Novel titanium-apatite hybrid scaffolds with spongy bone-like micro architecture intended for spinal application: In vitro and in vivo study.

Titanium alloy scaffolds with novel interconnected and non-periodic porous bone-like micro architecture were 3D-printed and filled with hydroxyapatite bioactive matrix. These novel metallic-ceramic hybrid scaffolds were tested in vitro by direct-contact osteoblast cell cultures for cell adhesion, proliferation, morphology and gene expression of several key osteogenic markers. The scaffolds were also evaluated in vivo by implanting them on transverse and spinous processes of sheep's vertebras and subsequent histology study. The in vitro results showed that: (a) cell adhesion, proliferation and viability were not negatively affected with time by compositional factors (quantitative MTT-assay); (b) the osteoblastic cells were able to adhere and to attain normal morphology (fluorescence microscopy); (c) the studied samples had the ability to promote and sustain the osteogenic differentiation, matrix maturation and mineralization in vitro (real-time quantitative PCR and mineralized matrix production staining). Additionally, the in vivo results showed that the hybrid scaffolds had greater infiltration, with fully mineralized bone after 6 months, than the titanium scaffolds without bioactive matrix. In conclusion, these novel hybrid scaffolds could be an alternative to the actual spinal fusion devices, due to their proved osteogenic performance (i.e. osteoinductive and osteoconductive behaviour), if further dimensional and biomechanical optimization is performed.

Synthetic open cell foams versus a healthy human vertebra: Anisotropy, fluid flow and µ-CT structural studies.

Commercial synthetic open-cell foams are an alternative to human cadaveric bone to simulate in vitro different scenarios of bone infiltration properties. Unfortunately, these artificial foams do not reproduce the anisotropic microstructure of natural bone and, consequently, their suitability in these studies is highly questionable. In order to achieve scaffolds that successfully mimic human bone, microstructural studies of both natural porous media and current synthetic approaches are necessary at different length scales. In this line, the present research was conducted to improve the understanding of local anisotropy in natural vertebral bone and synthetic bone-like porous foams. To attain this objective, small volumes of interest within these materials were reconstructed via micro-computed tomography. The anisotropy of the microstructures was analysed by means of both their main local histomorphometric features and the behaviour of an internal flow computed via computational fluid dynamics. The results showed that the information obtained from each of the micro-volumes of interest could be scaled up to understand not only the macroscopic averaged isotropic and/or anisotropic behaviour of the samples studied, but also to improve the design of macroscopic porous implants better fitting specific local histomorphometric scenarios. The results also clarify the discrepancies in the permeability obtained in the different micro-volumes of interest analysed. STATEMENT OF SIGNIFICANCE: A deep insight comparative study between the porous microstructure of healthy vertebral bone and that of synthetic bone-like open-cell rigid foams used in in vitro permeability studies of bone cement has been performed. The results obtained are of fundamental relevance to computational studies because, in order to achieve convergence values, the computation process should be limited to small computation domains or micro-volumes of interest. This makes the results specific spatial dependent and for this reason computation studies cannot directly capture the macroscopic average behaviour of an anisotropic porous structure such as the one observed in natural bones. The results derived from this study are also important because we have been able to show that the specific spatial information contained in only one healthy vertebra is enough to capture, from a geometric point of view, the same information of "specific surface area vs. porosity" - in other words, the same basic law - that can also be found in other human bones for different patients, even at different biological ages. This is an important finding that, despite the efforts made and the controversies formulated by other authors, should be studied more thoroughly with other bone species and tissues (healthy and/or diseased). Moreover, our results should help to understand that, with the extensive capabilities of current 3D printing technologies, there is an enormous potential in the design of biomimetic porous bone-like scaffolds for bone tissue engineering applications.

Are TiO2 nanoparticles safe for photocatalysis in aqueous media?

Although environmental and toxicity concerns are inherently linked, catalysis using photoactive nanoparticles and their hazardous potential are usually addressed independently. A toxicological assessment under the application framework is particularly important, given the pristine nanoparticles tend to change characteristics during several processes used to incorporate them into products. Herein, an efficient TiO2-functionalized macroporous structure was developed using widely adopted immobilization procedures. The relationships between photocatalysis, catalyst release and associated potential environmental hazards were assessed using zebrafish embryonic development as a proxy. Immobilized nanoparticles demonstrated the safest approach to the environment, as the process eliminates remnant additives while preventing the release of nanoparticles. However, as acute sublethal effects were recorded in zebrafish embryos at different stages of development, a completely safe release of TiO2 nanoparticles into the aquatic environment cannot be advocated.