Porous Silk Fibroin Membrane as a Potential Scaffold for Bone Regeneration

Abstract

The requirements for scaffolds for bone tissue engineering include appropriate chemistry, morphology and structure to promote cell adhesion and synthesis of new bone matrix. Silk fibroin (SF) represents an important biomaterial for biomedical application, due to its suitable mechanical properties, biodegradability, biocompatibility, and versatility in processing. Our group has developed a new method to obtain a porous SF membrane, and the study of its potential for use as a scaffold for bone regeneration was the aim of this study. Porous membranes were obtained from SF solution, through the compression of a material generated by phase separation. For in vitro calcification experiments, porous SF membrane samples were immersed in SBF at pH 7.4 placed in polyethylene flasks. The experiments were carried out for seven days, at 36.5±0.5 °C. After 48 and 96h, the solutions were changed for fresh SBF with the ion concentration 1.5-fold higher than that of the standard one, to accelerate the calcification process. The characterization of morphology and composition of samples was performed by using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS), respectively. The SEM micrographs indicated that the porous SF membranes presented calcium phosphate deposits after undergoing in vitro calcification. These results were confirmed by EDS spectra, which showed a stoichiometric molar Ca/P ratio ranging from 1.27 to 1.52. This fact may suggest that calcification deposits consisted of mixtures of HAP (Ca/P ratio = 1.67) and transient HAP precursor phases, such as octacalcium phosphate (Ca/P = 1.33) and dicalcium phosphate dehydrate (Ca/P = 1), indicating early stage mineralization. The porous silk fibroin membrane analysed in the current study is a promising material to be used as scaffolds for bone regeneration.