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1.
Implant Dent ; 28(3): 265-271, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31124823

RESUMEN

PURPOSE: The purpose of this study was to assess the effectiveness of a polyglycol dimethacrylate-based adhesive in preventing bacterial leakage through implant-abutment interfaces (IAIs). MATERIALS AND METHODS: After implant installation, the adhesive was applied in the experimental group (n = 10). None was applied in the control group (n = 10). Samples were collected from the inner walls of implants on days 0 and 90. The real-time polymerase chain reaction was used to detect bacterial DNA. RESULTS: All samples from the control group, versus 30% from the experimental group, harbored bacterial DNA on day 90. CONCLUSIONS: This polyglycol dimethacrylate-based adhesive may be used to seal the IAI. Further studies are warranted to verify its effectiveness over longer time periods.


Asunto(s)
Implantes Dentales , Filtración Dental , Pilares Dentales , Cementos Dentales , Diseño de Implante Dental-Pilar , Humanos , Polietilenglicoles
2.
Anaerobe ; 52: 50-58, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29857043

RESUMEN

Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum are strongly associated with periodontitis, and their evaluations are relevant to understand their role in the etiology and progression of periodontal diseases. In this study, the qualitative and quantitative detection of A. actinomycetemcomitans and F. nucleatum, as well as their genetic diversity, were evaluated in individuals with gingivitis, chronic periodontitis and periodontally healthy. In addition, the biotyping, serotyping, and prevalence of the ltx and cdt genes in A. actinomycetemcomitans were also determined. Subgingival biofilms obtained from gingivitis (70), periodontitis (75) and healthy (95) individuals were analyzed by cultures and PCR. Bacterial typing and presence of ltx and cdt genes in A. actinomycetemcomitans were also verified. DNA from A. actinomycetemcomitans and F. nucleatum was detected respectively, in 65.7% and 57.1% of gingivitis, 80% and 68% of periodontitis, and 57.8% and 37.8% of healthy. A. actinomycetemcomitans from gingivitis were biotypes I, II, IV, V, and X, and serotypes a, c, and e. In periodontitis, biotypes II, VI, and X, and serotypes a, b, and c were found. In healthy subjects, biotypes II and X, and serotypes b and c were found. The LTX and ltxA were observed in strains from gingivitis and periodontitis pockets. Subsequently, our data also showed no direct relationship between ltxA gene expression and leukotoxin gene 530-bp presence. On the other hand, cdt gene predominated during the inflammatory disease process. Our results strongly support a role of A. actinomycetemcomitans and F. nucleatum in advanced stage of periodontal disease.


Asunto(s)
Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Fusobacterium nucleatum/aislamiento & purificación , Enfermedades Periodontales/microbiología , Adulto , Aggregatibacter actinomycetemcomitans/clasificación , Aggregatibacter actinomycetemcomitans/genética , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Estudios Transversales , Exotoxinas/genética , Exotoxinas/metabolismo , Femenino , Fusobacterium nucleatum/clasificación , Fusobacterium nucleatum/genética , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven
3.
Immunology ; 149(4): 374-385, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27479869

RESUMEN

Periodontitis is a chronic inflammatory condition characterized by destruction of non-mineralized and mineralized connective tissues. It is initiated and maintained by a dysbiosis of the bacterial biofilm adjacent to teeth with increased prevalence of Gram-negative microorganisms. Nucleotide-binding oligomerization domain containing 1 (NOD1) is a member of the Nod-like receptors (NLRs) family of proteins that participate in the activation of the innate immune system, in response to invading bacteria or to bacterial antigens present in the cytoplasm. The specific activating ligand for NOD1 is a bacterial peptidoglycan derived primarily from Gram-negative bacteria. This study assessed the role of NOD1 in inflammation-mediated tissue destruction in the context of host-microbe interactions. We used mice with whole-genome deletion of the NOD1 gene in a microbe-induced periodontitis model using direct injections of heat-killed Gram-negative or Gram-negative/Gram-positive bacteria on the gingival tissues. In vitro experiments using primary bone-marrow-derived macrophages from wild-type and NOD1 knockout mice provide insight into the role of NOD1 on the macrophage response to Gram-negative and Gram-negative/Gram-positive bacteria. Microcomputed tomography analysis indicated that deletion of NOD1 significantly aggravated bone resorption induced by Gram-negative bacteria, accompanied by an increase in the numbers of osteoclasts. This effect was significantly attenuated by the association with Gram-positive bacteria. In vitro, quantitative PCR arrays indicated that stimulation of macrophages with heat-killed Gram-negative bacteria induced the same biological processes in wild-type and NOD1-deficient cells; however, expression of pro-inflammatory mediators was increased in NOD1-deficient cells. These results suggest a bone-sparing role for NOD1 in this model.


Asunto(s)
Aggregatibacter actinomycetemcomitans/inmunología , Resorción Ósea/inmunología , Encía/inmunología , Limosilactobacillus fermentum/inmunología , Macrófagos/fisiología , Proteína Adaptadora de Señalización NOD1/metabolismo , Enfermedades Periodontales/inmunología , Animales , Antígenos Bacterianos/inmunología , Resorción Ósea/microbiología , Células Cultivadas , Modelos Animales de Enfermedad , Encía/microbiología , Interacciones Huésped-Patógeno , Humanos , Mediadores de Inflamación/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína Adaptadora de Señalización NOD1/genética , Osteoclastos/patología , Enfermedades Periodontales/microbiología
4.
J Prosthet Dent ; 115(4): 428-36, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26597465

RESUMEN

STATEMENT OF PROBLEM: The longevity of dental implants depends on the absence of inflammation in the periimplant tissue. Similar to teeth, pathogenic bacteria can adhere on implant abutment surfaces and cause periimplant disease and consequently implant loss. PURPOSE: The purpose of this in vitro study was to evaluate the influence of physical and chemical properties of 2 common materials used as implant abutments, titanium (Ti) and zirconia (ZrO2), and the use of bovine enamel (BE) as a positive control on biofilm formation. MATERIAL AND METHODS: Biofilm formation was analyzed by growing Porphyromonas gingivalis and Fusobacterium nucleatum as monospecies and mixed species biofilms on the surfaces. The mean roughness (Ra) and surface free energy were evaluated for each material. Mature biofilm, formed after 7 days of incubation, was analyzed quantitatively and qualitatively by colony-forming unit and confocal laser scanning microscopy. RESULTS: The mean roughness in all disks was ≤0.21 µm and did not affect the bacterial adhesion. Titanium showed a greater degree of hydrophilicity compared with BE after 90 minutes of immersion in saliva. The surface free energy did not show differences, with the highest values for BE. Monospecies biofilms formed by P. gingivalis on Ti, and mixed species biofilm on ZrO2 exhibited small numbers of cells on disk surfaces. By confocal imaging, the mixed species biofilm appeared as a thin layer on ZrO2 surfaces. CONCLUSIONS: Material surfaces could have a significant impact on biofilm formation. ZrO2 implant abutment surfaces showed a decrease in anaerobic biofilm compared with Ti and BE.


Asunto(s)
Biopelículas , Diseño de Implante Dental-Pilar , Implantes Dentales/microbiología , Materiales Dentales/química , Adhesión Bacteriana , Humanos , Propiedades de Superficie , Titanio
5.
Anaerobe ; 18(3): 263-9, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22710412

RESUMEN

This present study evaluated the subgingival microbiota of the Cebus apella with different periodontal conditions kept by the Tufted Capuchin Monkey Procreation Center (São Paulo State University - UNESP) or free-ranging monkeys. For this purpose, clinical specimens of subgingival biofilm were collected from 52 monkeys, of both genders, 40 kept in captivity and 12 free-ranging monkeys. The primates were submitted to periodontal evaluation and biofilm samples were transferred to VMGA III transport medium and ultrapure water. The microbiota was cultivated in selective and non-selective culture media and microbial DNA was extracted and the presence of periodontal pathogens was evaluated using PCR and real-time PCR. The actinomycetes, fusobacteria, Campylobacter rectus, Eikenella corrodens, black-pigmented Gram-negative anaerobic rods, Tannerella forsythia, staphylococci and streptococci represent the predominantly detected microorganisms. Aggregatibacter actinomycetemcomitans, Dialister pneumosintes and Prevotella nigrescens were rarely observed, whereas Treponema denticola was not found. Populations of C. rectus, E. corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, T. forsythia and the total microbial load were significantly higher in animals with bone loss and, in smaller extension, in animals with gingival bleeding.


Asunto(s)
Cebus , Encía/microbiología , Gingivitis/veterinaria , Metagenoma , Enfermedades de los Monos/microbiología , Periodontitis/veterinaria , Actinobacteria/aislamiento & purificación , Actinobacteria/fisiología , Animales , Biopelículas , Enterobacteriaceae/aislamiento & purificación , Enterobacteriaceae/fisiología , Femenino , Fusobacterias/aislamiento & purificación , Fusobacterias/fisiología , Gingivitis/diagnóstico por imagen , Gingivitis/microbiología , Masculino , Periodontitis/diagnóstico por imagen , Periodontitis/microbiología , Porphyromonas gingivalis/aislamiento & purificación , Porphyromonas gingivalis/fisiología , Prevotella/aislamiento & purificación , Prevotella/fisiología , Radiografía , Staphylococcus/aislamiento & purificación , Staphylococcus/fisiología , Estadísticas no Paramétricas
6.
J Clin Periodontol ; 38(2): 131-41, 2011 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-21143491

RESUMEN

AIMS: periodontal disease (PD) and airway allergic inflammation (AL) present opposing inflammatory immunological features and clinically present an inverse correlation. However, the putative mechanisms underlying such opposite association are unknown. MATERIAL AND METHODS: Balb/C mice were submitted to the co-induction of experimental PD (induced by Actinobacillus actinomycetemcomitans oral inoculation) and AL [induced by sensitization with ovalbumin (OVA) and the subsequent OVA challenges], and evaluated regarding PD and AL severity, immune response [cytokine production at periodontal tissues, and T-helper transcription factors in submandibular lymph nodes (LNs)] and infection parameters. RESULTS: PD/AL co-induction decreased PD alveolar bone loss and periodontal inflammation while experimental AL parameters were unaltered. An active functional interference was verified, because independent OVA sensitization and challenge not modulate PD outcome. PD+AL group presented decreased tumour necrosis factor-α (TNF-α), interleukin (IL)-1ß, interferon-γ, IL-17A, receptor activator of nuclear factor κ-light-chain-enhancer of activated B cells ligand and matrix metalloproteinase (MMP)-13 levels in periodontal tissues, while IL-4 and IL-10 levels were unaltered by AL co-induction. AL co-induction also resulted in upregulated T-bet and related orphan receptor γ and downregulated GATA3 levels expression in submandibular LNs when compared with PD group. CONCLUSION: our results demonstrate that the interaction between experimental periodontitis and allergy involves functional immunological interferences, which restrains experimental periodontitis development by means of a skewed immune response.


Asunto(s)
Infecciones por Actinobacillus/inmunología , Citocinas/inmunología , Periodontitis/inmunología , Hipersensibilidad Respiratoria/inmunología , Infecciones por Actinobacillus/complicaciones , Infecciones por Actinobacillus/microbiología , Aggregatibacter actinomycetemcomitans/inmunología , Animales , Susceptibilidad a Enfermedades/inmunología , Fenómenos del Sistema Inmunológico , Mediadores de Inflamación/inmunología , Ratones , Ratones Endogámicos BALB C , Ovalbúmina , Periodontitis/complicaciones , Periodontitis/microbiología , Periodontitis/patología , Periodoncio/inmunología , Hipersensibilidad Respiratoria/inducido químicamente , Hipersensibilidad Respiratoria/complicaciones , Índice de Severidad de la Enfermedad , Linfocitos T/inmunología
7.
Braz J Microbiol ; 42(3): 1047-55, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24031721

RESUMEN

The aim of this study was to evaluate the occurrence of yeasts, pseudomonads and enteric bacteria in the oral cavity of patients undergoing radiotherapy (RT) for treatment of head and neck cancer. Fifty patients receiving RT were examined before, during and 30 days after RT. Saliva, mucosa, and biofilm samples were collected and microorganisms were detected by culture and polymerase chain reaction (PCR). The most prevalent yeasts in patients submitted to RT were Candida albicans, C. tropicalis, C. krusei, C. glabrata and C. parapsilosis. Citrobacter, Enterobacter, Enterococcus, Klebsiella, Proteus, and Pseudomonas were the most frequently cultivated bacteria. Before RT, targeted bacteria were cultivated from 22.2% of edentulous patients and 16.6% of dentate patients; 30 days after RT, these microorganisms were recovered from 77.8% edentulous and 46.8% dentate patients. By PCR, these microorganisms were detected from all edentulous patients, 78.1% of dentate patients. The presence of Gram-negative enteric roads and fungi was particularly frequent in patients presenting mucositis level III or IV. Modifications in the oral environment due to RT treatment seem to facilitate the colonization of oral cavity by members of family Enterobacteriaceae, genera Enterococcus and Candida.

8.
Res Vet Sci ; 136: 268-276, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33721714

RESUMEN

This objective of this study was to evaluate the use of tulathromycin on the timing of appearance and number of four indicator organisms representing the gastrointestinal microbial community, the incidence of diarrhea and a measure of the systemic inflammatory profile in Holstein heifers. Twenty-six Holstein heifer calves were distributed between receiving (ATB+) or not receiving (ATB-) tulathromycin at a dose of 2.5 mg/kg by 12 h of age. Samples from the calves were collected at six times during the neonatal period. Stool samples were used to determine the dry matter content and quantitative analysis of specific indicator bacterial populations. Samples of whole blood and serum were collected to determine the total number of neutrophils, the number of CD62L+ neutrophils, quantity of haptoglobin, and to allow for ex vivo measurement of reactive oxygen species. A higher frequency of diarrhea was detected in the ATB+ calves (84.6%) than ATB- (53.8%) on days 13-15 (P = 0.084). ATB- calves had a greater number of Bifidobacterium in stool on day 3-5 (P = 0.002), and on days 7-9 (P = 0.018). The ATB+ calves tended to have a higher number of Escherichia coli in stool on days 20-23 and days 27-30 (P = 0.052 and P = 0.072). Both the total number of neutrophils (P = 0.013) and the capacity for ROS production was higher in ATB- (P = 0.038) than ATB+ calves at all points tested. ATB+ calves had higher levels of haptoglobin (P = 0.032) on days 13-15. Administration of tulathromycin appeared to negatively impact the establishment of a normal microbiome and to modulate the development of innate immune function.


Asunto(s)
Antibacterianos/uso terapéutico , Profilaxis Antibiótica/veterinaria , Enfermedades de los Bovinos/tratamiento farmacológico , Diarrea/veterinaria , Disacáridos/uso terapéutico , Microbioma Gastrointestinal/efectos de los fármacos , Compuestos Heterocíclicos/uso terapéutico , Inflamación/veterinaria , Animales , Animales Recién Nacidos , Bacterias/efectos de los fármacos , Bovinos , Diarrea/tratamiento farmacológico , Heces/microbiología , Femenino , Inflamación/tratamiento farmacológico
9.
Clin Infect Dis ; 50(3): e11-8, 2010 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-20038241

RESUMEN

BACKGROUND: Periodontal diseases (PDs) are infectious diseases in which periodontopathogens trigger chronic inflammatory and immune responses that lead to tissue destruction. Recently, viruses have been implicated in the pathogenesis of PDs. Individuals infected with human T lymphotropic virus 1 (HTLV-1) present with abnormal oral health and a marked increased prevalence of periodontal disease. METHODS: In this study, we investigated the patterns of periodontopathogen infection and local inflammatory immune markers in HTLV-1-seropositive individuals with chronic periodontitis (CP/HTLV-1 group) compared with HTLV-1-seronegative individuals with chronic periodontitis (CP group) and periodontally healthy, HTLV-1-seronegative individuals (control group). RESULTS: Patients in the CP/HTLV-1 group had significantly higher values of bleeding on probing, mean probing depth, and attachment loss than patients in the CP group. The expression of tumor necrosis factor alpha and interleukin (IL) 4 was found to be similar in the CP and CP/HTLV-1 groups, whereas IL-12 and IL-17 levels trended toward a higher expression in the CP/HTLV-1 group. A significant increase was seen in the levels of IL-1beta and interferon gamma in the CP/HTLV-1 group compared with the CP group, whereas expression of the regulatory T cell marker FOXp3 and IL-10 was significantly decreased in the lesions from the CP/HTLV-1 group. Interestingly, similar frequency and/or load of periodontopathogens (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans) and frequency of viruses (herpes simplex virus 1, human cytomegalovirus, and Epstein-Barr virus) characteristically associated with PDs were found in the CP/HTLV and CP groups. CONCLUSIONS: HTLV-1 may play a critical role in the pathogenesis of periodontal disease through the deregulation of the local cytokine network, resulting in an exacerbated response against a standard periodontopathogen infection.


Asunto(s)
Bacterias Anaerobias/aislamiento & purificación , Enfermedad Crónica , Citocinas/biosíntesis , Bacterias Gramnegativas/aislamiento & purificación , Infecciones por HTLV-I/complicaciones , Infecciones por HTLV-I/inmunología , Virus Linfotrópico T Tipo 1 Humano/aislamiento & purificación , Periodontitis/microbiología , Adulto , Recuento de Colonia Microbiana , Femenino , Encía/patología , Herpesviridae/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Periodontitis/inmunología , Periodontitis/patología , Periodontitis/virología
10.
Braz J Microbiol ; 41(4): 1056-64, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24031586

RESUMEN

Chronic osteomyelitis of maxilla and mandible is rare in industrialized countries and its occurrence in developing countries is associated with trauma and surgery, and its microbial etiology has not been studied thoroughly. The aim of this investigation was to evaluate the microbiota associated with osteomyelitis of mandible or maxilla from some Brazilian patients. After clinical and radiographic evaluation, samples of bone sequestra, purulent secretion, and biopsies of granulomatous tissues from twenty-two patients with chronic osteomyelitis of mandible and maxilla were cultivated and submitted for pathogen detection by using a PCR method. Each patient harbored a single lesion. Bacterial isolation was performed on fastidious anaerobe agar supplemented with hemin, menadione and horse blood for anaerobes; and on tryptic soy agar supplemented with yeast extract and horse blood for facultative bacteria and aerobes. Plates were incubated in anaerobiosis and aerobiosis, at 37(o)C for 14 and 3 days, respectively. Bacteria were cultivated from twelve patient samples; and genera Actinomyces, Fusobacterium, Parvimonas, and Staphylococcus were the most frequent. By PCR, bacterial DNA was detected from sixteen patient samples. The results suggest that cases of chronic osteomyelitis of the jaws are usually mixed anaerobic infections, reinforcing the concept that osteomyelitis of the jaws are mainly related to microorganisms from the oral environment, and periapical and periodontal infections may act as predisposing factors.

11.
J Appl Oral Sci ; 28: e20190519, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32348444

RESUMEN

Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 µM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 µM (p<0.05) and 250 µM (p<0.01). The POH increased ROS production at both 10 µM and 100 µM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 µM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 µM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.


Asunto(s)
Antibacterianos/farmacología , Fusobacterium nucleatum/efectos de los fármacos , Macrófagos/efectos de los fármacos , Monoterpenos/farmacología , Porphyromonas/efectos de los fármacos , Especies Reactivas de Oxígeno/análisis , Animales , Arginasa/análisis , Productos Biológicos/farmacología , Proliferación Celular/efectos de los fármacos , Citometría de Flujo , Fusobacterium nucleatum/crecimiento & desarrollo , Expresión Génica , Lipopolisacáridos/farmacología , Macrófagos/metabolismo , Ratones , Pruebas de Sensibilidad Microbiana , Porphyromonas/crecimiento & desarrollo , Células RAW 264.7 , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los Resultados , Factores de Tiempo , Factor de Necrosis Tumoral alfa/análisis
12.
Braz J Microbiol ; 39(2): 257-61, 2008 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24031212

RESUMEN

The purpose of this study was to determine the prevalence of enteric bacteria and yeasts in biofilm of 80 HIV-positive patients with plaque-associated gingivitis or necrotizing periodontitis. Patients were subjected to extra, intra oral and radiographic examinations. The oral hygiene, bleeding on probing, gingival conditions, and attachment loss were evaluated. Clinical specimens were collected from gingival crevices or periodontal pockets, transferred to VMGA III, diluted and transferred to Sabouraud Dextrose agar with 100 µg/ml of chloramphenicol, peptone water, EVA broth, EMB agar, SS agar, Bile esculin agar and Brilliant green agar. Isolation of yeasts was carried out at room temperature, for 3-7 days; and for the isolation of enteric microorganisms plates were incubated at 37°C, for 24-48 h. The yeasts identification was performed according to the carbon and nitrogen assimilation, fermentation of carbohydrates and germ tube formation. Bacteria were identified according to their colonial and cellular morphologies and biochemical tests. Yeasts were identified as Candida albicans and its occurrence was more common in patients with CD4+ below 200/mm(3) and was affected by the extension of periodontal involvement (P = 0.0345). Enteric bacteria recovered from clinical specimens were identified as Enterobacter sakazakii, Enterobacter cloacae, Serratia liquefaciens, Klebsiella oxytoca and Enterococcus sp. Enterobacteriaceae and enterococci were detected in 32.5% of clinical samples from patients with necrotizing periodontitis. In conclusion, non-oral pathogenic bacteria and C. albicans were more prevalent in periodontal sites of HIV-positive patients with necrotizing periodontitis and chronic gingivitis.

13.
Braz J Microbiol ; 39(4): 658-63, 2008 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24031284

RESUMEN

Aggregatibacter actinomycetemcomitans is an important etiologic agent of the periodontitis and is associated with extra-oral infections. In this study, the detection of the ltxA gene as well as the ltx promoter region from leukotoxic A. actinomycetemcomitans isolated from 50 Brazilian patients with periodontitis and 50 healthy subjects was performed. The leukotoxic activity on HL-60 cells was also evaluated. Leukotoxic activity was determined using a trypan blue exclusion method. The 530 bp deletion in the promoter region was evaluated by PCR using a PRO primer pair. A. actinomycetemcomitans was detected by culture and directly from crude subgingival biofilm by PCR using specific primers. By culture, A. actinomycetemcomitans was detected in nine (18%) of the periodontal patients and one (2%) healthy subject. However, by PCR, this organism was detected in 44% of the periodontal patients and in 16% of the healthy subjects. It was verified a great discrepancy between PCR detection of the ltx operon promoter directly from crude subgingival biofilm and from bacterial DNA. Only one periodontal sample harbored highly leukotoxic A. actinomycetemcomitans. Moreover, biotype II was the most prevalent and no correlation between biotypes and leukotoxic activity was observed. The diversity of leukotoxin expression by A. actinomycetemcomitans suggests a role of this toxin in the pathogenesis of periodontal disease and other infectious diseases.

14.
Braz J Microbiol ; 46(4): 1141-5, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26691473

RESUMEN

Enterotoxigenic Bacteroides fragilis (ETBF) is an important part of the human and animal intestinal microbiota and is commonly associated with diarrhea. ETBF strains produce an enterotoxin encoded by the bft gene located in the B. fragilis pathogenicity island (BfPAI). Non-enterotoxigenic B. fragilis (NTBF) strains lack the BfPAI and usually show two different genetic patterns, II and III, based on the absence or presence of a BfPAI-flanking region, respectively. The incidence of ETBF and NTBF strains in fecal samples isolated from children without acute diarrhea or any other intestinal disorders was determined. All 84 fecal samples evaluated were B. fragilis-positive by PCR, four of them harbored the bft gene, 27 contained the NTBF pattern III DNA sequence, and 52 were considered to be NTBF pattern II samples. One sample was positive for both ETBF and NTBF pattern III DNA sequences. All 19 B. fragilis strains isolated by the culture method were bft-negative, 9 belonged to pattern III and 10 to pattern II. We present an updated overview of the ETBF and NTBF incidence in the fecal microbiota of children from Sao Paulo City, Brazil.


Asunto(s)
Toxinas Bacterianas/genética , Infecciones por Bacteroides/microbiología , Bacteroides fragilis/genética , Bacteroides fragilis/aislamiento & purificación , Heces/microbiología , Genotipo , Metaloendopeptidasas/genética , Animales , Infecciones por Bacteroides/epidemiología , Bacteroides fragilis/clasificación , Brasil/epidemiología , Niño , Preescolar , ADN Bacteriano/genética , Femenino , Humanos , Incidencia , Masculino , Tipificación Molecular , Reacción en Cadena de la Polimerasa
15.
Int J Mol Med ; 35(1): 211-7, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25384405

RESUMEN

In the present study, essential oils extracted from the leaves and flowers of Lippia alba (Mill.) N.E.Br. (L. alba) were analyzed for their antimicrobial activity and their effects on osteoclasts. The periodontal pathogens, Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans; ATCC 43717), Fusobacterium nucleatum (F. nucleatum; ATCC 25586) and Porphyromonas gingivalis (P. gingivalis); ATCC 33277) were used in antimicrobial activity assays for determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC), whereas Bacteroides fragilis (B. fragilis; ATCC 25285) was used as the control microorganism. Osteoclast (OC) apoptosis was assessed by TUNEL assay and Fas receptor expression was detected by immunocytochemistry. The analysis of antimicrobial activity revealed that P. gingivalis had the lowest MIC values, whereas A. actinomycetemcomitans had the highest. L. alba essential oils were found to be toxic to human cells, although the compounds, carvone, limonene and citral, were non-toxic and induced apoptosis in the OCs. This study demonstrates that L. alba has potential biotechnological application in dentistry. In fact periodontal disease has a multifactorial etiology, and the immune response to microbial challenge leads to osteoclast activation and the resorption of the alveolar bone, resulting in tooth loss.


Asunto(s)
Flores/química , Lippia/química , Aceites Volátiles/farmacología , Extractos Vegetales/farmacología , Hojas de la Planta/química , Antiinfecciosos/química , Antiinfecciosos/farmacología , Apoptosis/efectos de los fármacos , Células Cultivadas , Humanos , Aceites Volátiles/química , Osteoclastos/efectos de los fármacos , Extractos Vegetales/química
16.
J. appl. oral sci ; J. appl. oral sci;28: e20190519, 2020. tab, graf
Artículo en Inglés | LILACS, BBO - odontología (Brasil) | ID: biblio-1101254

RESUMEN

Abstract Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 μM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 μM (p<0.05) and 250 μM (p<0.01). The POH increased ROS production at both 10 μM and 100 μM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 μM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 μM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.


Asunto(s)
Animales , Ratones , Fusobacterium nucleatum/efectos de los fármacos , Especies Reactivas de Oxígeno/análisis , Porphyromonas/efectos de los fármacos , Monoterpenos/farmacología , Macrófagos/efectos de los fármacos , Antibacterianos/farmacología , Arginasa/análisis , Factores de Tiempo , Productos Biológicos/farmacología , Pruebas de Sensibilidad Microbiana , Expresión Génica , Lipopolisacáridos/farmacología , Reproducibilidad de los Resultados , Factor de Necrosis Tumoral alfa/análisis , Fusobacterium nucleatum/crecimiento & desarrollo , Especies Reactivas de Oxígeno/metabolismo , Porphyromonas/crecimiento & desarrollo , Proliferación Celular/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la Polimerasa , Citometría de Flujo , Células RAW 264.7 , Macrófagos/metabolismo
17.
Res Microbiol ; 155(10): 843-6, 2004 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-15567279

RESUMEN

Twenty Bacteroides fragilis group species isolated from children with and without diarrhea were analyzed. Antibiotic susceptibility was performed using an agar dilution method; beta-lactamase production was determined using a nitrocefin method, and plasmids were extracted using a commercial Miniprep System. MIC values ranged from 16 to 256 microg/ml for penicillin, 4-128 microg/ml for amoxicillin/clavulanic acid, 0.25-256 microg/ml for clindamycin, and 16-256 microg/ml for penicillin. beta-Lactamase was detected in all isolates. Only five isolates harbored plasmids varying from 7.8 to 1.8 kb. Loss of 6.4- and 3.8-kb plasmids in B. fragilis C68c was related to antibiotic resistance. Low molecular weight plasmids of 2.8-1.8 kb were stable. PCR amplification of cfiA and cepA genes was observed using total DNA, and the cfiA gene was also amplified from the 6.4-kb plasmid.


Asunto(s)
Bacteroides fragilis/enzimología , beta-Lactamasas/biosíntesis , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Infecciones por Bacteroides/microbiología , Bacteroides fragilis/efectos de los fármacos , Bacteroides fragilis/genética , Niño , ADN Bacteriano/química , ADN Bacteriano/genética , Diarrea/microbiología , Farmacorresistencia Bacteriana/genética , Humanos , Pruebas de Sensibilidad Microbiana , Plásmidos/genética , Reacción en Cadena de la Polimerasa , beta-Lactamasas/química , beta-Lactamasas/genética , beta-Lactamasas/metabolismo
18.
Rev Saude Publica ; 38(5): 723-8, 2004 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-15499445

RESUMEN

OBJECTIVE: To comparatively detect A. actinomycetemcomitans and F. nucleatum from periodontal and healthy sites. METHODS: Subgingival clinical samples from 50 periodontitis adult patients and 50 healthy subjects were analyzed. Both organisms were isolated using a trypticase soy agar-bacitracin-vancomycin (TSBV) medium and detected by PCR. Conventional biochemical tests were used for bacteria identification. RESULTS: A. actinomycetemcomitans and F. nucleatum were isolated in 18% and 20% of the patients, respectively, and in 2% and 24% of healthy subjects. Among A. actinomycetemcomitans isolates, biotype II was the most prevalent. Primer pair AA was 100% sensitive in the detection of A. actinomycetemcomitans from both subject groups. Primers ASH and FU were also 100% sensitive to detect this organism in healthy subject samples. Primer pair FN5047 was more sensitive to detect F. nucleatum in patients or in healthy samples than primer 5059S. Primers ASH and 5059S were more specific in the detection of A. actinomycetemcomitans and F. nucleatum, respectively, in patients and in healthy subject samples. CONCLUSIONS: PCR is an effective tool for detecting periodontal pathogens in subgingival samples, providing a faster and safer diagnostic tool of periodontal diseases. The method's sensitivity and specificity is conditioned by the choice of the set of primers used.


Asunto(s)
Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Fusobacterium necrophorum/aislamiento & purificación , Periodontitis/microbiología , Adulto , Aggregatibacter actinomycetemcomitans/clasificación , Aggregatibacter actinomycetemcomitans/genética , Técnicas de Tipificación Bacteriana , Cartilla de ADN/análisis , ADN Bacteriano/análisis , Femenino , Fusobacterium necrophorum/clasificación , Fusobacterium necrophorum/genética , Humanos , Masculino , Persona de Mediana Edad , Periodontitis/diagnóstico , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Análisis de Secuencia de ADN
19.
J Appl Oral Sci ; 22(2): 118-24, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24676582

RESUMEN

OBJECTIVES: Primary teeth work as guides for the eruption of permanent dentition, contribute for the development of the jaws, chewing process, preparing food for digestion, and nutrient assimilation. Treatment of pulp necrosis in primary teeth is complex due to anatomical and physiological characteristics and high number of bacterial species present in endodontic infections. The bacterial presence alone or in association in necrotic pulp and fistula samples from primary teeth of boys and girls was evaluated. MATERIAL AND METHODS: Necrotic pulp (103) and fistula (7) samples from deciduous teeth with deep caries of 110 children were evaluated. Bacterial morphotypes and species from all clinical samples were determined. RESULTS: A predominance of gram-positive cocci (81.8%) and gram-negative coccobacilli (49.1%) was observed. In 88 out of 103 pulp samples, a high prevalence of Enterococcus spp. (50%), Porphyromonas gingivalis (49%), Fusobacterium nucleatum (25%) and Prevotella nigrescens (11.4%) was observed. Porphyromonas gingivalis was detected in three out of seven fistula samples, Enterococcus spp. in two out of seven samples, and F. nucleatum, P. nigrescens and D. pneumosintes in one out of seven samples. CONCLUSIONS: Our results show that Enterococcus spp. and P. gingivalis were prevalent in necrotic pulp from deciduous teeth in boys from 2 to 5 years old, and that care of the oral cavity of children up to five years of age is important.


Asunto(s)
Fístula Dental/microbiología , Necrosis de la Pulpa Dental/microbiología , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/aislamiento & purificación , Diente Primario/microbiología , Factores de Edad , Distribución de Chi-Cuadrado , Niño , Preescolar , ADN Bacteriano/análisis , Cavidad Pulpar/microbiología , Femenino , Bacterias Gramnegativas/genética , Bacterias Grampositivas/genética , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Valores de Referencia , Factores Sexuales
20.
J Appl Oral Sci ; 22(6): 528-33, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25591021

RESUMEN

OBJECTIVE: In this study, the gingival conditions and the quantitative detection for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in pregnant women were determined. MATERIAL AND METHODS: Quantitative determinations of periodontal bacteria by using a SyBr green system in women during pregnancy were performed. Women at the 2nd and 3rd trimesters of pregnancy and non-pregnant women were included in this study. A. actinomycetemcomitans was observed in high numbers in women at the 2nd and 3rd trimesters of pregnancy with a significant difference (p<0.05). F. nucleatum and P. intermedia were also observed in high levels. RESULTS AND CONCLUSION: Our results show that pregnant women are more susceptible to gingivitis, and the presence of A. actinomycetemcomitans in subgingival biofilm might be taken into account for the treatment of periodontal disease.


Asunto(s)
Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Fusobacterium nucleatum/aislamiento & purificación , Encía/microbiología , Periodoncio/microbiología , Porphyromonas gingivalis/aislamiento & purificación , Prevotella intermedia/aislamiento & purificación , Adolescente , Adulto , Carga Bacteriana , Biopelículas/crecimiento & desarrollo , Femenino , Humanos , Estudios Longitudinales , Enfermedades Periodontales/microbiología , Índice Periodontal , Reacción en Cadena de la Polimerasa , Embarazo , Segundo Trimestre del Embarazo , Tercer Trimestre del Embarazo , Adulto Joven
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