Advanced biomimetic design strategies for porous structures promoting bone integration with additive-manufactured Ti6Al4V scaffolds

The natural bone structure exhibits a radial gradient with dense cortical bone externally and porous cancellous bone internally. However, previous studies proposing scaffold designs have predominantly focused on homogeneous porous structures. Moreover, no consensus exists on the optimal structure for gradient porous scaffolds. Our scaffold closely imitated the natural bone structure by incorporating a gradient of pillar diameters. Additionally, we introduced a inverse gradient structure and three uniform diameter pillar structures (400 mu m, 600 mu m, and 1000 mu m) for comparative analysis. Mechanical testing revealed that the compressive strength and elastic modulus of the Ti6Al4V porous scaffolds gradually increased with an increase in pillar diameter. In vitro experiments demonstrated that both the biomimetic gradient and inverse gradient scaffolds promoted osteogenic differentiation, with higher ALP activity (alkaline phosphatase) and osteogenesis-related gene expression compared to the uniform pillar structure scaffolds. The in vivo experiments confirmed these results, highlighting the superior ability of the biomimetic gradient Ti6Al4V porous scaffold to induce new bone formation. Based on our findings, we suggest that the optimal porous structure should have fine-diameter pillars (approximately 400 mu m) internally to enhance cell penetration, while coarse-diameter pillars (approximately 800 mu m) should be used externally to increase the cell attachment area and enhance mechanical strength. Overall, our study contributes to the field of bone tissue engineering by providing a biomimetic scaffold design that closely imitates the structure of natural bone and promotes osteogenic activity.