Diferencia en la expresión del ligando de receptor activador para el factor nuclear K B, en tejido tumoral frente a la infección por diferentes patógenos periodontales / Difference in receptor activator of nuclear factor K B expression in tumoral tissue by different periodontal pathogens

La enfermedad periodontal es una de las principales causas de pérdida dentaria. Clínicamente, esta patología, mediada por la desregulación del sistema inmune producto de una disbiosis ocurrida en el surco gingival, inicia con la inflamación de la encía y evoluciona con el daño irreversible de los tejidos que rodean el diente. El hueso alveolar es uno de los tejidos afectados esta patología, esto debido a la activación de osteoclastos por la sobreexpresión de la proteína RANKL en el huésped. El propósito de este trabajo es determinar el nivel de sobreexpresión de RANKL, en un modelo de células tumorales U2OS, frente a la infección con Porphyromonas gingivalis y Prevotella intermedia. Para identificar el nivel de RANKL, se definieron cuatro grupos: Un grupo control, no tratado; Grupo PG, tratado con P. gingivalis; Grupo PI, tratado con P. Intermedia; y un grupo PG+PI, tratado con ambas bacterias. El nivel relativo de la proteína RANKL fue determinado en el sobrenadante y en los extractos celulares de manera independiente, mediante la técnica Western blot. En sobrenadantes, el grupo PG mostró mayores niveles de RANKL comparados con PI (p < 0,05). En extractos celulares los niveles fueron mayores en el grupo PG+PI (p < 0,05). El grupo PI mostró los niveles más bajos de RANKL. La infección polimicrobiana resulta en una mayor expresión de RANKL en células tumorales U2OS, mientras que frente a la infección P. gingivalis, se observó mayor cantidad de RANKL soluble.

SUMMARY: Periodontal disease is one of the main causes of tooth loss. Clinically, this pathology, mediated by the deregulation of the immune system due to a dysbiosis occurred in the gingival sulcus, begins with the inflammation of the gum and evolves with the irreversible damage of the tissues that surround the tooth. Alveolar bone is one of the most affected tissues by this disease, due to the activation of osteoclasts by the upregulation of RANKL in the host. The aim of this study is to determine the increase of RANKL, in a U2OS tumor cells model, inoculated with Porphyromonas gingivalis and Prevotella intermedia. To identify the level of RANKL, four groups were defined: A control group, not treated; PG group, treated with P.gingivalis; PI group, treated with P. intermedia; and a PG+PI group, treated with both bacteria. The relative level of RANKL was determined in the supernatant and cell extracts independently, using the Western blot technique. In supernatants, the PG group showed higher RANKL levels compared to PI (p < 0.05). In cell extracts the levels were higher in the PG+PI group (p < 0.05.). The PI group showed the lowest levels of RANKL.Polymicrobial infection results in a greater expression of of soluble RANKL was observed.

Prevalencia de Porphyromonas gingivalis en un grupo de pacientes con aparatología fija de ortodoncia / Prevalence of Porphyromonas gingivalis in a group of patients with fixed orthodontic appliances

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)

Antioxidant and Inhibitory Capacity of Tomato Leaf Ethanolic Extract against Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans / Capacidad Antioxidante e Inhibitoria del Extracto Etanólico de la Hoja de Tomate contra Streptococcus mutans, Porphyromonas gingivalis y Candida albicans

Background: Tomato is a source of bioactive compounds, antimicrobials, and antioxidants. Tomato leaf preparations have been empirically used for anti-inflammatory, analgesic, antibiotic, and antiseptic purposes. However, research on the potential activity of tomato leaf extracts against oral microorganisms and in managing oropharyngeal infections is scarce. Objective: To investigate tomato leaf ethanolic extract's antioxidant and growth inhibitory capacity against common oral pathogenic microorganisms, namely, Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans.Methods: Ethanolic extracts were made from 'Chonto' tomato (Lycopersicon esculentum) leaves. The antimicrobial activity was measured with the microdilution technique using vancomycin and fluconazole as positive controls. The antioxidant capacity was measured with the ORAC assay using Trolox as a positive control. Results: We found a high percentage of growth inhibition (≥100%) against S. mutans and P. gingivalis at a concentration of 500 mg/L. However, the extract was ineffective in inhibiting the growth of C. albicans. Finally, we observed that the extract exerted a high antioxidant capacity (126%) compared to the positive control. Conclusions: This study provides new insights into the potential antimicrobial effect of tomato leaf extracts on common oral pathogenic bacteria, which may ultimately result in the development of new herbal products that might help prevent and treat oral infections, such as dental caries and periodontal disease. Our findings also support previous studies on the high antioxidant capacity of tomato leaf extracts

Antecedentes: El tomate es una fuente de compuestos bioactivos, antimicrobianos y antioxidantes. Las hojas de tomate se han utilizado empíricamente con fines antiinflamatorios, analgésicos, antibióticos y antisépticos. Sin embargo, los estudios sobre la actividad de los extractos de hojas de tomate contra los microorganismos orales y en el manejo de las infecciones orofaríngeas son escasos. Objetivo: Investigar la capacidad antioxidante del extracto etanólico de la hoja de tomate y su actividad inhibitoria de crecimiento contra microorganismos patógenos orales comunes, a saber, Streptococcus mutans, Porphyromonas gingivalis y Candida albicans.Métodos: Se realizaron extractos etanólicos a partir de hojas de tomate 'Chonto' (Lycopersicon esculentum). La actividad antimicrobiana se midió con la técnica de microdilución utilizando vancomicina y fluconazol como controles positivos. La capacidad antioxidante se midió con el ensayo ORAC utilizando Trolox como control positivo. Resultados: Encontramos un alto porcentaje de inhibición del crecimiento (≥100%) contra a S. mutans y P. gingivalis a una concentración de 500 mg/L. Sin embargo, el extracto fue ineficaz en la inhibición el crecimiento de C. albicans. Finalmente, observamos que el extracto tuvo una alta capacidad antioxidante (126%) en comparación con el control positivo. Conclusiones: Este estudio proporciona nuevos conocimientos sobre el posible efecto antimicrobiano de los extractos de hojas de tomate en bacterias patógenas orales comunes, lo cual puede resultar en el desarrollo de nuevos productos naturales que podrían ayudar a prevenir y tratar infecciones orales, como la caries dental y la enfermedad periodontal. Nuestros hallazgos también respaldan los estudios previos sobre la alta capacidad antioxidante de los extractos de hojas de tomate

Estresse e periodontite: avaliação da inervação simpática no periodonto e da resposta adrenérgica após infecção por Porphyromonas gingivalis / Stress and periodontitis: evaluation of sympathetic innervation in the periodontium and the influence of adrenergic signaling on Porphyromonas gingivalis infection

O estresse agrava a doença periodontal por vários mecanismos, sendo a estimulação do sistema nervoso simpático (SNS) um deles. A literatura mostra que a estimulação de receptores ß-adrenérgicos (ß-AR) aumenta a angiogênese em ossos longos, e a expansão microvascular agrava a periodontite. Ainda, catecolaminas aumentam a virulência de periodontopatógenos e agem na resposta imune. Assim, o objetivo deste trabalho foi avaliar: (1) a inervação simpática no periodonto e a influência da ativação do SNS na vascularização periodontal em camundongos e (2) a influência do sistema adrenérgico nos fatores de virulência de Porphyromonas gingivalis (Pg) e na resposta imunológica a este patógeno in vivo (Galleria mellonella). Na primeira parte, camundongos receberam injeção intraperitoneal de solução salina (PBS) ou isoproterenol (ISO; agonista não seletivo ß adrenérgico) por 1 mês, para detecção in situ de tirosina hidroxilase, neuropeptídeo Y, transportador de norepinefrina (NET) e endomucina em mandíbulas. Expressão de mRNA de Vegf-a, Il-1ß, Il-6, Adrb2 e Rankl foi quantificada 2 h após administração de ISO/PBS em mandíbula e tíbias, que serviram como controle positivo. Diferentemente das tíbias, não houve alteração na expressão dos genes analisados em mandíbula. Por outro lado, NET foi mais expresso no osso alveolar do que na tíbia, sendo detectado nos osteoblastos, osteócitos e células do ligamento. Embora o padrão de inervação e a expressão de Adrb2 sejam semelhantes entre mandíbula e tíbia, o tratamento com ISO não influenciou no número e área de vasos positivos para endomucina. Na segunda parte, investigamos a influência adrenérgica na resposta imune de G. mellonella durante infecção por Pg utilizando norepinefrina (NE; agonista α e ß adrenérgico) e ISO. Pg também foi cultivada na presença de ISO (PgISO) ou NE para avaliação da ação direta dos compostos na bactéria. ISO sistêmico protegeu as larvas da infecção por Pg, aumentando o número de hemócitos e reduzindo a contagem de células de Pg na hemolinfa, exclusivamente pelo ß-AR. Diferentemente, NE aumentou mortalidade, diminuiu o número de hemócitos. Apenas PgISO aumentou a morte das larvas, apesar de ambos, NE e ISO, terem aumentado a expressão de fatores de virulência na bactéria in vitro. ISO circulante, concomitante com PgISO, reduziu parcialmente a mortalidade das larvas. A influência do estresse na doença periodontal envolve diversas vias que alteram os dois pilares da periodontite (microbiota e sistema imune). No entanto, a ação na resposta do hospedeiro parece ser superior, uma vez que a estimulação ß-AR em osso alveolar saudável não alterou a produção de citocinas pró-inflamatórias ou microvascularização e a modulação da resposta imune em G. mellonella por compostos adrenérgicos foi mais importante para o desfecho da infecção que sua ação direta sobre a bactéria.

Stress aggravates periodontitis, and one possible mechanism is the activation of sympathetic nervous system (SNS). The literature shows that stimulation of ß-adrenergic receptors (ß-AR) induces angiogenesis in long bones, and microvasculature amplification was linked to periodontitis severity. Moreover, catecholamines increase the virulence of some periodontopathogenic bacteria in vitro and influences the innate immunity. Thus, the aim of this study was (1) evaluate the presence and influence of the SNS in the stimulation of periodontal vasculature, and (2) the influence of the adrenergic system on Porphyromonas gingivalis (Pg) virulence and on the immunological response to this pathogen in vivo (Galleria mellonella larvae). For the first part, mice received isoproterenol (ISO, a non-selective ß-AR agonist) or saline (PBS) for 1 month, for in situ analysis of tyrosine hydroxylase, neuropeptide Y, norepinephrine transporter (NET) and endomucin in the mandibles. Vegfa, Il-1ß, Il-6, Adrb2 and Rankl mRNA expression was assessed 2 hours after PBS/ISO treatment for mandibles and tibia, that served as positive control. We observed that, differently from the tibia, the expression of these genes did not alter on the mandible. However, NET expression was detected in osteoblasts, osteocytes, and periodontal ligament fibroblasts, and were higher expressed when compared to the tibias from the same animals. Although the pattern of sympathetic innervation and Adrb2 expression were similar between tissues, ISO treatment did not increase the area or number of endomucin+ vessels. For the second part, we addressed the adrenergic signaling influence on G. mellonella immune system during Pg infection using norepinephrine (NE, α- and ß-AR agonist), ISO and octopamine (insect's endogenous hormone). Pg was also cultivated in the presence of ISO (PgISO) or NE to investigate the direct action of the ligands on bacterial virulence. Systemic administration of ISO protected the larvae from Pg infection by increasing hemocyte density accompanied by reduction of Pg load in hemolymph, in a ß-AR manner. In contrast, NE increased mortality, with decreased hemocyte count and no influence on the other parameters. Only PgISO increased larvae death, despite of ISO and NE increased virulence in vitro. The concomitant injection of systemic ISO partially reversed the toxicity of the PgISO. The influence of stress on periodontitis involves different pathways, that alter the two pillars of disease's pathogenesis (microbiota and immune system). However, the influence on the host's inflammatory response seems to overcome the other players, since ß-AR activation on healthy alveolar bone didn't alter cytokines production or microvasculature. Besides, the modulation of innate immunity by adrenergic signaling in G. mellonella was more important for the disease's outcome than it's direct action on the bacteria.

Computer simulation of molecular docking between methylene blue and some proteins of Porphyromonas gingivalis / 北京大学学报(医学版)

OBJECTIVE@#To study the binding target of photosensitizer and bacteria in antimicrobial photodynamic therapy with computer-simulated target prediction and molecular docking research methods and to calculate the binding energy.@*METHODS@#The protein names of Porphyromonas gingivalis (Pg) were obtained and summarized in Uniprot database and RCSB PDB database; the structure diagrams of methy-lene blue were screened in SciFinder database, PubChem database, ChemSpider database, and Chemical Book, and ChemBioDraw software was used to draw and confirm the three-dimensional structure for target prediction and Cytoscape software was used to build a visual network diagram; a protein interaction network was searched and built between the methylene blue target and the common target of Pg in the String database; then we selected FimA, Mfa4, RgpB, and Kgp K1 proteins, used AutoDock software to calculate the docking energy of methylene blue and the above-mentioned proteins and performed molecular docking.@*RESULTS@#The target prediction results showed that there were 19 common targets between the 268 potential targets of methylene blue and 1 865 Pg proteins. The 19 targets were: groS, radA, rplA, dps, fabH, pyrG, thyA, panC, RHO, frdA, ileS, bioA, def, ddl, TPR, murA, lepB, cobT, and gyrB. The results of the molecular docking showed that methylene blue could bind to 9 sites of FimA protein, with a binding energy of -6.26 kcal/mol; with 4 sites of Mfa4 protein and hydrogen bond formation site GLU47, and the binding energy of -5.91 kcal/mol, the binding energy of LYS80, the hydrogen bond forming site of RgpB protein, was -5.14 kcal/mol, and the binding energy of 6 sites of Kgp K1 protein and the hydrogen bond forming site GLY1114 of -5.07 kcal/mol.@*CONCLUSION@#Computer simulation of target prediction and molecular docking technology can initially reveal the binding, degree of binding and binding sites of methylene blue and Pg proteins. This method provides a reference for future research on the screening of binding sites of photosensitizers to cells and bacteria.

Regulation of microRNA-126 on the polarization of human macrophages stimulated by Porphyromonas gingivalis lipopolysaccharide / 中华口腔医学杂志

Objective: To study the effect of microRNA-126 (miR-126) on the polarization of human monocyte-derived macrophages stimulated by Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS). Methods: Macrophages derived from human myeloid leukemia mononuclear cells were stimulated by Pg-LPS (5 mg/L) and by Pg-LPS (5 mg/L) after 24 h-transfection of miR-126 mimic or negative control RNA for 48 h, respectively. Real-time quantitative-PCR (qRT-PCR), enzyme-linked immunosorbent assay (ELISA) and Western blotting were conducted to detect the changes in miR-126, pro-inflammatory factor tumor necrosis factor-α (TNF-α), anti-inflammatory factors interleukin-10 (IL-10), inducible nitric oxide synthase (iNOS), arginase-1 (Arg-1) and M1 polarization-related pathways such as nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Results: Compared with non-LPS stimulation group (TNF-α: 1.000±0.020, iNOS: 1.125±0.064, miR-126: 1.004±0.113, IL-10: 1.003±0.053, Arg-1: 1.130±0.061), the mRNA levels of TNF-α (3.105±0.278) and iNOS (4.296±0.003) increased significantly (t=6.53, P=0.003; t=42.63, P<0.001, respectively), while miR-126, IL-10 and Arg-1 expressions (0.451±0.038, 0.545±0.004 and 0.253±0.017) decreased significantly (t=7.95, P=0.001; t=7.36, P=0.002; t=11.94, P<0.001, respectively) after Pg-LPS stimulated by human-derived macrophages for 48 h. The protein expression of iNOS, TNF-α, Arg-1 and IL-10 were consistent at mRNA levels. Meanwhile, the expressions of phospho-NF-κB p65 (p-p65), phospho-extracellular signal-regulated kinase (p-ERK) and phospho-p38 MAPK (p-p38) increased significantly, while the expression of Arg-1 decreased significantly. Compared with the negative controls (scramble RNA) (TNF-α: 1.141±0.197, iNOS: 1.173±0.115, IL-10: 1.032±0.138, Arg-1: 0.933±0.044), the mRNA levels of TNF-α (0.342±0.022) and iNOS (0.588±0.085) expressions significantly decreased (t=5.35, P=0.006; t=5.05, P=0.007), while IL-10 (1.786±0.221) and Arg-1 expressions (2.152±0.229) significantly increased (t=3.71, P=0.021; t=6.21, P=0.003) after Pg-LPS stimulation with miR-126 mimic transfection. The relative protein expressions of iNOS, p-p65, p-ERK and p-p38 significantly decreased (t=13.00, P<0.001; t=6.98, P=0.002; t=10.86, P<0.001; t=8.32, P=0.001), while the protein level of Arg-1 significantly increased (t=12.08, P<0.001). Conclusions: Pg-LPS could promote M1 polarization of macrophages. miR-126 might inhibit the effect of Pg-LPS on the M1 polarization of macrophages through down-regulating NF-κB and MAPK signaling pathways.

Effects of Porphyromonas gingivalis injected through tail vein on the expressions of biomarkers in neural stem cells and neurons of wild-type rats hippocampus / 中华口腔医学杂志

Objectives: To study the effects of Porphyromonas gingivalis (Pg) injected through tail vein on the molecular expression levels of biomarkers of neural stem cells (NSC) and neurons in the hippocampus of wild-type adult rats, and the effects on hippocampal neurogenesis. Methods: Eighteen male Sprague-Dawley (SD) rats were randomly divided into 3 groups based on the table of random numbers (n=6 in each group). In low-intensity group and high-intensity group, rats were injected intravenously through tail vein with 200 μl Pg ATCC33277 [1.0×103 and 1.0×108 colony forming unit (CFU), respectively] 3 times per week for 8 weeks. In the sham group, 200 μl of phosphate buffer saline (PBS) was given instead. Behavioral tests: the navigation and the exploration tests using Morris water maze (MWM) were applied to evaluate learning and memory ability of rats. Immunohistochemistry was performed to detect cells positively expressing nestin, doublecortin (DCX) and neuronal nuclei (NeuN) in the subgranular zone (SGZ) of rats in each group. Western blotting was used to evaluate the expression levels of nestin, DCX and NeuN in rat hippocampus. Results: Learning and memory abilities: on day 5 of navigation test, the lagency time was 22.83 (16.00, 38.34) s in the high-intensity group, significantly longer than the sham group [5.59 (5.41, 6.17) s] (t=-11.17, P<0.001). There were no significant differences between the low-intensity group [9.85 (8.75, 21.01) s] and the sham group (t=-6.83, P=0.080). Results in the exploration test showed that, in the high-intensity group, the number of fime crossing over the previous platform area within 60 s was 1.50 (1.00, 2.00), significantly less than the sham group [4.00 (2.75, 4.00)] (t=9.75, P=0.003); no significant differences between the low-intensity group [2.50 (2.00, 3.00)] and the sham one (t=4.50, P=0.382). Immunohistochemistry showed that the nestin+ cell density in the low-intensity group [(35.36±4.32) cell/mm2] and high-intensity group [(26.51±5.89) cell/mm2] were significantly lower than the sham group [(59.58±14.15) cell/mm2] (t=24.21, P=0.018; t=33.07, P=0.005); as for the mean absorbance of DCX+ cells, the low-intensity group (0.007±0.002) and the high-intensity group (0.006±0.002) were significantly lower than the sham group (0.011±0.001) (t=0.004, P=0.018; t=0.006, P=0.005); compared with the sham group [(1.13±0.14)×103 cell/mm2], the density of NeuN+ neurons in the high-intensity group [(0.75±0.08)×103 cell/mm2] was significantly reduced (t=0.38, P=0.017), and was not significantly changed in the low-intensity group [(0.88±0.19)×103 cell/mm2] (t=0.25, P=0.075). Western blotting results showed that, compared with the sham group, the expression levels of nestin, DCX, and NeuN were significantly reduced in the high-intensity group (t=0.74, P<0.001; t=0.18, P=0.014; t=0.35, P=0.008), but were not statistically changed in the low-intensity group (t=0.18, P=0.108; t=0.08, P=0.172; t=0.19, P=0.077). Conclusions: Pg injected through tail vein may reduce learning and memory abilities of wild-type rats, and may reduce the number of nestin, DCX, and NeuN-positive cells, and the protein expression levels of the above molecules in the hippocampus.

Nivel de higiene y prevalencia de Porphyromona gingivalis y Fusobacterium nucleatum en pacientes recuperados de SARS-CoV-2 / Hygiene level and prevalence of Porphyromona gingivalis and Fusobacterium nucleatum in recovered SARS-CoV-2 patients

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)

Actividad antimicrobiana de fracciones obtenidas del aceite esencial de Minthostachys mollis frente a patógenos orales / Antimicrobial activity of fractions obtained from essential oil of Minthostachys mollis against oral pathogens

Introducción: El aceite esencial de Minthostachys mollis ha demostrado poseer importantes propiedades antimicrobianas. Objetivo: Caracterizar químicamente las fracciones obtenidas del aceite esencial de Minthostachys mollis y evaluar la actividad antimicrobiana sobre Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Porphyromonas gingivalis y Candida albicans. Material y Métodos: Las fracciones de éter de petróleo, diclorometano y metanol del AE de M. mollis fueron caracterizadas químicamente por cromatografía de gases acoplada a espectrometría de masas. Las repeticiones del ensayo antimicrobiano se calcularon con el programa EPIDAT v.4.2. La actividad antimicrobiana se realizó por el método de difusión de disco y se calculó la concentración mínima inhibitoria por el método de microdilución. Los datos fueron analizados empleando la prueba ANOVA (p=0,05). Resultados: Los principales constituyentes de las fracciones de éter de petróleo, diclorometano y metanol fueron cis-Menthone (39,8 por ciento, thymol (31,2 por ciento) y α-Terpineol (43,6 por ciento), respectivamente. Todas las cepas fueron sensibles a las tres fracciones, aunque C. albicans fue la cepa más sensible, registrando halos de inhibición de 14,73±0,57 mm para la fracción de metanol, 20,91±0,55 mm para éter de petróleo y 20,38±0,58 mm para diclorometano, se encontraron diferencias significativas cuando se compararon frente a Clorhexidina al 0,12 por ciento y Nistatina (p<0,05). Las concentraciones mínimas inhibitorias de las fracciones variaron de 0,2 a 3,2 µg/mL. Conclusiones: Los principales constituyentes de las fracciones de éter de petróleo, diclorometano y metanol fueron cis-Menthone, thymol y α-Terpineol. Las fracciones de éter de petróleo y diclorometano fueron altamente efectivas para inhibir el crecimiento de S. mutans, L. acidophilus, E. faecalis, P. gingivalis y C. albicans(AU)

Introduction: The essential oil of Minthostachys mollis has demonstrated to have important antimicrobial properties. Objective: To chemically characterize the fractions obtained from the essential oil of Minthostachys mollis and to evaluate the antimicrobial activity against Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Porphyromonas gingivalis and Candida albicans. Material and Methods: The petroleum ether, dichloromethane and methanol fractions of the AE of M. mollis were chemically characterized by gas chromatography coupled to mass spectrometry. The repetitions of the antimicrobial test were calculated using the EPIDAT v.4.2 program. The antimicrobial activity was performed by the disk diffusion method and the minimum inhibitory concentration was calculated by the microdilution method. The data were analyzed using the ANOVA test (p=0.05). Results: The main constituents of the petroleum ether, dichloromethane and methanol fractions were cis-Menthone (39,8 percent), thymol (31,2 percent)) and α-Terpineol (43,6 percent)), respectively. All strains were sensitive to the three fractions, although C. albicans was the most sensitive strain, registering inhibition halos of 14,73±0.57 mm for the methanol fraction, 20,91±0.55 mm for petroleum ether and 20.38±0.58 mm for dichloromethane, finding significant differences when compared to 0,12 percent) Chlorhexidine and Nystatin (p<0,05). The minimum inhibitory concentrations of the fractions ranged from 0,2 to 3,2 µg/mL. Conclusions: The main constituents of the petroleum ether, dichloromethane and methanol fractions were cis-Menthone, thymol and α-Terpineol. The petroleum ether and dichloromethane fractions were highly effective in inhibiting the growth of S. mutans, L. acidophilus, E. faecalis, P. gingivalis, and Calbicans(AU)

Relación entre Porphyromonas gingivalis y diabetes mellitus tipo 2: revisión sistemática exploratoria / Relationship between Porphyromonas gingivalis and Type 2 Diabetes Mellitus: Scoping review

Objetivo: analizar la relación entre Porphyromonas gingivalis y diabetes mellitus tipo 2, mediante una revisión sistemática exploratoria de la literatura científica publicada entre los años 2000 y 2019. Métodos: se utilizaron los siguientes términos MeSH: Porphyromonas gingivalis, diabetes mellitus type 2, periodontal disease, non insulin dependent diabetes. Se obtuvieron 346 resultados, de los cuales se seleccionaron 41 por título, se excluyeron 11 posterior a la lectura del abstract e introducción y 19 después de la lectura del texto completo. Finalmente, se incluyeron 11 artículos. Resultados: el lipopolisacárido de Porphyromonas gingivalis y su fimbria tipo II se relacionan con una mayor producción de citoquinas proinflamatorias como IL-6 y TNF-α, las cuales afectan las vías de señalización de la glucosa y se relacionan con insulinoresistencia. La dipeptidil peptidasa 4 de Porphyromonas gingivalis puede participar en la degradación de incretinas, lo cual afecta la producción de insulina en el huésped y promueve estados de hiperglicemia. El interactoma de Porphyromonas gingivalis puede superponerse con genes involucrados en resistencia a la insulina y diabetes mellitus tipo 2. Conclusión: según la evidencia científica publicada existen factores de virulencia y mecanismos por los cuales la Porphyromonas gingivalis influye en el desarrollo de insulinorresistencia y diabetes mellitus tipo 2.

Objective: To analyze the relationship between Porphyromonas gingivalis and Diabetes Mellitus Type 2 by reviewing the scientific literature published between 2000 and 2019. Methods: The following MeSH terms were used: Porphyromonas gingivalis, Diabetes Mellitus type 2, periodontal disease, non-insulin dependent diabetes. We obtained 346 results, of which 41 were selected by title, 11 were excluded after reading the abstract and introduction and 19 after reading the full text. Finally, 11 articles were included. Results: Porphyromonas gingivalis lipopolysaccharide and its type II fimbria are associated with increased production of proinflammatory cytokines such as IL-6 and TNF-α, which affect glucose signaling pathways and are related to insulin resistance. Porphyromonas gingivalis dipeptidyl peptidase 4 (PgDPP4) may participate in incretin degradation which affects host insulin production and promotes hyperglycemic states. The Porphyromonas gingivalis interactome may overlap with genes involved in insulin resistance and type 2 diabetes mellitus. Conclusion: According to published scientific evidence, there are virulence factors and mechanisms by which Porphyromonas gingivalis influences the development of insulin resistance and type 2 Diabetes Mellitus.

Prevalencia de Prevotella spp y Porphyromona spp en periodontitis crónica / Prevalence of Prevotella spp and Porphyromona spp in chronic periodontitis

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)

Actividad antibacteriana y antifúngica del aceite esencial de Minthostachys mollis frente a microorganismos de la cavidad oral / Antibacterial and Antifungal Activity of the essential oil of Minthostachys mollis against oral microorganisms

Introducción: La Minthostachys mollises una planta aromática que crece en América Latina y produce aceites esenciales con acción antimicrobiana. Objetivo: Determinar la actividad del aceite esencial de Minthostachys mollis en diferentes concentraciones, comparado con doxiciclina y fluconazol frente a Porphyromonas gingivalis, Staphylococcus aureus y Candida albicans, a las 24, 48 y 72 horas. Métodos: Se realiza estudio experimental in vitro y longitudinal. Se prepararon 15 pocillos por subgrupo para evaluar el efecto inhibitorio de todas las concentraciones, dando un total de 360 pocillos. Por cromatografía de gases acoplada a espectrometría de masas se identificaron los componentes químicos del aceite esencial. Se analizó el efecto inhibitorio por el método de difusión de Kirby-Bauer en Agar Columbia y Agar Muller Hinton. El análisis estadístico se realizó mediante la prueba ANOVA y Tukey. Resultados: En el análisis químico se identificó principalmente pulegona (30,17 por ciento) y mentona (16,55 por ciento). Los halos de inhibición de Minthostachys mollis al 100 por ciento a las 24, 48 y 72 horas frente a la Porphyromonas gingivalis, midieron: 10,2 mm, 9,8 mm y 9,6 mm, respectivamente; frente al Staphylococcus aureus, midieron: 10,4 mm, 9,7 mm y 9,4 mm, respectivamente; y, por último, frente a Candida albicans midieron: 9,8 mm, 8,9 mm y 8,5 mm, respectivamente. Todas las concentraciones de Minthostachys mollis presentaron un efecto antimicrobiano significativamente menor que el fluconazol y la doxiciclina (p < 0,001). Conclusiones: El aceite esencial de Minthostachys mollis al 100 % presentó su mejor actividad inhibitoria frente al Staphylococcus aureus, la Porphyromonas gingivalis y la Candida albicans a las 24 horas. Sin embargo, este efecto antimicrobiano disminuye a medida que pasa el tiempo(AU)

Introduction: Minthostachys mollis is an aromatic plant species growing in Latin America which produces essential oils with antimicrobial activity. Objective: Determine the activity of essential oil from Minthostachys mollis at various concentrations as compared with doxycycline and fluconazole against Porphyromonas gingivalis, Staphylococcus aureus and Candida albicans at 24, 48 and 72 hours. Methods: An in vitro experimental longitudinal study was conducted. Fifteen wells were prepared per subgroup to evaluate the inhibitory effect of all concentrations, for a sum total of 360 wells. Chemical components of the essential oil were identified by gas chromatography-mass spectrometry. The inhibitory effect was analyzed with the Kirby-Bauer diffusion method in Mueller-Hinton and Columbia agar. Statistical analysis was based on ANOVA and Tukey's test. Results: Chemical analysis mainly found pulegone (30.17 percent) and menthone (16.55 percent). The inhibition halos of 100 percent Minthostachys mollis at 24, 48 and 72 hours against Porphyromonas gingivalis measured 10.2 mm, 9.8 mm and 9.6 mm, respectively, against Staphylococcus aureus they measured 10.4 mm, 9.7 mm and 9.4 mm, respectively, and against Candida albicans they measured 9.8 mm, 8.9 mm and 8.5 mm, respectively. The antimicrobial effect of Minthostachys mollis at all concentrations was significantly lower than that of fluconazole and doxycycline (p < 0.001). Conclusions: The essential oil from 100% Minthostachys mollis displayed its best inhibitory activity against Staphylococcus aureus, Porphyromonas gingivalis and Candida albicans at 24 hours. However, such antimicrobial effect decreases with the passing of time(AU)

Porphyromonas gingivalis exacerbates ulcerative colitis via Porphyromonas gingivalis peptidylarginine deiminase / 国际口腔科学杂志·英文版

Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.

Porphyromonas gingivalis infection promotes mitochondrial dysfunction through Drp1-dependent mitochondrial fission in endothelial cells / 国际口腔科学杂志·英文版

Porphyromonas gingivalis (P. gingivalis), a key pathogen in periodontitis, has been shown to accelerate the progression of atherosclerosis (AS). However, the definite mechanisms remain elusive. Emerging evidence supports an association between mitochondrial dysfunction and AS. In our study, the impact of P. gingivalis on mitochondrial dysfunction and the potential mechanism were investigated. The mitochondrial morphology of EA.hy926 cells infected with P. gingivalis was assessed by transmission electron microscopy, mitochondrial staining, and quantitative analysis of the mitochondrial network. Fluorescence staining and flow cytometry analysis were performed to determine mitochondrial reactive oxygen species (mtROS) and mitochondrial membrane potential (MMP) levels. Cellular ATP production was examined by a luminescence assay kit. The expression of key fusion and fission proteins was evaluated by western blot and immunofluorescence. Mdivi-1, a specific Drp1 inhibitor, was used to elucidate the role of Drp1 in mitochondrial dysfunction. Our findings showed that P. gingivalis infection induced mitochondrial fragmentation, increased the mtROS levels, and decreased the MMP and ATP concentration in vascular endothelial cells. We observed upregulation of Drp1 (Ser616) phosphorylation and translocation of Drp1 to mitochondria. Mdivi-1 blocked the mitochondrial fragmentation and dysfunction induced by P. gingivalis. Collectively, these results revealed that P. gingivalis infection promoted mitochondrial fragmentation and dysfunction, which was dependent on Drp1. Mitochondrial dysfunction may represent the mechanism by which P. gingivalis exacerbates atherosclerotic lesions.

MicroRNA-146a regulates the production of cytokines in lymphocytes stimulated by / 华西口腔医学杂志

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Detection of Oral Entamoeba Gingivalis and Trichomonas Tenax in Adult Quilombola Population with Periodontal Disease / Detección de Entamoeba Gingivalis oral y Trichomonas Tenax en población de quilombola adulta con enfermedad periodontal

Abstract: The objective was to analyze the periodontal condition severity and the occurrence of pathogenic microorganisms in the oral cavity of an adult population of an Afrodescent Community of northeastern Brazil. This is an observational and cross- sectional study performed through an oral clinical examination, using a standardized clinical record. For the subjects with periodontal disease, the bacterial biofilm was collected in a Petri dish containing 0.9% physiological solution to detect the presence of microorganisms Entamoeba gingivalis and Trichomonas tenax, and later observed under an optical microscope. Statistical analysis was performed by calculating the prevalence of periodontal disease and the frequency of the protozoa in the bacterial biofilm. Statistical significance of the relationships researched was verified by Fisher's exact test. It was evaluated 29 subjects pertaining to the Quilombola Patioba community, aged 35 to 44 years. The results showed that among the adults of the community, there was a high prevalence of periodontal disease (75.86%), being higher in the 1st and 6th sextants of the Community Periodontal Index (CPI). E. gingivalis positivity occurred in most sextants affected by gingivitis, while in the condition of periodontitis, this microorganism was not present in the 3rd, 4th and 6th sextants. In all sextants affected by periodontal disease, T. Tenax was observed when associated with gingivitis. It is worth mentioning the begging of the elaboration of health policies, social and professional commitment that foster a greater promotion of oral health and quality of life for the quilombolas of northeastern Brazil.

Resumen: El objetivo fue analizar la severidad de la condición periodontal y la aparición de microorganismos patógenos en la cavidad oral de una población adulta de una comunidad afrodescente del noreste de Brasil. Este es un estudio observacional y transversal realizado a través de un examen clínico oral, utilizando un registro clínico estandarizado. Para los sujetos con enfermedad periodontal, la biopelícula bacteriana se recogió en una placa de Petri que contenía una solución fisiológica al 0,9% para detectar la presencia de microorganismos Entamoeba gingivalis y Trichomonas tenax, y luego se observó bajo un microscopio óptico. El análisis estadístico se realizó calculando la prevalencia de la enfermedad periodontal y la frecuencia de los protozoos en la biopelícula bacteriana. La significación estadística de las relaciones investigadas se verificó mediante la prueba exacta de Fisher. Se evaluaron 29 sujetos pertenecientes a la comunidad Quilombola Patioba, de 35 a 44 años. Los resultados mostraron que entre los adultos de la comunidad, hubo una alta prevalencia de enfermedad periodontal (75.86%), siendo mayor en el sexto sexto y sexto del Índice periodontal comunitario (IPC). La positividad de E. gingivalis se produjo en la mayoría de los sextantes afectados por gingivitis, mientras que en la condición de periodontitis, este microorganismo no estaba presente en los sextantes tercero, cuarto y sexto. En todos los sextantes afectados por enfermedad periodontal, se observó T. Tenax cuando se asoció con gingivitis. Vale la pena mencionar el inicio de la elaboración de políticas de salud, compromiso social y profesional que promuevan una mayor promoción de la salud oral y la calidad de vida de las quilombolas del noreste de Brasil.

Salivary markers of oxidative stress and periodontal pathogens in patients with periodontitis from Santander, Colombia / Marcadores salivales de estrés oxidativo y agentes patógenos periodontales en pacientes con periodontitis en Santander, Colombia

Introduction: Periodontitis affects more than 20% of the Latin American population. Oxidative markers are associated with greater progression of periodontitis; therefore, its role in pathogenesis should be studied. Objective: To determine the prevalence of the main oral bacteria and viruses associated with periodontitis and estimate the total antioxidant capacity and lipid peroxidation in saliva from patients with periodontitis. Materials and methods: We conducted systemically a cross-sectional study in 101 healthy subjects, 87 of whom had been diagnosed with periodontitis (P), according to the criteria of the Centers of Disease Control and Prevention and the American Academy of Periodontology, and 14 without periodontal pockets as controls (C). In subgingival samples, major viruses and dental pathogenic bacteria were identified using PCR techniques. The levels of total antioxidant capacity and malon-di-aldehyde (MDA) were determined by spectrophotometry in samples of unstimulated saliva. Results: The mean of periodontal depth pocket and clinical attachment loss in patients with periodontitis was 5.6 ± 1.7 and 6.1 ± 3.1 mm, respectively. The most prevalent microorganisms were Aggregatibacter actinomycetemcomitans (32.5%) and Porphyromonas gingivalis (18.6%). The patients from rural areas showed a higher percentage of A . actinomycetemcomitans (urban: 17.9% vs. rural: 48.9%, p=0.0018). In patients with periodontitis, the frequency of EBV, HSV1 & 2, and HCMV genes was 2.3%. Periodontitis patients had higher levels of MDA (P: 2.1 ± 1.5; C: 0.46 ± 0.3 µmol/g protein; p=0.0001) and total antioxidant capacity (P: 0.32 ± 0.2; C: 0.15 ± 0.1 mM; p< 0.0036). Oxidative markers showed no modifications due to the presence of periodontopathic bacteria. Conclusions: Aggregatibacter actinomycetemcomitans was the most prevalent bacteria; its presence did not modify the levels of oxidative markers in the saliva of patients with periodontitis.

Introducción. La periodontitis afecta a más del 20 % de la población latinoamericana. La presencia de marcadores de estrés oxidativo se asocia con una mayor progresión de periodontitis, por lo que su rol en la patogenia debe estudiarse. Objetivo. Determinar la prevalencia de las principales bacterias y virus asociados con la periodontitis y estimar la capacidad antioxidante total y la peroxidación de lípidos en la saliva de los pacientes con periodontitis. Materiales y métodos. Se hizo un estudio transversal en 87 sujetos sanos diagnosticados con periodontitis (P) según los criterios de los Centers of Disease Control and Prevention y la American Academy of Periodontology y 14 sujetos sin enfermedad periodontal como grupo control (C). En las muestras subgingivales se identificaron los principales virus y bacterias mediante técnicas de PCR. Los niveles de capacidad antioxidante total y malon-di-aldehído (MDA) se establecieron mediante espectrofotometría en muestras de saliva no estimulada. Resultados. Las medias de profundidad del sondaje y del nivel de inserción clínico periodontal en pacientes con periodontitis fueron 5,6 ± 1,7 y 6,1 ± 3,1 mm, respectivamente. Los microorganismos más prevalentes fueron Aggregatibacter actinomycetemcomitans (32,5 %) y Porphyromonas gingivalis (18,6 %). Los pacientes de áreas rurales registraron un mayor porcentaje de A. actinomycetemcomitans (urbano: 17,9 % Vs. rural: 48,9 %; p=0,0018). La frecuencia de los genes EBV, HSV1 y 2, y HCMV fue de 2,3 %. En pacientes con periodontitis se evidenciaron mayores niveles de MDA (P: 2,1 ± 1,5; C: 0,46 ± 0,3 µmol/g proteína; p=0,0001) y capacidad antioxidante total (P: 0,32 ± 0,2; C: 0,15 ± 0,1 mM; p<0,0036). La presencia de bacterias periodontales patógenas no modificó los marcadores oxidativos. Conclusiones.Aggregatibacter actinomycetemcomitans fue el agente patógeno mas prevalente. Su presencia no modificó los niveles de marcadores oxidativos en la saliva de los pacientes con periodontitis.

In vitro homeostasis of rat oral epithelial cell cultures following withdrawal of periodontal pathogens

Abstract Inflammation of periodontal tissues is the consequence of interaction between periodontal pathogens and immune system. This is associated with increased expression of inflammatory cytokines, which may exert destructive effect to the periodontal tissues when released over long period. The aim of this study was to chronologically track the homeostasis of oral keratinocytes following removal of periodontal pathogens. This was done by investigating expression of selected inflammatory markers and integrity of epithelial monolayers in vitro. Rat oral keratinocytes were stimulated with heat-killed Fusobacterium nucleatum and Porphyromonas gingivalis over 7-days then bacteria were washed away and epithelial cells re-cultured for 3-days. Expression of IL-1β, IL-6, and IL-8 was measured by ELISA while transcription of tissue inhibitor of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase -8 (MMP-8) was measured by polymerase chain reaction before and after removal of bacteria. Integrity of epithelial sheet was investigated by using transepithelial electrical resistance. Data showed general downregulation of IL-1b, IL-6, and IL-8 associated with restoring transcription of TIMP-1 and MMP-8 to normal level following removal of bacteria from epithelial cultures. However, expression of IL-8 and MMP-8 remained significantly higher than unstimulated epithelial cells despite withdrawal of F. nucleatum and P. gingivalis respectively from oral keratinocytes cultures. In addition, integrity of epithelial barrier function remained compromised even after removal of P. gingivalis. Results suggest that even after three days following removal of periodontal pathogens, oral keratinocytes sustained persistent upregulation of certain inflammatory markers that could compromise integrity of epithelial barrier function.

Resumo A inflamação dos tecidos periodontais é a consequência da interação entre patógenos periodontais e o sistema imunológico. Isso está associado ao aumento da expressão de citocinas inflamatórias, que podem exercer efeito destrutivo nos tecidos periodontais quando liberadas por um longo período. O objetivo deste estudo foi rastrear cronologicamente a homeostase dos queratinócitos orais após a remoção dos patógenos periodontais. Isto foi feito através da investigação da expressão de marcadores inflamatórios selecionados e da integridade de monocamadas epiteliais in vitro. Os queratinócitos orais de rato foram estimulados com Fusobacterium nucleatum e Porphyromonas gingivalis destruídas pelo calor por 7 dias, depois as bactérias foram lavadas e as células epiteliais foram cultivadas novamente por 3 dias. A expressão de IL-1b, IL-6 e IL-8 foi medida por ELISA, enquanto a transcrição do inibidor tecidual de metaloproteinase-1 (TIMP-1) e matriz metalopeptidase-8 (MMP-8) foi medida por reação em cadeia da polimerase antes e após a remoção de bactérias. A integridade da folha epitelial foi investigada usando resistência elétrica transepitelial. Os dados mostraram uma regulação negativa geral de IL-1b, IL-6 e IL-8 associada à restauração da transcrição de TIMP-1 e MMP-8 para o nível normal após a remoção de bactérias de culturas epiteliais. No entanto, a expressão de IL-8 e MMP-8 permaneceu significativamente maior que as células epiteliais não estimuladas, apesar da retirada de F. nucleatum e P. gingivalis, respectivamente, das culturas de queratinócitos orais. Além disso, a integridade da função da barreira epitelial permaneceu comprometida mesmo após a remoção de P. gingivalis. Os resultados sugerem que, mesmo após três dias após a remoção dos patógenos periodontais, os queratinócitos orais sustentaram uma regulação positiva persistente de certos marcadores inflamatórios que poderiam comprometer a integridade da função da barreira epitelial.

Oral human cytomegalovirus prevalence and its relationships with periodontitis and Porphyromonas gingivalis in Japanese adults: a cross-sectional study

Abstract Objective This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. Methodology In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. Results HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). Conclusions Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease.