ABSTRACT
Introducción: la periodontitis es una enfermedad infecciosa multifactorial asociada a un biofilm de microorganismos patógenos. Objetivo: el objetivo del trabajo fue establecer la prevalencia de Porphyromonas gingivalis en pacientes con periodontitis y relacionarla con la severidad de la enfermedad. Material y métodos: participaron 45 pacientes, sistémicamente saludables, con edades entre 35 y 65 años. El grado de periodontitis se definió según los criterios de Papapanou y colaboradores. Como grupo control, se incluyeron 20 sujetos de ambos sexos sin periodontitis y sin enfermedades sistémicas. Se tomaron muestras de fluido gingival en dos sitios más profundos. Porphyromonas gingivalis se detectó por PCR (reacción en cadena de la polimerasa). Resultados: la frecuencia relativa de periodontitis fue de 13.3% grado I, 46.7% grado II y 40% grado III. El sexo masculino presentó periodontitis grado III 72.2% y grado II 52.3%. El grado I se registró con mayor frecuencia en el sexo femenino, 66.7%. La prevalencia de Porphyromonas gingivalis en la población con periodontitis fue de 44.4%. Se obtuvieron diferencias estadísticamente significativas entre los grados de severidad de periodontitis y la presencia de Porphyromonas gingivalis (p = 0.0002, α = 5%). Conclusión: la periodontitis predominó en el sexo masculino. La prevalencia de Porphyromonas gingivalis en la población con periodontitis crónica fue de 44.4% y su presencia está relacionada con la severidad (AU)
Introduction: periodontitis is a multifactorial infectious disease associated with a biofilm of pathogenic microorganisms. Objective: the objective of the work was to establish the prevalence of Porphyromonas gingivalis in patients with periodontitis and relate it to the severity of the disease. Material and methods: 45 systemically healthy patients, aged between 35 and 65 years old, participated. The degree of periodontitis was defined according to the criteria of Papapanou et al. As a control group, 20 patients of both sexes without periodontitis and without systemic diseases were included. Gingival fluid samples were taken from two deeper sites. Porphyromonas gingivalis was detected by PCR (polymerase chain reaction). Results: the relative frequency of periodontitis was 13.3% grade I, 46.7% grade II and 40% grade III. The male sex presented periodontitis grade III 72.2% and grade II 52.3%. Grade I was recorded more frequently in the female sex, 66.7%. The prevalence of Porphyromonas gingivalis in the population with periodontitis was 44.4%. Statistically significant differences were obtained between the degrees of severity of periodontitis and the presence of Porphyromonas gingivalis (p = 0.0002, α = 5%). Conclusion: periodontitis predominated in males. The prevalence of Porphyromonas gingivalis in the population with chronic periodontitis was 44.4% and its presence is related to severity (AU)
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Porphyromonas gingivalis/isolation & purification , Chronic Periodontitis/epidemiology , Argentina/epidemiology , Schools, Dental , Epidemiology, Descriptive , Cross-Sectional Studies , Age and Sex Distribution , CetrimoniumABSTRACT
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.
Subject(s)
Periodontal Diseases/microbiology , Bacteria, Anaerobic/physiology , Bone Resorption/microbiology , RANK Ligand/metabolism , Cells, Cultured , Blotting, Western , Porphyromonas gingivalis/physiology , Prevotella intermedia/physiology , Cell Line, Tumor , Electrophoresis , RANK Ligand/analysisABSTRACT
Objective: To detect and analyze the characteristics of oral microbiota in species composition, function and metabolism among caries, periodontitis and oral healthy individuals, hunting for the microbiome-derived biomarkers with specificity and sensitivity to estimate the occurrence of these two diseases. Methods: Saliva samples were collected from 10 patients with high caries risk [decayed-missing-filled teeth (DMFT)≥6, HC group] in Department of Endodontics, 10 patients with periodontitis of grade Ⅱ A-Ⅲ C (PG group) in Department of Periodontology and 10 oral healthy individuals (HH group) from School of Stomatology, The Fourth Military Medical University during from March 2022 to June 2022. A baseline examination was conducted on all participants, including their oral conditions of caries and periodontal health. Metagenomic sequencing (Illumina PE150 platform) and liquid chromatography-mass spectrometry were used to detect microorganisms and their metabolites in the samples respectively. The sequencing data were analyzed to obtain the information of microbial taxonomic composition, functional genes and metabolites in each group of samples. The basic oral conditions and saliva samples of subjects in each group were evaluated and collected by the same professional endodontist. Results: There were no significant difference in baseline characteristics such as age and sex among the subjects in each group (P>0.05). DMFT in HC group (9.0±1.7) was significantly higher than that in HH group (0) and PG group (0) (F=243.00, P<0.001). Sequencing data analysis showed that the taxonomic compositions of salivary microbiota in each group were mainly Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria at the phylum level, and Streptococcus, Neisseria, Rothia, Prevotella at the genus level. Differential analysis showed that, compared with the HH group, HC group and PG group had significant differences in taxonomic composition (P<0.05), and the most significant among them was Prevotella. At the species level, Prevotella pallens was the most significant change in HC group, and Porphyromonas gingivalis in PG group. Metabolite analysis showed that there were significant differences in metabolites between HC group and PG group. The results showed that, compared with the HH group, the most significant metabolite change was 3-hydroxy-1, 5-diphenylpentan-1-one in HC group (P=0.001) and N1 acetylspermine in PG group (P=0.002) respectively. Compared with the PG group, the metabolite of HC group with the most significant difference is D-glucosamine 6-phosphate (P=0.006). The metabolism gene function analysis showed that, the enrichment of carbohydrate metabolism related genes was highest in HC group, followed with HH group, and it was lowest in PG group. In addition, compared with the HH group, the abundance of functional genes related to glucose metabolism, such as ABC transporter and phosphotransferase system, were significantly decreased in PG group (P<0.05), but significantly increased in HC group (P<0.05). Conclusions: There is a significant correlation between the alternation of carbohydrate metabolism of salivary microbiota with the occurrence of caries and periodontitis. In the future, Prevotella pallens and 3-hydroxy-1, 5-diphenylpentan-1-one may be the potential biomarkers of caries; while Porphyromonas gingivalis and N1 acetylspermine work in the predictions of periodontitis.
Subject(s)
Humans , Saliva/microbiology , Dental Caries Susceptibility , Periodontitis/microbiology , Microbiota/genetics , Porphyromonas gingivalis/genetics , RNA, Ribosomal, 16S/geneticsABSTRACT
OBJECTIVE@#To describe the submucosal microbial profiles of peri-implantitis and healthy implants, and to explore bacteria that might be correlated with clinical parameters.@*METHODS@#In the present cross-sectional study, 49 patients were recruited. Each patient contributed with one implant, submucosal biofilms were collected from 20 healthy implants and 29 implants with peri-implantitis. DNA was extracted and bacterial 16S ribosomal RNA (16S rRNA) genes were amplified. Submucosal biofilms were analyzed using 16S rRNA sequencing at Illumina MiSeq platform. Differences between the groups were determined by analyzing α diversity, microbial component and microbial structure. The potential correlation between the bacteria with pocket probing depth (PPD) of peri-implant calculated by Spearman correlation analysis.@*RESULTS@#The α diversity of submucosal microbial of health group was significantly lower than that in peri-implantitis group (Chao1 index: 236.85±66.13 vs. 150.54±57.43, P < 0.001; Shannon index: 3.42±0.48 vs. 3.02±0.65, P=0.032). Principal coordinated analysis showed that the submucosal microbial structure had significant difference between healthy and peri-implantitis groups [R2=0.243, P=0.001, analysis of similarities (ANOSIM)]. Compared with healthy implants, relative abundance of periodontal pathogens were higher in peri-implantitis, including members of the red complex (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and some members of orange complex (Precotella intermedia, Eubacterium nodatum, Parvimonas micra), as well as some new periodontal pathogens, such as Fillifactor alocis, Fretibacterium fastidiosum, Desulfobulbus sp._HMT_041, and Porphyromonas endodontalis. Spearman correlation analysis revealed that the relative abundance of Treponema denticola (r=0.686, P < 0.001), Tannerella forsythia (r=0.675, P < 0.001), Fretibacterium sp. (r=0.671, P < 0.001), Desulfobulbus sp._HMT_041 (r=0.664, P < 0.001), Filifactor alocis (r=0.642, P < 0.001), Fretibacterium fastidiosum (r=0.604, P < 0.001), Porphyromonas gingivalis (r=0.597, P < 0.001), Porphyromonas endodontalis (r=0.573, P < 0.001) were positive correlated with PPD. While the relative abundance of Rothia aeria (r=-0.615, P < 0.001) showed negatively correlation with PPD.@*CONCLUSION@#Marked differences were observed in the microbial profiles of healthy implants and peri-implantitis. The members of red and orange complex as well as some new periodontal pathogens seem to play an important role in peri-implant disease. Compared with healthy implants, the submucosal microbial of peri-implantitis were characterized by high species richness and diversity.
Subject(s)
Humans , Peri-Implantitis/microbiology , Cross-Sectional Studies , RNA, Ribosomal, 16S/genetics , Bacterial Load , Porphyromonas gingivalis , Dental ImplantsABSTRACT
OBJECTIVE@#To explore the molecular mechanisms of Porphyromonas gingivalis infection-induced umbilical vein endothelial barrier dysfunction in vitro.@*METHODS@#Human umbilical vein endothelial cells (HUVECs) were cultured in vitro, and after the formation of the endothelial barrier, the cells were infected with P. gingivals at a multiplicity of infection (MOI). The transepithelial electrical resistance (TEER) of the cell barrier was measured, and FITC-dextran trans-endothelial permeability assay and bacterial translocation assay were performed to assess the endothelial barrier function. The expression levels of cell junction proteins including ZO-1, occludin and VE-cadherin in the cells were examined by qRT-PCR and Western blotting.@*RESULTS@#In freshly seeded HUVECs, TEER increased until reaching the maximum on Day 5 (94 Ωcm2), suggesting the formation of the endothelial barrier. P. gingivals infection caused an increase of the permeability of the endothelial barrier as early as 0.5 h after bacterial inoculation, and the barrier function further exacerbated with time, as shown by significantly lowered TEER, increased permeability of FITC-dextran (40 000/70 000), and increased translocation of SYTO9-E. coli cross the barrier. MTT assay suggested that P. gingivals infection did not significantly affect the proliferation of HUVECs (P>0.05), but in P. gingivalsinfected cells, the expressions of ZO-1, occludin and VE-cadherin increased significantly at 24 and 48 h after bacterial inoculation (P < 0.05).@*CONCLUSION@#P. gingivals may disrupt the endothelial barrier function by down-regulating the expressions of the cell junction proteins (ZO-1, occludin, VE-cadherin) and increasing the permeability of the endothelial barrier.
Subject(s)
Humans , Cadherins/metabolism , Escherichia coli/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Occludin , Porphyromonas gingivalis/metabolism , Umbilical Veins/metabolismABSTRACT
Bacteremia induced by periodontal infection is an important factor for periodontitis to threaten general health. P. gingivalis DNA/virulence factors have been found in the brain tissues from patients with Alzheimer's disease (AD). The blood-brain barrier (BBB) is essential for keeping toxic substances from entering brain tissues. However, the effect of P. gingivalis bacteremia on BBB permeability and its underlying mechanism remains unclear. In the present study, rats were injected by tail vein with P. gingivalis three times a week for eight weeks to induce bacteremia. An in vitro BBB model infected with P. gingivalis was also established. We found that the infiltration of Evans blue dye and Albumin protein deposition in the rat brain tissues were increased in the rat brain tissues with P. gingivalis bacteremia and P. gingivalis could pass through the in vitro BBB model. Caveolae were detected after P. gingivalis infection in BMECs both in vivo and in vitro. Caveolin-1 (Cav-1) expression was enhanced after P. gingivalis infection. Downregulation of Cav-1 rescued P. gingivalis-enhanced BMECs permeability. We further found P. gingivalis-gingipain could be colocalized with Cav-1 and the strong hydrogen bonding between Cav-1 and arg-specific-gingipain (RgpA) were detected. Moreover, P. gingivalis significantly inhibited the major facilitator superfamily domain containing 2a (Mfsd2a) expression. Mfsd2a overexpression reversed P. gingivalis-increased BMECs permeability and Cav-1 expression. These results revealed that Mfsd2a/Cav-1 mediated transcytosis is a key pathway governing BBB BMECs permeability induced by P. gingivalis, which may contribute to P. gingivalis/virulence factors entrance and the subsequent neurological impairments.
Subject(s)
Animals , Rats , Bacteremia/metabolism , Blood-Brain Barrier/microbiology , Caveolin 1/metabolism , Gingipain Cysteine Endopeptidases/metabolism , Permeability , Porphyromonas gingivalis/pathogenicity , Transcytosis , Virulence Factors/metabolismABSTRACT
OBJECTIVE@#To investigate the effect of Porphyromonas gingivalis (Pg) infection on IFNGR1 palmitoylation and biological behaviors of esophageal squamous cell carcinoma (ESCC) cells and the clinical implications.@*METHODS@#The expression levels of IFNGR1 protein in ESCC cell lines KYSE30 and KYSE70 were detected using Western blotting at 24 and 48 h after Pg infection, and 2-BP was used to detect IFNGR1 palmitoylation in the cells. KYSE70 cells with wild-type IFNGR1 (IFNGR1-WT cells) and with IFNGR1-C122A palmitoylation site mutation induced by site-specific mutagenesis (IFNGR1-C122A cells) were both infected with Pg, and the changes in palmitoylation of IFNGR1-C122A were analyzed using immunofluorescence and Click-iT assays. The changes in proliferation, migration and invasion ability of the infected cells were evaluated using plate cloning assay, scratch assay and Transwell assay, and IFNGR1 co-localization with lysosomal marker LAMP2 was dected using immunofluorescence assay. Immunohistochemistry was used to detect Pg infection and IFNGR1 protein expression in 50 ESCC tissues, and their correlation with the clinicopathological characteristics and survival outcomes of the patients was analyzed.@*RESULTS@#Pg infection down-regulated the protein expression of IFNGR1 in ESCC and promoted IFNGR1 palmitoylation at site 122. In IFNGR1-WT cells, Pg infection significantly enhanced cell proliferation, migration and invasion (P < 0.05). Similarly, Pg also significantly promoted proliferation, migration and invasion of IFNGR1-C122A cells, but to a lesser extent as compared with the wild-type cells (P < 0.05). Immunofluorescence assay showed that Pg and ZDHHC3 promoted IFNGR1 degradation within the lysosome. Immunohistochemical studies of the ESCC tissue samples showed a negative correlation between IFNGR1 and Pg expression, and a reduced IFNGR1 expression was correlated with a poorer survival outcome of the patient.@*CONCLUSION@#Pg infection enhances IFNGR1 palmitoylation to promote progression of ESCC, and elimination of Pg and inhibiting IFNGR1 palmitoylation may effectively control ESCC progression.
Subject(s)
Humans , Esophageal Neoplasms , Porphyromonas gingivalis , Lipoylation , Esophageal Squamous Cell Carcinoma , LysosomesABSTRACT
OBJECTIVES@#This study aimed to explore the functions and potential regulatory targets of local macrophages in nonalcoholic fatty liver combined with Porphyromonas gingivalis (P. gingivalis)infection.@*METHODS@#Single-cell RNA sequencing was used to analyze the phenotypes and functional changes in various cells in the liver tissue of nonalcoholic steatohepatitis (NASH) mice fed with P. gingivalis. Real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay, and immunofluorescence staining were applied to observe the inflammation and expression levels of macrophage antigen presenting functional markers in the NASH liver. Oil red staining was performed to observe the accumulation of local adipose tissue in the NASH liver. Results were verified through RT-PCRand RNA sequencing using P. gingivalis-lipopolysaccharide treated mouse peritoneal macrophages.@*RESULTS@#In comparison with healthy livers with Kupffer cells, the NASH liver combined with P. gingivalis infection-related macrophages showed significant heterogeneity. C1qb, C1qc, Mafb, Apoe, and Cd14 were highly expressed, but Cd209a, H2-Aa, H2-Ab1, and H2-DMb1, which are related to the antigen presentation function, were weakly expressed. Further in vivo and in vitro investigations indicated that the activation and infiltration of these macrophages may be due to local P. gingivalis-lipopolysaccharide accumulation.@*CONCLUSIONS@#P. gingivalis-lipopolysaccharide induces a local macrophage immunotolerance phenotype in nonalcoholic fatty liver, which may be the key mechanism of periodontitis pathogen infection that promotes NASH inflammation and pathogenesis. This study further clarifies the dysfunction and regulatory mechanisms of macrophages in the pathogenesis of P. gingivalis-infected NASH, thereby providing potential therapeutic targets for its clinical treatment.
Subject(s)
Mice , Animals , Non-alcoholic Fatty Liver Disease/pathology , Kupffer Cells/pathology , Porphyromonas gingivalis , Lipopolysaccharides/metabolism , Inflammation/pathology , Macrophages/metabolism , Mice, Inbred C57BLABSTRACT
While several previous studies have indicated the link between periodontal disease (PD) and myocardial infarction (MI), the underlying mechanisms remain unclear. Autophagy, a cellular quality control process that is activated in several diseases, including heart failure, can be suppressed by Porphyromonas gingivalis (P.g.). However, it is uncertain whether autophagy impairment by periodontal pathogens stimulates the development of cardiac dysfunction after MI. Thus, this study aimed to investigate the relationship between PD and the development of MI while focusing on the role of autophagy. Neonatal rat cardiomyocytes (NRCMs) and MI model mice were inoculated with wild-type P.g. or gingipain-deficient P.g. to assess the effect of autophagy inhibition by P.g. Wild-type P.g.-inoculated NRCMs had lower cell viability than those inoculated with gingipain-deficient P.g. This study also revealed that gingipains can cleave vesicle-associated membrane protein 8 (VAMP8), a protein involved in lysosomal sensitive factor attachment protein receptors (SNAREs), at the 47th lysine residue, thereby inhibiting autophagy. Wild-type P.g.-inoculated MI model mice were more susceptible to cardiac rupture, with lower survival rates and autophagy activity than gingipain-deficient P.g.-inoculated MI model mice. After inoculating genetically modified MI model mice (VAMP8-K47A) with wild-type P.g., they exhibited significantly increased autophagy activation compared with the MI model mice inoculated with wild-type P.g., which suppressed cardiac rupture and enhanced overall survival rates. These findings suggest that gingipains, which are virulence factors of P.g., impair the infarcted myocardium by cleaving VAMP8 and disrupting autophagy. This study confirms the strong association between PD and MI and provides new insights into the potential role of autophagy in this relationship.
Subject(s)
Mice , Rats , Animals , Porphyromonas gingivalis , Gingipain Cysteine Endopeptidases , Autophagosomes , Myocardium , Periodontal Diseases , Heart RuptureABSTRACT
Introducción: Porphyromonas gingivalis es un microorganismo presente en las periodontitis, productor de la enzima peptidil arginina desminasa, inductora de la citrulinación de proteínas que convierte en antígenos, y que son reconocidos por los anticuerpos antipéptido cíclico citrulinados, marcadores específicos de la artritis reumatoide. Estudios clínicos y epidemiológicos relacionan el hábito de fumar con la periodontitis y la artritis reumatoide. Objetivo: Evaluar la asociación entre el hábito de fumar, la periodontitis crónica y la artritis reumatoide. Métodos: Se realizó un estudio observacional, analítico, de corte transversal, de casos y controles de pacientes con diagnóstico de artritis reumatoide tratados en el Centro de Reumatología y pacientes atendidos por medicina interna en el Hospital Clínico Quirúrgico 10 de octubre de La Habana, en el periodo entre septiembre del 2017 y mayo del 2019. Se estudiaron las variables edad, sexo, hábito de fumar y estado periodontal evaluado a través del índice de enfermedad periodontal de Russell y el nivel de inserción clínica. Para identificar la asociación entre variables se empleó la prueba de ji al cuadrado y el odds ratio. Se respetaron las legislaciones éticas. Resultados: En el estudio prevaleció el grupo de 35 a 44 años y el sexo femenino. El hábito de fumar predominó en los pacientes artríticos, con manifiesto incremento de la prevalencia y gravedad de la enfermedad periodontal. Conclusiones: El hábito de fumar incrementó el riesgo de periodontitis crónica en ambos grupos, y con menos intensidad de riesgo en la artritis reumatoide.
Introduction: Porphyromonas gingivalis is a microorganism present in periodontitis, producer of the enzyme peptidyl arginine deminase that induces citrullination of proteins, turning them into antigens, which are recognized by anti-citrullinated cyclic peptide antibodies, specific markers of rheumatoid arthritis. Clinical and epidemiological studies link smoking with periodontitis and rheumatoid arthritis. Objective: To evaluate the association between smoking, the presence of chronic periodontitis and rheumatoid arthritis. Methods: An observational, analytical, cross-sectional study of cases and controls of patients with a diagnosis of rheumatoid arthritis treated at the Rheumatology Center and patients treated by Internal Medicine in 10 de Octubre Surgical- Clinic Hospital in Havana, between September 2017 and May 2019. The variables were: age, sex, smoking habit and periodontal status evaluated through the Russell Periodontal Disease Index and Level of Clinical Insertion. For the association and relationship between variables, the chi square and the odds ratio were used. Ethical legislation was respected. Results: In the study the group of 35 to 44 years old and the female sex prevailed. Smoking prevailed in arthritic patients with a remarkable increase in the prevalence and severity of periodontal disease. Conclusions: Smoking increased the risk of chronic periodontitis in both groups with less intensity of risk in rheumatoid arthritis.
Subject(s)
Female , Adult , Arthritis, Rheumatoid/complications , Smoking/adverse effects , Porphyromonas gingivalis/pathogenicity , Chronic Periodontitis/complicationsABSTRACT
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)
Subject(s)
Humans , Male , Female , Child , Adolescent , Orthodontic Brackets/adverse effects , Porphyromonas gingivalis/isolation & purification , Dental Plaque/microbiology , Orthodontic Appliances, Fixed/adverse effects , Epidemiology, Descriptive , Cross-Sectional Studies , Gingival Crevicular Fluid/microbiology , Observational Study , Mexico , Molecular Biology/methodsABSTRACT
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
Subject(s)
Humans , Antioxidants , Streptococcus mutans , Candida albicans , Porphyromonas gingivalis , Solanum lycopersicum , EthanolABSTRACT
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.
Subject(s)
Animals , Male , Rats , Biomarkers/metabolism , Hippocampus/metabolism , Nestin/metabolism , Neural Stem Cells/metabolism , Neurons/metabolism , Porphyromonas gingivalis/metabolism , Rats, Sprague-Dawley , Tail/metabolismABSTRACT
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.
Subject(s)
Humans , Cell Polarity , Interleukin-10/metabolism , Lipopolysaccharides/pharmacology , Macrophage Activation , Macrophages/drug effects , MicroRNAs/metabolism , NF-kappa B/metabolism , Porphyromonas gingivalis , RNA, Messenger/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
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.
Subject(s)
Computer Simulation , Methylene Blue , Molecular Docking Simulation , Photosensitizing Agents , Porphyromonas gingivalisABSTRACT
La periodontitis afecta en Uruguay a más de 340 mil individuos. Para optimizar su tratamiento además del mantenimiento profesional de la salud periodontal, del control de placa por el paciente, y del control de otros factores de riesgo como las enfermedades sistémicas, estrés, hábito tabáquico y susceptibilidad genética, se administran frecuentemente antibióticos como coadyuvantes. Entre los periodontopatógenos más prevalentes Porphyromonas gingivalis es el más estudiado a nivel molecular, pero se encontró poca información para la región acerca de su sensibilidad a los antibióticos. Dado el compromiso etiológico en la enfermedad periodontal de Porphyromonas gingivalis, es relevante la determinación de su resistencia antimicrobiana in vitro para un tratamiento racional considerando la temporalidad de los resultados. Objetivo general: Contribuir al conocimiento de la infección por Porphyromonas gingivalis y al uso racional de antibióticos en el tratamiento de la periodontitis. Objetivos específicos: caracterizar la población estudiada según factores de riesgo modificables y no modificables para periodontitis, cultivar e identificar Porphyromonas gingivalis de la bolsa periodontal de la población, determinar la concentración inhibitoria mínima (CIM) a los antibióticos y la producción de betalactamasa, analizar las concentraciones en el fluido gingival y su relación con la CIM, establecer la relación riesgo -beneficio- costo de los diferentes tratamientos antibióticos. Métodos: se realizó la toma de muestras de la microbiota del surco gingival a los pacientes que reunieron los criterios de inclusión y firmaron consentimiento informado.Las muestras se incubaron en diferentes medios microbiológicos específicos, para identificar el microorganismo diana, determinar su sensibilidad a la amoxicilina, asociación amoxicilina ácido clavulánico, metronidazol, tetraciclina, clindamicina, ciprofoxacina y moxifloxacina. Resultados: de un total de 29 pacientes en un rango de 23 a 78 años y con bolsas periodontales iguales o mayores a 4 mm, se obtuvieron 12 aislamientos de Porphyromonas gingivalis. Estos aislamientos se realizaron en pacientes que tenían bolsas periodontales con una profundidad mayor a 4 mm, con una edad comprendida entre 41 a 70 años. Si bien el 48% de la población era fumadora, sólo el 29% presentó aislamientos. Todos los pacientes diabéticos presentaron Porphyromonas gingivalis. Los 12 aislamientos fueron betalactamasa positivos y por lo tanto resistentes a las penicilinas G y V, aminopenicilinas, amoxipenicilinas, ureidopenicilinas y carboxipenicilinas. El rango de CIM fue: amoxicilina de 8-16 mg.L-1, asociación amoxicilina ácido clavulánico 0.25/0.125-1/0.5 mg.L-1, metronidazol de 0,25- 1 mg.L-1, tetraciclina 0.25-0.5 mg.L-1, clindamicina 0.25-0.5 mg.L-1, ciprofloxacina de 1-2 mg.L-1y moxifloxacina 0.064-0.25 mg.L-1. El parámetro farmacocinética/farmacodinamia (PK/PD) en el fluido gingival indica actividades antimicrobianas insuficientes para amoxicilina-ácido clavulánico y satisfactoria para el resto de los antibióticos a los que la bacteria es sensible in vitro. En orden decreciente, la mejor relación riesgo-beneficio-costo fue para el metronidazol seguido de clindamicina, la moxifloxacina y la tetraciclina. Conclusiones: el metronidazol y clindamicina podrían ser los más beneficiosos como coadyuvantes para la disminución de la carga del periodonto patógeno en el fluido crevicular. En segundo lugar podrían usarse tetraciclina y moxifloxacina debido a su menor seguridad.Este es el primer estudio nacional que aporta datos sobre proporción de Porphyromonas gingivalis en periodontitis por método de cultivo, de su sensibilidad in vitro a los antimicrobianos y de la actividad antimicrobiana de estos últimos en el fluido crevicular.
Subject(s)
Humans , Periodontitis/drug therapy , Bacteroidaceae Infections , Porphyromonas gingivalis , Drug Resistance, Bacterial , Epidemiologic Studies , Epidemiology, Descriptive , Prospective StudiesABSTRACT
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.
Subject(s)
Animals , Mice , Periodontitis , Stress, Psychological , Sympathetic Nervous System , Porphyromonas gingivalis , Virulence FactorsABSTRACT
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)
Subject(s)
Humans , Male , Female , Adult , Oral Hygiene , Fusobacterium nucleatum , Porphyromonas gingivalis , SARS-CoV-2 , DNA , Oral Hygiene Index , Periodontal Index , Polymerase Chain Reaction , Dental Plaque/microbiology , Electrophoresis, Agar Gel , Age and Sex Distribution , Gingivitis/epidemiology , MexicoABSTRACT
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)
Subject(s)
Humans , Male , Female , Oils, Volatile/therapeutic use , Microbial Sensitivity Tests , Enterococcus faecalis , Porphyromonas gingivalis , Lactobacillus acidophilus , Analysis of Variance , Chromatography, GasABSTRACT
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.