In vitro antifungal susceptibility of Candida spp. isolates from patients with chronic periodontitis and from control patients

Superinfection by Candida can be refractory to conventional periodontal treatments in specific situations, such as in immunocompromised patients. In these cases, the systemic therapy with antifungal drugs could be indicated. The aim of this study was to analyse antifungal susceptibility of Candida spp. strains isolated from chronic periodontitis patients and from control individuals. A total of 39 C. albicans isolates, 9 C. tropicalis, 2 C. glabrata and 5 Candida spp. from control individuals and 30 C. albicans, 3 C. tropicalis and 2 C. glabrata from periodontitis patients were tested. In the control group, 1 isolate of C. glabrata was resistant to ketoconazole and 1 Candida spp. was resistant to amphotericin B, ketoconazole and miconazole. Among the isolates of periodontitis group, 1 (3.33%) C. albicans isolate was resistant to flucytosine and ketoconazole. According to the obtained results, it could be concluded that fluconazole was the most effective drug against the several Candida species studied. There were not expressive differences in the susceptibility of isolates from periodontitis patients or from control individuals.

Systemic use of metronidazole in the treatment of chronic periodontitis: a pilot study using clinical, microbiological, and enzymatic evaluation

The aim of the present parallel, double-blind investigation was to evaluate the effect of using systemic metronidazole alone or associated to scaling and root planing on adult chronic periodontal disease, monitored at baseline, 30, 60 and 90 days. Twelve subjects were divided into three groups: the first group (Group I - 22 sites) was submitted to scaling and root planing (SRP) alone; the second group (Group II - 30 sites) received SRP and 250 mg of metronidazole (3 times a day for 10 days), and the third group (Group III - 31 sites) was treated with metronidazole alone. The clinical parameters evaluated were probing depth (PD), clinical attachment level (CAL), plaque index (PlI), gingival index (GI) and bleeding upon probing (BP). Microbiological (BANA test) and enzymatic (Pocket Watch) tests were also performed. All three proposed treatments produced significant improvements in clinical conditions of subjects, from baseline, 30, 60 and 90-day period, except for clinical attachment level. The results obtained by microbiological and enzymatic tests did not show statistical differences among the groups for the 90-day period (r = 0.7924 and r = 0.7757, respectively). In relation to clinical parameters, statistical differences among groups were observed only for the gingival index (p = 0.0261) between Groups I and II, and probing depth (p = 0.0124) between Group I and the others. We conclude that the use of systemic metronidazole did not produce additional effects on the microbiological conditions of these patients with chronic periodontal disease.

Occurrence of Actinobacillus actinomycetemcomitans in patients with chronic periodontitis, aggressive periodontitis, healthy subjects and children with gingivitis in two cities of the state of São Paulo, Brazil

The aim of this study was to determine the frequency of isolation of Actinobacillus actinomycetemcomitans (Aa) in 100 patients with chronic periodontitis, 14 patients with aggressive periodontitis, 142 pre-school children with gingivitis and 134 periodontally healthy subjects. Samples of subgingival plaque were taken using sterilized paper points introduced into periodontal pockets or gingival crevice for 60 seconds and inoculated on TSBV agar, which was incubated under anaerobiosis at 37°C, for 4 days. Microbial identification was performed through biochemical methods and morphocellular and morphocolonial analysis. Aa was detected in 40.3% of healthy subjects, 68% of patients with chronic periodontitis, 92.86% of patients with aggressive periodontitis and 40.14% of children with gingivitis. The rate of recovery of Aa in the tested human groups proved to be higher than previously reported and in agreement with participation of this facultative anaerobe as a member of native microbiota of the periodontium and its relation with aggressive and chronic periodontitis in Brazil.

Transmission of Aggregatibacter actinomycetemcomitans between Brazilian women with severe chronic periodontitis and their children

This study evaluated the transmission of Aggregatibacter actinomycetemcomitans (Aa) in women with severe chronic periodontitis and their children. Thirty women (mean age = 36.1±6.0 years) who were mothers of at least one child aged 7 to 16 years were enrolled. In order to investigate mother-child transmission of Aa, the children were also evaluated when their mothers were colonized by the bacterium. Subgingival plaque samples of each woman were collected from 3 sites (mean probing depth of 7.3±1.2 mm and mean clinical attachment level of 7.9±1.5 mm) and pooled in reduced transport fluid (RTF). These samples were processed, inoculated onto TSBVagar selective medium and incubated at 37°C in microaerophilic atmosphere for 5 days. Aa was identified on the basis of colony morphology, Gram staining, catalase and oxidase reactions. Aa was found in 8 out of 30 women. Therefore, 8 children from these women (mean age= 12 ± 3.7 years) were evaluated, but Aa was found only in 2 of them. Aa strains of the two mother-child pairs were evaluated by arbitrarily-primed polymerase chain reaction (AP-PCR), although it was not found similarity between the amplitypes of each pair. No Aa transmission was found between Brazilian women with severe chronic periodontitis and their children.

Phenotypic characterization of Candida spp. isolates from chronic periodontitis patients

Aim: Several typing methods for Candida spp. have been suggested in the literature in order to distinguish isolates for studies about the virulence or infection routes of these microorganisms and, in particular, for epidemiological purposes. The aim of this study was to establish a comparison between the phenotypic profile of oral Candida isolates from periodontitis patients and control individuals. Methods: The morphotyping and biotyping of 35 C. albicans isolates obtained from chronic periodontitis patients and 48 isolates from control individuals were performed. For morphotyping, the isolates were plated on malt extract agar and incubated for 10 days. Sixteen different morphotypes were observed for C. albicans, the most frequently observed being 0000 and 0001. Results: Biotype 0000 (complete absence of fringe) was most prevalent among the isolates obtained from periodontitis patients compared to those from control individuals, with statistical significance. Biotyping revealed 5 different biotypes with higher prevalence of the biotype 357 among the isolates from control and periodontitis groups. Conclusions: The results obtained by biotyping of the isolates did not permit to differentiate a characteristic model related to periodontal disease, whilst the morphotype 0000 was most frequently isolated from periodontitis patients.

Variation in the CXCR1 gene (IL8RA) is not associated with susceptibility to chronic periodontitis

Background: The chemokine receptor 1 CXCR-1 (or IL8R-alpha) is a specific receptor for the interleukin 8 (IL-8), which is chemoattractant for neutrophils and has an important role in the inflammatory response. The polymorphism rs2234671 at position Ex2+860G > C of the CXCR1 gene causes a conservative amino acid substitution (S276T). This single nucleotide polymorphism (SNP) seemed to be functional as it was associated with decreased lung cancer risk. Previous studies of our group found association of haplotypes in the IL8 and in the CXCR2 genes with the multifactorial disease chronic periodontitis. In this study we investigated the polymorphism rs2234671 in 395 Brazilian subjects with and without chronic periodontitis. Findings. Similar distribution of the allelic and genotypic frequencies were observed between the groups (p > 0.05). Conclusions: The polymorphism rs2234671 in the CXCR1 gene was not associated with the susceptibility to chronic periodontitis in the studied Brazilian population. © 2011 Scarel-Caminaga et al; licensee BioMed Central Ltd.

Association between IL8 haplotypes and pathogen levels in chronic periodontitis

Chronic periodontitis (CP) is considered to be a multifactorial disease influenced by microbial and genetic factors. The aim of the present study was to investigate whether the genetic susceptibility to CP in individuals with the IL8 ATC/TTC haplotype is associated with subgingival levels of periodontopathogens. Sixty-five individuals, grouped according to the presence (n = 28) or absence (n = 37) of the IL8 haplotype, were evaluated. After clinical periodontal evaluation, each group was subdivided according to the presence (CP) or absence (H) of periodontitis. Four subgingival samples were obtained from CP and two samples per subject from H patients. The levels and proportions of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were analyzed using quantitative real-time polymerase chain reaction (q-PCR). No differences were found in the proportion of periodontopathogenic bacteria between groups with the presence or absence of the IL8 haplotype. However, in the CP groups, the levels of periodontopathogens were significantly higher in the individuals without the IL8 haplotype than in the individuals with the IL8 haplotype. These results suggest that periodontal destruction may occur in patients who are considered to be genetically susceptible to CP with a lower microbial challenge because of the presence of the IL8 ATC/TTC haplotype than in patients without this haplotype. © 2013 Springer-Verlag Berlin Heidelberg.

Interleukin 4 haplotypes of susceptibility to chronic periodontitis are associated with IL-4 protein levels but not with clinical outcomes of periodontal therapy

Different IL4 haplotypes were associated to susceptibility to/or protection against chronic periodontitis (CP). The aim of this study was to investigate if individuals carrying different haplotypes would present differences in clinical periodontal parameters and in the IL-4 levels at baseline, 45 and 90 days after non-surgical periodontal therapy. 62 patients were subdivided: genetically protected without CP (PH), genetically protected with CP (PCP), genetically susceptible with CP (SCP), genetically susceptible without CP (healthy) (SH). Clinical examination and gingival crevicular fluid (GCF) collection were performed for all patients, and IL-4 levels were measured by ELISA. At baseline, higher values for plaque index (PI, p = 0.013), gingival index (GI, p = 0.005) were observed for the SCP group in comparison to the PCP group but not after the completion of periodontal therapy. 45 and 90 days after the non-surgical therapy, PCP demonstrated significantly higher IL-4 levels than the SCP (p = 0.000002). Correlation analysis showed different results between clinical parameters and IL-4 production or GCF volume for groups with different genetic loads. The IL4 gene which was previously associated with susceptibility to CP was related with differences in the IL-4 protein levels in the GCF. However, independent of genetic carriage, individuals responded similarly to this therapy. © 2013 American Society for Histocompatibility and Immunogenetics.

Subgingival biodiversity in subjects with uncontrolled type-2 diabetes and chronic periodontitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Periodontopathogens levels and clinical response to periodontal therapy in individuals with the interleukin-4 haplotype associated with susceptibility to chronic periodontitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Peripheral Blood Mononuclear Phagocytes From Patients With Chronic Periodontitis Are Primed for Osteoclast Formation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expression of Protease Activated Receptor-1 in Chronic Periodontitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Haplotypes of susceptibility to chronic periodontitis do not influence MMP-8 levels or the outcome of non-surgical periodontal therapy

Introduction: Chronic periodontitis (CP) is a multifactorial condition, presenting immunoinflammatory reaction, in which a myriad of molecules including cytokines and matrix metalloproteinases (MMPs) interplays, making the system extremely intricate. There is scarce information regarding interconnections of biological influence among IL-4, IL-8 and MMP-8, mainly considering genetic polymorphisms, and also, whether this can influence the outcome of periodontal therapy. Previously, we reported that variants in the interleukin 4 (IL4) and interleukin 8 (IL8) genes were associated with CP in Brazilians. The aim of this study was to investigate, in individuals with different genetic backgrounds with regard to the IL4 or IL8 haplotypes, differences in the immunological levels of MMP-8 in gingival crevicular fluid (GCF) before and after non-surgical periodontal treatment. A total of 141 patients participated of this study, classified as susceptible or not to CP, according to the presence of haplotypes formed by polymorphysms in the IL4 or IL8 genes. All individuals received non-surgical periodontal therapy and follow–up continued for 45 days. The GCF samples were collected at baseline and on the 45th day. The MMP-8 levels were determined by ELISA. Results: No association was found between genetic backgrounds and MMP-8 levels in GCF or the outcome of non-surgical periodontal therapy. Conclusions: In this longitudinal clinical study, the presence of IL4 or IL8 haplotypes previously associated with CP did not influence the outcome of non-surgical periodontal therapy and the MMP-8 levels in the GCF. Additional studies are necessary to determine the mechanisms by which the IL4 or IL8 haplotypes affect individual susceptibility to CP.

In vitro antifungal susceptibility of candida albicans isolates from patients with chronic periodontitis and diabetes

Alterations that lead to deficiency of the immune system, such as diabetes mellitus, may promote proliferation of Candida albicans and selection of strains which have greater ability to adhere and to penetrate the host tissues. Recent studies indicate an increase of the antifungal resistance of C. albicans isolates in periodontal pockets, suggesting that the oral cavity could be a reservoir of resistant yeast to antifungal agents. Moreover, oral cavity can act as a reservoir of certain pathogens that may cause systemic infections. The periodontal pocket is an ecological niche suitable to host microorganisms that could act as opportunistic pathogens. The aim of this study is to contribute to the understanding of resistance to conventional antifungal against C. albicans isolates from patients with periodontitis and diabetes. The determination of the minimal inhibitory concentrations (MIC) was evaluated according to M27S3 of the CLSI (2008), with modifications. The results showed that 48.8% of the studied strains were resistant to one or more antifungals and 6.6% were resistant to fluconazole and voriconazole. These results suggest an increasing resistance to conventional antifungal agents among Candida species, suggesting that the oral cavity could host pathogen fungi.