Human Coronary Artery Endothelial Cell Response to Porphyromonas gingivalis W83 in a Collagen Three-Dimensional Culture Model.

P. gingivalis has been reported to be an endothelial cell inflammatory response inducer that can lead to endothelial dysfunction processes related to atherosclerosis; however, these studies have been carried out in vitro in cell culture models on two-dimensional (2D) plastic surfaces that do not simulate the natural environment where pathology develops. This work aimed to evaluate the pro-inflammatory response of human coronary artery endothelial cells (HCAECs) to P. gingivalis in a 3D cell culture model compared with a 2D cell culture. HCAECs were cultured for 7 days on type I collagen matrices in both cultures and were stimulated at an MOI of 1 or 100 with live P. gingivalis W83 for 24 h. The expression of the genes COX-2, eNOS, and vWF and the levels of the pro-inflammatory cytokines thromboxane A2 (TXA-2) and prostaglandin I2 (PGI2) were evaluated. P. gingivalis W83 in the 2D cell culture increased IL-8 levels at MOI 100 and decreased MCP-1 levels at both MOI 100 and MOI 1. In contrast, the 3D cell culture induced an increased gene expression of COX-2 at both MOIs and reduced MCP-1 levels at MOI 100, whereas the gene expression of eNOS, vWF, and IL-8 and the levels of TXA2 and PGI2 showed no significant changes. These data suggest that in the collagen 3D culture model, P. gingivalis W83 induces a weak endothelial inflammatory response.

Biomarkers for the severity of periodontal disease in patients with obstructive sleep apnea:IL-1 ß, IL-6, IL-17A, and IL-33.

Objective: This study aims to compare the salivary and gingival crevicular fluid (GCF) concentrations of five cytokines: IL-1ß, IL-6, IL-17A, IL-33, and Tumor Necrosis Factor-alpha (TNF-α) in patients with OSA and their association with periodontitis. Methods: Samples of saliva and GCF were obtained from 84 patients classified into four groups according to periodontal and OSA diagnosis: G1(H) healthy patients, G2(P) periodontitis and non-OSA patients, G3(OSA) OSA and non-periodontitis patients, and G4(P-OSA) periodontitis and OSA patients. The cytokines in the samples were quantified using multiplexed bead immunoassays. Data were analyzed with the Kruskal-Wallis test, Dunn's multiple comparisons test, and the Spearman correlation test. Results: Stage III periodontitis was the highest in patients with severe OSA (69%; p=0.0142). Similar levels of IL-1ß and IL-6 in saliva were noted in G2(P) and G4(P-OSA). The IL-6, IL-17A and IL-33 levels were higher in the GCF of G4(P-OSA). There was a significant positive correlation between IL-33 in saliva and stage IV periodontitis in G4(P-OSA) (r s  = 0.531). The cytokine profile of the patients in G4(P-OSA) with Candida spp. had an increase of the cytokine's levels compared to patients who did not have the yeast. Conclusions: OSA may increase the risk of developing periodontitis due to increase of IL-1ß and IL-6 in saliva and IL-6, IL-17A and IL-33 in GCF that share the activation of the osteoclastogenesis. Those cytokines may be considered as biomarkers of OSA and periodontitis.

Cryptic Oral Microbiota: What Is Its Role as Obstructive Sleep Apnea-Related Periodontal Pathogens?

Periodontitis has been commonly linked to periodontopathogens categorized in Socransky's microbial complexes; however, there is a lack of knowledge regarding "other microorganisms" or "cryptic microorganisms", which are rarely thought of as significant oral pathogens and have been neither previously categorized nor connected to illnesses in the oral cavity. This study hypothesized that these cryptic microorganisms could contribute to the modulation of oral microbiota present in health or disease (periodontitis and/or obstructive sleep apnea (OSA) patients). For this purpose, the presence and correlation among these cultivable cryptic oral microorganisms were identified, and their possible role in both conditions was determined. Data from oral samples of individuals with or without periodontitis and with or without OSA were obtained from a previous study. Demographic data, clinical oral characteristics, and genera and species of cultivable cryptic oral microorganisms identified by MALDI-TOF were recorded. The data from 75 participants were analyzed to determine the relative frequencies of cultivable cryptic microorganisms' genera and species, and microbial clusters and correlations tests were performed. According to periodontal condition, dental-biofilm-induced gingivitis in reduced periodontium and stage III periodontitis were found to have the highest diversity of cryptic microorganism species. Based on the experimental condition, these findings showed that there are genera related to disease conditions and others related to healthy conditions, with species that could be related to different chronic diseases being highlighted as periodontitis and OSA comorbidities. The cryptic microorganisms within the oral microbiota of patients with periodontitis and OSA are present as potential pathogens, promoting the development of dysbiotic microbiota and the occurrence of chronic diseases, which have been previously proposed to be common risk factors for periodontitis and OSA. Understanding the function of possible pathogens in the oral microbiota will require more research.

Patients with obstructive sleep apnea can favor the predisposing factors of periodontitis by the presence of P. melaninogenica and C. albicans, increasing the severity of the periodontal disease.

Objective: The aim of this study was to analyze the cultivable oral microbiota of patients with obstructive sleep apnea (OSA) and its association with the periodontal condition. Methods: The epidemiology profile of patients and their clinical oral characteristics were determined. The microbiota was collected from saliva, subgingival plaque, and gingival sulcus of 93 patients classified into four groups according to the periodontal and clinical diagnosis: Group 1 (n = 25), healthy patients; Group 2 (n = 17), patients with periodontitis and without OSA; Group 3 (n = 19), patients with OSA and without periodontitis; and Group 4 (n = 32), patients with periodontitis and OSA. Microbiological samples were cultured, classified, characterized macroscopically and microscopically, and identified by MALDI-TOF-MS. The distribution of complexes and categories of microorganisms and correlations were established for inter- and intra-group of patients and statistically evaluated using the Spearman r test (p-value <0.5) and a multidimensional grouping analysis. Result: There was no evidence between the severity of OSA and periodontitis (p = 0.2813). However, there is a relationship between the stage of periodontitis and OSA (p = 0.0157), with stage III periodontitis being the one with the highest presence in patients with severe OSA (prevalence of 75%; p = 0.0157), with more cases in men. The greatest distribution of the complexes and categories was found in oral samples of patients with periodontitis and OSA (Group 4 P-OSA); even Candida spp. were more prevalent in these patients. Periodontitis and OSA are associated with comorbidities and oral conditions, and the microorganisms of the orange and red complexes participate in this association. The formation of the dysbiotic biofilm was mainly related to the presence of these complexes in association with Candida spp. Conclusion: Periodontopathogenic bacteria of the orange complex, such as Prevotella melaninogenica, and the yeast Candida albicans, altered the cultivable oral microbiota of patients with periodontitis and OSA in terms of diversity, possibly increasing the severity of periodontal disease. The link between yeasts and periodontopathogenic bacteria could help explain why people with severe OSA have such a high risk of stage III periodontitis. Antimicrobial approaches for treating periodontitis in individuals with OSA could be investigated in vitro using polymicrobial biofilms, according to our findings.

Viability and Adhesion of Periodontal Ligament Fibroblasts on a Hydroxyapatite Scaffold Combined with Collagen, Polylactic Acid-Polyglycolic Acid Copolymer and Platelet-Rich Fibrin: A Preclinical Pilot Study.

Background: Conventional periodontal therapy relies on bone regeneration strategies utilizing scaffolds made of diverse materials, among which collagen, to promote cell adhesion and growth. Objective: To evaluate periodontal ligament fibroblast (HPdLF) cell adhesion and viability for periodontal regeneration purposes on hydroxyapatite scaffolds containing collagen (HAp-egg shell) combined with polylactic acid−polyglycolic acid copolymer (PLGA) and Platelet-Rich Fibrin (PRF). Methods: Four variations of the HAp-egg shell were used to seed HPdLF for 24 h and evaluate cell viability through a live/dead assay: (1) (HAp-egg shell/PLGA), (2) (HAp-egg shell/PLGA + collagen), (3) (HAp-egg shell/PLGA + PRF) and (4) (HAp-egg shell/PLGA + PRF + collagen). Cell adhesion and viability were determined using confocal microscopy and quantified using central tendency and dispersion measurements; significant differences were determined using ANOVA (p < 0.05). Results: Group 1 presented low cell viability and adhesion (3.70−10.17%); groups 2 and 3 presented high cell viability and low cell adhesion (group 2, 59.2−11.1%, group 3, 58−4.6%); group 4 presented the highest cell viability (82.8%) and moderate cell adhesion (45%) (p = 0.474). Conclusions: The effect of collagen on the HAp-egg shell/PLGA scaffold combined with PRF favored HPdLF cell adhesion and viability and could clinically have a positive effect on bone defect resolution and the regeneration of periodontal ligament tissue.

Expression of CD44 in previously grafted alveolar bone / Expresión del marcador óseo CD44 en hueso alveolar previamente injertado

Objetive: To determine the expressions of the bone surface marker CD44 in samples of alveolar bone previously regenerated with allograft, xenograft, and mixed, using the technique of guided bone regeneration. Material and Methods: This exploratory study was approved by the institutional research and ethics committee. By means of intentional sampling and after obtaining informed consent for tissue donation, 20 samples of alveolar bone previously regenerated with guided bone regeneration therapy with particulate bone graft and membrane were taken during implant placement. The samples were stained with hematoxylin-eosin for histological analysis, and by immunohistochemistry for the detection of CD44. Results: Sections with hematoxylin-eosin showed bone tissue with the presence of osteoid matrix and mature bone matrix of usual appearance. Of the CD44+ samples, 80% were allograft and 20% xenograft. The samples with allograft-xenograft were negative. There were no differences in the intensity of CD44 expression between the positive samples. The marker was expressed in osteocytes, stromal cells, mononuclear infiltrate, and some histiocytes. Eighty percent of the CD44+ samples and 100% of the samples in which 60 or more cells were labelled corresponded to allografts (p=0.000). A total of 67% of the samples from the anterior sector, and 40% from the posterior sector were CD44+ (p=0.689). Conclusion: This study shows for the first time that guided bone regeneration using allografts is more efficient for the generation of mature bone determined by the expression of CD44, compared to the use of xenografts and mixed allograft-xenograft, regardless of the regenerated anatomical area.

Objetivo: Determinar la expresión del marcador de membrana óseo CD44 en muestras de hueso alveolar previamente regenerado con aloinjerto, xenoinjerto y mezcla mediante la técnica de regeneración ósea guiada. Material y Métodos: Con aval del Comité de Investigación y Ética, se realizó un estudio exploratorio. Por muestreo intencional y firma de consentimiento informado de donación, se tomaron durante la colocación del implante, 20 muestras de hueso alveolar previamente regenerado con terapia de regeneración ósea guiada con injerto óseo particulado y membrana. Las muestras fueron teñidas con hematoxilina-eosina para el análisis histológico y por inmunohistoquímica para la detección del CD44. Resultados: : Los cortes con hematoxilina-eosina mostraron tejido óseo con presencia de matriz osteoide y matriz ósea madura de aspecto usual. De las muestras CD44+, 80% fueron de aloinjerto y 20% de xenoinjerto. Las muestras con aloinjerto-xeoninjerto fueron negativas. No hubo diferencias en la intensidad de la expresión del CD44 entre las muestras positivas. El marcador se expresó en osteocitos, células estromales, infiltrado mononuclear y algunos histiocitos. El 80% de las muestras CD44+ y el 100% de las muestras con marcación de 60 o más células correspondían a aloinjertos (p=0,000). El 67% de las muestras del sector anterior y el 40% del sector posterior fueron CD44+ (p=0,689). Conclusión: Este estudio muestra por primera vez que la regeneración ósea guiada usando aloinjertos, es más eficiente para la generación de hueso maduro determinado por la expresión de CD44, comparado con el uso de xenoinjertos y mezcla de aloinjerto-xenoinjerto, independientemente del sector anatómico regenerado.

Integrated genomic epidemiology and phenotypic profiling of Clostridium difficile across intra-hospital and community populations in Colombia.

Clostridium difficile, the causal agent of antibiotic-associated diarrhea, has a complex epidemiology poorly studied in Latin America. We performed a robust genomic and phenotypic profiling of 53 C. difficile clinical isolates established from diarrheal samples from either intrahospital (IH) or community (CO) populations in central Colombia. In vitro tests were conducted to evaluate the cytopathic effect, the minimum inhibitory concentration of ten antimicrobial agents, the sporulation efficiency and the colony forming ability. Eleven different sequence types (STs) were found, the majority present individually in each sample, however in three samples two different STs were isolated. Interestingly, CO patients were infected with STs associated with hypervirulent strains (ST-1 in Clade-2). Three coexistence events (two STs simultaneously detected in the same sample) were observed always involving ST-8 from Clade-1. A total of 2,502 genes were present in 99% of the isolates with 95% of identity or more, it represents a core genome of 28.6% of the 8,735 total genes identified in the set of genomes. A high cytopathic effect was observed for the isolates positive for the two main toxins but negative for binary toxin (TcdA+/TcdB+/CDT- toxin production type), found only in Clade-1. Molecular markers conferring resistance to fluoroquinolones (cdeA and gyrA) and to sulfonamides (folP) were the most frequent in the analyzed genomes. In addition, 15 other markers were found mostly in Clade-2 isolates. These results highlight the regional differences that C. difficile isolates display, being in this case the CO isolates the ones having a greater number of accessory genes and virulence-associated factors.

Inflamatory response in pregnant women with high risk of preterm delivery and its relationship with periodontal disease: a pilot study.

Periodontal disease and its inflammatory response have been related to adverse outcomes in pregnancy such as preterm birth, preeclampsia and low birth weight. This study analyzed systemic inflammatory response in patients with high risk of preterm delivery and its relationship to periodontal disease. A pilot study was conducted for a case and control study, on 23 patients at risk of preterm delivery and 23 patients without risk of preterm delivery as controls. Exclusion criteria were patients who had received periodontal treatment, antibiotic or antimicrobial agents within the past three months, or with infections or baseline diseases such as diabetes or hypercholesterolemia. All patients underwent periodontal assessment, laboratory tests (complete blood count, lipid profile, baseline glycemia) and quantification of cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α and TNF-γ). Higher levels of pro-inflammatory cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α and TNF-γ) were found in patients with chronic periodontitis than in patients with gingivitis or periodontal health. These cytokines, in particular IL-2, IL-10 and TNF-α, were higher in patients at high risk of preterm delivery. Patients with high risk of preterm delivery had higher severity of periodontal disease as well as higher levels of the pro-inflammatory markers IL-2, IL-4, IL-6, IL-10, TNF-α and TNF-γ.

La enfermedad periodontal y su repuesta inflamatoria ha sido relacionada con desenlaces adversos del embarazo como el parto pretérmino, preeclampsia y bajo peso al nacer. La presente investigación analizó la respuesta inflamatoria sistèmica en pacientes embarazadas con alto riesgo de parto pretérmino y su relación con la enfermedad periodontal. Se realizó un estudio piloto de casos y controles, en el cual se contó con 23 pacientes que presentaban riesgo de parto pretérmino como casos y 23 pacientes sin riesgo de parto pretérmino como controles. Fueron excluidas las pacientes que hubieran recibido tratamiento periodontal, antibióticos o antimicrobianos en los últimos tres meses, que tuvieran infecciones, o enfermedades de base como diabetes o hipercolesterolemia. A todas las pacientes se les hicieron valoración periodontal, exámenes de laboratorio (cuadro hemático, perfil lipídico, glucemia basal) y cuanti-ficación de citocinas (IL-2, IL-4, IL-6, IL-10, TNF-α e TNF-γ). En las pacientes con periodontitis crónica se encontraron niveles más elevados en las citocinas proinflamatorias (IL-2, IL-4, IL-6, IL-10, TNF-α e TNF-γ) en comparación con las pacientes con gingivitis o sanas periodontales. Estas citocinas se encontraron más elevadas en las pacientes con alto riesgo de parto pretérmino, en especial la IL-2, IL-10 y TNF-α. Las pacientes con alto riesgo de parto pretérmino presentaron mayor severidad de la enfermedad periodontal y adicionalmente niveles aumentados de los marcadores pro inflamatorios IL-2, IL-4, IL-6, IL-10, TNF-α e TNF-γ.

Inflammatory response in pregnant women with high risk of preterm delivery and its relationship with periodontal disease: a pilot study / Respuesta inflamatoria en pacientes embarazadas con alto riesgo de parto pretérmino y su relación con la enfermedad periodontal. Estudio piloto

Periodontal disease and its inflammatory response have been related to adverse outcomes in pregnancy such as preterm birth, preeclampsia and low birth weight. This study analyzed systemic inflammatory response in patients with high risk of preterm delivery and its relationship to periodontal disease. A pilot study was conducted for a case and control study, on 23 patients at risk of preterm delivery and 23 patients without risk of preterm delivery as controls. Exclusion criteria were patients who had received periodontal treatment, antibiotic or antimicrobial agents within the past three months, or with infections or baseline diseases such as diabetes or hypercholesterolemia. All patients underwent periodontal assessment, laboratory tests (complete blood count, lipid profile, baseline glycemia) and quantification of cytokines (IL2, IL4, IL6, IL10, TNFα and INFγ). Higher levels of proinflammatory cytokines (IL2, IL4, IL6, IL10, TNFα and INFγ) were found in patients with chronic periodontitis than in patients with gingivitis or periodontal health. These cytokines, in particular IL2, IL10 and TNFα, were higher in patients at high risk of preterm delivery. Patients with high risk of preterm delivery had higher severity of periodontal disease as well as higher levels of the proinflammatory markers IL2, IL4, IL6, IL10, TNFα and INFγ (AU)

La enfermedad periodontal y su repuesta inflamatoria ha sido relacionada con desenlaces adversos del embarazo como el parto pretérmino, preeclampsia y bajo peso al nacer. La presente investigación analizó la respuesta inflamatoria sistémica en pacientes embarazadas con alto riesgo de parto pretérmino y su relación con la enfermedad periodontal. Se realizó un estudio piloto de casos y controles, en el cual se contó con 23 pacientes que presentaban riesgo de parto pretérmino como casos y 23 pacientes sin riesgo de parto pretérmino como controles. Fueron excluidas las pacientes que hubieran recibido tratamiento periodontal, antibióticos o antimicrobianos en los últimos tres meses, que tuvieran infecciones, o enfermedades de base como diabetes o hipercolesterolemia. A todas las pacientes se les hicieron valoración periodontal, exámenes de laboratorio (cuadro hemático, perfil lipídico, glucemia basal) y cuanti ficación de citocinas (IL2, IL4, IL6, IL10, TNFα e INFγ). En las pacientes con periodontitis crónica se encontraron niveles más elevados en las citocinas proinflamatorias (IL2, IL4, IL6, IL10, TNFα e INFγ) en comparación con las pacientes con gingivitis o sanas periodontales. Estas citocinas se encontraron más elevadas en las pacientes con alto riesgo de parto pretérmino, en especial la IL2, IL10 y TNFα. Las pacientes con alto riesgo de parto pretérmino presentaron mayor severidad de la enfermedad periodontal y adicional mente niveles aumentados de los marcadores pro inflamatorios IL2, IL4, IL6, IL10, TNFα e INFγ (AU)

Comparación de la síntesis de interleucina- 1β por monocitos y linfocitos B estimulados con lipopolisacárido en pacientes conenfermedad periodontal / Interleukin-1β production by peripheral blood monocytes and B lymphocytes in patients with periodontal disease

Antecedentes: la interleucina-1β- (IL-1β) se considera un mediador en la pérdidaósea en enfermedad periodontal. No es claro si existen diferencias en la producciónde IL-1b ex vivo por linfocitos B (LB) y monocitos (Mo) de sangre periféricaentre pacientes sanos y con enfermedad periodontal. Objetivo: cuantificar laproducción de IL-1β por LB y Mo de sangre periférica de individuos con enfermedadperiodontal y sin esta en presencia de lipopolisacárido (LPS). Métodos: setomaron células de cinco individuos con diagnóstico sano/gingivitis, cinco conperiodontitis crónica (PC) y cinco con periodontitis agresiva (PAg). Los Mo y LB desangre periférica se purificaron con anticuerpos anti-CD14 y anti-CD19 marcadoscon perlas magnéticas. La pureza de las poblaciones celulares fue confirmadapor citometría de flujo. Las células se estimularon con LPS y la producción deIL-1b se cuantificó por ELISA. Resultados: las concentraciones basales de IL-1bproducidas por LB fueron menores en sujetos con PC comparados con sujetoscon PAg (p = 0,03) e individuos sanos (p = 0,0079). En presencia del estímulo, seincrementaron sin encontrarse diferencias entre los grupos. La IL-1b producidapor los Mo a nivel basal y frente al estímulo fue similar entre los sujetos, peromayor comparada con la liberada por los LB (p = 0,0079 en PC y p = 0,00571 en PAg)en pacientes con periodontitis. Conclusiones: la producción de IL-1b por LB enel estado basal está significativamente disminuida en pacientes con PC, comparadocon sanos y PAg, pero una vez estimulados con LPS, la respuesta es similaren los tres grupos. La capacidad de los LB para producir IL-1b en respuesta alestímulo con LPS es limitada comparada con la de los Mo...

loss in patients with periodontal disease. It is unclear whether there are differencesin the production of IL-1 b ex vivo by B cells and monocytes from peripheralblood between healthy patients and with periodontal disease. Objective:To quantify the production of IL-1b by monocytes and peripheral blood B cellsof individuals with and without periodontal disease in the presence of lipopolysaccharide(LPS). Methods: Monocytes and peripheral blood B cells from 5healthy/gingivitis, 5 chronic periodontitis (CP) and 5 aggressive periodontitis(AgP) patients were purified with anti-CD14 and anti-CD19 marked with magneticbeads. The purity of the cell populations was confirmed by flow citometry. Cellswere stimulated with LPS and the production of IL-1b was quantified by ELISA.Results: Basal levels of IL-1b produced by B lymphocytes were lower in subjectswith chronic periodontitis than in healthy (p = 0.0079) and aggressive periodontitispatients (p = 0.03). In presence of LPS, the IL-1b levels are increased withno differences between groups. IL-1b produced by monocytes at baseline andwith LPS, was similar between subjects, but higher compared to that releasedby B lymphocytes (p = 0.0079 in chronic periodontitis, p = 0.00571 in aggressiveperiodontitis) in the periodontitis groups. Conclusions: The production of IL-1bby LB at baseline is significantly reduced in CP patients compared to healthy andAgP, but once stimulated with LPS, the response is similar in the three groups.The ability of LB to produce IL-1b in response to stimulation with LPS is lowercompared with monocytes...

Efecto de la Calendula officinalis en la proliferación del fibroblasto gingival humano / Effect of Calendula officinalis on the proliferation of human gingival fibroblast

Antecedentes: la Calendula officinalis tiene múltiples propiedades terapéuticas, dentro de las que se encuentra su capacidad para mejorar procesos de cicatrización. Por ello resulta interesante estudiar su posible valor para tratar patologías orales. Objetivo: observar los efectos que producen tres presentaciones de C. officinalis en diferentes concentraciones en la proliferación de los fibroblastos gingivales humanos. Método: a fibroblastos gingivales humanos (FGH) en quinto pase, obtenidos de explantes de donantes sanos, se les realizaron estímulos con extracto etanólico de C. officinalis (EEC), tintura de C. officinalis (TC) y enjuague K-Trix® en concentraciones de 750, 500, 150, 100 μg/ml y se observó la proliferación de los FGH a las 12, 24 y 48 horas en relación con un grupo de células sin estimular, utilizando la prueba MTA con Cell Titer 96®. Resultados: el mayor efecto proliferativo se logró con el EEC en concentraciones de 750 y 500 μg/ml a las 12 horas. La TC a 100 y 150 μm/ml a las 48 horas inhibió el crecimiento, mientras que el enjuague K-Trix® no tuvo efecto en la proliferación de los FGH en ninguna concentración en ningún tiempo. Conclusión: la proliferación de los FGH depende de la presentación de la C. officinalis, del tiempo y de la concentración.

Background: Calendula officinalis has multiple therapeutic properties such as improving the process of cicatrization. Therefore, it is interesting its possible value and use to treat oral pathologies. Objective: To observe the effect of three presentations of C. officinalis in different concentrations on the proliferation of the human gingival fibroblasts. Methods: Human Gingival Fibroblasts (FGH) in fifth pass were obtained from healthy donor explants and exposed to stimuli with ethanolic extract of C. officinalis (EEC), tincture of C. officinalis (TC) and K-Trix® rinsing in concentrations of 750, 500, 150, 100 μg/ml. Proliferation of the FGH was observed at 12, 24 and 48 hours in relation with a group of cells without getting any stimulation, using the MTA test with Cell Titer 96®. Results: The major proliferative effect was achieved by the EEC in concentrations of 750 and 500 μg/ml at 12 hours. The tincture of 100 and 150 μg/ml C. officinalis at 48 hours generated inhibition in cell growth whereas the rinsing K-Trix® did not have effect on the proliferation of the FGH in any concentration at any time. Conclusion: The proliferation of the FGH depends on the presentation of the C. officinalis, time, and concentration.

Cytokines produced by CD4+ T cells against a synthetic GTF-I (1301-1322) peptide of Streptococcus mutans in naturally sensitized humans

Streptococcus mutans (S. mutans) es el principal agente etiologico de la caries dental. Las proteínas PAc y glucosiltransferasas (GTFs) son factores de virulencia de este microorganismo relacionados con su fisiopatogenia y han sido usados en investigaciones de una vacuna para la caries dental. El objetivo de este estudio fue observar si GTFs (1301-1322) tiene la capacidad de activar las células T CD4+ en CMSP de humanos naturalmente sensibilizados, identificar el tipo de respuesta y establecer su relación con la caries dental. 30 individuos clasificados en los siguientes 3 grupos, fueron estudiados: caries activa (AC), historia de caries (HC) y libres de caries (H). Muestras de sangre fueron tomadas de cada individuo. La estimulación antígeno específico y la citometría de flujo fueron usadas para determinar las células productoras de citoquina IFN-y (citoquinas tipo 1) e IL-13 (citoquinas tipo 2). Se encontró respuesta de memoria celular frente a GTF-I (1301-1322) en humanos naturalmente sensibilizados. Tres tipos de respuesta fueron detectados: TH0, TH1 y NR. Se encontró un mayor porcentaje de LTCD4+ productores de IFN-y que de IL-13 (p=0.006). No se encontraron diferencias estadísticamente significativas para las otras variables estudiadas para los tres grupos (p<0.05). Se concluye que la respuesta inmune celular específica frente al péptido sintético GTF-I (1301-1322) de S, mutans, no es diferente entre los individuos sensibilizados naturalmente, resistentes a caries y con caries.

Comparación de técnicas de disgregación mecánica, enzimática y mecano-enzimática en la extracción de poblaciones linfocitarias y macrófagos en tejidos con periodontitis crónica avanzada / Comparison of mechanical, enzymatic and mechanical-enzymatic dissagregation tecniques in extraction of lymphocytary populations and macrophagues in tissues with chronic advanced periodontitis

ANTECEDENTES: En este estudio se compararon tres técnicas de disgregación en la recuperación de poblaciones linfocitarias y macrófagos en tejidos con periodontitis crónica avanzada. OBJETIVO: comparar, por citometría de flujo,el recuento de poblaciones linfocitarias y macrófagos de biopsias de tejidos periodontalmente afectados, con tres procesos previos de disgregación celular (mecánica, enzimática con colagenasa y mecano-enzimática) para observar cuál de éstos es más efectivo en la recuperación celular y produce menos alteraciones en los marcadores de superficie. MÉTODOS: Se realizó un estudio descriptivo comparativo donde se eligió una muestra de 11 biopsias de tejidos con periodontitis crónica avanzada.El procedimiento consistió en tomar las biopsias por medio de biseles internos que se colocaban en un Eppendorf con el medio de transporte. En el grupo 1 se realizó la técnica de disgregación mecánica Medimachine®, en el grupo 2 técnica enzimática (colagenasa 2%), y el en grupo 3 técnica mecano-enzimática (Medimachine® y colagenasa 2%). Posteriormente las muestras fueron analizadas por un citómetro de flujo FACSAria (Becton Dickinson San José California). RESULTADOS: No se encontraron diferencias estadísticamente significativas entre las tres técnicas de disgregación en la recuperación de poblaciones linfocitarias; sin embargo, hubo una mayor recuperación de macrófagos con la técnica mecano-enzimática comparada con la mecánica, siendo estas diferencias estadísticamente significativas.CONCLUSIONES: Las tres técnicas estudiadas presentaron resultados comparables en cuanto a recuperación y daño a los marcadores de superficie,por lo cual, por tiempo y costo, se aconseja realizar una técnica mecánica con Medimachine®.

BACKGROUND: Three disaggregation techniques for the recovery of lymphocyte and macrophage populations from tissues with advanced chronic periodontitis were analyzed. OBJECTIVE: Compare by flow cytometry the recovery of lymphocyte and macrophage populations from biopsies of periodontal tissues with periodontitis by using three processes of cellular disaggregration (mechanical, enzymatic with colagenase and mechanical-enzymatic) to observe which of these is more effective and produces fewer alterations in the surface markers. METHODS: A comparative descriptive study was carried out with a sample of 11 biopsies of tissues taken from sites with advanced chronic periodontitis. The procedure consisted of taking the biopsies by means of internal bevels and placing them in an Eppendorf for transportation. In group 1, the mechanical technique of disaggregation (Medimachine®) was carried out. In group 2, the enzymatic technique (colagenase 2%) was used. In group 3, the mechanical-enzymatic technique (Medimachine® and colagenase 2%) was perfomred. Later on, the samples were analyzed with a FACS-Aria cytometer (Becton Dickinson San José California). RESULTS: There were not statistically significant differences among the three disaggregation techniques for the recovery of lymphocyte populations. However, there was a bigger recovery of macrophages with the mechanical-enzymatic technique when compared to the other two techniques, being statistically significant. CONCLU SIONS: The three techniques studied produce similar results in the recovery and damage to the surface markers; hence, considering time and cost constrains, it is advisable to carry out the mechanical technique with Medimachine®.

Principios de inmunidad pasiva y activa: usos y aplicabilidad / Pinciples of passive and active immunity: usefulness and applicability

Cuando un huésped está inmunizado, tiene mecanismos específicos de reconocimiento y defensa ante una sustancia extraña en particular (antígeno), ya sea de manera natural o artificial. Ésta inmunidad puede ser lograda en forma pasiva o activa. Si los productos de defensa se originan en un sitio endógeno por medio de la estimulación antigénica del propio sistema inmune del sujeto, la inmunidad es activa. Si por otro lado, los anticuerpos o células son producidos en un donador, por efecto de la estimulación antigénica, y luego son transferidas a un receptor, la inmunidad conferida es pasiva.