Triply periodic minimal surface microarchitectures (TPMS) were developed by mathematicians and evolved in all kingdoms of living organisms. Renowned for their lightweight yet robust attributes, TPMS structures find application in diverse fields, such as the construction of satellites, aircrafts, and electric vehicles. Moreover, these microarchitectures, despite their intricate geometric patterns, demonstrate potential for application as bone substitutes, despite the inherent gothic style of natural bone microarchitecture. Here, we produced three TPMS microarchitectures, D-diamond, G-gyroid, and P-primitive, by 3D printing from hydroxyapatite. We explored their mechanical characterization and, further, implanted them to study their bone augmentation and osteoconduction potential. In terms of strength, the D-diamond and G-gyroid performed significantly better than the P-primitive. In a calvarial defect model and a calvarial bone augmentation model, where osteoconduction is determined as the extent of bony bridging of the defect and bone augmentation as the maximal vertical bone ingrowth, the G-gyroid performed significantly better than the P-primitive. No significant difference in performance was observed between the G-gyroid and D-diamond. Since, in real life, the treatment of bone deficiencies in patients comprises elements of defect bridging and bone augmentation, ceramic scaffolds with D-diamond and G-gyroid microarchitectures appear as the best choice for a TPMS-based scaffold in bone tissue engineering.
Composite chitosan fibers filled with chitin nanofibrils (CNF) were obtained by the wet spinning method. The paper discusses the mechanical properties of such type fibers and their hemocompatibility, as well as the possibility of optimizing these properties by adding chitin nanofibrils. It was shown that low CNF concentration (about 0.5%) leads to an increase in fiber tensile strength due to the additional orientation of chitosan macromolecules. At the same time, with an increase in the content of CNF, the stability of the mechanical properties of composite fibers in a humid medium increases. All chitosan fibers, except 0.5% CNF, showed good hemocompatibility, even on prolonged contact with human blood. The addition of chitin nanofibers leads to decrease in hemoglobin molecules sorption due to the decline in optical density at wavelengths of 414 nm and 540 nm. Nevertheless, the hemolysis of fibers was comparable or even lesser that carbon hemosorbent, which is actively used in clinical practice.
The early phase of bone healing is a complex and poorly understood process. With additive manufacturing, we can generate a specific and customizable library of bone substitutes to explore this phase. In this study, we produced tricalcium phosphate-based scaffolds with microarchitectures composed of filaments of 0.50 mm in diameter, named Fil050G, and 1.25 mm named Fil125G, respectively. The implants were removed after only 10 days in vivo followed by RNA sequencing (RNAseq) and histological analysis. RNAseq results revealed upregulation of adaptive immune response, regulation of cell adhesion, and cell migration-related genes in both of our two constructs. However, significant overexpression of genes linked to angiogenesis, regulation of cell differentiation, ossification, and bone development was observed solely in Fil050G scaffolds. Moreover, quantitative immunohistochemistry of structures positive for laminin revealed a significantly higher number of blood vessels in Fil050G samples. Furthermore, µCT detected a higher amount of mineralized tissue in Fil050G samples suggesting a superior osteoconductive potential. Hence, different filament diameters and distances in bone substitutes significantly influence angiogenesis and regulation of cell differentiation involved in the early phase of bone regeneration, which precedes osteoconductivity and bony bridging seen in later phases and as consequence, impacts the overall clinical outcome.
Hemorrhage occurring during and after surgery still remains one of the biggest problems in medicine. Although a large number of hemostatic products have been created, there is no universal preparation; thus, the development of new materials is an urgent task. The aim of this research is to increase hemostatic properties of chitosan by introducing chitin nanofibrils (ChNF). The blood absorbance by ChNF-containing chitosan sponges and time-until-arrest of bleeding were studied. Non-woven materials containing 0.5% of ChNF and materials without chitin were obtained. The studies of ζ-potential showed that the material containing 0.5% ChNF had relatively a high positive charge, but efficiencies of both materials for hemorrhage arrest were comparable to those of commercial hemostatic products (Surgicel and TachoComb). To investigate the interaction between the materials and living organism, histological studies and optical microscopy studies were conducted after implantation of fibers. Despite bioinertness of fibers, implantation of non-woven materials led to formation of significant granulomas.
The functionalization of bone substitutes with exosomes appears to be a promising technique to enhance bone tissue formation. This study investigates the potential of exosomes derived from bone marrow mesenchymal stromal cells (BMSCs) to improve bone healing and bone augmentation when incorporated into wide open-porous 3D-printed ceramic Gyroid scaffolds. We demonstrated the multipotent characteristics of BMSCs and characterized the extracted exosomes using nanoparticle tracking analysis and proteomic profiling. Through cell culture experimentation, we demonstrated that BMSC-derived exosomes possess the ability to attract cells and significantly facilitate their differentiation into the osteogenic lineage. Furthermore, we observed that scaffold architecture influences exosome release kinetics, with Gyroid scaffolds exhibiting slower release rates compared to Lattice scaffolds. Nevertheless, in vivo implantation did not show increased bone ingrowth in scaffolds loaded with exosomes, suggesting that the scaffold microarchitecture and material were already optimized for osteoconduction and bone augmentation. These findings highlight the lack of understanding about the optimal delivery of exosomes for osteoconduction and bone augmentation by advanced ceramic scaffolds.
The chitosan fibers have been obtained by coagulation method from an aqueous solution of 2% acetic acid into an ethanol–alkaline mixture. The influence of the chitosan molecular mass, the...