Small Molecules Accelerate Skin Wound Healing: Shikonin Efficacy and Mechanism of Action in Mice / Las Moléculas Pequeñas Aceleran la Curación de Heridas en la Piel: Eficacia de Shikonin y Mecanismo de Acción en Ratones

SUMMARY: The objective of this study was to investigate the therapeutic wound healing potential and molecular mechanisms of shikonin as small molecules in vitro. A mouse burn model was used to explore the potential therapeutic effect of shikonin; we traced proliferating cells in vivo to locate the active area of skin cell proliferation. Through the results of conventional pathological staining, we found that shikonin has a good effect on the treatment of burned skin and promoted the normal distribution of skin keratin at the damaged site. At the same time, shikonin also promoted the proliferation of skin cells at the damaged site; importantly, we found a significant increase in the number of fibroblasts at the damaged site treated with shikonin. Most importantly, shikonin promotes fibroblasts to repair skin wounds by regulating the PI3K/AKT signaling pathway. This study shows that shikonin can effectively promote the proliferation of skin cell, and local injection of fibroblasts in burned skin can play a certain therapeutic role.

El objetivo de este trabajo fue investigar el potencial terapéutico de cicatrización de heridas y los mecanismos moleculares de la shikonina como moléculas pequeñas in vitro. Se utilizó un modelo de quemaduras en ratones para explorar el posible efecto terapéutico de la shikonina; Rastreamos las células en proliferación in vivo para localizar el área activa de proliferación de células de la piel. A través de los resultados de la tinción para patología convencional, encontramos que la shikonina tiene un buen efecto en el tratamiento de la piel quemada y promueve la distribución normal de la queratina de la piel en el sitio dañado. Al mismo tiempo, la shikonina también promovió la proliferación de células de la piel en el sitio dañado. Es importante destacar que encontramos un aumento significativo en la cantidad de fibroblastos en el sitio dañado tratado con shikonina. Lo más importante es que la shikonina promueve la función reparadora de fibroblastos en las heridas de la piel regulando la vía de señalización PI3K/ AKT. Este estudio muestra que la shikonina puede promover eficazmente la proliferación de células de la piel y que la inyección local de fibroblastos en la piel quemada puede desempeñar un cierto papel terapéutico.

Ayudas diagnósticas y diagnóstico diferencial de la enfermedad de Pompe

Introducción: La enfermedad de Pompe (EP) o glucogenosis tipo II es una enfermedad autosómica recesiva causada por mutaciones en el gen GAA que codifica para la proteína alfa-1,4-glucosidasa. Su deficiencia lleva a un almacenamiento anormal de glucógeno en los lisosomas de varias células, a través de los diferentes tejidos, lo que causa un compromiso musculoesquelético predominante. Contenidos: Los fenotipos de la enfermedad dependen de las variantes genéticas y de los niveles de la actividad enzimática residual. La enfermedad se presenta como EP de inicio infantil, EP de inicio tardío y EP intermedio, por lo que es de suma importancia su diagnóstico temprano, por medio de estudios moleculares como la secuenciación de Sanger y la secuenciación de nueva generación. Conclusiones: Se ha demostrado, mediante diferentes estudios, que las variaciones genéticas pueden diferir entre etnias, y es importante su caracterización molecular para determinar el tratamiento más adecuado, de acuerdo con el estado del material inmunológico de reacción cruzada (CRIM).

Introduction: Pompe disease (PD) or Glycogenosis Type II is a rare autosomal recessive disease caused by mutations in the GAA gene that codes for the alpha-1,4-glucosidase protein. Its deficiency leads to abnormal glycogen storage in the lysosomes of various cells throughout the different tissues causing a predominant musculoskeletal compromise. Contents: The phenotypes of the disease depend on the genetic variants and the levels of residual enzyme activity, presenting as infantile-onset PD, late-onset PD, and intermediate PD; Therefore, early diagnosis of the disease through molecular studies such as Sanger sequencing and new generation sequencing is of utmost importance. Conclusions: It has been shown through different studies that genetic variations can vary between ethnic groups and the molecular characterization of the variants is important to determine the most appropriate treatment depending on the state of the cross-reactive immunological material (CRIM)

Tumor inflamatorio miofibroblástico gastrointestinal. Reporte de caso / Gastrointestinal myofibroblastic inflammatory tumor. Case report

Introducción. El tumor miofibroblástico inflamatorio es una enfermedad proliferativa rara, de etiología incierta, caracterizada por la proliferación de miofibroblastos epitelioides o fusionados mezclados con células inflamatorias, predominantemente mononucleares. En general se considera una lesión benigna, aunque en algunos casos esta neoplasia ha mostrado un comportamiento agresivo en cuanto a recidiva local y metástasis. El tratamiento definitivo es la resección quirúrgica completa. Caso clínico. Paciente de 67 años con dos meses de evolución de fiebre y masa abdominal, en quien se realizó una tomografía computarizada de abdomen que identificó una lesión de aspecto infiltrativo tumoral, comprometiendo la grasa retroperitoneal en la transcavidad de los epiplones. Por vía percutánea se tomó una biopsia que informó un pseudotumor inflamatorio retroperitoneal. Fue llevado a cirugía radical abdominal, con patología quirúrgica final que describió un tumor miofibroblástico inflamatorio de compromiso multifocal, adherido a la serosa del estómago e intestino delgado, sin compromiso muscular. Discusión. El tumor inflamatorio miofibroblástico es una entidad rara, de etiología por esclarecer y difícil diagnóstico. Presentamos el caso clínico de un paciente con tumor inflamatorio miofibroblástico gastrointestinal.Conclusión. Se describe el caso clínico de un paciente con un tumor inflamatorio miofibroblástico gastrointestinal, de presentación rara en nuestro medio. Es importante la comparación con casos similares para poder hacer conclusiones útiles en la práctica clínica

Introduction. Inflammatory myofibroblastic tumor is a rare proliferative disease of uncertain etiology, characterized by the proliferation of epithelioid or fused myofibroblasts mixed with predominantly mononuclear inflammatory cells. In general, it is considered a benign lesion, although in some cases this neoplasm has shown aggressive behavior in terms of local recurrence and metastasis. The definitive treatment is complete surgical resection. Clinical case. A 67-year-old patient with a two-month history of fever and an abdominal mass underwent a computed tomography scan of the abdomen that identified an infiltrative tumor, compromising the retroperitoneum fat in the lesser cavity. A biopsy was taken percutaneously, which reported a retroperitoneal inflammatory pseudotumor. He was taken to radical abdominal surgery, with final surgical pathology describing an inflammatory myofibroblastic tumor with multifocal involvement attached to the serosa of the stomach and small intestine without muscle involvement. Discussion. Inflammatory myofibroblastic tumor is a rare entity, of unknown etiology and difficult to diagnose. We present a clinical case of gastrointestinal myofibroblastic inflammatory tumor to better understand this entity.Conclusion. The clinical case of a patient with a gastrointestinal myofibroblastic inflammatory tumor, a rare presentation in our environment, is described. Comparison with similar cases is important to draw useful conclusions in clinical practice

Wound healing effect of a one-week Aloe Vera mouthwash: a pilot study / Efeito de reparação tecidual de uma semana de enxaguante bucal de Aloe Vera: um estudo piloto

Aloe Vera, a perennial Liliaceae plant, has medical, cosmetic, and wound-healing properties. Aloe vera has antioxidant, anti-cancer, anti-diabetic, and regenerative effects. Glucommannan increases collagen synthesis and aids healing after ginivectomy treatment. Natural mouthwashes may offer gingival wound healing efficacy with reduced side-effects when compared to Chlorhexidine. Objective: the objective of this clinical study was to compare the effects on wound healing of a one-week Aloe vera mouthwash with chlorhexidine mouthwash before gingivectomy surgical therapy. Material and Methods:a total of 45 individuals experiencing inflammatory gingival enlargement were included in the study. They underwent professional mechanical plaque removal and were then randomly divided into three groups. In group I, comprising 15 patients, participants were advised to utilize 100% Aloe vera juice as a mouthwash twice daily. Group II, also consisting of 15 patients, was instructed to use Chlorhexidine (0.2%) mouthwash twice daily. The Control group, which consisted of 15 patients, was recommended to use a placebo mouth rinse in addition to mechanical plaque removal. During the second visit, which occurred one week after the initial visit, the enlarged gingival tissue was surgically removed through scalpel gingivectomy. Immunohistochemical (IHC) analysis was performed on the excised tissue to measure the l evels of fibroblast growth factor-2. Results: when compared to the control group, Aloe vera showed significant differences regarding the expression of fibroblast growth factor-2(FGF-2), and highly significant differences in angiogenesis. At the same time, there were substantial differences in angiogenesis w ith no significant differences in the expression of FGF2 between Chlorhexidine and control groups. Conclusion: aloe vera has exhibited potential wound-healing effects as i t s ignificantly affected the IHC expression of FGF2 and angiogenesis when used as an adjunct to plaque control before gingivectomy surgical therapy (AU)

Aloe Vera, uma planta perene de Liliaceae, tem propriedades médicas, cosméticas e cicatrizantes. Aloe vera tem efeitos antioxidantes, anticancerígenos, antidiabéticos e regenerativos. O glucomanano aumenta a síntese de colágeno e auxilia na cicatrização a pós o tratamento de gengivectomia. Enxaguatórios bucais naturais podem oferecer efi cácia na reparação de feridas gengivais com efeitos colaterais reduzidos quando comparados à clorexidina. Objetivo:O objetivo deste estudo clínico foi comparar os efeitos na cicatrização de feridas de uma semana de enxaguatório bucal de Aloe vera com clorexidina antes da terapia cirúrgica de gengivectomia. Material e Métodos: um total de 45 indivíduos com aumento gengival inflamatório foram incluídos no estudo. Eles foram submetidos à remoção mecânica profissional da placa e foramdivididos aleatoriamente em três grupos. No grupo I, composto por 15 pacientes, os participantes foram orientados a utilizar 100% de suco de Aloe vera como enxaguante bucal duas vezes ao dia. O grupo II, também composto por 15 pacientes, foi instruído a usar enxaguante bucal com clorexidina (0,2%) duas vezes ao dia. O grupo controle, composto por 15 pacientes, foi recomendado o uso de enxaguatório bucal placebo além da remoção mecânica da placa. Durante a segunda visita, que ocorreu uma semana após a visita inicial, o tecido gengival aumentado foi removido cirurgicamente por meio de gengivectomia com bisturi. A análise imuno-histoquímica (IHC) foi realizada no tecido excisado para medir os níveis do fator de crescimento de fibroblastos-2 (FGF-2). Resultados: quando comparado ao grupo controle, o Aloe vera apresentou diferenças significativas em relação àexpressão do FGF-2, e diferenças altamente significativas na angiogênese. Ao mesmo tempo, houve diferenças substanciais na angiogênese, sem diferenças significativas na expressão de FGF-2 entre a clorexidina e os grupos controle. Conclusão: Aloe vera exibiu potenciais efeitos de cicatrização de feridas, pois afetou significativamente a expressão IHC de FGF-2 e a angiogênese quando usada como adjuvante no controle de placa antes da terapia cirúrgica de gengivectomia (AU)

Lymphangioleiomyomatosis: circulating levels of FGF23 and pulmonary diffusion / Linfangioleiomiomatose: níveis circulantes de FGF23 e difusão pulmonar

ABSTRACT Objective: Lymphangioleiomyomatosis (LAM) is a rare, destructive disease of the lungs with a limited number of determinants of disease activity, which are a critical need for clinical trials. FGF23 has been implicated in several chronic pulmonary diseases. We aimed to determine the association between serum FGF23 levels and pulmonary function in a cohort of patients with LAM. Methods: This was a descriptive single-center study in which subjects with LAM and controls with unreported lung disease were recruited. Serum FGF23 levels were measured in all subjects. Clinical data, including pulmonary function testing, were retrospectively obtained from electronic medical records of LAM subjects. Associations between FGF23 levels and clinical features of LAM were explored via nonparametric hypothesis testing. Results: The sample comprised 37 subjects with LAM and 16 controls. FGF23 levels were higher in the LAM group than in the control group. In the LAM group, FGF23 levels above the optimal cutoff point distinguished 33% of the subjects who had nondiagnostic VEGF-D levels. Lower FGF23 levels were associated with impaired DLCO (p = 0.04), particularly for those with isolated diffusion impairment with no other spirometric abnormalities (p = 0.04). Conclusions: Our results suggest that FGF23 is associated with pulmonary diffusion abnormalities in LAM patients and elicit novel mechanisms of LAM pathogenesis. FGF23 alone or in combination with other molecules needs to be validated as a biomarker of LAM activity in future clinical research.

RESUMO Objetivo: A linfangioleiomiomatose (LAM) é uma doença rara e destrutiva dos pulmões com um número limitado de determinantes da atividade da doença, que são uma necessidade crítica para ensaios clínicos. O FGF23 já foi implicado em várias doenças pulmonares crônicas. O nosso objetivo foi determinar a associação entre os níveis séricos de FGF23 e a função pulmonar em uma coorte de pacientes com LAM. Métodos: Estudo descritivo unicêntrico no qual foram recrutados indivíduos com LAM e controles com doenças pulmonares não declaradas. Os níveis séricos de FGF23 foram medidos em todos os indivíduos. Os dados clínicos, incluindo testes de função pulmonar, foram obtidos retrospectivamente a partir dos prontuários eletrônicos dos indivíduos com LAM. As associações entre os níveis de FGF23 e as características clínicas da LAM foram exploradas por meio do teste de hipóteses não paramétrico. Resultados: A amostra incluiu 37 indivíduos com LAM e 16 controles. Os níveis de FGF23 foram mais altos no grupo LAM do que no grupo controle. No grupo LAM, níveis de FGF23 acima do ponto de corte ideal distinguiram 33% dos indivíduos com níveis não diagnósticos de VEGF-D. Níveis mais baixos de FGF23 estavam associados à DLCO comprometida (p = 0,04), particularmente naqueles com comprometimento isolado da difusão e sem outras alterações espirométricas (p = 0,04). Conclusões: Nossos resultados sugerem que o FGF23 está associado a alterações na difusão pulmonar em pacientes com LAM e potencialmente indicam novos mecanismos de patogênese da LAM. O FGF23 isoladamente ou em combinação com outras moléculas precisa ser validado como um biomarcador da atividade da LAM em futuras pesquisas clínicas.

The effect of scaffold hydroxyapatite derived from portunus pelagicus shell on the expression of fibroblast growth factor-2 (fgf-2) and bone morphogenetic proteins-2 (bmp -2) in the extraction sockets of cavia cobaya / El efecto del scaffold de hidroxiapatita derivada de portunus pelagicus sobre la expresión del factor de crecimiento de fibroblastos-2 (FGF-2) y las proteínas morfogenéticas óseas-2 (BMP-2) en los alvéolos dentarios de cavia cobaya

Objetive: To determine the expression of Fibroblast Growth Factor (FGF)-2 and Bone Morphogenetic Protein (BMP)-2 after application of scaffold hydroxyapatite from Rajungan crab shell (Portunus pelagicus) in the tooth extraction socket of Cavia cobaya. Material and Methods: This study used a post-test only control group design with 28 Cavia cobaya separated into two groups, control and treatment group. The left mandibular incisor was extracted, and socket preservation was conducted. A hydroxyapatite graft derived from crab shells was mixed with gelatin and eventually turned into a scaffold, which was afterward put into the extraction socket. After 7 days and 14 days, each group was terminated and examined using immunohistochemical staining to observe the expression of FGF-2 and BMP-2. One-Way Anova and Tukey HSD were used to examine the research data. Results: FGF-2 and BMP-2 expressions were observed higher in the group that received hydroxyapatite scaffold at the post-extraction socket than those in the group that did not receive hydroxyapatite scaffold. Conclusion: The application of a hydroxyapatite scaffold from Rajungan crab shell (Portunus pelagicus) to the tooth extraction socket can increase FGF-2 and BMP-2 expression.

Objetivo: Determinar la expresión del factor de crecimiento de fibroblastos (FGF)-2 y la proteína morfogenética ósea (BMP)-2 después de la aplicación de hidroxiapatita de andamio de caparazón de cangrejo Rajungan (Portunus pelagicus) en el alvéolo de extracción dental de Cavia cobaya. Material y Métodos: Este estudio utilizó un diseño de grupo de control solo posterior a la prueba con 28 Cavia cobaya separados en dos grupos, grupo de control y grupo de tratamiento. Se extrajo el incisivo mandibular izquierdo y se realizó la preservación del alvéolo. Un injerto de hidroxiapatita derivado de caparazones de cangrejo se mezcló con gelatina y se convirtió en un andamio, que luego se colocó en el alvéolo de extracción. Después de 7 días y 14 días, se terminó cada grupo y se examinó mediante tinción inmunohistoquímica para observar la expresión de FGF-2 y BMP-2. Se utilizaron One-Way Anova y Tukey HSD para examinar los datos de la investigación. Resultados: Las expresiones de FGF-2 y BMP-2 se observaron más altas en el grupo que recibió la estructura de hidroxiapatita en el alvéolo posterior a la extracción que en el grupo que no recibió la estructura de hidroxiapatita. Conclusión: La aplicación de un andamio de hidroxiapatita de caparazón de cangrejo Rajungan (Portunus pelagicus) al alvéolo de extracción dental puede aumentar la expresión de FGF-2 y BMP-2.

Immune challenges upregulate the expression of cannabinoid receptors in cultured human odontoblasts and gingival fibroblasts / Los desafíos inmunológicos regulan la expresión de los receptores cannabinoides en cultivos de odontoblastos y fibroblastos gingivales humanos

ABSTRACT Odontoblasts and gingival fibroblasts play essential roles in the physiological and pathological processes of dental tissue. Cannabinoid receptors (CB1 and CB2) are involved in analgesia by modulating the función of calcium channels that inhibit the synthesis of some neurotransmitters. A better understanding of the physiology of these receptors would provide the possibility of using them as therapeutic targets in controlling dental pain. The aim of this study was to evaluate the presence and activity of cannabinoid receptors in human odontoblast-like cells (OLC) and human gingival fibroblasts (HGF). CB1 and CB2 transcription was analyzed by real-time PCR, proteins were detected by immunofluorescence, and functional cannabinoid receptors were evaluated by measuring intracellular calcium concentration after stimulation with cannabidiol (CBD) and pre-treatment with a CB1 antagonist, a CB2 inverse agonist and a TRPV1 antagonist. Transcripts for CB1 and CB2 were found in both odontoblasts and gingival fibroblasts. Cannabidiol induced an increase in [Ca2+]i in both cells types, but surprisingly, pre-treatment with selective cannabinoid antagonists attenuated this effect, suggesting a functional communication between specific cannabinoid receptors and other CBD target receptors. In conclusion, human odontoblasts and gingival fibroblasts express functional CB1 and CB2 cannabinoid receptors, which could be modulated to improve the treatment of pain or dental sensitivity.

RESUMEN Los odontoblastos y los fibroblastos gingivales desempeñan funciones esenciales en los procesos fisiológicos y patológicos de los tejidos dentales. Los receptores cannabinoides (CB1 y CB2) participan en la analgesia mediante la modulación de la función de canales de calcio que inhiben la síntesis de algunos neurotransmisores. Un mejor conocimiento de su fisiología abre la posibilidad de utilizar estos receptores como dianas terapéuticas en el control del dolor dental. Este trabajo tuvo como objetivo evaluar la presencia y la actividad de los receptores cannabinoides en células humanas similares a los odontoblastos (OLC) y en fibroblastos gingivales humanos (HGF). Se analizó la transcripción de CB1 y CB2 por PCR en tiempo real, la detección de las proteínas por inmunofluorescencia y se evaluaron los receptores cannabinoides funcionales midiendo las concentraciones de calcio intracelular, tras la estimulación con cannabidiol (CBD) y el pretratamiento con un antagonista de CB1, un agonista inverso de CB2 y un antagonista de TRPV1. Se encontraron mensajeros para CB1 y CB2 tanto en odontoblastos como en fibroblastos gingivales. El cannabidiol indujo un aumento de la [Ca2+]i en ambos tipos de células, pero sorprendentemente el pretratamiento con antagonistas cannabinoides selectivos atenuó este efecto, lo que sugiere una comunicación funcional entre receptores cannabinoides específicos y otros receptores diana del CBD. En conclusión, los odontoblastos humanos y los fibroblastos gingivales expresan receptores cannabinoides CB1 y CB2 funcionales, que podrían ser modulados para mejorar el tratamiento del dolor o la sensibilidad dental.

Mitochondrial energy metabolism in baby hamster kidney (BHK-21/C13) cells treated with karnozin extra® / Metabolismo energético mitocondrial en células de riñón de hámster bebé (BHK-21 / C13) tratadas con karnozin extra®

SUMMARY: Carnosine is known as a natural dipeptide, which inhibits the proliferation of tumor cells throughout its action on mitochondrial respiration and cell glycolysis. However, not much is known about its effects on the metabolism of healthy cells. We explored the effects of Karnozin EXTRA® capsule with different concentrations of L-carnosine, on the cell viability and the expressions of intermediate filament vimentin (VIM) and superoxide dismutase (SOD2) in normal fibroblasts BHK-21/C13. Furthermore, we investigated its action on the energy production of these cells. Cell viability was quantified by the MTT assay. The Clark oxygen electrode (Oxygraph, Hansatech Instruments, England) was used to measure the "intact cell respiration rate", state 3 of ADP-stimulated oxidation, maximum oxidation capacity and the activities of complexes I, II and IV. Results showed that Karnozin EXTRA® capsule in concentrations of 2 and 5 mM of L-carnosine did not induce toxic effects and morphological changes in treated cells. Our data revealed a dose-dependent immunofluorescent signal amplification of VIM and SOD2 in the BHK-21/C13 cell line. This supplement substantially increased the recorded mitochondrial respiration rates in the examined cell line. Due to the stimulation of mitochondrial energy production in normal fibroblasts, our results suggested that Karnozin EXTRA® is a potentially protective dietary supplement in the prevention of diseases with altered mitochondrial function.

RESUMEN: La carnosina se conoce como dipéptido natural, que inhibe la proliferación de células tumorales a través de su acción sobre la respiración mitocondrial y la glucólisis celular. Sin embargo, no se sabe mucho de sus efectos sobre el metabolismo de las células sanas. Exploramos los efectos de la cápsula Karnozin EXTRA® con diferentes concentraciones de L-carnosina, sobre la viabilidad celular y las expresiones de vimentina de filamento intermedio (VIM) y superóxido dismutasa (SOD2) en fibroblastos normales BHK-21 / C13. Además, estudiamos su acción sobre la producción de energía de estas células. La viabilidad celular se cuantificó mediante el ensayo MTT. Se utilizó el electrodo de oxígeno Clark (Oxygraph, Hansatech Instruments, Inglaterra) para medir la "tasa de respiración de células intactas", el estado 3 de oxidación estimulada por ADP, la capacidad máxima de oxidación y las actividades de los complejos I, II y IV. Los resultados mostraron que la cápsula de Karnozin EXTRA® en concentraciones de 2 y 5 mM de L- carnosina no indujo efectos tóxicos ni cambios morfológicos en las células tratadas. Nuestros datos revelaron una amplificación de señal inmunofluorescente dependiente de la dosis de VIM y SOD2 en la línea celular BHK-21 / C13. Este suplemento aumentó sustancialmente las tasas de respiración mitocondrial registradas en la línea celular examinada. Debido a la estimulación de la producción de energía mitocondrial en fibroblastos normales, nuestros resultados sugirieron que Karnozin EXTRA® es un suplemento dietético potencialmente protector en la prevención de enfermedades con función mitocondrial alterada.

The antioxidant activity of lemongrass leaves extract against fibroblasts oxidative stress / Atividade antioxidante do extrato das folhas de capim-limão contra o estresse oxidativo em fibroblastos

Objective : The purpose of this research is to assess the antioxidant activity of lemongrass leaves extract in terms of lowering ROS generation and its effect on the viability and proliferation of fibroblasts under oxidative stress. Material and Methods: The antioxidant activity was measured using the DPPH method and the ROS assay was carried out by fluorescent H2DCFDA staining. Viability and proliferation assays were performed using the Cell Counting Kit-8 (CCK-8) and was read at 450 nm using microplate reader. The groups were divided into 8, namely fibroblasts without treatment (comparison group), fibroblast induced by H2O2 (negative control), fibroblast with H2O2 then treated with ascorbic acid (positive control), and fibroblast with H2O2 then treated with lemongrass leaves extract at various concentrations (10, 20, 30, 40, and 50 ppm). Results: The results showed that the antioxidant activity of lemongrass leaves extract had an IC value of 64.17 ppm. ROS production were reduced by the LgLE of all concentrations if compared with negative control (p=0.819). LgLE can maintained the fibroblast viability with 10 ppm of LgLE was the most optimum concentration (p<0.05). LgLE can induced the proliferation of fibroblast, with the most effective was at 24 h of observation (p<0.05). Conclusion: Lemongrass leaves extract has a strong antioxidant activity that can reduce oxidative stress and increase the viability and proliferation of fibroblasts with the optimum concentration is at 10 ppm. (AU)

Objetivo: O intuito deste estudo foi determinar a ação antioxidante do extrato das folhas de capim-limão no que se refere a diminuição da produção de espécies reativas do oxigênio (EROS) e o seu efeito na viabilidade e proliferação de fibroblastos submetidos à estresse oxidativo. Material e Métodos: A atividade antioxidante foi medida utilizando o método de DPPH e o ensaio de EROS foi realizado pela coloração fluorescente de H2DCFDA. Os ensaios de proliferação e viabilidade foram realizados utilizando-se o kit de contagem de células CCK-8 em microplacas de leitura à 450nm. Os grupos foram divididos em 8: Fibroblastos sem tratamento (grupo controle), Fibroblastos tratados com H2O2 (controle negativo), Fibroblastos tratados com H2O2 e extrato da folha de capim-limão em concentrações variadas (10, 20, 30, 40 e 50 ppm). Resultados: Os resultados mostraram que a atividade antioxidante do extrato de capim-limão teve uma IC50 (com o numeroal subscrito) com valor de 64.17ppm. A produção de ROS foi reduzida pelo tratamento com o extrato em todas as concentrações testadas quando comparado ao grupo controle negativo (p=0.819). O extrato manteve a viabilidade dos fibroblastos, sendo 10ppm a concentração menos tóxica (p<0.05). LgLE pôde induzir a proliferação de fibroblastos, sendo que a melhor eficiencia foi após 24h de observação (p<0.05). Conclusão: O extrato das folhas de capim-limão apresentam forte atividade antioxidante reduzindo o estresse oxidativo e aumentando a viabilidade e proliferação de fibroblastos, sendo a concentração ótima de 10ppm. (AU)

Marcadores FGF-23 y Klotho en enfermedad renal crónica y su implicación cardiovascular, una revisión sistemática / FGF-23 and Klotho markers in chronic kidney disease and its cardiovascular implication, a systematic review

La principal causa de mortalidad en enfermedad renal crónica (ERC), en el 80% de pacientes se da por enfermedad cardiovascular asociada, los parámetros bioquímicos clásicos del metabolismo óseomineral aún no logran explicar la progresión patológica por tanto, se ha empezado a estudiar nuevos marcadores con relación al daño cardiovascular, donde se ha encontrado al marcador FGF-23 y su correceptor Klotho, implicados en la génesis del daño cardiovascular y enfermedad óseomineral asociada al fosforo, que en conjunto causan remodelamiento cardiovascular, lo que ha empezado aclarecer aún más esta dinámica fisiopatológica. Esta revision busca conocer la implicación de los marcadores FGF-23 y Klotho en ERC y el riesgo cardiovascular asociado y para ello realizó una revision sistemática de literatura, indagando en bases biomédicas como COCHRANE, Embase, LILACS, Scielo, Pub-Med, EBSCO, Hinari, Sociedades médicas, así como tesauros MeSH propios de esta investigación.

The main cause of mortality in chronic kidney disease (CKD), in 80% of patients, is due to associated cardiovascular disease, the classic biochemical parameters of bone-mineral metabolism will not yet be able to explain the pathological progression, therefore, new markers have begun to be studied in relation to cardiovascular damage, where the marker FGF-23 and its co-receptor Klotho have been found, involved in the genesis of cardiovascular damage and bone-mineral disease associated with phosphorus, which together cause cardiovascular remodeling , which has begun to further clarify this pathophysiological dynamic.This review seeks to know the implication of the FGF-23 and Klotho markers in CKD and the associated cardiovascular risk and for this purpose, a systematic review of the literature was carried out, investigating biomedical bases such as COCHRANE, Embase, LILACS, Scielo, Pub-Med, EBSCO, Hinari, Medical Societies, as well as MeSH thesauri specific to this research.

Comparación de métodos histoquímicos para el análisis histológico de muestras de encía humana / Comparison of histochemical methods for the histological analysis of human gingival samples

Introducción: Las técnicas de coloración histológica son útiles en el análisis ultraestructural de muestras tisulares, incluyendo el tejido gingival. Objetivo: Comparar la utilidad de tres métodos histoquímicos (hematoxilina-eosina, Masson Goldner y rojo sirio) en la identificación de elementos celulares y otros constituyentes del tejido gingival. Métodos: Estudio experimental in vitro que comprendió el análisis de tejidos gingivales de donantes sanos sin signos de inflamación gingival y con indicación de cirugía periodontal. Las muestras de encía se obtuvieron mediante gingivectomía, se procesaron e incluyeron en parafina, posteriormente se realizaron cortes con un micrótomo y se depositaron en portaobjetos de adhesión con polisina. Las muestras se agruparon y fueron teñidas con hematoxilina-eosina, Masson Goldner y rojo sirio, finalmente fueron visualizadas en un microscopio óptico Leica DM 750®. La lectura de los hallazgos fue realizada por patólogos orales. Resultados: La coloración hematoxilina-eosina evidencia elementos celulares y extracelulares del tejido epitelial y conectivo. Núcleos de color azul violeta, citoplasmas rosados, fibras de colágeno de matiz rosa claro, arteriolas y vénulas con túnica adventicia, media e íntima diferenciadas. La coloración Masson Goldner diferencia núcleos de coloración púrpura y citoplasma fucsia, presenta especificidad en identificar fibras de colágeno con tonalidad verde, distribuidas densa, homogénea y paralelamente en el tejido conectivo gingival. La tinción rojo sirio, permitió identificar las fibras de colágeno de color rosa brillante, mientras que el tejido epitelial y los vasos sanguíneos fueron de color amarillo. Conclusión: Cada coloración histológica evaluada en el presente trabajo tiene cierta afinidad y sensibilidad por estructuras celulares y componentes de la matriz extracelular específica. Su empleo es útil en el estudio de tejidos gingivales y podrían contribuir en el análisis de biopsias gingivales(AU)

Introduction: Histological staining techniques are useful in the ultrastructural analysis of tissue samples, including gingival tissue. Objective: Compare the usefulness of three histochemical methods (hematoxylin-eosin, Masson-Goldner and sirius red) for identification of cellular elements and other constituents of gingival tissue. Methods: An in vitro experimental study was conducted which included the analysis of gingival tissue from healthy donors without gingival inflammation signs and indication of periodontal surgery. The gum samples were obtained by gingivectomy, processed with paraffin, cut with a microtome and placed on Polysine adhesion slides. The samples were grouped, stained with hematoxylin-eosin, Masson Goldner and sirius red, and visualized under a Leica DM 750® microscope. Reading of the findings was performed by oral pathologists. Results: Hematoxylin-eosin staining found cellular and extracellular elements of the epithelial and connective tissue: violet-blue nuclei, pink cytoplasms, light rose collagen fibers, and arterioles and venules with differentiated tunica adventitia, media and intima. Masson-Goldner staining differentiated purple nuclei and fuchsia cytoplasm. It displayed specificity identifying green collagen fibers with dense, homogeneous and parallel distribution in the gingival connective tissue. Sirius red staining allowed identification of bright rose collagen fibers, whereas epithelial tissue and blood vessels were yellow. Conclusion: Each of the histological staining methods evaluated in the study shows a certain affinity with and sensitivity to cellular structures and components of the specific extracellular matrix. All three are useful for the study of gingival tissue and could contribute to the analysis of gingival biopsies(AU)

Efeitos do microagulhamento associado a radiofrequência no tratamento de flacidez abdominal em mulheres: Estudo experimental / Effects of microneedling associated radiofrequency in the treatment of abdominal sagging in women: Experimental study

A flacidez tissular abdominal é uma disfunção dermatológica que incomoda principalmente as mulheres. A radiofrequência e o microagulhamento são recursos utilizados para minimizar essa flacidez. Objetivo: Investigar os efeitos do microagulhamento associado a radiofrequência na flacidez tissular abdominal. Métodos: Trata-se de um estudo experimental, controlado e randomizado, com amostra de 20 mulheres, faixa etária entre 18 e 35 anos, dispostas em dois grupos: Grupo 1 (G1) foi aplicada 1 sessão de microagulhamento, após 15 dias reavaliação utilizando a plicometria e perimetria e Grupo 2 (G2) 1 sessão de microagulhamento, após 15 dias realizaram-se 4 sessões de radiofrequência com intervalo de 1 dia entre as sessões. Resultados: O G2 apresentou diminuição de flacidez do músculo reto abdominal direito apresentando p = 0,009, flanco direito p = 0,001 e flanco esquerdo p = 0,004, assim como a redução da circunferência abdominal. A avaliação de satisfação corporal do G2 teve escore final p = 0,029. Conclusão: O microagulhamento associado a radiofrequência promoveram uma melhora clínica da flacidez tissular abdominal e flancos. (AU)

Regenerative potential of basic fibroblast growth factor contained in biodegradable gelatin hydrogel microspheres applied following vocal fold injury: Early effect on tissue repair in a rabbit model / Potencial regenerativo do fator básico de crescimento de fibroblastos contido em microesferas biodegradáveis de hidrogel de gelatina aplicadas após lesão de prega vocal: efeito inicial no reparo tecidual em modelo de coelho

Abstract Introduction Postoperative dysphonia is mostly caused by vocal fold scarring, and careful management of vocal fold surgery has been reported to reduce the risk of scar formation. However, depending on the vocal fold injury, treatment of postoperative dysphonia can be challenging. Objective The goal of the current study was to develop a novel prophylactic regenerative approach for the treatment of injured vocal folds after surgery, using biodegradable gelatin hydrogel microspheres as a drug delivery system for basic fibroblast growth factor. Methods Videoendoscopic laryngeal surgery was performed to create vocal fold injury in 14 rabbits. Immediately following this procedure, biodegradable gelatin hydrogel microspheres with basic fibroblast growth factor were injected in the vocal fold. Two weeks after injection, larynges were excised for evaluation of vocal fold histology and mucosal movement. Results The presence of poor vibratory function was confirmed in the injured vocal folds. Histology and digital image analysis demonstrated that the injured vocal folds injected with gelatin hydrogel microspheres with basic fibroblast growth factor showed less scar formation, compared to the injured vocal folds injected with gelatin hydrogel microspheres only, or those without any injection. Conclusion A prophylactic injection of basic fibroblast growth factor -containing biodegradable gelatin hydrogel microspheres demonstrates a regenerative potential for injured vocal folds in a rabbit model.

Resumo Introdução A disfonia pós-operatória é causada principalmente por cicatrizes nas pregas vocais. Tem sido relatado que o manejo cuidadoso da cirurgia das pregas vocais reduz o risco de formação de cicatriz. No entanto, a depender da lesão da prega vocal, o tratamento da disfonia pós-operatória pode ser desafiador. Objetivo Desenvolver uma nova abordagem regenerativa profilática para o tratamento de pregas vocais lesionadas após a cirurgia, com microesferas biodegradáveis de hidrogel de gelatina como sistema de administração de medicamentos para o Fator Básico de Crescimento de Fibroblastos (bFGF). Método A cirurgia laríngea videoendoscópica foi feita para criar lesão nas pregas vocais em 14 coelhos. Imediatamente após esse procedimento, microesferas biodegradáveis de hidrogel de gelatina com bFGF foram injetadas na prega vocal. Duas semanas após a injeção, as laringes foram excisadas para avaliação da histologia das pregas vocais e do movimento da mucosa. Resultados A presença de função vibratória deficiente foi confirmada nas pregas vocais lesionadas. A histologia e a análise de imagem digital demonstraram que as pregas vocais lesionadas injetadas com microesferas de hidrogel de gelatina com bFGF apresentaram menor formação de cicatriz, em comparação com as pregas vocais lesionadas injetadas apenas com microesferas de hidrogel de gelatina ou aquelas sem injeção. Conclusão Uma injeção profilática de microesferas biodegradáveis de hidrogel de gelatina com bFGF demonstra um potencial regenerativo para pregas vocais lesionadas em um modelo de coelho.

Alteración del colágeno en la Hipertrofia Gingival en pacientes con ortodoncia: caracterización histológica e inmunohistoquímica / Collagen alteration in gingival hypertrophy in orthodontic patients: histological and immunohistochemical characterization

Resumen Introducción: La hipertrofia gingival (HG) es el aumento del volumen de la encía asociado a ciertas enfermedades sistémicas, hereditarias (idiopático), ingesta de algunos medicamentos o a factores locales como el tratamiento ortodóntico, capaz de provocar cambios histológicos en el tejido conectivo gingival. Objetivo: Describir las características histológicas e identificar el colágeno tipo I y tipo III en tejidos gingivales de sujetos con hipertrofia gingival portadores de ortodoncia. Materiales y método: Se diseñó un estudio de casos y controles que incluyó el análisis de biopsias de tejido gingival de 12 pacientes sometidos a cirugías periodontales. La muestra se dividió en dos grupos: individuos sanos (control; n= 6) y pacientes con HG portadores de ortodoncia (pacientes; n= 6). Las muestras fueron procesadas e incluidas en parafina. Las tinciones Masson-Goldner y rojo sirius/verde rápido fueron empleadas. El colágeno tipo I y tipo III fueron identificados mediante inmunohistoquímica con anticuerpos monoclonales. Resultado: En los pacientes con HG portadores de ortodoncia se observó un epitelio hiperplásico y tejido conectivo denso con abundantes fibras de colágeno distribuidos aleatoriamente. La inmunodetención de colágeno tipo I indicó la presencia de abundantes fibras desorganizadas y el colágeno tipo III fue inmunolocalizado subyacente a la membrana basal, vasos sanguíneos y toda la extensión del tejido conectivo de los pacientes con HG con tratamiento ortodóntico. Conclusión: La acumulación de fibras de colágeno, particularmente del colágeno tipo I y tipo III, son hallazgos histológicos que caracterizan la HG en pacientes portadores de ortodoncia. Futuros estudios son necesarios para dilucidar el fenotipo de los fibroblastos gingivales y la probable pérdida homeostática entre la producción y degradación de colágeno en esta patología.

Abstract Introduction: Gingival hypertrophy (GH) is the increase in the volume of the gingiva associated with certain systemic, hereditary (idiopathic) diseases, the intake of some medications or local factors such as orthodontic treatment, capable of causing histological changes in the gingival connective tissue. Objective: To describe the histological characteristics and identify type I and type III collagen in gingival tissues of subjects with gingival hypertrophy wearing orthodontics. Method: A case-control study was designed that included the analysis of gingival tissue biopsies from 12 patients submitted to periodontal surgeries. The sample was divided into two groups: healthy individuals (Control; n= 6) and patients with GH wearing orthodontics (Patients; n= 6). The samples were processed and embedded in paraffin. Masson-goldner and sirius red/fast green stains were used. Type I and type III collagen were identified by immunohistochemistry with monoclonal antibodies. Result: A hyperplastic epithelium and dense connective tissue with abundant randomly distributed collagen fibers were observed in patients with orthodontic GH. Immunodetention of type I collagen indicated the presence of abundant disorganized fibers and type III collagen was inmunolocalized underlying the basement membrane, blood vessels and the entire extension of the connective tissue of patients with GH orthodontic. Conclusion: The accumulation of collagen fibers, particularly type I and type III collagen, are histological findings that characterize GH in orthodontic wearers. Future studies are necessary to elucidate the phenotype of gingival fibroblasts and the probable homeostatic loss between collagen production and degradation in this pathology.

Fibroma de fibroblastos gigantes: presentación de 122 casos / Giant Cell fibroma: presentation of 122 cases

El fibroma de fibroblastos gigantes (FFG), conocido también como fibroma de células gigantes, es una pápula o nódulo asintomático localizado en la encía, paladar y lengua, es del mismo color que la mucosa adyacente, de base sésil o pediculada, con superficie lisa o papilar que por lo general mide menos de 1 cm. Objetivo: Identificar las características demográficas e histopatológicas de los casos de FFG de un laboratorio privado de patología bucal en la Ciudad de México. Material y métodos: Se obtuvieron los datos de edad, sexo, diagnóstico presuntivo y definitivo de 122 casos de FFG de 2004 a 2019 con un total de 7,681 muestras. Se describe su distribución por edad, sexo y localización. Resultados: El rango de edad obtenido es de 1 a 84 años, con un promedio de X = 38 años, se presenta con mayor frecuencia en la segunda década de vida, con una razón de 1.6:1 mujer a hombre. La localización más frecuente es en lengua (46%); sin embargo, sólo 49.1% de los estudios especificaban este dato. Conclusión: La importancia de esta lesión es que clínicamente se parece a otras patologías de tejido fibroso, por lo tanto, se debe tener presente al FFG como diagnóstico diferencial (AU)

Giant cell fibroma (GCF) is an asymptomatic papule or nodule that is similar in color to the surrounding mucosa, with a sessile or pedunculated base. It is usually less than 1 cm in size and it features a smooth or papillary surface. Objective: To identify the demographic and histopathological characteristics of GCF cases in a private oral pathology laboratory in Mexico City. Material and methods: Data on age, sex, as well as on presumptive and definitive diagnosis of 122 GCF cases were obtained from 2004 to 2019 with a total of 7,681 samples. Its distribution by age, sex and localization is described. Results: The age range obtained is from 1 to 84 years, with a mean age of 38 years. Frequently during the second decade of life, the female to male ratio is 1.6:1. The most frequent location is the tongue (46%), however, only 60 of 122 studies specified this data. Conclusion: Since this lesion clinically resembles another fibrous tissue pathology, it is warranted to have prior knowledge on its clinical characteristics, as GCF should be regarded as a differential diagnosis (AU)

Comparison of primary keloid fibroblast cultivation methods and the characteristics of fibroblasts cultured from keloids, keloid-surrounding tissues, and normal skin tissues / Comparación de los métodos de cultivo de fibroblastos queloides primarios y las características de los fibroblastos cultivados a partir de queloides, tejidos circundantes queloides y tejidos cutáneos normales

SUMMARY: The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surrounding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and collagenase digestion, were deployed and compared. Expression levels of the keloid-related genes α -SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two methods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to generate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts correlated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several key points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts and in vitro primary keloid fibroblasts.

RESUMEN: La identificación de un cultivo de fibroblastos queloides primarios, siempre ha sido una medida fundamental para estudiar los mecanismos de la enfermedad queloide. La calidad del cultivo de células primarias puede afectar directamente los resultados de otros experimentos. Este estudio se realizó para investigar las condiciones óptimas de crecimiento, incluido el tiempo óptimo de almacenamiento y el tiempo de tratamiento con colagenasa, para modelos de cultivo celular in vitro y los métodos adecuados para la separación epidermis-dermis en diferentes tejidos. Se recogieron de los pacientes tejidos queloides, tejidos circundantes queloides y tejidos cutáneos normales, para cultivo primario de fibroblastos. Se implementaron y compararon dos métodos, explante de tejido y digestión con colagenasa. Los niveles de expresión de los genes relacionados con queloides α -SMA, Col1 y Col3 se evaluaron en células cultivadas usando ambos métodos, para verificar las cualidades de las células primarias. Se realizó un análisis comparativo entre los dos métodos y entre los tres tejidos diferentes utilizados. La contaminación de bacterias y lípidos se minimizó inmediatamente después de que se procesaron las muestras. Se requirieron varios métodos de eliminación de la epidermis y diferentes tiempos de digestión con colagenasa en los tejidos para generar resultados óptimos. Los resultados de la PCR en tiempo real mostraron que los niveles de expresión de ARNm de genes relacionados con queloides en fibroblastos cultivados se correlacionaban con su perfil de expresión in vivo, como se informó en estudios anteriores. Los resultados de este studio indicaron varios puntos clave en el cultivo de fibroblastos queloides primarios y han demostrado la correlación en la expresión génica entre fibroblastos queloides in vivo y fibroblastos queloides primarios in vitro.

The action of subconjunctival cyclophosphamide in rabbits / Ação da ciclofosfamida subconjuntival em coelhos

ABSTRACT Objective: To observe clinically, in rabbits, the side effects of topical injection of subconjunctival cyclophosphamide, studying its role as an antifibrotic drug. Methods: Prospective study in 20 albino rabbits of New Zealand race. All rabbits were treated with cyclophosphamide, 10mg/ml in a volume of 0.3 ml, in the left eye through subconjunctival injection. They were evaluated for 1, 7, 30, and 60 days after the procedure. All the animals were examined for the detection of ocular reactions such as necrosis, hyperemia, chemosis, secretion, opacity, and iritis. Other side effects as changes in the behavior, in the feed, and the water consumption were also evaluated. Results: It was observed that from the 20 rabbits studied, three rabbits (15%) showed side effects only at the 24 hours analysis. One rabbit (5%) presented hyperemia, one rabbit (5%) had hyperemia associated with iritis, and one rabbit (5%) presented hyperemia associated with secretion. These reactions were not observed at 1, 7, 30, and 60 days. Conclusion: Cyclophosphamide subconjunctival injection induces minor side effects on the conjunctiva of rabbits such as hyperemia, associated with iritis and secretion.

RESUMO Objetivo: Observar clinicamente os efeitos colaterais de injeção subconjuntival de ciclofosfamida, pensando em sua ação como um agente antifibrótico. Métodos: Estudo prospectivo realizado com 20 coelhos albinos da raça Nova Zelândia. Todos os coelhos foram submetidos a 0,3 ml de injeção subconjuntival de ciclofosfamida 10mg/ml no olho esquerdo e foram avaliados de acordo com os efeitos locais no primeiro dia após a injeção, 7, 30 e 60 dias. Foram examinados para detecção de reações oculares como necrose, hiperemia, quemose, secreção, opacidade corneana, irite além de alterações comportamentais e variação no consumo de água e alimentação. Resultados: Dos 20 coelhos estudados, apenas 3 apresentaram reações oculares e somente na leitura de 24 horas. Um coelho (5%) apresentou hiperemia, 1 coelho (5%) apresentou hiperemia associada a presença de irite e 1 coelho (5%) apresentou hiperemia associada a presença de secreção. As reações não foram mais observadas durante os exames de 7, 30 e 60 dias. Conclusão: A ciclofosfamida subconjuntival causou poucos efeitos colaterais na conjuntiva dos coelhos. Os únicos efeitos encontrados foram hiperemia, irite e secreção.

Estudo comparativo da inibição da proliferação de fibroblastos in vitro na conjuntiva utilizando mitomicina C e ciclofosfamida / Comparative study of inhibition of fibroblasts proliferation in vitro in the conjunctiva using mitomycin and ciclophosphamide

RESUMO Objetivo: Avaliar a inibição da proliferação de fibroblastos in vitro das conjuntivas obtidas através de exérese de pterígios de pacientes utilizando mitomicina C (MMC) e ciclofosfamida (CF). Métodos: Os pterígios foram retirados de 7 pacientes e submetidos a cultivo celular. Após o cultivo, 3 fragmentos de dimensões iguais deste material foram colhidos de áreas adjacentes do pterígio removido de cada paciente. Eles foram randomicamente selecionados de tal forma que: um fragmento de cada paciente foi exposto: ao meio de cultura (grupo controle), a MMC e a CF por igual período de tempo nas concentrações de 0,4 mg/ml e 10 mg/ml respectivamente. Após este período realizou-se a contagem celular de fibroblastos destes 3 grupos. Cada grupo continha 7 fragmentos. Resultados: Com a utilização da MMC tivemos uma taxa de 95% da inibição da proliferação dos fibroblastos, enquanto com a CF 100%. Conclusões: Ambas as drogas apresentaram elevada taxa da inibição da proliferação de fibroblastos, porém a CF apresentou inibição maior que a MMC.

Abstract Objective: To evaluate the inhibition of fibroblast proliferation in vitro of conjunctiva obtained by excision of pterygium from patients using mitomycin (MMC) and cyclophosphamide (CF). Methods: Pterygiums were removed from 7 patients and subjected to cell culture. After cell cultivation, 3 fragments of equal dimensions of these tissues were collected from adjacent areas of each patient removed pterygium. They were randomly selected in such a way that one fragment of each patient was exposed to: the culture medium (group control), to MMC and to CF for an equal period of time at concentrations of 0,4 mg/dl and 10 mg/dl respectively. After this period, the fibroblast cell count of these groups were performed. Each group had seven fragments. Results: With the use of MMC we had a 95% rate of inhibition of fibroblast proliferation, while with CF 100%. Conclusion: Both drugs showed a high rate of inhibition of fibroblast proliferation, but CF showed greater inhibition than MMC.

Diferencias fenotípicas de fibroblastos gingivales en sujetos con hiperplasia gingival idiopática frente a sujetos periodontalmente sanos: estudio piloto / Phenotypic differences of gingival fibroblasts in subjects with idiopathic gingival hyperplasia versus periodontally healthy subjects: a pilot study

Objetivo: identificar, describir y diferenciar las características fenotípicas de los fibroblastos gingivales (FGs) en pacientes con hiperplasia gingival idiopática (HGI) e individuos periodontalmente sanos. Métodos: los FGs fueron aislados a partir de tejido gingival de individuos periodontalmente sanos (n=2) y pacientes con HGI (n=2). Los FGs se cultivaron en el medio DMEM (Dulbecco's Modified of Eagle Medium) a 37°C con 5% de CO2. La identificación y localización de la actina, vimentina y mitocondrias en FGs fue realizada y evaluada microscópicamente mediante inmunofluorescencia con anticuerpos monoclonales. La capacidad de migración de los FGs en los pacientes con HGI e individuos sanos también fue estudiada. Resultados: todos los FGs fueron mononucleares, fusiformes y con prolongaciones citoplasmáticas visibles. La faloidina permitió identificar una densa red de actina en los FGs de pacientes con HGI, contrariamente a los FGs de individuos periodontalmente sanos. La vimentina y mitocondrias fueron identificadas en los FGs de individuos sanos y pacientes con HGI sin ninguna alteración en su expresión y localización. La migración de la monocapa de los FGs indicó una actividad de migración celular importante en los FGs de los pacientes con HGI, en relación a los FGs de los individuos periodontalmente sanos. Conclusión: los FGs de pacientes con HGI conservan características fenotípicas celulares similares a los FGs de individuos periodontalmente sanos. Sin embargo, los FGs de pacientes con HGI simulan tener una mayor capacidad migratoria que amerita ser explorada en futuros trabajos de investigación.

Objective: To identify and to describe the phenotypic characteristics of gingival fibroblasts from patients with idiopathic gingival hyperplasia (IGH) and periodontally healthy individuals. Methods: Gingival fibroblasts (GFs) were isolated from gingival tissue from periodontally healthy individuals (n=2) and patients with IGH (n=2). The GFs were grown in DMEM (Dulbecco's Modified of Eagle Medium) at 37°C with 5% CO2. The identification and location of actin, vimentin and mitochondria in GFs were performed and evaluated microscopically by immunofluorescence with monoclonal antibodies. The migration capacity of GFs from IGH and healthy individuals was also studied. Results: All the GFs were mononuclear, fusiform and with visible cytoplasmic extensions. The phalloidin allowed to identify a dense actin network in the GFs of patients with IGH, contrary to the GFs of periodontally healthy individuals. Vimentin and mitochondria were identified in the GFs of healthy individuals and patients with IGH without any alteration in their expression and location. Monolayer migration of GFs indicates significant cell migration activity in the GFs of patients with IGH in relation to the GFs of periodontally healthy individuals. Conclusion: GFs from patients with IGH retain cellular phenotypic characteristic similar to GFs from periodontally healthy individuals. However, the GFs of patients with IGH simulate having a greater migratory capacity that deserves to be explored in future research works.