Feasibility of Using H₃PO₄/H₂O₂ in the Synthesis of Antimicrobial TiO₂ Nanoporous Surfaces

Ti6Al4V alloys are the primary materials used for clinical bone regeneration and restoration; however, they are substantially susceptible to biomaterial-related infections. Therefore, in the present work, we applied a controllable and stable oxidative nanopatterning strategy by applying H₃PO₄, a weaker dissociating acid, as a substitute for H₂SO₄ in the classical piranha reaction. The results suggest that our method acted as a concomitant platform to develop reproducible diameter-controlled TiO₂ nanopores (NPs). Interestingly, our procedure illustrated stable temperature reactions without exothermic responses since the addition of mixture preparation to the nanopatterning reactions. The reactions were carried out for 30 min (NP14), 1 h (NP7), and 2 h (NP36), suggesting the formation of a thin nanopore layer as observed by Raman spectroscopy. Moreover, the antimicrobial activity revealed that NP7 could disrupt active microbial colonization for 2 h and 6 h. The phenotype configuration strikingly showed that NP7 does not alter the cell morphology, thus proposing a disruptive adhesion pathway instead of cellular lysis. Furthermore, preliminary assays suggested an early promoted osteoblasts viability in comparison to the control material. Our work opens a new path for the rationale design of nanobiomaterials with "intelligent surfaces" capable of decreasing microbial adhesion, increasing osteoblast viability, and being scalable for industrial transfer.