It has not been previously possible to observe bone formation in undecalcified sections with titanium implants at high magnification because of the difficulty in sectioning bone together with implants. A method for examining the bone-implant interface in undecalcified sections is described in which implants are left in situ and confocal laser scanning microscopy (CLSM) is used to examine both the implant surface and adjacent bone. Pulsing of animals at different times with the fluorescent dyes calcein and alizarin red permitted assessment of temporal patterns of bone formation by CLSM. Reflectivity of the polished implant surface permitted accurate assessment of the position of the implant relative to labeled bone. The analysis showed that bone first formed as thin processes towards and across the implant surface, followed by further bone formation behind these processes. The interface between calcified bone tissue and the implant surface was characterized by a 10-microm space. The CLSM technique enabled detailed observations of new bone formation at the titanium implant interface.
The bactericidal potency of ozone was examined for exploring the potential of dental application of ozone gel. Treatment with10 ppm ozone gel with quenching after 3 h and treatment with 100-ppm ozone gel with immediate quenching showed antimicrobial effects in eight aerobic strains, namely, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candidaalbicans, Methicillin-resistant S. aureus (MRSA), S. epidermidis, Klebsiella pneumoniae, and Streptococcus mutans. For five of these strains (E. coli, P. aeruginosa, C. albicans, K. pneumoniae, and S. mutans), the number of colony forming units (CFUs) were below the detection limit after treatment with 10 ppm ozone gel with quenching after 3 h. For Bacillus subtilis, an antimicrobial effect was observed after 3 h of treatment with 100 ppm of ozone gel. In the case of anaerobic bacteria, bactericidal effect was observed for five strains, namely, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Eikenella corrodens, by using ozone gel and ozone cream. Of these, the number of CFUs for three strains (P. gingivalis, P. intermedia, and F. nucleatum) was below the detection limit. These results suggest that the ozone gel can be clinically useful in oral surgery for implant treatment because of its instantaneous antimicrobial effects, and that it can be used against a variety of bacterial strains.
fetal bovine serum (FBS), 100 units/ml penicillin G, and 100 µg/ml streptomycin at 37ºC in a 5% CO 2 and 95% air humidified incubator. HGFs used in this study were obtained from volunteers after appropriate informed consent was obtained. The Ethics Committee of Osaka Dental University approved the study (protocol 110778). HGFs isolated from adherent gingival tissue on the extracted teeth of patients with chronic periodontal were cultured on collagen-coated plates in medium. Reagents The following materials and antibodies were purchased: 100 ppm Ozone gel (VMC Co., Ltd.); lipopolysaccharide (LPS) from Porphyromonas gingivalis (P. gingivalis) (InvivoGen, San Diego, CA, USA); anti-Interleukin (IL)-6 and biotinylated anti-IL-6 antibodies (eBiosciences, San Diego, CA, USA); anti-IL-8 and biotinylated anti-IL-8 antibodies (R&D Systems, Minneapolis, MN, USA); biotinylated anti-collagen type I antibody (Rockland, Limerick, PA, USA); and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) (Sigma-Aldrich, St. Louis, MO, USA); BrdU (5-bromo-2'deoxyuridine) Cell Proliferation Assay kit (Millipore, Billerica, MA, USA). DNA synthesis and MTT assays For analysis of DNA synthesis, HGFs (1 × 10 4 /cm 2) were cultured in DMEM containing 0.5% FBS (0.5% DMEM) for 24 h. The cells were
The medical use of ozone has been based on its antibacterial and oxidative characteristics. Currently ozone is being discussed in dentistry as a possible alternative oral antiseptic agent. In this study, we examined the hemostatic effect of water and gel contains aqueous ozone in animal testing. The mean of bleeding time using ozonated water and ozonated gel were observed for significant difference compared with no treatment. 0.5 ppm diluted ozonated water shortened bleeding time the same as 4.0 ppm ozonated water. These results suggest that ozonated water and gel which contains aqueous ozone show hemostatic ability which is almost equal to the Bosmin solution and Liquid Thrombin.
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