RESUMEN
AIM: To compare the subgingival microbiota of patients receiving supportive periodontal care (SPC) with and without subgingival instrumentation, over 2 years. MATERIALS AND METHODS: This study was a randomized clinical trial that included 62 participants (50.97 ± 9.26 years old; 40 females) who completed non-surgical periodontal therapy. Participants were randomly assigned to receive oral prophylaxis with oral hygiene instructions alone (test) or in combination with subgingival instrumentation (control) during SPC. Pooled subgingival biofilm samples were obtained from four sites per patient at SPC baseline and at 3, 6, 12, 18, and 24 months. Real-time polymerase chain reaction was used for absolute quantification of Eubacteria and the target bacteria Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. Data were analysed using generalized estimating equations, taking into consideration the clustering of observations within individuals. RESULTS: No significant differences were found between the experimental groups regarding the mean counts of Eubacteria and target bacteria, as well as the periodontal parameters at the sampled sites. Although significant variability in bacterial counts was present during SPC, all counts after 2 years were not statistically different from those at baseline. Bacterial counts were associated with the presence of plaque, bleeding on probing, mean probing depth ≥3 mm, and follow-up period. CONCLUSIONS: SPC with or without subgingival instrumentation can result in comparable subgingival microbiological outcomes. CLINICAL TRIAL REGISTRATION: clinicaltrials.gov: NCT01598155 (https://clinicaltrials.gov/study/NCT01598155?intr=supragingival%20control&rank=4#study-record-dates).
Asunto(s)
Porphyromonas gingivalis , Tannerella forsythia , Treponema denticola , Humanos , Femenino , Masculino , Persona de Mediana Edad , Treponema denticola/aislamiento & purificación , Porphyromonas gingivalis/aislamiento & purificación , Tannerella forsythia/aislamiento & purificación , Biopelículas , Adulto , Higiene Bucal , Carga Bacteriana , Estudios de Seguimiento , Índice Periodontal , Encía/microbiología , Raspado Dental/instrumentación , Raspado Dental/métodos , Placa Dental/microbiología , Periodontitis Crónica/microbiología , Periodontitis Crónica/terapiaRESUMEN
OBJECTIVES: This double-blind, randomized, placebo-controlled clinical trial evaluated the adjuvant effects of Bifidobacterium lactis HN019 on the treatment of plaque-induced generalized gingivitis. MATERIALS AND METHODS: Sixty patients were submitted to professional supragingival scaling and prophylaxis. They were randomly assigned to test (probiotic lozenges containing B. lactis HN019, n = 30) or control (placebo lozenges, n = 30) groups. Lozenges were consumed twice a day for 8 weeks. Bleeding on probing (BoP), Gingival Index (GI), Plaque Index (PI), probing depth (PD), and clinical attachment level (CAL) were evaluated at baseline and after 2 and 8 weeks. Gingival crevicular fluid (GCF) was collected at baseline and at 8 weeks for analysis of the inflammatory mediators IL-1ß, IL-1α, IL-8, MCP-1, and MIP-1ß. Data were statistically analyzed (p < 0.05). RESULTS: After 8 weeks, both groups showed reduction in the percentage of PI, with no significant difference between groups (p = 0.7423). The test group presented a lower percentage of BoP and a higher percentage of sites with GI ≤ 1 when compared with the control group at the end of the study (p < 0.0001). At 8 weeks, the test group had a greater number of patients without generalized gingivitis than the control group (20 and 11 patients, respectively; p < 0.05). The test group presented significantly lower levels of IL-1α, IL-1ß, and MCP-1 in GCF than the control group at the end of the study (p < 0.05). CONCLUSION: The adjunct use of B. lactis HN019 promotes additional clinical and immunological benefits in the treatment of generalized gingivitis. CLINICAL RELEVANCE: B. lactis HN019 can be an efficient and side-effect-free adjunct strategy in the treatment of generalized gingivitis.
Asunto(s)
Bifidobacterium animalis , Placa Dental , Gingivitis , Placa Aterosclerótica , Humanos , Gingivitis/terapia , Raspado Dental , Placa Dental/terapia , Placa Dental/microbiología , Administración Oral , Líquido del Surco GingivalRESUMEN
BACKGROUND: Despite the body of evidence supporting the clinical benefits of metronidazole (MTZ) and amoxicillin (AMX) in the treatment of young patients with periodontitis, the microbiological outcomes of this antibiotic protocol have been less explored. This study evaluated the microbiological effects of adjunctive MTZ+AMX in the treatment of young patients with periodontitis. METHODS: Subjects with periodontitis Stages III or IV and ≤30 years old were randomly allocated to receive scaling and root planing (SRP) with placebo (n = 15) or with MTZ (400 mg) and AMX (500 mg) three times a day for 14 days (n = 15). Nine subgingival biofilm samples per subject (three samples from each probing depth (PD) category: ≤3, 4-6, and ≥7 mm) were collected at baseline and 3-, 6-, and 12-months post-treatment and individually analyzed for 40 bacterial species by checkerboard DNA-DNA hybridization. RESULTS: Thirty subjects (15/group) with mean ages 27.6 ± 3.5 (control) and 26.8 ± 3.9 (test) were included. At 12 months post-therapy, the antibiotic group harbored lower proportions of red complex (1.3%) than the placebo group (12.5%) (p < 0.05). SRP + MTZ+AMX was more effective than mechanical treatment in reducing levels/proportions of several pathogens and increasing proportions of Actinomyces species (p < 0.05). Levels/proportions of Aggregatibacter actinomycetemcomitans were only reduced in the antibiotic group (p < 0.05). This group also exhibited greater reduction in the number of sites with PD ≥5 mm and higher percentage of subjects reaching the clinical end point for treatment (≤4 sites with PD ≥5 mm) than the control group (p < 0.05). CONCLUSION: SRP+MTZ+AMX allowed for establishing a long-term healthier subgingival biofilm community and periodontal clinical condition, than SRP only.
Asunto(s)
Placa Dental , Periodontitis , Humanos , Adulto Joven , Adulto , Metronidazol/uso terapéutico , Amoxicilina/uso terapéutico , Terapia Combinada , Placa Dental/microbiología , Antibacterianos/uso terapéutico , Periodontitis/tratamiento farmacológico , Raspado Dental/métodos , Aplanamiento de la Raíz/métodos , ADN/uso terapéutico , Resultado del TratamientoRESUMEN
Introducción: existen diversos patógenos que pueden afectar no sólo la salud periodontal, sino también la salud general de los pacientes. Objetivo: determinar la Porphyromonas gingivalis (PG) en el primer molar superior derecho de adolescentes, de entre 12 y 18 años, con al menos un mes de tratamiento de ortodoncia con aparatología fija. Material y métodos: se realizó un estudio observacional, descriptivo, transversal de casos en un grupo de 26 adolescentes con tratamiento de ortodoncia, compuesto de brackets metálicos, tubos o bandas, arcos NiTi termoactivos, módulos, cadenas o ligaduras; sin importar sexo, edad, tiempo de tratamiento o maloclusión. Se formaron dos pares de grupos 1 y 2 (15 mujeres y 11 hombres), A y B (13 mujeres y 13 hom- bres) comparando los resultados obtenidos entre los grupos. Resulta- dos: dentro del grupo 1 y 2 la detección molecular de microorganismos arroja que 80% fueron positivas a la PG, 58.33% presenta maloclusión y en promedio 89% de las pacientes son positivas a PG. La detección molecular del grupo A y B indica que 54.54% fueron positivos a PG, mientras que 83.3% presenta maloclusión y en promedio 47% son positivos a PG. Conclusión: la explicación de los eventos moleculares que se desencadenan en la cavidad oral y los sistemas afectados por PG contribuyen a la prevención de complicaciones al tener una mejor comprensión de los fenómenos infecciosos (AU)
Introduction: there are various pathogens that can affect not only periodontal health, but also the general health of patients. Objective: to determine Porphyromonas gingivalis (PG) in the upper right first molar of adolescents, between 12 and 18 years old, with at least one month of orthodontic treatment with fixed appliances. Material and methods: a cross-sectional descriptive observational study of cases was carried out in a group of 26 adolescents with orthodontic treatment, consisting of metal brackets, tubes or bands, thermoactive NiTi archwires, modules, chains or ligatures; regardless of sex, age, treatment time or malocclusion. Two pairs of groups 1 and 2 (15 women and 11 men), A and B (13 women and 13 men) were formed, comparing the results obtained between the groups. Results: within group 1 and 2, the molecular detection of microorganisms shows that 80% were positive for PG, 58.33% presented malocclusion and an average of 89% of patients were positive for PG. The molecular detection of group A and B indicates that 54.54% were positive for PG while 83.3% presented malocclusion and on average 47% were positive for PG. Conclusion: the explanation of the molecular events that are triggered in the oral cavity and the systems affected by PG contribute to the prevention of complications by having a better understanding of the infectious phenomena (AU)
Asunto(s)
Humanos , Masculino , Femenino , Niño , Adolescente , Soportes Ortodóncicos/efectos adversos , Porphyromonas gingivalis/aislamiento & purificación , Placa Dental/microbiología , Aparatos Ortodóncicos Fijos/efectos adversos , Epidemiología Descriptiva , Estudios Transversales , Líquido del Surco Gingival/microbiología , Estudio Observacional , México , Biología Molecular/métodosRESUMEN
Introducción: el biofilm dental microbiano es el precursor de diversas enfermedades orales, una de ellas la caries, ésta representa la enferme- dad oral más significativa a nivel mundial, con una incidencia de 1.76 billones de niños afectados. Las nanopartículas de plata (AgNPs) se están usando como alternativa para el control y prevención del biofilm dental, ya que poseen propiedades antimicrobianas contra bacterias relacionadas a estas enfermedades. Sin embargo, no hay estudios que evalúen este comportamiento en pacientes pediátricos. Objetivo: eva- luar la actividad antimicrobiana de las AgNPs en bacterias de aislados clínicos tomados de pacientes pediátricos. Material y métodos: se tomó muestra del biofilm dental de 22 pacientes pediátricos, el efecto micro- biológico se evaluó mediante ensayos microbiológicos estandarizados internacionalmente por triplicado, usando dos diferentes tamaños de AgNPs. Resultados: los dos tamaños de AgNPs mostraron inhibición bacteriana, sin embargo, sólo se vio una diferencia estadísticamente significativa entre el género (p < 0.05), además, en general, hubo una correlación positiva significativa en relación a la concentración de las AgNPs y la velocidad del crecimiento bacteriano (p < 0.05). Conclusión: las AgNPs se pueden considerar como una alternativa para la prevención del biofilm dental y de esta manera para el control de diferentes enfermedades orales (AU))
Introduction: dental biofilm is the precursor of oral diseases, one of them dental caries, this represents the most significant oral disease worldwide with an incidence of 1.76 billion affected children. Silver nanoparticles (AgNPs) are being used as an alternative for the control and prevention of dental biofilm since they have antimicrobial properties against bacteria related to these diseases. However, there are no studies evaluating this behavior in pediatric patients. Objective: to evaluate the antimicrobial activity of AgNPs in bacteria from clinical isolates taken from pediatric patients. Material and methods: a sample of dental biofilm was taken from 22 pediatric patients, the microbiological effect was evaluated by international standardized microbiological tests in triplicate, using two different sizes of AgNPs. Results: the two sizes of AgNPs showed bacterial inhibition, however, only a statistically significant difference was seen between gender (p < 0.05), in addition, in general, there was a significant positive correlation in relation to the concentration of AgNPs and the speed bacterial growth (p < 0.05). Conclusion: AgNPs can be considered as an alternative for the prevention of dental biofilm and thus for the control of different oral diseases (AU)
Asunto(s)
Humanos , Masculino , Femenino , Preescolar , Niño , Caries Dental/prevención & control , Placa Dental/prevención & control , Nanopartículas/uso terapéutico , Crecimiento Bacteriano , Atención Dental para Niños/métodos , Medios de Cultivo , Placa Dental/microbiología , Distribución por Edad y SexoRESUMEN
Objetivo: actualizar la información sobre la disbiosis bacteriana oral y su efecto en enfermedades bucales. Material y métodos: se realizó una revisión bibliográfica detallada, donde la búsqueda de artículos comenzó desde el 2014 con trabajos de investigación relacionados con el tema. Se aplicaron palabras clave para facilitar y delimitar el tema. En los resultados obtenidos se observa información específica de disbiosis bacteriana y los problemas y enfermedades que causan en la cavidad bucal. Conclusión: la cavidad oral es un ecosistema muy complejo e interactivo donde se desarrollan variedades de hábitats que establecen relaciones entre los microorganismos en los distintos medios bucales. Por lo general, el cuerpo humano vive en simbiosis con dichas bacterias, esta relación hospedador-huésped es producto de años de evolución y convivencia para poder tolerar a dichas especies y por medio de años de investigación, determinar a los agentes patógenos y a los simbióticos, lo que permitirá en un futuro tener enfoques terapéuticos y científicos, para así solucionar, mejorar y evitar problemas relacionados con la salud (AU)
Objective: this review aimed to update the information on oral bacterial dysbiosis and its effect on oral diseases. Material and methods: a detailed literature review was performed, where the search for articles began in 2014 with research papers related to the topic. Keywords were applied to facilitate and delimit the topic. The results obtained show specific information on bacterial dysbiosis and the problems and diseases they cause in the oral cavity. Conclusion: the oral cavity is a very complex and interactive ecosystem where a variety of habitats develop and establish relationships between microorganisms in different oral environments. Generally, the human body lives in symbiosis with these bacteria, this host-guest relationship is the product of years of evolution and coexistence to be able to tolerate these species and through years of research to determine the pathogens and symbiotics, which will allow in the future to have therapeutic and scientific approaches, to solve, improve and avoid health-related problems (AU)
Asunto(s)
Humanos , Infecciones Bacterianas/complicaciones , Disbiosis/etiología , Enfermedades de la Boca/microbiología , Bacilos Grampositivos/patogenicidad , Bacilos y Cocos Aerobios Gramnegativos/patogenicidad , Placa Dental/microbiología , Interacciones Microbiota-Huesped , Boca/microbiologíaRESUMEN
It has been hypothesised that oral bacteria can migrate, through the blood, from the mouth to the arterial plaques, thus exacerbating atherosclerosis. This study compared bacteria present in the peripheral blood of individuals with and without coronary artery disease (CAD). RNA sequences obtained from blood were downloaded from GEO (GSE58150). Eight patients with coronary artery calcification (CAC) scoring > 500 and eight healthy individuals were analysed. After conducting quality control, the sequences were aligned to the hg38 reference genome using Hisat2. Bacterial taxa were analysed by inputting the unmapped sequences into Kraken. Ecological indices were calculated using Vegan. The package DESeq2 was used to compare the counts of bacteria per standard rank between groups. A total of 51 species were found only in patients with CAD and 41 were exclusively present in healthy individuals. The counts of one phylum, one class, three orders, two families and one genus were significantly different between the analysed groups (p < 0.00032, FDR < 10%), including the orders Cardiobacteriales, Corynebacteriales and Fusobacteriales. Twenty-three bacterial species belonging to the subgingival plaque bacterial complexes were also identified in the blood of individuals from both the groups; Fusobacterium nucleatum was significantly less frequent in patients with CAD (p = 0.0012, FDR = 4.8%). Furthermore, the frequency of another 11 bacteria differed significantly among patients with CAD than that among healthy individuals (p < 0.0030, FDR < 10%). These bacteria have not been previously reported in patients with atherosclerosis and periodontitis. The presence of members of the subgingival plaque bacterial complexes in the blood of patients with CAC supports the hypothesis that the periodontopathogens can be disseminated through the blood flow to other body parts where they may enhance inflammatory processes that can lead to the development or exacerbation of atherosclerosis.
Asunto(s)
Sangre/microbiología , Enfermedad de la Arteria Coronaria/microbiología , Placa Dental/microbiología , Enfermedades Periodontales/complicaciones , Estudios de Casos y Controles , Femenino , Humanos , Persona de Mediana Edad , Enfermedades Periodontales/microbiologíaRESUMEN
BACKGROUND: Following human immunodeficiency virus-1 (HIV-1) infection and antiretroviral therapy, the development of periodontal disease was shown to be favored. However, the influence of HIV-1 infection on the periodontal microbiota after non-surgical periodontal debridement (NSPD) needs a broad comprehension. This work aimed to compare the subgingival microbiological content of patients infected with HIV-1 and controls (non-infected) with periodontitis undergoing NSPD. METHODS: The bacterial profile of subgingival biofilm samples of patients with HIV-1 (n = 18) and controls (n = 14) with periodontitis was assessed using 16S rRNA gene sequencing. The samples were collected at baseline, 30, and 90 days after NSPD. The taxonomic analysis of gingival microbiota was performed using a ribosomal RNA database. The microbiota content was evaluated in the light of CD4 cell count and viral load. RESULTS: Both HIV and control groups showed similar stages and grades of periodontitis. At baseline, the HIV group showed higher alpha diversity for both healthy and periodontal sites. Streptococcus, Fusobacterium, Veillonella and Prevotella were the predominant bacterial genera. A low abundance of periodontopathogenic bacteria was observed, and the NSPD induced shifts in the subgingival biofilm of patients with HIV-1, leading to a microbiota similar to that of controls. CONCLUSIONS: Different subgingival microbiota profiles were identified-a less diverse microbiota was found in patients infected with HIV-1, in contrast to a more diverse microbiota in controls. NSPD caused changes in the microbiota of both groups, with a greater impact on the HIV group, leading to a decrease in alpha diversity, and produced a positive impact on the serological immune markers in patients infected with HIV-1. Control of periodontitis should be included as part of an oral primary care, providing the oral health benefits and better control of HIV-1 infection.
Asunto(s)
Placa Dental , Infecciones por VIH , VIH-1 , Periodontitis , Humanos , VIH-1/genética , ARN Ribosómico 16S/genética , Desbridamiento Periodontal , Placa Dental/microbiología , Periodontitis/microbiología , BacteriasAsunto(s)
Humanos , Masculino , Femenino , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Periodontitis/microbiología , Placa Dental/microbiología , Diabetes Mellitus Tipo 2/complicaciones , Control Glucémico/métodos , Argentina , Hemoglobina Glucada , Estudios Transversales , Interpretación Estadística de Datos , Bacterias Gramnegativas/aislamiento & purificaciónRESUMEN
Introducción: El SARS-CoV-2 afecta el sistema respiratorio en diferentes grados. La cavidad oral es el lugar más colonizado por bacterias, por lo tanto, al no tener una adecuada higiene pueden presentarse diferentes enfermedades secundarias, lo que ha causado alerta en el gremio odontológico, ya que puede contribuir a complicaciones posteriores en los pacientes. Material y métodos: El estudio fue conformado por 47 pacientes voluntarios recuperados de SARS-CoV-2, residentes de Montemorelos, Nuevo León, México, donde fueron atendidos en Bucalia Dent, consultorio dental. Después del consentimiento informado de cada paciente, se realizó una historia clínica para conocer los síntomas, enfermedades sistémicas, ausencia de dientes y nivel de inflamación gingival de acuerdo al índice de Loe y Silness. A continuación, se tomó una muestra de biofilm microbiano (placa dentobacteriana), la cual se suspendió en una solución buffer de fosfato, posteriormente fue llevada al Centro de Investigación y Desarrollo en Ciencias de la Salud (CIDICS), Monterrey, N.L, México. Se extrajo DNA y se purificó, después se realizó PCR para detectar los patógenos orales; la PCR se visualizó en gel de agarosa (1.5%) por tinción de bromuro de etidio. Resultados: Se detectó 80.85% Porphyromona gingivalis y 68.09% Fusobacterium nucleatum en pacientes recuperados de SARS-CoV-2; 23.4% presentaron inflamación leve de acuerdo al índice de Loe y Silness, 54.5% fueron masculinos y 45.5% femeninos. Por otro lado, 36.4% de los pacientes con inflamación leve tenían de cuatro a seis dientes ausentes. En estos pacientes se detectó 18.18% únicamente con Fusobacterium nucleatum y 27.27% sólo con Porphyromona gingivalis; el sexo masculino tuvo predisposición en 66.6% y el femenino en 33.33%. Se observó infección con los dos patógenos presentes en 45.45%; y 60% de estos pacientes fueron masculinos. Conclusiones: Los pacientes recuperados de SARSCoV- 2 analizados en esta investigación mostraron mala higiene oral y alta prevalencia de los patógenos mencionados altamente relacionados a inflamación gingival o enfermedad periodontal, lo que nos indica que es indispensable la intervención del odontólogo al finalizar el periodo de infección de cada paciente (AU)
Introduction: SARS-CoV-2 affects the respiratory system to different degrees. The oral cavity is a colonized place by bacterias, therefore, by not having good hygiene, different secondary diseases can occur; this has caused an alert in the dental industry, since it can contribute to later complications in patients. Material and methods: The study was conducted in 47 SARS-CoV-2 recovered volunteers from the Montemorelos city of the Nuevo León state, Mexico, who were attended at the Bucalia Dent dental clinic. An informed consent was obtained from each of the patients, then their clinical history was documented in order to know the symptoms, previous systemic diseases, absence of teeth and degree of gingival inflammation, as suggested by Loe and Silness. Subsequently, a dental plaque sample was taken from all patients, which was suspended in a phosphate buffered solution and shipped to The Center for Research and Development in Health Sciences (CIDICS), Monterrey, NL, Mexico for storage. DNA extraction and purification was performed and PCR was carried out for the oral pathogens detection. All PCR products were visualized on 1.5% agarose gel by ethidium bromide staining. Results: Porphyromona gingivalis and Fusobacterium nucleatum were detected in 80.85% and 68.09% of SARS-CoV-2 recovered patients, respectively. 23.4% showed mild inflammation based on the Loe and Silness criteria, 54.5% were male and 45.5% female. On the other hand, 36.4% of patients with mild inflammation had between 4 to 6 missing teeth. A single infection by Fusobacterium nucleatum was detected in 18.18% and by Porphyromona gingivalis in 27.27%; the male sex had a predisposition with 66.66% and 33.33% female; coinfection of both pathogens was observed in 45.45% where 60% were male. Conclusions: SARS-CoV-2 recovered patients show poor oral hygiene and a high prevalence of oral pathogens related to the development of inflammatory gingival or periodontal disease, this suggests the need for an odontological clinical intervention at the end of the course of infection or disease caused by SARS-CoV-2 (AU)
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Higiene Bucal , Fusobacterium nucleatum , Porphyromonas gingivalis , SARS-CoV-2 , ADN , Índice de Higiene Oral , Índice Periodontal , Reacción en Cadena de la Polimerasa , Placa Dental/microbiología , Electroforesis en Gel de Agar , Distribución por Edad y Sexo , Gingivitis/epidemiología , MéxicoRESUMEN
Las enfermedades del periodonto tienen una etiopatogenia compleja y puede considerarse multifactorial. El factor etiológico esencial en la patología inflamatoria periodontal es la biopelícula dental y cuando el desequilibrio entre el huésped y los microorganismos cambia la complejidad de la flora. Ciertas bacterias como Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens y Treponema spp., han sido comúnmente relacionadas con la periodontitis crónica y son consideradas como indicadores de riesgo para la progresión de dicha enfermedad. El objetivo de este trabajo fue establecer la prevalencia de Prevotella spp y Porphyromona spp en los distintos estadios de periodontitis crónicas. Material y métodos: Se estudiaron 48 pacientes sistémicamente saludables con diagnóstico de periodontitis crónica. Se completó el consentimiento informado, se realizó historia clínica y examen periodontal. El estado periodontal se clasificó en distintos grados de severidad: leve, moderada y severa. Se tomaron muestras de dos sitios con mayor profundidad de sondaje con conos de papel absorbente estériles y se transportaron en un medio prerreducido. Para el aislamiento de Prevotella spp se utilizó agar Brucella más sangre ovina al 5%, hemina, vitamina K al que se agregaron vancomicina y kanamicina; Porphyromonas sp se aisló en el mismo medio con el agregado de bacitracina y colistina. Se sembraron 10 µl de muestra entera y las placas fueron incubadas en jarras de anaerobiosis por 5 a 7 días a 37ºC. Resultados: los distintos grados de periodontitis correspondieron a un 17% periodontits leve, 57% moderada y 26% severa. En el total de pacientes se determinó la presencia de Prevotella spp en el 54% de los casos y un 12,5% de Porphyromona spp. Conclusión: De los pacientes estudiados con periodontits crónica, un 52% correspondió al sexo masculino, un 57% de los casos correspondieron a periodontitis moderada. Se aisló Prevotella sp en todos los estadios de periodontitis crónica y Porphyromonas sp sólo en periodontitis severas (AU)
Periodontal diseases have a complex etiopathogenesis and can be considered multifactorial. The essential etiological factor in periodontal inflammatory pathology is the dental biofilm and when the imbalance between the host and the microorganisms changes the complexity of the flora. Certain bacteria such as Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens and Treponema spp., Have been commonly related to chronic periodontitis and are considered as risk indicators for the progression of said disease. The objective of this work was to establish the prevalence of Prevotella spp and Porphyromonas spp in the different stages of chronic periodontitis. Forty eight systemically healthy patients diagnosed with chronic periodontitis were studied. Informed consent was completed, a medical history and periodontal examination was carried out. The periodontal state was classified into different degrees of severity: mild, moderate and severe. Samples were taken from two sites with greater depth of probing with sterile absorbent paper cones and transported in a prereduced medium. For the isolation of Prevotella spp, Brucella agar plus 5% sheep blood, hemin, vitamin K to which vancomycin and kanamycin were added. For Porphyromonas spp, the same medium was used and bacitracin and colistin were added. 10 µl of the whole sample was seeded and the plates were incubated in anaerobic jars for 5 to 7 days at 37 ° C. Different degrees of periodontitis corresponded to 17% mild periodontitis, 57% moderate and 26% severe. In the total number of patients, the presence of Prevotella spp was determined in 54% of the cases and 12.5% of Porphyromona spp. Of the patients studied with chronic periodontitis, 52% corresponded to the male sex, 57% of the cases corresponded to moderate periodontitis. Prevotella spp was isolated in all stages of chronic periodontitis and Porphyromonas sp only in severe periodontitis (AU)
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Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Porphyromonas gingivalis/aislamiento & purificación , Prevotella intermedia/aislamiento & purificación , Periodontitis Crónica/microbiología , Recuento de Colonia Microbiana , Factores de Riesgo , Medios de Cultivo , Placa Dental/microbiología , Distribución por Edad y SexoRESUMEN
The aim of this study was to evaluate changes in periodontal status and maxillary buccal bone by considering clinical and tomographic parameters during the first year of orthodontic expansion with Invisalign® aligners. Upper first (1PM) and upper second (2PM) premolars of 19 patients with orthodontic expansion requirement treated with Invisalign® aligners were evaluated. Plaque index (PI), gingival index (GI), probing pocket depth (PPD), clinical attachment level (CAL) and cone beam tomographic (CBCT) records were collected at 76 sites before starting treatment (T0) and at 12 months (T1). Bone height was measured from cementoenamel junction (CEJ) to the crest cortical bone (CC). Bone thickness was measured at two levels: 4 mm (CEJ+4) and 6 mm (CEJ+6) apical to the CEJ. A descriptive analysis was made of the variations of bone thickness and height in a series of cases. The average expansion was 1.93 mm for 1PM and 167 mm for 2PM. Arithmetic mean of distance CEJ-CC in 1PM was 3.05 mm at T0, and remained at 3.05 mm at T1. Arithmetic mean of distance CEJ-CC in 2PM was 2.06 mm at T0 and 2.31 at T1. Post-expansion, most of the analyzed sites (86%) exhibited a bone thickness of ≥0.5 mm. The greatest variations between T0 and T1 were observed at the level of 1PM CEJ+ 4 and 2PM CEJ+ 6. The minimal changes in the clinical records (GI, PI, PPD and CAL) between T0 and T1 were compatible with the maintenance of gingivalperiodontal health. Invisalign® for expansion movements did not produce substantial changes in the evaluated periodontal clinical parameters or in the bone measurements. Removable appliances reduce plaque retentive factors and favor adequate oral hygiene.
El objetivo de este estudio fue evaluar los cambios en el estado periodontal y hueso facial maxilar a través de parámetros clínicos y tomográficos durante la expansión ortodóncica con alineadores Invisalign® en el primer año de tratamiento. Se evaluaron los primeros (1PM) y segundos (2PM) premolares superiores pertenecientes a 19 pacientes con requerimiento de expansión ortodóncica tratados con alineadores Invisalign®. Se registraron los índices de placa (IP), índice gingival (IG), profundidad al sondaje (PS) y nivel de inserción (NI) y registros tomográficos de haz cónico (CBCT) en 76 sitios antes de comenzar el tratamiento (T0) y a los 12 meses (T1). Se midió la altura ósea desde el límite amelocementario (LAC) hasta la cortical de la cresta (CC) y el espesor en dos niveles; a 4 mm (LAC+4) y a 6 mm (LAC+6) hacia apical del LAC. Se realizó un análisis descriptivo de las variaciones de la altura y espesor óseo en una serie de casos. La expansión promedio para 1PM fue de 1,93 mm y para 2PM fue de 1,67 mm. La media aritmética de LAC-CC en primeros premolares fue de 3,05 mm en T0 y se mantuvo el valor de 3,05 mm en T1. La media aritmética de LAC-CC en segundos premolares fue de 2,06 mm en T0 y 2,31 en T1. Post expansión, la mayoría de los sitios (86%) analizados exhibieron un espesor óseo ≥0,5 mm. Las mayores variaciones entre T0 y T1 se observaron a nivel de 1PM CEJ+4 y 2PM CEJ+6. Los registros clínicos (PI, GI, PPD y CAL) evidenciaron mínimos cambios entre T0 y T1, compatibles con el mantenimiento de la salud gíngivo-periodontal. El uso de Invisalign ® para movimientos de expansión no produjo cambios sustanciales en los parámetros clínicos periodontales evaluados ni en las mediciones óseas. La aparatología removible reduce los factores retentivos de placa bacteriana y facilita una adecuada higiene oral.
Asunto(s)
Placa Dental/etiología , Maloclusión/terapia , Maxilar/diagnóstico por imagen , Salud Bucal , Aparatos Ortodóncicos Removibles/efectos adversos , Técnicas de Movimiento Dental/efectos adversos , Tomografía Computarizada de Haz Cónico , Placa Dental/microbiología , Índice de Placa Dental , Estado de Salud , Humanos , Técnicas de Movimiento Dental/instrumentaciónRESUMEN
OBJECTIVES: This study aims to describe the oral microbiome diversity and prevalence of ARGs in periodontal health and disease. BACKGROUND: The human oral cavity harbors a complex microbial community known as the oral microbiome. These organisms are regularly exposed to selective pressures, such as the usage of antibiotics, which drive evolution and acquisition of antibiotic resistance genes (ARGs). Resistance among oral bacteria jeopardizes not only antibiotic therapy for oral infections, but also extra-oral infections caused by bacterial translocation. METHODS: We carried out a cross-sectional investigation. Saliva and subgingival plaque samples were collected during a clinical exam. 16S rRNA gene sequencing was performed to assess microbial diversity. Resistance genes were identified through PCR assays. RESULTS: Of the 110 participants, only 22.7% had healthy periodontium, while the majority was diagnosed with gingivitis (55.4%) and chronic periodontitis (21.8%). The composition of the oral microbiota differed from healthy and diseased samples, being Streptococcus spp. and Rothia spp. predominant in periodontal disease. Regarding ARGs, 80 (72.7%) samples were positive for at least one of genes screened, erm being the most frequent variant (58.2%), followed by blaTEM (16.4%), mecA (2.7%), pbp2b and aac(6 ') (1.8%). Neither genes coding resistance to carbapenems nor metronidazole were detected. CONCLUSIONS: Our findings indicate that there are no significant differences in terms of taxonomic enrichment between healthy and diseased oral microbiomes. However, samples retrieved from healthy patients had a more diverse microbial community, whereas diseased samples have lower taxonomic diversity. We have also identified clinically relevant ARGs, providing baseline information to guide antibiotic prescription in dentistry.
Asunto(s)
Bacterias/aislamiento & purificación , Farmacorresistencia Bacteriana/genética , Microbiota , Boca/microbiología , Adolescente , Adulto , Bacterias/genética , Proteínas Bacterianas/genética , Estudios Transversales , Placa Dental/microbiología , Placa Dental/patología , Femenino , Gingivitis/diagnóstico , Gingivitis/microbiología , Humanos , Masculino , Persona de Mediana Edad , Periodontitis/diagnóstico , Periodontitis/microbiología , Periodoncio/patología , ARN Ribosómico 16S/química , ARN Ribosómico 16S/metabolismo , Saliva/microbiología , Streptococcus/genética , Streptococcus/aislamiento & purificación , Adulto JovenRESUMEN
ABSTRACT The aim of this study was to evaluate changes in periodontal status and maxillary buccal bone by considering clinical and tomographic parameters during the first year of orthodontic expansion with Invisalign® aligners. Upper first (1PM) and upper second (2PM) premolars of 19 patients with orthodontic expansion requirement treated with Invisalign® aligners were evaluated. Plaque index (PI), gingival index (GI), probing pocket depth (PPD), clinical attachment level (CAL) and cone beam tomographic (CBCT) records were collected at 76 sites before starting treatment (T0) and at 12 months (T1). Bone height was measured from cementoenamel junction (CEJ) to the crest cortical bone (CC). Bone thickness was measured at two levels: 4 mm (CEJ+4) and 6 mm (CEJ+6) apical to the CEJ. A descriptive analysis was made of the variations of bone thickness and height in a series of cases. The average expansion was 1.93 mm for 1PM and 167 mm for 2PM. Arithmetic mean of distance CEJ-CC in 1PM was 3.05 mm at T0, and remained at 3.05 mm at T1. Arithmetic mean of distance CEJ-CC in 2PM was 2.06 mm at T0 and 2.31 at T1. Post-expansion, most of the analyzed sites (86%) exhibited a bone thickness of ≥0.5 mm. The greatest variations between T0 and T1 were observed at the level of 1PM CEJ+ 4 and 2PM CEJ+ 6. The minimal changes in the clinical records (GI, PI, PPD and CAL) between T0 and T1 were compatible with the maintenance of gingivalperiodontal health. Invisalign® for expansion movements did not produce substantial changes in the evaluated periodontal clinical parameters or in the bone measurements. Removable appliances reduce plaque retentive factors and favor adequate oral hygiene.
RESUMEN El objetivo de este estudio fue evaluar los cambios en el estado periodontal y hueso facial maxilar a través de parámetros clínicos y tomográficos durante la expansión ortodóncica con alineadores Invisalign® en el primer año de tratamiento. Se evaluaron los primeros (1PM) y segundos (2PM) premolares superiores pertenecientes a 19 pacientes con requerimiento de expansión ortodóncica tratados con alineadores Invisalign®. Se registraron los índices de placa (IP), índice gingival (IG), profundidad al sondaje (PS) y nivel de inserción (NI) y registros tomográficos de haz cónico (CBCT) en 76 sitios antes de comenzar el tratamiento (T0) y a los 12 meses (T1). Se midió la altura ósea desde el límite amelocementario (LAC) hasta la cortical de la cresta (CC) y el espesor en dos niveles; a 4 mm (LAC+4) y a 6 mm (LAC+6) hacia apical del LAC. Se realizó un análisis descriptivo de las variaciones de la altura y espesor óseo en una serie de casos. La expansión promedio para 1PM fue de 1,93 mm y para 2PM fue de 1,67 mm. La media aritmética de LAC-CC en primeros premolares fue de 3,05 mm en T0 y se mantuvo el valor de 3,05 mm en T1. La media aritmética de LAC-CC en segundos premolares fue de 2,06 mm en T0 y 2,31 en T1. Post expansión, la mayoría de los sitios (86%) analizados exhibieron un espesor óseo ≥0,5 mm. Las mayores variaciones entre T0 y T1 se observaron a nivel de 1PM CEJ+4 y 2PM CEJ+6. Los registros clínicos (PI, GI, PPD y CAL) evidenciaron mínimos cambios entre T0 y T1, compatibles con el mantenimiento de la salud gíngivo-periodontal. El uso de Invisalign ® para movimientos de expansión no produjo cambios sustanciales en los parámetros clínicos periodontales evaluados ni en las mediciones óseas. La aparatología removible reduce los factores retentivos de placa bacteriana y facilita una adecuada higiene oral.
Asunto(s)
Humanos , Aparatos Ortodóncicos Removibles/efectos adversos , Técnicas de Movimiento Dental/efectos adversos , Salud Bucal , Placa Dental/etiología , Maloclusión/terapia , Maxilar/diagnóstico por imagen , Técnicas de Movimiento Dental/instrumentación , Índice de Placa Dental , Estado de Salud , Placa Dental/microbiología , Tomografía Computarizada de Haz CónicoRESUMEN
Objetivo: El objetivo de este estudio fue determinar la contaminación bacteriana de los conos de gutapercha de tipo beta (ß) en los tiempos 0, 24, 47 y 72 horas de las diferentes proveedurías de la Clínica Odontológica de la Universidad Científica del Sur (Lima, 2020). Materiales y métodos: Se obtuvo 16 conos de gutapercha tipo beta (ß) de empaques cerrados bajo medidas asépticas, los cuales fueron colocados en viales con 2 ml de caldo BHI y, posteriormente, fueron sembrados en agar BHI, así como en medios selectivos agar manitol salado y agar MacConkey. Pasadas las 24 horas de incubación a 37 °C, se realizó la lectura de las placas y el conteo de UFC. El mismo procedimiento se realizó para los tiempos 24, 48 y 72 horas, lo que dio un total de 64 conos de gutapercha tipo ß. Resultados: Se observó que el nivel de contaminación bacteriana fue el mismo tanto entre las distintas proveedurías como a las 0, 24, 48 y 72 horas. Solo se hallaron diferencias estadísticamente significativas (p = 0,044) entre los distintos tiempos de la proveeduría número 5. Finalmente, todas las muestras sometidas a la prueba de la coagulasa arrojaron resultados negativos. Conclusión: Los conos de gutapercha de tipo beta (ß) se contaminaron por igual producto de su almacenamiento y manipulación, independientemente de la proveeduría en la que permanecieron. (AU)
Objective: The purpose of this study was to determinate the bacterial contamination of Beta (ß) gutta-percha cones at 0, 24, 47 and 72 hours of the different supplies of the Universidad Científica del Sur, Lima 2020. Materials and Methods: 16 ß-type gutta-percha cones were obtained of sealed packages under aseptic measurements, they were placed in vials with 2ml BHI and subsequently planted in BHI agar plates as well as in selective medias as Salted mannitol agar and MacConkey agar after 24 hours of incubation at 37 ° the plates were read and count in CFU, the same procedure was performed for the other times evaluated 24, 48 and 72 hours, giving a total of 64 ß-type gutta-percha cones. Results: It was observed that the level of bacterial contamination was the same among the different supplies in all the establish times of in this study 0, 24, 48 and 72 hours. Therefore, there were no significant differences in the level of bacterial contamination between the supplies. On the other hand, only statistically significant differences (p = 0.044) were found between the different times of the supply number 5. Finally, all the samples submitted to the coagulase test had a negative result. Conclusion: The gutta-percha cones of type ß were contaminated equally regardless of the supply in which they were stored or manipulated. (AU)
Asunto(s)
Humanos , Infecciones por Bacterias Grampositivas , Infecciones por Bacterias Gramnegativas , Coagulasa , Placa Dental/microbiología , GutaperchaRESUMEN
The antimicrobial photodynamic therapy (aPDT) has stood out as an alternative and promising method of disinfection and has been exploited for the treatment of oral bacteria. In this study, we evaluate in vitro the action of aPDT, mediated by methylene blue, chlorin-e6, and curcumin against clinical subgingival plaques that were resistant to metronidazole. The sensitivity profile of the samples to metronidazole was analyzed by the agar dilution method. Cell viability in the planktonic and biofilm phase was assessed by CFU / mL. The composition of the biofilm was evaluated by the checkboard DNA-DNA Hibrydization technique. Photosensitizers internalization was qualitatively assessed by confocal fluorescence microscopy (CLSM). The aPDT mediated by the three photosensitizers tested was able to reduce the totality of the planktonic microbial load and partially reduce the biofilm samples. The analysis performed by CLSM showed that the photosensitizers used in the application of aPDT were able to permeate the interior of the biofilm. The aPDT has been shown to be useful in a supportive and effective approach to the treatment of periodontal disease.
Asunto(s)
Antibacterianos/farmacología , Placa Dental/microbiología , Farmacorresistencia Bacteriana/efectos de los fármacos , Metronidazol/farmacología , Fotoquimioterapia/métodos , Biopelículas/efectos de los fármacos , Clorofilidas , Curcumina/farmacología , Humanos , Azul de Metileno/farmacología , Pruebas de Sensibilidad Microbiana , Enfermedades Periodontales/tratamiento farmacológico , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Porfirinas/farmacologíaRESUMEN
Objetivos: A cavidade oral pode atuar como reservatório de vários patógenos de importância clínica, incluindo bacilos Gram-negativos (BGN) e Enterococcus spp. Essas espécies podem ainda aumentar em uma condição disbiótica, como ocorre nas doenças periodontais. Assim, este estudo teve como objetivo determinar a prevalência, susceptibilidade antimicrobiana e presença de fatores de virulência de BGN e enterococos isolados do biofilme subgengival de indivíduos com diferentes condições periodontais, correlacionando esses achados com parâmetros clínicos e composição da microbiota subgengival. Métodos: Na análise dos BGN, amostras de biofilme subgengival foram obtidas de indivíduos com Saúde Periodontal (SP, n=81), Gengivite (G, n=74) e Periodontite (P, n=207); para Enterococcus spp., amostras foram coletadas de 139 indivíduos com SP, 103 com G e 305 com P. As amostras foram cultivadas em meio seletivo e as colônias isoladas e identificadas por MALDI-ToF. A susceptibilidade antimicrobiana foi determinada por disco-difusão (CLSI); já os genes de virulência de enterococos e genes codificadores de ESBL e carbapenemases em BGN foram pesquisados por PCR. A produção de ESBL e carbapenemases por BGN foi avaliada pelo teste de sinergia de disco duplo e hidrólise de imipenem por espectrofotometria, respectivamente. A microbiota subgengival desses indivíduos foi determinada pelo Checkerboard. Diferenças entre os grupos foram avaliadas pelos testes de KruskalWallis, Mann-Whitney e Qui-quadrado. Resultados: BGN foram isolados em 36.2% das amostras, com maior prevalência (p<0,001) em pacientes com P (46.4%) em comparação com SP (22.2%) e G (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%) e Enterobacter asburiae (10.7%) foram as espécies mais predominantes. Resistência/sensibilidade reduzida a ≥ 1 antimicrobiano foi encontrada em 60% dos BGN, mas apenas 4.6% eram multirresistentes. Altas taxas de resistência (>40%) foram observadas na família Enterobacteriacea para cefoxitina, cefalotina, amoxicilina- clavulanato e cefazolina. Uma única cepa de K. pneumoniae apresentou resistência/sensibilidade reduzida ao imipenem, embora o fenótipo ESBL e a detecção dos genes codificadores de beta-lactamases foram negativos. Enterococcus spp. foram isolados em 7.4% de todas as amostras, 53.7% eram E. faecalis. Essas espécies foram mais predominantes na P (9.8%) e G (7.8%) do que na SP (2.2%, p< 0,05); entretanto não houve correlação com os níveis de gravidade da P. Altas taxas de resistência/susceptibilidade reduzida foram observadas para ciprofloxacina, eritromicina e rifampicina. Os fatores de virulência mais predominantes incluíram ace, asa e esp, todos relacionados à formação de biofilme e colonização. F. nucleatum foi mais prevalente na microbiota de indivíduos enterococos +. Por outro lado, Dialister pneumosintes foi pouco detectado em indivíduos portadores de enterococos bopD+. Estreptococos orais foram prevalentes (>70%) na microbiota de pacientes que apresentavam enterococos suceptíveis à doxiciclina (p<0,05), frequentemente bopD- e esp- (p<0,01). Conclusão: Uma prevalência elevada de BGN da família Enterobacteriacea com resistência a cefalosporinas e penicilina é observada na microbiota subgengival de indivíduos com P. Enterococcus spp., principalmente E. faecalis são pouco frequentes na microbiota subgengival associada à SP, porém aumentam significativamente nas doenças periodontais. Os mesmos apresentam diversos genes de virulência compatíveis com destruição tecidual, bem como resistência a antimicrobianos de uso na clínica periodontal, o que pode limitar uma resposta terapêutica favorável. (AU)
Background/Aim: The oral cavity can act as a reservoir for several pathogens of clinic importance, including Gram-negative bacilli (GNB) and Enterococcus spp. These species may increase even more in a dysbiotic condition as seen in periodontal diseases. Thus, this study aimed to determine the prevalence, antimicrobial susceptibility and virulence factors of GNB and enterococci isolated from subgingival biofilm of individuals with different periodontal conditions, correlating these findings with clinical parameters and the composition of the subgingival microbiot. Methods: For GNB analysis, subgingival biofilm was obtained from individuals with periodontal health (PH, n=81), gingivitis (G, n=74) and periodontitis (P, n=207), whereas for enterococci isolation samples were taken from 139 patients with PH, 103 with G, and 305 with P. Samples were cultivated in selective media and isolated colonies were identified by MALDI-ToF. Antimicrobial susceptibility was determined by CLSI disk diffusion, whereas virulence genes by PCR. Production of ESBL and carbapenemases were evaluated by double disk synergy test and spectrophotometric detection of imipenem hydrolysis, respectively, and ESBL and carbapenemase encoding genes were surveyed by PCR. The subgingival microbiota was determined by checkerboard. Differences among groups were examined by Chi-square, Kruskal-Wallis or Mann-Whitney tests. Results: GNB were isolated from 36.2% of all samples, with a significantly greater prevalence (p<0.001) in P patients (46.4%) compared to PH (22.2%) and G (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%) and Enterobacter asburiae (10.7%) were the most predominant species. Resistance/reduced sensitivity to ≥ 1 antimicrobial was found in 60% of GNB, but only 4.6% were multidrug resistant. High resistance rates (>40%) were seen in the Enterobacteriaceae family for cefoxitin, cephalotin, amoxicillin-clavulanate, and cefazolin. One strain of K. pneumoniae showed resistance/reduced sensitivity to imipenem, although the ESBL-phenotype and PCR targeting beta-lactamase encoding genes were negative. Enterococcus spp. were isolated from 7.4% of all samples; 53.7% were E. faecalis. Enterococci were more predominant in P (9.8%) and G (7.8%) samples than PH (2.2%; p<0.05), however there were no associations with distinct levels of disease severity. High rates of low susceptibility/resistance were seen for ciprofloxacin, erythromycin and rifampicin. Predominant virulence factors included ace, asa and esp, all related to colonization and biofilm formation. F. nucleatum was prevalent in the microbiota of enterococci+ individuals. In contrast, lower frequency of Dialister pneumosintes was found in patients carrying bopD+ enterococci. Oral streptococci were prevalent (>70%) in the microbiota of patients carrying enterococci susceptible to doxycycline (p<0.05), which were also frequently bopD- and esp- (p<0.01). Conclusion: A high prevalence of GNB of the Enterobacteriacea family, resistant to cephalosporins and penicillins is observed in the subgingival microbiota of patients with P, Enterococcus spp., mainly E.faecalis are not commonly detected in the healthy-related subgingival microbiota, however their frequency increases significantly in patients with periodontal diseases. These species carry several genes related to tissue destruction, as well as resistance to antimicrobials routinely used in the periodontal clinic, which may hinder a successful therapeutic response. (AU)
Asunto(s)
Humanos , Enfermedades Periodontales/microbiología , Infecciones por Bacterias Gramnegativas/epidemiología , Enterococcus/aislamiento & purificación , Farmacorresistencia Bacteriana , Placa Dental/microbiología , Microbiota , Prevalencia , Infecciones por Bacterias Gramnegativas/microbiología , Factores de VirulenciaRESUMEN
O presente estudo teve como objetivo avaliar, in vitro, fatores de virulência e susceptibilidade ao fluconazol, associado ou não a Clorexidina, de Candida spp. isoladas de diferentes sítios da cavidade bucal de crianças hospitalizadas em Unidade de Terapia Intensiva (UTI) comparando com isolados de crianças não hospitalizadas. Para isso, foram utilizados isolados de Candida spp., previamente identificados e estocados, oriundos de 30 pacientes de UTI (G1) e 30 pacientes saudáveis (G2), com idade entre 01 e 13 anos, pareados por sexo e idade. A coleta dos espécimes clínicos foi realizada através de esfregaço de mucosa (swab) e raspagem de biofilme dental supragengival, e a identificação das espécies de Candida se deu por espectrometria de massa (MALDI-TOF). Todos os isolados de Candida (n=60, sendo 46 G1 e 14 G2 (C.albicans n=15, C. tropicalis n=5, C. guilliermondii n=32, C. parapsilosis n=5, C. glabrata n=2, C. kefyr n=1) foram então selecionados para a avaliação de fatores de virulência como, formação de biofilme, produção de fosfolipase e protease, susceptibilidade ao fluconazol (FLZ) e clorexidina (CHX); e, verificação de sinergismo, checkerboard, entre as duas drogas. Os resultados foram analisados através do SPSS versão 20 e comparados por meio dos testes Qui-quadrado, Fisher e Razão de chance (95% IC), com nível de significância de 95% (p≤0,05). Tanto os isolados de G1 (97,8%) quanto os de G2 (100%) produziram biofilme (χ2, p=0,76). Quanto a produção de fosfolipase, somente 3 (6,5%) isolados do G1 (todos de C. albicans) foram produtores, sendo um isolado de mucosa (16,7%) e dois em biofilme (40%); No G2, isso aconteceu de modo similar (1 isolado produtor) (χ2, p=0,66). Já em relação a produção de protease, 42 isolados (91,3%), dos quais 100% dos isolados de C. guilliermondii, seguida de C. albicans (81,8%), produziram esta enzima em G1, sendo esta frequência significativamente maior do que a encontrada no G2 com 9 isolados (64,3%) (χ2, p=0,025). Não houve diferença significativa quando comparamos os fatores de virulência entre os isolados oriundos de diferentes sítios, nos dois grupos. Cinquenta e um (85%) isolados foram resistentes ao FLZ, sendo 38 (82,6%) no G1 e 13 (92,9%) no G2 (χ2, p=0,47), sendo a frequência de isolados resistentes similar entre os sítios (mucosa x biofilme dental) (χ2, p=0,40). Foi observado que os poucos isolados resistentes a Clorexidina (n=2; 1 C. tropicalis do G1 e 1 C. albicans do G2), foram sensíveis a combinação FLZ e CHX, mostrando um sinergismo entre as duas drogas. Conclui-se que os pacientes internados em UTI apresentam maior freqüência de isolados de Candida spp. produtores de protease e que isolados de biofilme dental são tão virulentos quanto os de mucosa nesses pacientes. Ainda, a combinação de fluconazol e clorexidina foi eficiente no controle do crescimento de Candida spp. (AU)
The present study aimed to evaluate, in vitro, virulence and susceptibility factors to fluconazole, associated or not with Chlorhexidine, from Candida spp. isolated from different sites in the oral cavity of children hospitalized in an Intensive Care Unit (ICU) compared with isolates from children not hospitalized. For this, Candida spp. Isolates, previously identified and stored, were used, coming from 30 ICU patients (G1) and 30 healthy patients (G2), aged between 01 and 13 years, matched by sex and age. The collection of clinical specimens was carried out through mucosa smear (swab) and scraping of supragingival dental biofilm, and Candida species were identified by mass spectrometry (MALDI-TOF). All Candida isolates (n = 60, 46 G1 and 14 G2 (C.albicans n = 15, C. tropicalis n = 5, C. guilliermondii n = 32, C. parapsilosis n = 5, C. glabrata n = 2, C. kefyr n = 1) were then selected for the evaluation of virulence factors such as biofilm formation, phospholipase and protease production, susceptibility to fluconazole (FLZ) and chlorhexidine (CHX); and, synergism check, checkerboard, between the two drugs. The results were analyzed using SPSS version 20 and compared using the Chi-square, Fisher and odds ratio tests (95% CI), with a 95% significance level (p≤0.05) Both G1 (97.8%) and G2 isolates (100%) produced biofilm (χ2, p = 0.76). Regarding the production of phospholipase, only 3 (6.5%) isolates from G1 (all of C. albicans) were producers, one isolated from the mucosa (16.7%) and two in biofilm (40%); In G2, this happened in a similar way (1 producer isolate) (χ2, p = 0.66). Regarding protease production, 42 isolates (91.3%), of which 100% of the isolates of C. guilliermondii, followed by C. albicans (81.8%), produced this enzyme in G1, this frequency being significantly greater than that found in G2 with 9 isolates (64.3%) (χ2, p = 0.025). There was no significant difference when comparing the virulence factors between the isolates from different sites, in the two groups. Fifty-one (85%) isolates were resistant to FLZ, 38 (82.6%) in G1 and 13 (92.9%) in G2 (χ2, p = 0.47), the frequency of resistant isolates being similar between sites (mucosa x dental biofilm) (χ2, p = 0.40). It was observed that the few isolates resistant to chlorhexidine (n = 2; 1 C. tropicalis from G1 and 1 C. albicans from G2), were sensitive to the combination of FLZ and CHX, showing a synergism between the two drugs. It is concluded that patients admitted to the ICU have a higher frequency of Candida spp. protease producers and that dental biofilm isolates are as virulent as mucosal ones in these patients. Also, the combination of fluconazole and chlorhexidine was effective in controlling the growth of Candida spp. (AU)
Asunto(s)
Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Candida/patogenicidad , Unidades de Cuidado Intensivo Pediátrico , Fluconazol/normas , Factores de Virulencia , Técnicas In Vitro , Estudios de Casos y Controles , Placa Dental/microbiologíaRESUMEN
ABSTRACT The aim of this study was to describe the microbiological profile of HIV patients under highly active antiretroviral treatment (HAART). This crosssectional study comprised 32 HIV patients with periodontal disease (PD) who had been under HAART for more than 6 months. Information about the patients' medical history was obtained from clinical records. Clinical dental examination was performed by a calibrated researcher using standard dental instruments to determine probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP). A total 4,765 periodontal sites were evaluated, 125 of which were also studied microbiologically. Subgingival biofilm samples were obtained using sterile paper points; one set was used for microbiological culture studies and the other for endpoint PCR. Statistical analysis was performed using KruskalWallis and posthoc DunnBonferroni contrast tests. All participants were on HAART at the time of the study, and 90.6% had a viral load below 50 copies/mm3. Prevalence of periodontally active sites was low in the study population. Microbiological studies: Black pigmented anaerobic bacteria and fusiform CFU counts were significantly higher in samples from sites with BOP and PD ≥4mm (p 0.020 and p 0.005, respectively). Molecular Assays: Detection of Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) and Treponema denticola (p 0.015) was significantly more frequent at sites with BOP and PD ≥4mm. Conclusions: The patients living with HIV/AIDS under HAART studied here had low prevalence of clinical periodontal disease signs. However, significant detection of P. gingivalis, T. denticola, and T. forsythia in periodontal active sites, and the involvement of these microorganisms as potential HIV reactivators, show the importance of creating awareness among dental health professionals of the need for close dental and periodontal monitoring in HIV patients.
RESUMEN El objetivo de este estudio fue describir el perfil microbiológico del biofilm subgingival de los pacientes con VIH bajo tratamiento antirretroviral de alta actividad (TARGA). El estudio comprendió a 32 pacientes VIH seropositivos con enfermedad periodontal (EP) que se encontraran en tratamiento con TARGA por más de 6 meses. Los antecedentes médicos de los pacientes se obtuvieron de las historias clínicas. El examen clínico instrumental (profun didad de sondaje (PS), nivel de inserción clínico (NIC) y sangrado al sondaje (SS)) fue realizado con instrumental odontológico estándar por un investigador calibrado. De este modo, se evaluaron un total de 4.765 sitios periodontales de los cuales 125 fueron estudiados microbiológicamente. Las muestras de biope lícula subgingival se obtuvieron empleando conos de papel estéril. Las muestras se emplearon en estudios microbiológicos y moleculares por PCR de punto final. El análisis estadístico se realizó según KruskalWallis y pruebas de contrastes posthoc de DunnBonferroni. El 90,6% de la población en estudio presentó carga viral inferior a 50 copias/mm3. La prevalencia de sitios periodontales activos fue baja (1%). Los recuentos de bacterias anaerobias estrictas pigmentadas de negro y fusiformes fueron significativamente más altos en muestras de sitios periodontales con SS positivo y PS ≥4 mm (p 0.020 y p 0.005). La detección molecular de Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) y Treponema denticola (p 0.015) fue significativamente mayor en los sitios con SS y PS ≥4mm. La prevalencia del 1% de enfermedad periodontal en el grupo de pacientes estudiados fue menor a la esperada, sin embargo; la detección significativa de P. gingivalis, T. denticola y T. forsythia en sitios periodontales activos y su potencial participación como agentes reactivadores del VIH, nos alerta de la importancia de crear conciencia en los profesionales de la salud (médicos y odontólogos) acerca de la necesidad de un monitoreo minucioso del estado periodontal de pacientes con características semejantes a las descriptas en la muestra poblacional estudiada.