• Restorative Dentistry and Endodontics
• /
• v.23 no.1
• /
• pp.328-338
• /
• 1998

Porphyromonas endodontalis, an anaerobic Gram negative cocobacillus which was known to be associated with the infected root canals and periapical lesions, is very difficult to culture and to detect by the traditional method in that it requires much time to induce the specific black pigmentation, and it is very sensitive to oxygen and the antibiotics added in the culture medium. In this study, the nucleotide sequences of the 'probe h' (0.73kb), one of the specific DNA probes top. endodontalis (ATCC 35406) which had been developed by our department, was determined and then a pair of primers for PCR amplification was fabricated to identify P. endodontalis. The plasmids containing 'probe h' were purified by $Wizard^{TM}$ Midipreps DNA Purification System (Promega Corp.), and the nucleotide sequences of the 'probe h' were determined by the dideoxy chain termination method using TaqTrack Sequencing System (Promega Corp.) and detected by fluorescent labelling method. The sense/antisense PCR primers were designed with computer software (Lasergene, DNASTAR Ind. PCR was done with a programmable GeneAmp PCR System 2400 (Perkin Elmer-Cetus Co.). Each sample containing the whole genomic DNA of P. endodontalis and other black-pigmented Bacteroides was itailly denatured at $94^{\circ}C$ for 5 min and then subjected to 30 cycles, each of them consisting of 60s at $94^{\circ}C$, 60s at $60^{\circ}C$, and 90s. at $72^{\circ}C$. The amplified DNA was resolved electrophoretically in a 1.0 % agarose gel in 1X TAE buffer, stained with EtBr, and photographed on a UV transilluminator. The results were as follows : 1. The nucleotide sequences of 'probe h' (743 base pairs) were obtained by dideoxy chain termination method, and from that results the specific primers to P. endodontalis (ATCC 35406), 'Primer H1/ Primer H2', were designed. 2. It has been found that 'Primer H1/H2' could detect P. endodontalis (ATCC 35406) using PCR. 3. The PCR system with this primers may be a powerful technique to amplify the specific sequences of 'probe h' of P. endodontalis (ATCC 35406) that produce distinct identification of it from other black-pigmented Bacteroides, and this could help us to determine the nature of periapical disease.

• Journal of Periodontal and Implant Science
• /
• v.52 no.5
• /
• pp.422-434
• /
• 2022

Purpose: The purpose of this study was to evaluate and compare the healing patterns of 2-mm and 4-mm proximal infrabony defects adjacent to dental implants in canine mandibles. Methods: Four male beagles were used. Two groups were created: a 2-mm group (n=4) and a 4-mm group (n=4) depending on the horizontal dimension of proximal infrabony defects adjacent to implants. Bone healing patterns between the 2 groups were evaluated and compared at 8 and 16 weeks using radiographic, histological, histomorphometric, and fluorescent labelling analyses. Results: According to microcomputed tomography, the median bone volume fraction, bone mineral density, and the percentage of radiographic distance from the defect bottom to the most coronal bone-to-implant contact (radio-mcBIC) were 32.9%, 0.6 g/cm³, and 73.7% (8 weeks) and 45.7%, 0.7 g/cm³, and 76.0% (16 weeks) in the 2-mm group and 57.7%, 0.8 g/cm³, and 75.7% (8 weeks) and 50.9%, 0.8 g/cm³, and 74.7% (16 weeks) in the 4-mm group, respectively. According to histomorphometry, the median bone area fraction, mcBIC and the percentage of BIC amounted to 36.7%, 3.4 mm, and 58.4% (8 weeks) and 49.2%, 3.4 mm, and 70.2% (16 weeks) in the 2-mm group and 50.0%, 3.0 mm, and 64.8% (8 weeks) and 55.7%, 3.0 mm, and 69.6% (16 weeks) in the 4-mm group, respectively. No statistically significant differences were found between the groups for any variables (P>0.05). Conclusions: The proximal defects that measured 2 mm and 4 mm showed similar healing patterns at 8 and 16 weeks, and the top of bone formation in the defects was substantially limited to a maximum of 1.6 mm below the implant shoulder in both groups.