Fabrication of hierarchically porous silk fibroin-bioactive glass composite scaffold via indirect 3D printing: Effect of particle size on physico-mechanical properties and in vitro cellular behavior.

In order to regenerate bone defects, bioactive hierarchically scaffolds play a key role due to their multilevel porous structure, high surface area, enhanced nutrient transport and diffusion. In this study, novel hierarchically porous silk fibroin (SF) and silk fibroin-bioactive glass (SF-BG) composite were fabricated with controlled architecture and interconnected structure, by combining indirect three-dimensional (3D) inkjet printing and freeze-drying methods. Further, the effect of 45S5 Bioactive glass particles of different sizes (<100 nm and 6 μm) on mechanical strength and cell behavior was investigated. The results demonstrated that the hierarchical structure in this scaffold was composed of two levels of pores in the order of 500-600 μm and 10-50 μm. The prepared SF-BG composite scaffolds utilized by nano and micro particles possessed mechanical properties with a compressive strength of 0.94 and 1.2 MPa, respectively, in dry conditions. In a wet condition, the hierarchically porous scaffolds did not exhibit any fluctuation after compression load cell and were incredibly flexible, with excellent mechanical stability. The SF-BG composite scaffold with nanoparticles presented a significant 50% increase in attachment of human bone marrow stem cells in comparison with SF and SF-BG scaffold with microparticles. Moreover, SF-BG scaffolds promoted alkaline phosphatase activity as compared to SF scaffolds without BG particles on day 14. In brief, the 3D porous silk fibroin-based composites containing BG nanoparticles with excellent mechanical properties are promising scaffolds for bone tissue regeneration in high load-bearing applications.