish gelatin hydrogels, in the aim of developing bioinspired ECM analogues. NDPS-loaded FG fibers were obtained by electrospinning. The biocomposites were obtained through the enzymatic crosslinking of gelatin in the presence of NDPs-loaded FG fibers. The mechanical behavior was assessed at different preparative stages (precursors, fibrous mesh, biocomposite scaffolds).
Nano-apatite and gelatin-alginate hydrogel microparticles have been prepared by a one-step synthesis combined with electrostatic bead generation, for the reconstruction of bone defects. Based on the analysis of bone composition, architecture and embryonic intramembranous ossification, a bio-inspired fabrication has been developed. Accordingly, the mineral phase has been in situ synthesized, calcifying the hydrogel matrix while the latter was crosslinked, finally generating microparticles that can assemble into a bone defect to ensure interconnected pores. Although nano-apatite-biopolymer composites have been widely investigated, microstructural optimization to provide improved distribution and stability of the mineral is rarely achieved. The optimization of the developed method progressively resulted in two types of formulations (15P and 7.5P), with 15 and 7.5 (wt%) phosphate content in the initial precursor. The osteolytic potential was investigated using differentiated macrophages. A commercially available calcium phosphate bone graft substitute (Eurocer 400) was incorporated into the hydrogel, and the obtained composites were in vitro tested for comparison. The cytocompatibility of the microparticles was studied with mouse osteoblast-like cell line MC3T3-E1. Results indicated the best in vitro performance have been obtained for the sample loaded with 7.5P. Preliminary evaluation of biocompatibility into a critical size (3 mm) defect in rabbits showed that 7.5P nanocomposite is associated with newly formed bone in the proximity of the microparticles, after 28 days.
In this work, we focused on the potential of biogenous mineral from Cuttlefish bone for inducing biomimetic mineralization with further biomedical applications. We demonstrate the formation of strong and flexible hydrogels via physical immobilization of cuttlefish bone powder in the presence of bovine gelatin and alginate. The properties of hydrogels loaded with biogenous mineral, with a commercial bone substitute, with commercial nano-hydroxyapatite and in situ generated hydroxyapatite. In vitro cell adhesion tests demonstrated that the new developed hydrogels showed a promising cell adhesion.
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