In vitro attachment of osteoblasts on contaminated rough titanium surfaces treated by Er:YAG laser

Abstract

Microbial contamination of implant surfaces inhibits formation of new osseous tissues. Biocompatibility of sandblasted large grid (SLA) surface, after previous in vitro cocultivation with Porphyromonas gingivalis and concomitant Er:YAG laser irradiation of microorganisms, was tested by attachment of newly cultured osteoblasts. A total of 36 customized titanium cubes with SLA surface were placed into human osteoblast culture for 14 days. After removal of 1 control cube, 35 other cubes were contaminated with precultured P. gingivalis (ATCC33277) and incubated in broth medium for 1 week. Ablation was carried out on 32 cubes. Each side was treated for 23.5 s with a pulsed, water-cooled laser beam. After irradiation, cubes were again placed into fresh osteoblast culture for 2 weeks. One randomly selected single side per cube was analyzed by scanning electron microscope in 22 cubes. On other 10 cubes, vitality of attached cells was tested with ethidiumbromide staining by fluorescence microscopy. Three negative controls revealed constantly adherent P. gingivalis, and no osteoblasts were detectable after P. gingivalis contamination on the surfaces. Laser-treated specimens showed newly attached osteoblasts, extending over 50-80% of the surface. Positive control cube (without bacterial contamination) showed over 80% cell coverage of the surface. Vitality of widely stretched osteoblasts was confirmed by FITC staining. Our results indicate that Er:YAG laser was effective in removing P. gingivalis and cell compounds, offering an acceptable surface for new osteoblast attachment.