Keratinocytes protect soft-tissue integration of dental implant materials against bacterial challenges in a 3D-tissue infection model

摘要

The soft-tissue seal around dental implants protects the osseo-integrated screw against bacterial challenges. Surface properties of the implant material are crucial for implant survival against bacterial challenges, but there is no adequate in vitro model mimicking the soft-tissue seal around dental implants. Here, we set up a 3D-tissue model of the soft-tissue seal, in order to establish the roles of oral keratinocytes, gingival fibroblasts and materials surface properties in the protective seal. To this end, keratinocytes were grown on membrane filters in a transwell system, while fibroblasts were adhering to TiO2 surfaces underneath the membrane. In absence of keratinocytes on the membrane, fibroblasts growing on the TiO2 surface could not withstand challenges by commensal streptococci or pathogenic staphylococci. Keratinocytes growing on the membrane filters could withstand bacterial challenges, but tight junctions widened to allow invasion of bacteria to the underlying fibroblast layer in lower numbers than in absence of keratinocytes. The challenge of this bacterial invasion to the fibroblast layer on the TiO2 surface negatively affected tissue integration of the surface, demonstrating the protective barrier role of keratinocytes. Streptococci caused less damage to fibroblasts than staphylococci. Importantly, the protection offered by the soft-tissue seal appeared sensitive to surface properties of the implant material. Integration by fibroblasts of a hydrophobic silicone rubber surface was affected more upon bacterial challenges than integration of more hydrophilic hydroxyapatite or TiO2 surfaces. This differential response to different surface-chemistries makes the 3D-tissue infection model presented a useful tool in the development of new infection-resistant dental implant materials.