Shinji Iyama

Study of bone formation around dense hydroxyapatite implants using light microscopy, image processing and confocal laser scanning microscopy

The bone reaction to hydroxyapatite (HA) implants inserted transcortically and extending into the medullary canal of rat tibiae was quantitatively assessed using light microscopy, confocal laser scanning microscopy and an image processing system. Sixty-five male rats (6 weeks old) were divided into two groups, 60 for histological observation and image analysis and five for time-labelling. In the histological observation, control sections of 168 days showed a few bone trabeculae in the fatty bone marrow, and Ti implants had become gradually encapsulated with a thick bone tissue layer; however, HA implants became almost completely encapsulated with a thin bone tissue layer during the 168 day experimental period. Histometrical analysis of the percent bone contact revealed that Ti implants showed a continuous increasing curve, and HA implants showed rapid increase in the initial healing period up to 14 days, with 96% bone contact reaching a plateau at 84 days after operation. There was a significant difference in the percent of bone contact between Ti and HA implants throughout the experimental period. Confocal laser scanning microscopic observations revealed the presence of calcein at the 14th day and only slight alizarin colour layer in the bone tissue at the 28th day, both indicating bone formation. These findings suggest that the activity of bone formation was higher at the 14th day than at the 28th day. Also, the percentage of bone contact of HA is superior to titanium throughout the experimental period, and the ascending patterns of both implants are quite different to each other.

Long-term evaluation of bone-titanium interface in rat tibiae using light microscopy, transmission electron microscopy, and image processing

We conducted a 2-year histologic and histometric evaluation of the tibial bone-titanium (Ti) implant interface in male rats. Thirty male 6-week-old rats were used in this study. They were divided into two groups: 15 for day 28 and 15 for day 730. Microscopic observation at day 28 revealed that the newly formed bone around the implant almost surrounded the implant, but fibroblastlike cells were interposed in some histologic sections. At day 730, in contrast, such cells were rarely seen, and the bone around the implant presented a lamellar structure. Transmission electron microscopic observation at day 28 disclosed mature or poorly mineralized bone near the implant; however, an electron-dense amorphous zone about 50 nm in thickness was interposed between the bone and Ti. In places slender cells were interposed between the bone and Ti. The amorphous zone was also observed at the cell-Ti interface. At day 730, a poorly mineralized layer remained in some areas between the mature bone and the titanium, and the interposed amorphous zone was still observed. Occasionally, a 200-nm-thick layer, thought to be cell remnant, was seen. As calculated in an image-processing, system analysis, the percent bone contact and the thickness and area of the surrounding bone for the Ti implant at day 28 were 43.6%, 30.4 microns, and 0.10 mm2, respectively, and those at day 730 were 89.9%, 53.5 microns, and 0.19 mm2, respectively. In summary, although the passage of time may affect bone maturity, interfacial cells remain at the bone-Ti interface as a uniform layer together with unmineralized bone.

Histopathologic Observations of Tissue Reaction to Three Kinds of Membrane Placed on the Bone Defects around Implants.

The purpose of this study was to investigate the tissue reaction to three kinds of membranes, Gore-Tex Augmentation Material (GTAM), Vicryl, and Dura Mater, placed on bone defects around the titanium implants. Nine adult dogs weighing 15kg were used in this study. The dogs were sacrificed either 2, 4, or 12 weeks after placement. Vicryl was exposed 2 weeks after placement. Histologically, it was found that Vicryl was completely resorbed, Dura Mater was replaced with connective tissue at 12 weeks, and bone defects using GTAM and Dura Mater but not Vicryl were partially filled with newly-formed bone tissue. Active bone formation was observed in the space between membranes and pre-existing bone tissue.