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Bone tissue regeneration strategies have incorporated the use of natural polymers, such as hydroxyapatite (nHA), chitosan (CH), gelatin (GEL), or alginate (ALG). Additionally, platelet concentrates, such as platelet-rich fibrin (PRF) have been suggested to improve scaffold biocompatibility. This study aimed to develop scaffolds composed of nHA, GEL, and CH, with or without ALG and lyophilized PRF, to evaluate the scaffold's properties, growth factor release, and dental pulp stem cells (DPSC), and osteoblast (OB) derived from DPSC viability. Four scaffold variations were synthesized and lyophilized. Then, degradation, swelling profiles, and morphological analysis were performed. Furthermore, PDGF-BB and FGF-B growth factors release were quantified by ELISA, and cytotoxicity and cell viability were evaluated. The swelling and degradation profiles were similar in all scaffolds, with pore sizes ranging between 100 and 250 µm. FGF-B and PDGF-BB release was evidenced after 24 h of scaffold immersion in cell culture medium. DPSC and OB-DPSC viability was notably increased in PRF-supplemented scaffolds. The nHA-CH-GEL-PRF scaffold demonstrated optimal physical-biological characteristics for stimulating DPSC and OB-DPSC cell viability. These results suggest lyophilized PRF improves scaffold biocompatibility for bone tissue regeneration purposes.
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Alginatos , Supervivencia Celular , Quitosano , Pulpa Dental , Durapatita , Gelatina , Osteoblastos , Fibrina Rica en Plaquetas , Células Madre , Andamios del Tejido , Humanos , Pulpa Dental/citología , Quitosano/química , Quitosano/farmacología , Gelatina/química , Fibrina Rica en Plaquetas/química , Fibrina Rica en Plaquetas/metabolismo , Andamios del Tejido/química , Células Madre/efectos de los fármacos , Células Madre/citología , Células Madre/metabolismo , Supervivencia Celular/efectos de los fármacos , Durapatita/química , Durapatita/farmacología , Alginatos/química , Alginatos/farmacología , Osteoblastos/efectos de los fármacos , Osteoblastos/citología , Adhesión Celular/efectos de los fármacos , Ingeniería de Tejidos/métodos , Células CultivadasRESUMEN
OBJECTIVE: To identify the characteristics of the oral microbiota and the relationship of the dental caries and periodontal status in patients aged 0 to 18 years with non-syndromic cleft lip and palate (CLP). MATERIALS AND METHODS: A systematic review of the literature was carried out. Five databases were consulted, including publications in English, Spanish and Portuguese. The evaluations of the quality of the observational studies and the experimental studies were carried out with the Newcastle-Ottawa scale and CONSORT guidelines, respectively. The risk of bias of the studies was determined using Rev Manager 5.4, and 5 publications were meta-analyzed. RESULTS: The cariogenic microbiota of children and adolescents with cleft lip and palate was similar to that of children without clefts, although with higher counts of Streptococcus mutans and Lactobacillus spp. The periodontopathogenic microbiota was related to the presence of Campylobacter spp, Fusobacterium spp, Fusobacterium nucleatum, Prevotella intermedia/nigrescens, Parvimonas micra and Porphyromonas gingivalis, considered microorganisms with high pathogenic capacity. Heterogeneity was shown in relation to the microbiota and the type of fissure, presenting numerous microorganisms associated with the pre- and post-surgical condition (cheilorrhaphy and palatorrhaphy) such as Staphylococcus aureus, Streptococcus beta hemolyticus, Klebsiella pneumoniae and Klebsiella oxytoca, Moraxella catarrhalis, Candida spp, Candida albicans, Candida krusei and Candida tropicalis. The meta-analysis revealed that patients with cleft lip and palate were 2.03 times more likely to have caries than the control group (p<0.005). CONCLUSION: In the microbiota, there was a great diversity of microorganisms that can vary according to the type of fissure and surgical interventions predisposing patients to a greater probability of dental caries, it is important to take into account the technique used to describe the oral microbiota in order to be able to compare the different studies. CLINICAL RELEVANCE: Studying the microbiota and the relationship of dental caries and periodontal status in children and adolescents with cleft lip and palate can facilitate the comprehensive care of patients with these conditions.
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Labio Leporino , Fisura del Paladar , Caries Dental , Microbiota , Niño , Humanos , AdolescenteRESUMEN
Saliva samples are important for diagnosis, because they are noninvasive and easy to acquire. The objective of this cross-sectional study was to investigate the value saliva samples have in detecting SARS-CoV-2 in comparison to nasal swabs and a new system named CovidCheck. A standard methodology identified the virus in 185 nasopharyngeal swabs and saliva samples revealing a sensitivity, specificity and positive and negative predictive values of 82,100,100 and 94.67%, respectively for saliva samples. Viral presence in saliva samples with the standard methodology in comparison to the CovidCheck system was evaluated in 67 samples with sensitivity, specificity and positive and negative predictive values of 68, 81, 68 and 81%, respectively. In conclusion, our results highlight the usefulness saliva samples have in detecting respiratory viral infections. However, presence of viral inhibitors and viral load in saliva, and the patient's clinical status should be considered as they might affect amplifying systems results.
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COVID-19 , Nasofaringe , SARS-CoV-2 , Saliva , Sensibilidad y Especificidad , Humanos , Saliva/virología , COVID-19/diagnóstico , SARS-CoV-2/aislamiento & purificación , Estudios Transversales , Nasofaringe/virología , Carga Viral , Masculino , Femenino , Adulto , Persona de Mediana Edad , AncianoRESUMEN
Objective: To investigate the frequencies and bacterial load of three species of periodontal bacteria in samples from oropharyngeal cancer patients versus healthy individuals. Study design: This is a case-control study based on biopsies collected from tumor tissues obtained from patients with oropharyngeal squamous cell carcinoma between 2016 and 2017 and shed oral mucosal epithelial cells that were collected from controls using the Cepimax® brush, carrying out several brushings towards the posterior third edge of the tongue and the cheek. Porphyromonas gingivalis, Tannerella forsythia and Prevotella intermedia detection and absolute quantification was determined through q-PCR. Statistical analysis included a U- test, X 2 , Fisher's exact test, odds ratio (OR) and Conditional logistic regression analysis and unconditional regression analysis (p < 0.05). Results: A total of 48 donors older than 55 years old participated in this study. The population was distributed into 24 patients (cases) and 24 controls. A robust association was established in cases and controls with significance regarding Prevotella intermedia (OR: 15.00) and Porphyromonas gingivalis (OR:11.00). In the comparison between the amount of each bacteria in the groups, P. intermedia showed a higher bacterial load in oropharyngeal cancer patients (p = 0.04). However, multivariate analysis adjusted to the presence of different bacteria and the diverse confounding variables did not reveal significant differences for oropharyngeal cancer association. Conclusion: P. gingivalis and P. intermedia were detected more frequently in the group of patients with cancer. The bivariate analysis of the bacterial load evidenced significant differences for Prevotella intermedia, suggesting that it could be associated with oropharyngeal cancer.
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Resumen Objetivo: Comparar la expresión de mRNA y proteínas de SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 y HAND1 en pacientes con lesión intra-epitelial cervical de bajo y alto grado, con posterior progresión o regresión. Material y Método: Se realizó análisis de expresión de genes mediante RT-PCR y análisis de expresión de proteínas por inmunohistoquímica. El análisis estadís tico fue realizado con las pruebas: Wilcoxon, coeficiente de correlación de Spearman e índice de concordancia. Las muestras fueron pareadas en momento 1 y momento 2. Resultados: SFRP1 mostró tendencia de mayor expresión de mRNA en lesión intra-epitelial de bajo grado (momento 2) Vs. alto grado (momento 1). La expresión de proteínas por inmunohistoquímica de SFRP1 en casos de progresión (83,3 %) mostró disminución en su graduación (p = 0,0313*); los demás genes en estudio no tuvieron cambios estadísticamente significativos. Discusión: SFRP1 mostró comportamiento ajustado a resultados de estudios previos donde se encontró hipermetilado en lesiones intra-epiteliales de alto grado; su subexpresión por hipermetilación se reportó en otros canceres, proceso que colabora con su silenciamiento y transición epitelial-mesenquimatosa del cáncer de cuello uterino. Conclusiones. SFRP1 es potencial biomarcador en lesiones preneoplásicas del cuello uterino asociadas al virus de papiloma humano.
Abstract Objective. The aim of this work was to compare the expression of mRNA and proteins of SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 and HAND1 in patients with low and high grade cervical intraepithelial lesion, with subsequent progression or regression. Material and Methods: Gene expression analysis was conducted through RT-PCR and protein expression analysis was performed by immunohistochemistry. The statistics analysis were Wilcoxon test, Spearman's correlation coefficient and concordance index. The samples were paired during moment 1 (initial patient diag nosis) and moment 2 (follow-up histological diagnosis). Results: SFRP1 showed a trend of higher mRNA expression in low-grade intra-epithelial lesions (moment 2) Vs. high-grade (moment 1). The expression of proteins by immunohistochemistry of SFRP1 in progression cases (83.3%) showed a decrease in its graduation (p = 0.0313*); the other genes under study had no statistically significant. Discussion: SFRP1 showed a biological behavior adjusted to the results of previous studies where hypermethylation was found in high-grade intra-epithelial lesions; its subexpression by hypermethylation has been reported in other cancers, a process that collaborates with its silencing and epithelial-mesenchymal tran sition of cervical. Conclusions. SFRP1 is a potential biomarker in preneoplastic lesions of the cervix associated with human papillomavirus.
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Humanos , Femenino , Adulto , Papiloma , Sondas de ADN de HPV , Virus , Proteínas , Neoplasias del Cuello Uterino , Progresión de la Enfermedad , AlphapapillomavirusRESUMEN
Human papilloma virus (HPV) is a DNA virus associated with the development of cervical, penile, anal, vulvar, and oral cancers. In recent years, there has been an increase in oral cancer, which could be due to changes in sexual behavior in the general population. In México, there is scarce information on this regard, which prompted us to study HPV infection prevalence in the oral cavity of an indigenous community from the municipality of Siltepec, Chiapas, Mexico. Oral samples from 198 individuals were obtained with cytobrush for virus detection by nested PCR, using MY09/MY11 and GP5+/GP6+ primers, and positive samples were sequenced for HPV genotyping. We observed 12.1% HPV infection prevalence, which depended on gender, number of sexual partners, lack of using condoms, and oral sex practices. In contrast, no significant association between HPV infection and tobacco or alcohol consumption was detected. Furthermore, sequencing analyzes were performed where HPV-13 (21/24), -16 (2/24), -32 (1/24), -81 (1/24), and -83 (1/24) were evidenced and HPV-16 European/Asian and Asian/American E6 variants identified. These results demonstrated an important prevalence of HPV infection in the oral cavity of a Mexican indigenous community, where the predominant genotypes were associated with benign pathologies, and showed that high-risk genotype variants derived from different lineages.
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Boca/virología , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , Adulto , Anciano , Estudios Transversales , Femenino , Genotipo , Neoplasias de Cabeza y Cuello/virología , Humanos , Pueblos Indígenas/estadística & datos numéricos , Masculino , México/epidemiología , México/etnología , Persona de Mediana Edad , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/virología , Prevalencia , Conducta Sexual , Parejas Sexuales , Carcinoma de Células Escamosas de Cabeza y Cuello/virologíaRESUMEN
Antecedentes: Los factores de crecimiento utilizados en salud se pueden obtener de una fuente autóloga de primera generación llamada plasma rico en plaquetas (PRP). La diversidad de protocolos para prepararlos genera resultados variables en cuanto al tiempo entre la activación del PRP y sus efectos sobre la proliferación y viabilidad celular. Objetivo: Evaluar proliferación y viabilidad celular de fibroblastos de ligamento periodontal y osteoblastos tratados con PRP en diferentes concentraciones y tiempos de aplicación. Métodos: Se cultivaron líneas celulares de fibroblastos y osteoblastos y se preparó el PRP de sangre venosa de un adulto sano mediante centrifugación, seguido de activación con CaCl2 al 10 %. El efecto sobre la proliferación de las líneas celulares tras la aplicación de PRP y plasma pobre en plaquetas al 1 %, al 3 % y al 5 % se evaluó a las 0, 12, 24, 48 y 72 horas después de su activación mediante MTS. El grupo control consistió en cultivos sin tratamiento. Los datos se analizaron mediante las pruebas de Chi cuadrado, Fischer y McNemar. Resultados: Se observó un aumento de la viabilidad en células tratadas con PRP 24 horas después de su activación en una concentración del 5%. El ensayo de viabilidad celular mostró diferencias estadísticamente significativas entre el grupo experimental y el grupo control (p = 0,05). Conclusión: Los cultivos de fibroblastos y osteoblastos mostraron una tendencia a mayor viabilidad 24 horas después de a la activación con PRP al 5 %.
Background : Growth factors used in health treatments can be obtained from a first-generation source called platelet-rich plasma. The variety of protocols to prepare PRP produces variable results regarding PRP activation time and its effects on cell proliferation and viability. Purpose: To evaluate proliferation and cell viability of periodontal ligament fibroblasts and osteoblasts stimulated with PRP in several concentrations and times after PRP activation. Methods: An in vitro study was carried out using periodontal ligament fibroblast and osteoblast cell cultures. PRP from venous blood of a healthy adult was prepared through centrifugation and activated with 10% CaCl2. The effect on cell proliferation after application of 1%, 3%, and 5% PRP and platelet-poor plasma was evaluated at 0, 12, 24, 48, and 72 hours after activation through MTS. The control group consisted of culture that did not receive any treatment. Data were analyzed using Chi square, Fisher, and McNemar tests. Results: The cell viability assay showed statistically significant differences between the experimental and the control groups. Cell viability increased in cells treated with 5% PRP 24 hours after activation (p = 0.05). Conclusions: Fibroblast and osteoblast cell lines tended to be more viable 24 hours after activation with 5% PRP.
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Humanos , Ligamento Periodontal , Proliferación Celular , Osteoblastos , Técnicas In Vitro , Plasma Rico en PlaquetasRESUMEN
Resumen Introducción y objetivo: Los tejidos dentales contienen células madre mesenquimales con diversas capacidades de diferenciación. El objetivo de este estudio fue comparar la capacidad de osteodiferenciación de células madre de pulpa dental y ligamento periodontal cultivadas con diferentes medios de osteodiferenciación. Materiales y métodos: Se obtuvieron 24 muestras de tejido pulpar y 30 de ligamento periodontal de terceros molares, se aislaron y se cultivaron con 5 condiciones diferentes de osteoinducción. La osteodiferenciación fue evaluada mediante RT-PCR y Rojo Alizarin. Resultados: Las células aisladas fueron caracterizadas mediante citometría de flujo. Se observó una mayor expresión de Colágeno tipo I para células de pulpa dental, mientras que las células de ligamento periodontal expresaron mayores niveles de RUNX2. Las células Madre de ligamento periodontal presentaron mejor perfil de osteodiferenciación cuando fueron cultivadas en el medio 5, mientras que las células madre extraídas de la pulpa dental presentaron mejores características de osteodiferenciación en el medio de cultivo 4. La única diferencia entre estos dos medios fue la concentración de 50uM de Ácido Ascórbico para el medio 5 y 200uM de Ácido Ascórbico para el medio 4. Conclusiones: Las células obtenidas de pulpa dental y ligamento periodontal mostraron características de células madre. La capacidad de osteodiferenciación de estas células fue dependiente de las concentraciones de los sustratos aplicados al cultivo.
Introduction and objective: Dental tissues have mesenchymal stem cells with capacities to differentiate in several tissues. The aim of this research was to compare the ability of osteodifferentiation of stem cells isolated of dental pulp and periodontal ligament cultured with different media. Materials and methods: 24 samples of pulp tissue and 30 samples of periodontal ligament were obtained of third molars. Stem cells isolated by the explant method were subjected to 5 osteodifferentiation conditions. Osteodifferentiation was evaluated by RT-PCR and Alizarin Red. Results: The isolated cells were characterized as stem cells by flow cytometer. Collagen type I showed the highest expression in dental pulp Stem Cells while Runx2 was increased in periodontal ligament Stem Cells. Medium 5 showed the best conditions for osteodifferentiation of periodontal ligament Stem cells while Dental pulp Stem Cells showed better osteogenic profile in medium 4. The unique difference between mediums 4 and 5 was the concentration of Ascorbic Acid 50um in medium 5 and 200um in medium 4). Conclusions: The cells obtained both pulp and ligament showed features of Stem Cells. The osteogenic capacity of cells was depending of the concentrations of the substrates applied.
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Objetivos: comparar cuatro métodos de restauración del ADN en plasma y láminas cérvico-uterinas como una herramienta para mejorar la calidad de la muestra. Métodos: a 20 muestras de plasma sanguíneo y 20 muestras de láminas citológicas, se les realizó aislamiento de ADN mediante kit comercial y fenol-cloroformo. A todas las muestras se les realizó un tratamiento pre-PCR con cuatro diferentes tipos de actividad de ADN polimerasa: 1. Exonucleasa y endonucleasa 5'-3'. 2. Exonucleasa 5'-3'. 3. Klenow, y 4. Klenow más ligasa. Los diferentes métodos se evaluaron mediante PCR en tiempo real con el gen ALU. Resultados: todos los métodos de restauración mejoran la calidad del ADN en los dos tipos de muestras. El método 3 mostró mejores resultados en plasma y en lámina, incrementando la concentración del ADN de 0,0022 ng/µL a 0,6474 ng/µL en láminas de citología y de 0,0039 ng/µL a 0,435 ng/µL en plasma sanguíneo. Conclusiones: ADN de las muestras de plasma y lámina al ser tratadas con un proceso de restauración aumenta la calidad del ADN en comparación a las muestras no tratadas.
Objetives: To compare four methods of restoration of DNA in plasma and PAP smears as a tool to improve the quality of the samples. Methods: 20 blood samples and 20 PAP smears samples, we performed DNA isolation by commercial kit and phenol-chloroform respectively. Then all samples underwent a pre-PCR treatment with four different types of activity DNA polymerase: 1. Exonuclease and endonuclease 5'-3'. 2. Exonuclease 5'-3'. 3. Klenow, and 4. Klenow more ligase. Different restoration methods were evaluated quantitatively by real-time PCR with gene ALU. Results: All restoration methods improve the quality of DNA in both types of samples. However, the 3th method showed better results in both plasma and PAP smears, increasing the concentration of DNA from 0.0022 ng/mL to 0.6474 ng/mL in PAP smears and 0.0039 ng/mL to 0.435 ng/mL in blood plasma. Conclusions: DNA from plasma samples and PAP smears to be treated with a restoration process increases the quality of DNA compared to untreated samples.
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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.
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Calendula/inmunología , Fibroblastos , Microbiología , Cicatrización de Heridas , Proliferación CelularRESUMEN
El virus de papiloma humano (vph) es el principal factor de riesgo asociado al cáncer cervical, y es la causa de muerte más común por cáncer entre las mujeres en Colombia. Por tanto, crece el interés a nivel mundial y nacional por mejorar las estrategias de control y diagnóstico de la infección; incluyendo técnicas de diagnóstico molecular que identifiquen y diferencien tipos virales específicos para así tener mejor entendimiento de la dinámica del virus en la historia natural de la infección por vph. En el presente trabajo se detectó el vph en 363 pacientes diagnosticadas con lesiones asc-us o lsil en su citología, pertenecientes al programa de tamizaje de cáncer de cérvix de la eps Sanitas. Sólo a 302 de estas muestras se les pudo realizar genotipificación por Reverse Line Blot, de éstas el 20,5% (62/302 pacientes) fueron positivas para vph; los tipos virales de alto riesgo estuvieron presentes en el 82,2% y los de bajo riesgo, en el 17,8%. Por primera vez se realiza un acercamiento a la descripción de tipos virales específicos, encontrados en muestras con diagnóstico citológico de asc-us o lsil en Bogotá.
Human papillomavirus (hpv) is the principal risk factor associated with cervical cancer, the most common malignancy among women in Colombia. Therefore, a growing concern globally and nationally to improve strategies to control and diagnosis of infection, including molecular diagnostic techniques to identify and differentiate specific hpv types thus have a better understanding of the dynamics of the virus in history natural infection. In the present work was performed hpv detection and genotyping in 302 of 363 patients who normally attended the screening program of the eps Sanitas, and were subsequently diagnosed with asc-us lesions. We found hpv in 20.5% (62/302 patients) divided into 82.2% for high-risk viral types and 17.8% for low risk. For the first time was a description of specific viral types found in samples of asc-us in Bogotá.
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Humanos , Sondas de ADN de HPV , Infecciones por Papillomavirus , Neoplasias del Cuello UterinoRESUMEN
El ADN libre en sangre incrementa con algunas condiciones patológicas y ciertos estados fisiológicos. Varios reportes en la literatura han resaltado que el ADN libre en plasma o suero tiene potencial clínico como una posible herramienta para el pronóstico de cáncer en humanos. Sin embargo, hasta el momento no se tienen valores de referencia de individuos sanos con un tamaño de muestra representativo y tampoco se han descrito valores para poblaciones específicas como la bogotana. Es por ello que en el presente estudio se cuantificó la concentración de ADN libre en personas sanas de la población bogotana y así se estableció un rango normal o valor de referencia, adicionalmente se analizó la relación entre los niveles de ADN libre y las características como edad y género. La concentración de ADN libre en la población bogotana fue de 0,72 ng/μL y no se encontraron diferencias significativas entre las edades y los géneros.