Design and 3D Printing of Personalized Hybrid and Gradient Structures for Critical Size Bone Defects.

Treating critical-size bone defects with autografts, allografts, or standardized implants is challenging since the healing of the defect area necessitates patient-specific grafts with mechanically and physiologically relevant structures. Three-dimensional (3D) printing using computer-aided design (CAD) is a promising approach for bone tissue engineering applications by producing constructs with customized designs and biomechanical compositions. In this study, we propose 3D printing of personalized and implantable hybrid active scaffolds with a unique architecture and biomaterial composition for critical-size bone defects. The proposed 3D hybrid construct was designed to have a gradient cell-laden poly(ethylene glycol) (PEG) hydrogel, which was surrounded by a porous polycaprolactone (PCL) cage structure to recapitulate the anatomical structure of the defective area. The optimized PCL cage design not only provides improved mechanical properties but also allows the diffusion of nutrients and medium through the scaffold. Three different designs including zigzag, zigzag/spiral, and zigzag/spiral with shifting the zigzag layers were evaluated to find an optimal architecture from a mechanical point of view and permeability that can provide the necessary mechanical strength and oxygen/nutrient diffusion, respectively. Mechanical properties were investigated experimentally and analytically using finite element analysis (FEA), and computational fluid dynamics (CFD) simulation was used to determine the permeability of the structures. A hybrid scaffold was fabricated via 3D printing of the PCL cage structure and a PEG-based bioink comprising a varying number of human bone marrow mesenchymal stem cells (hBMSCs). The gradient bioink was deposited inside the PCL cage through a microcapillary extrusion to generate a mineralized gradient structure. The zigzag/spiral design for the PCL cage was found to be mechanically strong with sufficient and optimum nutrient/gas axial and radial diffusion while the PEG-based hydrogel provided a biocompatible environment for hBMSC viability, differentiation, and mineralization. This study promises the production of personalized constructs for critical-size bone defects by printing different biomaterials and gradient cells with a hybrid design depending on the need for a donor site for implantation.

'Cookies on a tray': Superselective hierarchical microstructured poly(l-lactide) surface as a decoy for cells.

In this research we developed a micro-sized hierarchical structures on a poly(l-lactide) (PLLA) surface. The obtained structures consist of round-shaped protrusions with a diameter of ~20 µm, a height of ~3 µm, and the distance between them ~30 µm. We explored the effect of structuring PLLA to design a non-cytotoxic material with increased roughness to encourage cells to settle on the surface. The PLLA films were prepared using the casting melt extrusion technique and were modified using ultra-short pulse irradiation - a femtosecond laser operating at λ = 1030 nm. A hierarchical microstructure was obtained resembling 'cookies on a tray'. The cellular response of fibro- and osteoblasts cell lines was investigated. The conducted research has shown that the laser-modified surface is more conducive to cell adhesion and growth (compared to unmodified surface) to such an extent that allows the formation of highly-selectively patterns consisting of living cells. In contrast to eukaryotic cells, the pathogenic bacteria Staphylococcus aureus covered modified and unmodified structures in an even, non-preferential manner. In turn, adhesion pattern of eukaryotic fungus Saccharomyces boulardii resembled that of fibro- and osteoblast cells rather than that of Staphylococcus. The discovered effect can be used for fabrication of personalized and smart implants in regenerative medicine.

Melt Electrowriting of Complex 3D Anatomically Relevant Scaffolds.

The manufacture of fibrous scaffolds with tailored micrometric features and anatomically relevant three-dimensional (3D) geometries for soft tissue engineering applications remains a great challenge. Melt electrowriting (MEW) is an advanced additive manufacturing technique capable of depositing predefined micrometric fibers. However, it has been so far inherently limited to simple planar and tubular scaffold geometries because of the need to avoid polymer jet instabilities. In this work, we surmount the technical boundaries of MEW to enable the manufacture of complex fibrous scaffolds with simultaneous controlled micrometric and patient-specific anatomic features. As an example of complex geometry, aortic root scaffolds featuring the sinuses of Valsalva were realized. By modeling the electric field strength associated with the MEW process for these constructs, we found that the combination of a conductive core mandrel with a non-conductive 3D printed model reproducing the complex geometry minimized the variability of the electric field thus enabling the accurate deposition of fibers. We validated these findings experimentally and leveraged the micrometric resolution of MEW to fabricate unprecedented fibrous aortic root scaffolds with anatomically relevant shapes and biomimetic microstructures and mechanical properties. Furthermore, we demonstrated the fabrication of patient-specific aortic root constructs from the 3D reconstruction of computed tomography clinical data.