[Establishment of animal model of bacterial microleakage at implant-abutment interface]

Objective: To study the bacterial microleakage at the interface between dental implant and abutment in rats. Methods: Under aseptic conditions, suspension of 0.25 μl of Porphyromonas gingivalis (Pg) (10(9) CFU/ml) was added into the customized implant. After the abutment was connected, the suspension was cultured in an Ep (eppendorf) tube containing 1 ml brain heart infusion (BHI) culture medium. After 7 days and 14 days, the liquid in the Ep tube was taken and inoculated, and the growth of bacteria was observed. Six male SD rats with 12 implants were divided into experimental group (4 implants), negative control group (4 implants) and blank control group (4 implants). All 6 rats had two implants implanted in their bilateral upper jaws. During the second operation, suspension of 0.25 μl Pg (10(9) CFU/ml) was added to the inner part of the implant of the experimental group, culture solution of 0.25 μl was added to the control group and nothing was added to the blank control group. The amount of Pg and total bacteria in each group were evaluated by quantitative real-time PCR (qPCR). The inflammatory cell infiltrate in the peri-implant mucosa was evaluated histomorphometrically. Results: The in vitro model directly verified the presence of bacterial microleakage at implant-abutment interface (IAI), and the animal model confirmed the existence of microleakage through the infiltrate of inflammatory cells near the micro-gap in the experimental group indirectly. In vitro experiments found that Pg had penetrated from the implant within a week by observation and culture. In animal study, the presence of 10(2)-10(4) Pg was detected in the experimental group and it was not detected in the negative control group and the blank control group. At the same time, under the light microscope, in the experimental group, there were inflammatory cells aggregation in the connective tissue around the micro-gap and the density of inflammatory cells gradually decreased from the micro-gap to coronal and the apical of the connective tissue, while there were only scattered inflammatory cells in the connective tissue around the blank control group and the negative control group. In the experimental group, inflammatory cells density in area of 0.25-0.50 mm, 0-0.25 mm coronal to the micro-gap and 0-0.25 mm, 0.25-0.50 mm apical to the mico-gap was respectively, 976 (655), 1 673 (1 245), 2 267 (819) and 895 (162) cells/mm(2),which was significantly more than the blank control group in the corresponding position [respectively 201 (180), 321 (351), 309 (236) and 218 (272) cells/mm(2)] (P<0.05). Conclusions: Pg in the dental implants of rats can be found in the microleakage through implant-abutment interface, and cause the soft tissue inflammation around the implant, and the inflammation has certain distribution characteristics.