These findings indicate that ossification proceeds at different modes around the titanium implant in rat maxilla, depending on the nature of the recipient bones and the dimension of the gap between the implant and osteotomy margin.
The present study aimed to establish a titanium implantation model using rat maxillae as well as demonstrate the chronological tissue responses to implantation. Pure titanium implants were inserted in the upper first molar extraction sites of Wistar rats 1 month after tooth extraction. The animals were sacrificed at 1 to 30 days postimplantation, and prepared tissue specimens were processed for light microscopy. The removal of implants from tissue blocks was done using 2 methods: mechanical removal or a cryofracture technique. In the early stages, peri-implant tissues showed severe damage to the oral epithelium and collagen bundles with significant inflammatory cell infiltration. The peri-implant epithelium grew apically along the implant by 10 days postimplantation, and regenerated to show a similar feature of junctional epithelium seen in normal rats at 15 days postimplantation, at which time no signs of inflammation were observed. The regenerated collagen bundles in the connective tissue were arranged circumferentially to the implants in the horizontal sections. New bone formation first appeared around the implants at 5 days postimplantation, covering the entire perimeter of implants by 30 days postimplantation. Scanning electron microscopic observations of the surface texture of the removed implants suggest the probability of an adhesive mechanism between the implants and the peri-implant epithelium and/or the alveolar bone. These findings indicate that this experimental model is useful for detailed analysis of peri-implant tissue because of its easy implantation procedure.
Tissue responses to titanium implantation with two different surface conditions in our established implantation model in rat maxillae were investigated by light and transmission electron microscopy and by histochemistry for tartrate-resistant acid phosphatase (TRAPase) activity. Here we used two types of implants with different surface qualities: titanium implants sandblasted with Al2O3 (SA-group) and implants coated with hydroxyapatite (HA-group). In both groups, bone formation had begun by 5 days postimplantation when the inflammatory reaction had almost disappeared in the prepared bone cavity. In the SA-group, however, the bone formation process in the bone cavity was almost identical to that shown in our previous report using smooth surfaced implants (Futami et al. 2000): new bone formation, which occurred from the pre-existing bone toward the implant, was preceded by active bone resorption in the lateral area with a narrow gap, but not so in the base area with a wide gap. In the HA-group, direct bone formation from the implant toward the pre-existing bone was recognizable in both lateral and base areas. Many TRAPase-reactive cells were found near the implant surface. On the pre-existing bone, new bone formation occurred with bone resorption by typical osteoclasts. Osseointegration around the implants was achieved by postoperative day 28 in both SA- and HA-groups except for the lateral area, where the implant had been installed close to the cavity margin. These findings indicate that ossification around the titanium implants progresses in different patterns, probably dependent on surface properties and quality.
The response of nerve fibres in the peri-implant epithelium to titanium implantation was investigated with an experimental model using rat maxilla and immunohistochemical techniques. The latter employed antibodies to protein gene product 9.5 (PGP9.5), and to calcitonin gene-related peptide (CGRP). In control rats without an implantation, a dense innervation of PGP9.5- and CGRP-positive nerve fibres was recognized throughout the junctional epithelium, as has been previously reported. A titanium-implantation induced a remarkable inflammatory reaction, as well as the destruction of covering epithelial cells. By 3-5 days post-implantation, inflammatory reaction showed a tendency to disappear, and the peri-implant epithelium showed proliferation and down-growth along the implant. At this stage, no nerve fibres were found around the peri-implant epithelium. At 10 days, a few nerve fibres reached the basal cell layers of the peri-implant epithelium, and entered it 15 days after implantation when the peri-implant epithelial cells showed morphological features roughly resembling those of normal junctional epithelial cells. At the complete osseointegration stage (days 20-30), the PGP9.5- and CGRP-positive nerve fibres, thin and beaded in appearance, were found distributed in the peri-implant epithelium. After 20 days, the numerical density of the intraepithelial nerves in the peri-implant epithelium appeared the same as, or less than, that in the normal junctional epithelium. These findings indicate that the peri-implant epithelium shows the same innervation as that in normal junctional epithelium, and that the intraepithelial nerve fibres in the peri-implant epithelium might have diverse functions, which have been suggested in the literature.
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