Porphyromonas gingivalis, one of the causative agents of adult periodontitis, attaches and forms biofilms on substrata of Streptococcus gordonii. Coadhesion and biofilm development between these organisms requires the interaction of the short fimbriae of P. gingivalis with the SspB streptococcal surface polypeptide. In this study we investigated the structure and binding activities of the short fimbriae of P. gingivalis. Electron microscopy showed that isolated short fimbriae have an average length of 103 nm and exhibit a helical structure with a pitch of ca. 27 nm. Mfa1, the major protein subunit of the short fimbriae, bound to SspB protein, and this reaction was inhibited by purified recombinant Mfa1 and monospecifc anti-Mfa1 serum in a dose-dependent manner. Complementation of a polar Mfa1 mutant with the mfa1 gene restored the coadhesion phenotype of P. gingivalis. Hence, the Mfa1 structural fimbrial subunit does not require accessory proteins for binding to SspB. Furthermore, the interaction of Mfa1 with SspB is necessary for optimal coadhesion between P. gingivalis and S. gordonii.
This study was conducted to examine the oral condition and the salivary and microbiological parameters associated with dental caries in 62 children with cerebral palsy, who came from households of low socioeconomic status (Study Group). This group had mixed (6 to 11 years old) and permanent (11 to 16 years old) dentition. Dental examinations were performed to measure dental caries, plaque index, salivary levels of mutans streptococci and lactobacilli, salivary flow rate, pH of stimulated saliva, and buffer capacity of saliva. A group of 67 non-handicapped children from similar socioeconomic backgrounds also were examined using these parameters (Control Group). Data were analyzed statistically by non-parametric tests and by correlation. The results showed that children with cerebral palsy who had permanent dentitions had a higher mean decayed, missing and filled surfaces index, as well as a higher plaque index for both sexes. Microbiological examination revealed higher levels of mutans streptococci among Study Group subjects with mixed dentition than in the Control Group. Also, lactobacillus counts were higher in the Study Group, regardless of sex or dentition. With respect to salivary flow rate, pH and buffering capacity, lower mean values were obtained for the Study Group.
Porphyromonas gingivalis, one of the causative agents of adult periodontitis, develops biofilm microcolonies on substrata of Streptococcus gordonii but not on Streptococcus mutans. P. gingivalis genome microarrays were used to identify genes differentially regulated during accretion of P. gingivalis in heterotypic biofilms with S. gordonii. Thirty-three genes showed up-or downregulation by array analysis, and differential expression was confirmed by quantitative reverse transcription-PCR. The functions of the regulated genes were predominantly related to metabolism and energy production. In addition, many of the genes have no current known function. The roles of two upregulated genes, ftsH (PG0047) encoding an ATP-dependent zinc metallopeptidase and ptpA (PG1641) encoding a putative tyrosine phosphatase, were investigated further by mutational analysis. Strains with mutations in these genes developed more abundant biofilms with S. gordonii than the parental strain developed. ftsH and ptpA may thus participate in a regulatory network that constrains P. gingivalis accumulation in heterotypic biofilms. This study provided a global analysis of P. gingivalis transcriptional responses in an oral microbial community and also provided insight into the regulation of heterotypic biofilm development.Periodontal diseases are a group of infections characterized by destruction of the supporting structures of the teeth. Porphyromonas gingivalis is a gram-negative anaerobe that is an important pathogen in severe manifestations of these diseases (15,41). With regard to the disease process, the primary ecological niche of P. gingivalis is in the subgingival area, where toxic products, such as proteases, can readily access the periodontal tissues. However, initial colonization of the oral cavity by P. gingivalis involves attachment to sites remote from the subgingival area, including the supragingival tooth surface (28,39,43,47,57,58). Indeed, introduction of P. gingivalis into the mouths of human volunteers results in localization almost exclusively on supragingival surfaces (39). The bacterial inhabitants of the supragingival tooth surface comprise a complex multispecies biofilm (42), and numerous in vitro studies have demonstrated the ability of P. gingivalis to attach to common constituents of the supragingival biofilm, including Actinomyces species and oral streptococci (12, 35). The molecular basis of P. gingivalis adhesion to Streptococcus gordonii has been investigated in some detail and has been shown to be multivalent (3,4,23,24,45). The P. gingivalis long fimbriae (FimA) bind to glyceraldehyde-3-phosphate dehydrogenase present on the streptococcal surface (27). In addition, the P. gingivalis short fimbriae (Mfa) engage the streptococcal SspA/B (antigen I/II) adhesins (33) through an approximately 80-amino-acid binding epitope of SspA/B termed BAR (11). Coadhesion mediated through these effectors is required for P. gingivalis to accumulate in a heterotypic biofilm with S. gordonii (23).In contrast to the synergistic rela...
Abstract. The purpose of this study was to assess photodynamic antimicrobial chemotherapy (PACT) via irradiation, using a low power laser associated with a photosensitization dye, as an alternative to remove cariogenic microorganisms by drilling. Remaining dentinal samples in deep carious lesions on permanent molars (n = 26) were treated with 0.01% methylene blue dye and irradiated with a low power laser (InGaAIP -indium gallium aluminum phosphide; λ = 660 nm; 100 mW; 320 Jcm − 2 ; 90 s; 9J). Samples of dentin from the pulpal wall region were collected with a micropunch before and immediately after PACT and kept in a transport medium for microbiological analysis. Samples were cultured in plates of Brucella blood agar, Mitis Salivarius Bacitracin agar and Rogosa SL agar to determine the total viable bacteria, mutans streptococci and Lactobacillus spp. counts, respectively. After incubation, colony-forming units were counted and microbial reduction was calculated for each group of bacteria. PACT led to statistically significant reductions in mutans streptococci (1.38 log), Lactobacillus spp. (0.93 log), and total viable bacteria (0.91 log). This therapy may be an appropriate approach for the treatment of deep carious lesions using minimally invasive procedures. C 2011 Society of Photo-Optical Instrumentation Engineers (SPIE).
Background and Objective Although previous studies revealed the potential use of probiotics in the control of periodontitis, little is known about their interactions with gingival epithelial cells (GECs). Since GECs comprise the first defense in the subgingival microenvironment, the aim of this study was to evaluate the effect of probiotic lactobacilli and bifidobacteria strains on OBA‐9 cells challenged with Porphyromonas gingivalis. Methods Immortalized human GECs (OBA‐9) were challenged with live P. gingivalis (strains W83 and ATCC33277) and co‐infected with one of 12 tested probiotic strains at a multiplicity of infection (MOI) of 1:1000 for 2 hours. Bacterial adhesion and invasion were determined by antibiotic exclusion analysis and CFU counting. OBA‐9 viability was assessed by MTT assay, and levels of inflammatory mediators (TNF‐α, IL‐1β, and CXCL8) in the supernatants were determined by ELISA. The expression of genes encoding Toll‐like receptors (TLR2, TLR4) was evaluated by RT‐qPCR. Results Both strains of P. gingivalis were able to adhere and invade OBA‐9 cells, with significant loss in cell viability, increase in the levels of TNF‐α and IL‐1β, and upregulation of TLR4. However, co‐infection with probiotics attenuated these effects in P. gingivalis challenged GECs. Most probiotics maintained OBA‐9 viability and reduced pathogens adhesion and invasion. Furthermore, probiotics were able to adhere to GECs, which was enhanced for most strains in the presence of P. gingivalis. The synthesis of IL‐1β and TNF‐α by P. gingivalis in challenged GECs was reduced in co‐culture with most of the tested probiotics, whereas the secretion of CXCL8 increased, and TLR4 was downregulated. Conclusion Probiotics can alter the interaction of GECs with P. gingivalis by modulating the pathogen's ability to adhere and invade these cells, as well as by regulating the innate immune response. Such properties are strain‐specific and may indicate the most efficient probiotics to control periodontitis.
The data suggested an association between P. gingivalis genotype fimA IV and disease severity in smoker-chronic periodontitis subjects.
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