Characterization of bacterial flora in persistent apical periodontitis lesions.

INTRODUCTION: Microorganisms are able to survive and induce persistent infection in periapical tissues. The aim of this study was to investigate the composition of the microflora of persistent apical periodontitis lesions. METHODS: Twenty apical lesion samples were obtained from 20 patients with chronic apical periodontitis by root end surgery and processed using aerobic or anaerobic culture techniques. All isolated strains were identified by 16S ribosomal DNA sequence analysis. RESULTS: Seventy-four strains were isolated, belonging to 31 bacterial species obtained from the 20 apical lesions that were isolated. The majority of the strains were facultative anaerobes (51.6%). Propionibacterium acnes, Staphylococcus epidermidis, Pseudomonas aeruginosa and Fusobacterium nucleatum were isolated from 16.2, 9.5, 6.8 and 5.4% of the samples, respectively. Fifteen samples harboured more than one species. The predominant association was P. acnes, S. epidermidis and F. nucleatum. CONCLUSION: The microbiota of persistent apical periodontitis lesions is composed by diverse types of microorganisms with biofilm-forming capacity, including P. acnes, S. epidermidis and F. nucleatum.

Stimulation of Fusobacterium nucleatum biofilm formation by Porphyromonas gingivalis.

BACKGROUND/AIMS: Bacterial infection is a major cause of periapical periodontitis. Eradication of these microorganisms from apical lesions is essential to the success of endodontic treatment. The aim of this study was to clarify the molecular interaction between Fusobacterium nucleatum, Porphyromonas gingivalis and other microorganisms associated with periapical periodontitis. METHODS: Microorganisms isolated from periapical lesions were inoculated into type-I collagen-coated polystyrene microtiter plates and maintained at 37 degrees C under anaerobic conditions for 2 days, after which, the quantity of organized biofilm on the plates was evaluated by crystal violet staining. Growth enhancement via soluble factor was evaluated by separated coculture using a 0.4-mum membrane filter. RESULTS: F. nucleatum exhibited strong adherence to type-I collagen-coated polystyrene microplates. Biofilm formation by F. nucleatum was significantly enhanced by P. gingivalis. It was complemented by compartmentalized coculture with P. gingivalis. Enhancement of biofilm formation by P. gingivalis was only slightly reduced by inactivation of its autoinducer-2-producing gene luxS. CONCLUSION: The results suggest that P. gingivalis enhances biofilm formation by F. nucleatum by releasing diffusible signaling molecules other than autoinducer-2.

Thymic epithelial cells responsible for impaired generation of NK-T thymocytes in Alymphoplasia mutant mice.

We have previously shown that the generation of an NK1.1+TCRalphabeta+ (NK-T) cell population is severely impaired in an alymphoplasia mutant (aly/aly) mouse strain and the defect resides in the thymic environment. In the present study, to elucidate the thymic stromal component(s) that affects the development of NK-T cells, radiation bone marrow chimeras were established with the aly/aly mouse as a donor and either the beta2 microglobulin knockout (beta2m-/-) or the CD1d1-/- mouse that also lacks the NK-T cell population as a recipient. A normal population of NK-T cells with a typical NK-T phenotype and functions was detected in both the thymus and the spleen of these chimeras. These findings indicated that a radiation-resistant CD1(-) component of the thymus supported generation of functional NK-T cells from aly/aly precursors. Furthermore, transfer of an intact medullary thymic epithelial cell line into aly/aly thymus significantly induced the generation of NK-T cells in the thymus. These findings suggest that CD1 molecules of bone marrow-derived cells and the medullary epithelial cells acted in concert in the generation of the NK-T cell population and that a function(s) of the medullary thymic epithelial cells other than direct presentation of CD1 molecules to the NK-T precursors is indispensable for the development of NK-T cells.

Pertussis toxin-sensitive signal controls the trafficking of thymocytes across the corticomedullary junction in the thymus.

We investigated a role of chemokines in thymocyte trafficking. Genes encoding stromal cell-derived factor-1 and its receptor CXCR4 were detected in the cortex by in situ hybridization. Early immigrant cells did not express CXCR4, whereas their descendant CD44+CD25+CD4-CD8- cells did. CXCR4 expression was down-modulated when CD4+CD8+ double-positive cells became CD4+CD8- or CD4-CD8+ single-positive (SP) cells. Positively selected CD69+CD3intermediate cells gained CCR4, of which ligand, thymus activation-regulated chemokine, was expressed in the medulla. At the next developmental stage, CD69-CD3high cells lost CCR4 but gained CCR7. These results suggest that thymocytes use different chemokines along with their development. Blockade of chemokine receptor-mediated signaling by pertussis toxin perturbed the normal distribution of SP cells and resulted in the accumulation of SP cells in the cortex. Thus, a pertussis toxin-sensitive event controls the trafficking of SP cells across the corticomedullary junction.

[Histopathological studies of periodontal tissue reactions following apical plugging with autogenous dentin chips].

After treating the infected canals, we investigated to determine the influence of autogenous dentin chips accidentally applied as filling in the root apexes. Materials were 15 mandibular premolars and molars obtained from healthy, mature dogs. According to standard procedures, after pulp extirpation, canals were temporarily filled with sandarac cotton pellets and left otherwise untreated for 4 weeks. At this time, the degree of infection was ascertained. After root-canal enlargement, root canals were filled with No. 55 gutta-percha points; and a sealer (Neotriozinc Paste, AH26, or Sealapex) was applied. The animals were sacrificed at the conclusion of either a short term (2 weeks) or a long term (16 weeks), and histological studies were performed. Conclusions In short-term specimens, no formation of new hard tissue was observed at the apical dentino-cemental junction; and inflammatory changes in the periapical soft tissue were remarkable. In long-term specimens, hard-tissue formation had resulted in apical closure in 6 out of 8 instances; and periapical inflammatory changes had decreased. Relations between dentin-chip density and histopathological conditions were as follows: In most of the specimens that were evaluated as good, dentin chips were very densely applied. Insufficient numbers of dentin chips produced poor results. In other words, dense applications of dentin chips result in good prognoses. Consequently, when root-canal enlargement has been adequately performed, application of dentin chips to the apex of infected canals stimulates hard-tissue formation resulting in biological apical closure. In infected canals, however, the degree to which dentin chips are infected can be a major factor.