Physicochemical properties and cytocompatibility assessment of non-degradable scaffolds for bone tissue engineering applications.

Bone is a dynamic tissue with an amazing but yet limited capacity of self-healing. Bone is the second most transplanted tissue in the world and there is a huge need for bone grafts and substitutes which lead to a decrease in bone banks donors. In this study, we developed three-dimensional scaffolds based on Ti6Al4V, ZrO2 and PEEK targeting bone tissue engineering applications. Experimental mechanical compressive tests and finite element analyses were carried out to study the mechanical performance of the scaffolds. Overall, the scaffolds presented different hydrophilicity properties and a reduced elastic modulus when compared with the corresponding solid materials which can in some extension minimize the phenomenon of stress shielding. The ability as a scaffold material for bone tissue regeneration applications was evaluated in vitro by seeding human osteosarcoma (SaOS-2) cells onto the scaffolds. Then, the successful culture of SaOS-2 cells on developed scaffolds was monitored by assessment of cell's viability, proliferation and alkaline phosphatase (ALP) activity up to 14 days of culturing. The in vitro results revealed that Ti6Al4V, ZrO2 and PEEK scaffolds were cytocompatible allowing the successful culture of an osteoblastic cell line, suggesting their potential application in bone tissue engineering. Statement of Significance. The work presented is timely and relevant since it gathers both the mechanical and cellular study of non-degradable cellular structures with the potential to be used as bone scaffolds. This work allow to investigate three possible bone scaffolds solutions which exhibit a significantly reduced elastic modulus when compared with conventional solid materials. While it is generally accepted that the Ti6Al4V, ZrO2 and PEEK are candidates for such applications a further study of their features and their comparison is extremely important for a better understanding of their potential.

Micro-CT based finite element modelling and experimental characterization of the compressive mechanical properties of 3-D zirconia scaffolds for bone tissue engineering.

The present study aims at developing a computational framework with experimental validation to determine the mechanical properties of zirconia foams for bone tissue engineering. A micro-CT based finite element model that allows characterizing the mechanical property of such cellular structures is developed. Micro-CT images are filtered to vanish noises and smooth boundaries before constructing 3D zirconia foams using an adaptive Body-Centered Cubic background lattice. In addition to micro-CT images, the local material property at the scaffold struts is measured using a micro-indentation test, which shows a considerable difference with that of common zirconia owing to the manufacturing process. The computational model also takes the plastic deformation of material into account employing the Voce law, a nonlinear isotropic hardening law, as well as Von-mises yield criterion. Zirconia foams with different pore sizes are manufactured using the replica method and their mechanical properties determined experimentally. Such experimental outcomes are to validate and demonstrate the capability of the developed model, which can be used for pre-operational evaluations and preclinical tests of zirconia scaffolds. The stress magnitude and distribution within the scaffold as well as plastic strains and flow stress of the zirconia scaffold are computed and analysed. Using the proposed approach, a deep insight into the association of macroscopic behaviour of the scaffold to microscopic features, e.g. strut waviness, Plateau border, thickness variation of cells, irregularity, microstructural variability, imperfections and strut's material property associated with to the manufacturing procedure, can be gained.

Gellan Gum-based luminal fillers for peripheral nerve regeneration: an in vivo study in the rat sciatic nerve repair model.

Peripheral nerve injuries (PNI) resulting in a gap to be bridged between the transected nerve ends are commonly reconstructed with autologous nerve tissue, but there is a need for valuable alternatives. This experimental work considers the innovative use of the biomaterial Gellan Gum (GG) as a luminal filler for nerve guidance channels made from chitosan with a 5% degree of acetylation. The engineered constructs should remodel the structural support given to regenerating axons by the so-called bands of Büngner. Four different GG formulations were produced by combining varying amounts of High-Acyl GG (HA-GG) and Methacrylated GG (MA-GG). The effective porosity of the freeze-dried networks was analysed by SEM and micro-CT 3D reconstructions, while the degradation and swelling abilities were characterized in vitro for up to 30 days. The metabolic activity and viability of immortalized Schwann cells seeded onto the freeze-dried networks were also evaluated. Finally, the developed hydrogel formulations were freeze-dried within the chitosan nerve guides and implanted in a 10 mm rat sciatic nerve defect. Functional and histomorphological analyses after 3, 6, and 12 weeks in vivo revealed that although it did not result in improved nerve regeneration, the NGC25:75 formulations could provide a basis for further development of GG scaffolds as luminal fillers for hollow nerve guidance channels.

Biomechanical and cellular segmental characterization of human meniscus: building the basis for Tissue Engineering therapies.

OBJECTIVE: To overcome current limitations of Tissue Engineering (TE) strategies, deeper comprehension on meniscus biology is required. This study aims to combine biomechanical segmental analysis of fresh human meniscus tissues and its correlation with architectural and cellular characterization. METHOD: Morphologically intact menisci, from 44 live donors were studied after division into three radial segments. Dynamic mechanical analysis (DMA) was performed at physiological-like conditions. Micro-computed tomography (CT) analysis of freeze-dried samples assessed micro-structure. Flow cytometry, histology and histomorphometry were used for cellular study and quantification. RESULTS: Anterior segments present significantly higher damping properties. Mid body fresh medial meniscus presents higher values of E' compared to lateral. Cyclic loads influence the viscoelastic behavior of menisci. By increasing the frequency leads to an increase in stiffness. Conversely, with increasing frequencies, the capacity to dissipate energy and damping properties initially decrease and then rise again. Age and gender directly correlate with higher E' and tan δ. Micro-CT analysis revealed that mean porosity was 55.5 (21.2-89.8)% and 64.7 (47.7-81.8)% for freeze-dried lateral and medial meniscus, respectively. Predominant cells are positive for CD44, CD73, CD90 and CD105, and lack CD31, CD34 and CD45 (present in smaller populations). Histomorphometry revealed that cellularity decreases from vascular zone 1 to zone 3. Anterior segments of lateral and medial meniscus have inferior cellularity as compared to mid body and posterior ones. CONCLUSION: Menisci are not uniform structures. Anterior segments have lower cellularity and higher damping. Cyclic loads influence viscoelastic characteristics. Future TE therapies should consider segmental architecture, cellularity and biomechanics of fresh tissue.