Histochemical and confocal laser scanning microscopy study of the bone-titanium interface: an experimental study in rabbits.

The aim of our study was an analysis of the presence of an unmineralized bone matrix between mineralized bone and titanium screws in rabbit tibiae. A microscopical analysis, using a histochemical technique, was performed on the titanium-bone interface of commercially pure titanium implants placed in rabbit tibiae and harvested after 2 months. Thin ground sections of the specimens were prepared by the cutting-grinding system and stained using the von Kossa method for calcium salts and basic fuchsin for osteoid. The microscopical and morphometrical evaluation showed that bone covered about 40% (+/- 7.5%) of all implants. Mineralized bone was, however, in direct contact with the titanium surface on only about 10% of the implant, while in the remaining 30% the mineralized bone was separated from the implant by an unmineralized tissue. This basophilic, probably osteoid matrix, could represent the medium that allows the biochemical exchanges between bone and cells under the influence of the implant. A small, optically translucent gap (1-5 microns), probably an artifact, was present in some areas between titanium and bone. Confocal laser scanning microscopy (CLSM) in a fluorescent mode showed the presence at the interface of a fluorescent material. Results from our study showed that light microscopy of thin ground sections allowed a good analysis of the real nature of the titanium-bone interface. Moreover, this double staining technique showed the presence of an unmineralized bone matrix at most of the bone-titanium interface.