3D printed scaffolds of biosilica and spongin from marine sponges: Analysis of genotoxicity and cytotoxicity for bone tissue repair

Abstract

Biosilica (BS) and spongin (SPG) from marine sponges are highlighted for their potential to promote bone regeneration. Moreover, additive manufacturing, specifically 3D printing, is introduced as a technology for producing bone grafts with optimized interconnected porous structures, allowing for better cell attachment, proliferation, and differentiation. Thus, the aims of this study were to characterize the BS and BS/SPG 3D printed scaffolds and to evaluate the biological effects in vitro. The physicochemical characteristics of BS and BS/SPG 3D printed scaffolds were analyzed by SEM, FTIR, porosity, evaluation of mass loss, and pH measurement. For in vitro analysis, the cellular viability of the MC3T3-E1 cell lineage was assessed using the AlamarBlue® assay and SEM, while genotoxicity was evaluated through the micronucleus assay. SEM analysis revealed distinct features: the presence of spicules in BS, the fibrillar structure of SPG, and material degradation over the immersion period. FTIR indicated peaks corresponding to silicon oxide in BS samples and carbon oxide and amine in SPG samples. BS-SPG scaffolds exhibited higher porosity, while BS scaffolds displayed greater mass loss. pH measurements indicated a significant decrease induced by BS, which was mitigated by SPG over the experimental periods. In vitro studies demonstrated the biocompatibility and non-cytotoxicity of scaffold extracts. The micronucleus (MN) test further confirmed the absence of cytotoxicity in the samples. These findings suggest that 3D printed BS and BS/SPG scaffolds may possess desirable morphological and physicochemical properties, indicating in vitro biocompatibility.