Antimicrobial potential and osteoblastic cell growth on electrochemically modified titanium surfaces with nanotubes and selenium or silver incorporation

Titanium nanotube surfaces containing silver, zinc, and copper have shown antimicrobial effects without decreasing osteoblastic cell growth. In this in-vitro study we present first results on the biological evaluation of surface modifications by incorporating selenium and silver compounds into titanium-dioxide (TiO₂) nanotubes by electrochemical deposition. TiO₂-nanotubes (TNT) and Phosphate-doped TNT (pTNT) were grown on the surface of Ti6Al4V discs by anodization. Hydroxyapatite (HA), selenium (Se) and silver (Ag) compounds were incorporated by electrochemical deposition. Colony forming units of Staphylococcus epidermidis (DSM 3269) were significantly decreased in SepTNT (0.97 ± 0.18 × 10⁶ CFU/mL), SepTNT-HA (1.2 ± 0.39 × 10⁶ CFU/mL), AgpTNT (1.36 ± 0.42 × 10⁶ CFU/mL) and Ag₂SepTNT (0.999 ± 0.12 × 10⁶ CFU/mL) compared to the non-modified control (2.2 ± 0.21 × 10⁶ CFU/mL). Bacterial adhesion was calculated by measuring the covered area after fluorescence staining. Adhesion was lower in SepTNT (37.93 ± 12%; P = 0.004), pTNT (47.3 ± 6.3%, P = 0.04), AgpTNT (24.9 ± 1.8%; P < 0.001) and Ag₂SepTNT (14.9 ± 4.9%; P < 0.001) compared to the non-modified control (73.7 ± 11%). Biofilm formation and the growth of osteoblastic cells (MG-63) was observed by using Crystal Violet staining. Biofilm formation was reduced in SepTNT (22 ± 3%, P = 0.02) and Ag₂SepTNT discs (23 ± 11%, P = 0.02) compared to the non-modified control (54 ± 8%). In comparison with the non-modified control the modified SepTNT-HA and pTNT surfaces showed a significant higher covered area with osteoblastic MG-63-cells. Scanning electron microscope (SEM) images confirmed findings regarding bacterial and osteoblastic cell growth. These findings show a potential synergistic effect by combining selenium and silver with titanium nanotubes.