Optimized composition of nanocomposite scaffolds formed from silk fibroin and nano-TiO2 for bone tissue engineering

• Preparation a novel fibroin/nano-TiO2 scaffolds
• Formation of directional pores in scaffolds.
• Improving the bioactivity of fibroin/nano-TiO2 scaffolds
• Improving the biocompatibility of fibroin/nano-TiO2 scaffolds
• Optimizing the composition of fibroin/nano-TiO2 scaffold

Natural silk fibroin (SF) polymer has biomedical and mechanical properties as a biomaterial for bone tissue engineering scaffolds. Freeze-dried porous nanocomposite scaffolds were prepared from silk fibroin and titanium dioxide (TiO2) nanoparticles as a bioactive reinforcing agent by a phase separation method. In order to fabricate SF/TiO2 scaffolds, 5, 10, 15 and 20 wt% of the TiO2 were added to the SF. The phase structure, functional groups and morphology of the scaffolds were evaluated using X-ray diffraction, Fourier transform infrared spectroscopy and scanning electron microscopy techniques, respectively. Porosity of the scaffolds was measured by Archimedes' Principle. In addition, mechanical properties of prepared scaffolds were evaluated by measuring the compressive strength and compressive modulus. The bioactivity property of these scaffolds was examined for 7, 14, 21 and 28 days immersion in simulated body fluid (SBF) at 37 °C and the in vitro degradation was studied by incubation in phosphate buffered saline (PBS) at 37 °C and pH 7.4 for up to 30 days. Moreover, the scaffolds' biocompatibility was evaluated by seeding and culture of SaOS-2 osteoblast-like cells and assessment of their proliferation with MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Results showed that the prepared scaffolds had directional porosity and the reduction of porosity in composite scaffolds with higher contents of TiO2 nanoparticles resulted to an improvement of the mechanical strength. The macroporous structures with open interconnected and directional pores were successfully obtained without applying any porogen or inorganic solvent. The bioactivity of these scaffolds was confirmed by scanning electron microscopy (SEM) showing surface crystallization of the apatite layer proportional to the duration of immersion in the SBF and the degradation rate of scaffolds were increased by increasing the TiO2 content. The osteoblast-like cells showed a high attachment and proliferation on these scaffolds and their viability was increased with increasing the SF content. Finally, an optimum composition of SF/TiO2 nanocomposite scaffolds was selected.