Cancer-associated fibroblasts contribute to cancer metastasis and apoptosis resistance in human ovarian cancer via paracrine SDF-1a

Background Cancer-associated fibroblasts (CAFs), one of the main members of stromal cells in tumor microenvironment are proposed to play a central role in promoting tumor metastasis. It is unclear whether and how CAFs mediates tumor metastasis or chemoresistance in human ovarian cancer. Methods CAFs were extracted from human ovarian cancer tissues (OCs) of patients with different kinds of histological types. Results We found that CAFs showed more aggressive potency than those tumor cells, both of which were isolated from the same ovarian cancer specimen. Moreover, when co-cultured with CAFs, cell migration abilities of ovarian cancer cells (SKOV3, OVCAR3 and HEY) were significantly increased. Next, we preliminarily detected a higher CAFs density in sections of metastatic lesions than those in primary tumor site of primary OCs clinically. However, no significant difference of stromal derived factors-1α (SDF-1α) production from CAFs was found between primary and metastatic lesions. Additionally, in contrast with tumor cells, CAFs exhibited obvious apoptosis resistance when treated with cisplatin. Furthermore, we found that cisplatin-induced cytotoxicity and apoptosis were significantly inhibited by co-cultured with recombinant human SDF-1α in SKOV3 in a time and dose-dependent manner, and this effect was suppressed by the CXCR4 antagonist AMD3100. Conclusions CAFs might be involved in the malignant metastasis in human ovarian cancer through promoting cell migration in tumor cells. And their resistance to cytotoxic agents might be mediated by paracrine SDF-1α/CXCR4 signaling in ovarian cancer (AU)

Efecto del uso de secretoma de células madre mesenquimales sobre heridas cutáneas. Análisis comparativo in vitro e in vivo de tres tipos de secretoma / Effects of mesenchymal stem cell secretome on skin wounds. In vitro and in vivo comparative analysis of three types of secretome

Introducción y objetivo: Los avances en el campo de la Ingeniería Tisular han promovido el desarrollo de sustitutos de piel y su aplicación en el campo de la Cirugía Plástica. Uno de los principales inconvenientes de la bioingeniería de la piel es la necesidad de obtener una gran cantidad de células viables en periodos cortos de tiempo, lo que conlleva que el proceso de biofabricación sea largo y complejo. En este estudio se pretende valorar los efectos de la aplicación de tres tipos diferentes de secretoma derivados de células madre mesenquimales humanas en cultivos de fibroblastos con el objetivo de favorecer la escalabilidad de los procesos de fabricación de piel artificial. Material y método: Se evaluaron los efectos a las 24, 48 y 72 horas sobre la viabilidad, la proliferación y la migración celular de fibroblastos humanos tras la aplicación de dos concentraciones (C1 y C2) de tres tipos diferentes de secretoma derivado de células madre mesenquimales humanas. Los resultados in vitro fueron contrastados en un modelo in vivo. Resultados: El uso de secretoma derivado de células madre mesenquimales mejoró los protocolos de cultivo de fibroblastos actualmente disponibles. El uso de secretoma se asoció a un aumento de la proliferación y migración celular manteniendo cifras altas de viabilidad. Los datos fueron especialmente positivos para el secretoma de células madre mesenquimales de pulpa dental y de tejido adiposo. Conclusiones: El efecto de la aplicación de secretoma procedentes de células madre mesenquimales permite mantener cifras de viabilidad celular elevadas, además de incrementar el ritmo de proliferación de fibroblastos. Los estudios in vivo e in vitro no evidenciaron efectos adversos a corto plazo. (AU)

Background and objective: Advances in Tissue Engineering promoted the development of skin substitutes. One of the main drawbacks of skin bioengineering is the requirement of a considerable quantity of viable cells in short periods of time, which leads to a challenging biofabrication process. This article aims to analyse the effects of the application of three different types of human mesenchymal stem cell-derived secretome in fibroblast cultures. The final goal is to achieve new strategies to promote the scalability of artificial skin manufacturing processes. Methods: The effects of the three different types of human mesenchymal stem cell-derived secretome were analyzed at 24, 48 and 72 hours. To study cell viability, cell proliferation and cell migration we exposed human fibroblasts to different secretome concentrations (C1 and C2). An in vivo study was proposed to corroborate in vitro results. Results: The use of mesenchymal stem cell-derived secretome improved the currently available fibroblast culture protocols. The use of secretome was associated with increased cell proliferation and cell migration while maintaining high viability values. Data were especially positive when secretome from dental pulp mesenchymal stem cells and adipose tissue mesenchymal stem cells were applied. Conclusions: The application of mesenchymal stem cell-derived secretome maintained high cell viability data and increased fibroblast proliferation. In vivo and in vitro studies showed no short-term adverse effects. (AU)

Orientin inhibits the progression of fibroblast-like synovial cells in rheumatoid arthritis by regulating MAPK-signaling pathway

Background Natural compounds are found to play an essential role in diverse inflammatory diseases, including rheumatoid arthritis (RA). Orientin, a flavonoid compound, is closely related to diverse pathological processes. Nevertheless, the role of orientin in RA is still unknown. Methods The cell viability was tested through cell counting kit 8 (CCK-8) assay, and the number of cell colonies was calculated via colony formation assay. In addition, flow cytometry assay was employed to detect apoptosis rate in human RA fibroblast-like synoviocytes (RA-FLS). Besides, Transwell assay was introduced to determine cell migratory and invasive abilities. Moreover, the level of cytokines (IL-8, IL-1β, and IL-6) was estimated with quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent serologic assay. Furthermore, western blotting analysis was used to test the protein levels of cleaved-caspase-3, Bax, BCL-2, matrix metalloproteinase (MMP)-2, MMP-9, phosphorylated c-Jun N-terminal kinase, p-P38, and phospho-extracellular signal-related kinase. Results Orientin inhibited cell viability, migration as well as invasion in a concentration- dependent manner in human RA-FLS. Additionally, treatment of orientin facilitated apoptosis and decreased the secretion of cytokines induced by tumor necrosis factor alpha (TNF-α) in human RA-FLS. Moreover, orientin inactivated mitogen-activated protein kinase (MAPK)-related signaling pathway, notably in human RA-FLS. Conclusion These findings confirmed that orientin inhibited human RA-FLS development and decreased TNF-α-induced inflammatory factors, at least partly, by modulating MAPK-signaling pathway, which implied that orientin might be an effective agent for treating RA (AU)

Coristoma óseo lingual. Nuevo caso de una entidad extremadamente infrecuente / Lingual osseous choristoma. New case of an extremely uncommon entity

Los osteomas constituyen un grupo de tumores óseos benignos que se localizan habitualmente en el cráneo y los huesos de la región facial, siendo rara su presencia en los tejidos blandos de la cavidad oral. También se les conoce como coristomas óseos, ya que están compuestos por tejido óseo maduro de aspecto normal. En la cavidad oral, la localización más frecuente es el tercio posterior de la lengua y suele aparecer en mujeres con edades comprendidas entre la segunda y tercera década de vida. A pesar de que la mayoría de los pacientes presentan un curso asintomático, no es infrecuente que su motivo de consulta sea la sensación de cuerpo extraño y disfagia. Afortunadamente, los osteomas linguales tienen buen pronóstico, ya que una vez realizada la extirpación no hay descritos casos de recurrencia. Debido a su insólita presentación, con menos de 100 casos publicados en la literatura, se decide presentar este caso clínico. (AU)

Osteomas are rare benign tumors that can be located on the craniomaxillofacial skeleton, being unusual its presence in the soft tissues of the oral cavity. They are also known as osseous choristomas, as they consist of normal matured bone tissue. Osteomas of the tongue occur more frequently in women in their second or third decade and the most frequent location in the oral cavity is the posterior third of the tongue. Most patients are asymptomatic, though other forms of presentation could be complaining of the sensation of having a foreign body or, even, dysphagia. The fact that less than 100 cases have been reported in the literature, has motivated the presentation of this clinical case. (AU)

CITED2 alleviates lipopolysaccharide-induced inflammation and pyroptosis in - human lung fibroblast by inhibition of NF-kB pathway

Background: Pneumonia, a severe infectious respiratory disease, is one of the leading causes of mortality and morbidity in children. Cbp/P300 interacting transactivator with Glu/Asp‑rich carboxy‑terminal domain 2 (CITED2) functions as a transcription cofactor, and plays critical roles in the development of embryonic and extra‑embryonic tissues, including fetal lung matu‑ration. The present study investigates the role of CITED2 in infantile pneumonia.Methods: The human fetal lung fibroblasts (MRC‑5 and WI‑38) were treated with lipopolysac‑charides to induce cytotoxicity, and the cell viability was detected by MTT. Inflammation was evaluated by ELISA, and western blot was used to investigate the pyroptosis.Results: CITED2 was down‑regulated in lipopolysaccharide‑treated MRC‑5/WI ‑38 cells. The over‑expression of CITED2 protected MRC‑5 and WI‑38 cells from lipopolysaccharide‑ induced cytotoxicity by increasing the cell viability and decreasing LDH expression. CITED2 reduced the expression of TNF‑α, IL‑ 6, IL‑ 1β in lipopolysaccharide‑treated MRC‑5/WI ‑38 cells. Lipopolysaccharide stimulated pyroptosis in MRC‑5 and WI‑38 cells through the up‑regulation of NL+RP3, GSDMD‑N, caspase‑1, IL ‑1β and IL‑18. However, CITED2 down‑regulated the expression of NLRP3, GSDMD‑N, caspase‑1, IL ‑1β, and IL‑18 protein in lipopolysaccharide‑treated MRC‑5/WI ‑38 cells. CITED2 also down‑regulated the protein expression of p‑p65 in lipopolysaccharide‑ treated MRC‑5/WI ‑38 cells.Conclusion: CITED2 exhibited anti‑inflammatory effect on lipopolysaccharide‑treated human lung fibroblasts and reduced pyroptosis through inactivation of NF‑κB pathway (AU)

Cytokine-driven positive feedback loop organizes fibroblast transformation and facilitates gastric cancer progression

BackgroundGastric cancer (GC) is a malignancy that belongs to one of the most common leading causes of cancer death. Cancer-associated fibroblasts (CAFs) promote the GC cells’ malignant behavior. It is still unknown how GC converts normal fibroblasts (NFs) to CAFs.MethodsGC cells were co-cultured with NFs. Bioinformatics was used to analyze the genes and signaling pathways that were changed in fibroblast. RT-PCR, western blot, and Elisa assays were used to detect the expression of cytokines in fibroblast and condition medium. Western blot and immunofluorescence demonstrated activation of relevant pathways in CAFs-like cells. Transwell, scrape, colony formation, and CCK-8 assays were performed to reveal the feedback effect of CAFs-like cells on GC cells.ResultsGC promoted the conversion of NFs to CAFs by secreting Interleukin 17A (IL-17). It included both morphological and molecular marker changes. This process was achieved by activating the nuclear factor-κB (NF-κB) pathway. On the other hand, CAFs cells could secrete C-X-C Motif Chemokine Ligand 8 (IL-8, IL-8), which promoted the malignant phenotype of GC cells. In this way, a feedback loop of mutual influence was constructed in the GC and tumor microenvironment (TME).ConclusionsOur research proved a novel model of GC-educated NFs. GC-IL-17-fibroblast-IL-8-GC axis might be a potential pathway of the interaction between GC and TME.

Cancer-associated fibroblasts in colorectal cancer

Colorectal cancer (CRC) is one of the leading causes of mortality among cancers. Many aspects of this cancer are under investigation to find established markers of diagnosis, prognosis, and also potential drug targets. In this review article, we are going to discuss the possible solution to all these aims by investigating the literature about cancer-associated fibroblasts (CAFs) involved in CRC. Moreover, we are going to review their interaction with the tumor microenvironment (TME) and vitamin D and their role in tumorigenesis and metastasis. Moreover, we are going to expand more on some markers produced by them or related to them including FAP, a-SMA, CXCL12, TGF- β, POSTN, and β1-Integrin. Some signaling pathways related to CAFs are as follows: FAK, AKT, activin A, and YAP/TAZ. Some genes related to the CAFs which are found to be possible therapeutic targets include COL3A1, JAM3, AEBP1 and, CAF-derived TGFB3, WNT2, and WNT54.

MicroRNAs sequencing of plasma exosomes derived from patients with atrial fibrillation: miR-124-3p promotes cardiac fibroblast activation and proliferation by regulating AXIN1

MicroRNAs (miRNAs) play an important role in the pathogenesis of atrial fibrillation (AF). Exosomal miRNAs may develop as promising biomarkers for AF. To explore significant exosomal miRNAs in AF, plasma exosomes were extracted from 3 patients with AF and 3 patients with sinus rhythm (SR), respectively. Differential expression of exosomal miRNAs were screened by high-throughput sequencing analysis and verified by qRT-PCR from 40 patients with AF and 40 patients with SR. The target genes prediction, biological function, and signaling pathways analysis were conducted by miRanda software, gene ontology (GO), and KEGG analysis. The results showed that there were 40 differently expressed exosomal miRNAs from AF patients compared with SR patients, of which 13 miRNAs were upregulated and 27 miRNAs were downregulated. qRT-PCR validation demonstrated that miR-124-3p, miR-378d, miR-2110, and miR-3180-3p were remarkably upregulated, while miR-223-5p, miR-574-3p, miR-125a-3p, and miR-1299 were downregulated. To explore the function of miR-124-3p associated with AF, plasma exosomes derived from AF patients were co-incubated with rat myocardial fibroblasts. The expression of miR-124-3p was upregulated in myocardial fibroblasts. The viability and proliferation of myocardial fibroblasts were elevated by transfecting with miR-124-3p overexpression plasmids using CCK8 and immunofluorescence-staining methods. AXIN1 was verified to be the target of miR-124-3p by luciferase assay in vitro. Expression of AXIN1 was reduced, while β-catenin, Collagen 1, and α-SMA were increased in myocardial fibroblasts with miR-124-3p overexpression. In conclusion, these findings suggested that circulating exosomal miRNAs may serve as novel biomarkers for AF, and miR-124-3p promotes fibroblast activation and proliferation through regulating WNT/β-catenin signaling pathway via AXIN1. (AU)

Los glicosaminoglicanos se encuentran implicados en la adherencia de Candida albicans y Malassezia spp. a queratinocitos, pero no a fibroblastos dérmicos / Glycosaminoglycans Are Involved in the Adhesion of Candida albicans and Malassezia Species to Keratinocytes But Not to Dermal Fibroblasts

Antecedentes y objetivo Las micosis superficiales son algunas de las enfermedades más comunes en todo el mundo, siendo los agentes causales más frecuentes las levaduras de los géneros Malassezia y Candida, comensales habituales de la piel que pueden actuar como patógenos oportunistas. El objetivo de este trabajo es investigar si los glicosaminoglicanos (GAG) de las células epiteliales son utilizados por estos microrganismos como receptores de adhesión a las mismas. Materiales y métodos Se utilizaron cultivos de queratinocitos y fibroblastos dérmicos. La participación de los GAG en la adhesión de Candida albicans (C. albicans) y Malassezia spp. se estudió mediante inhibición específica de la síntesis de estas moléculas empleando rodamina B o genisteína. También se analizó mediante digestión enzimática in situ empleando liasas específicas. Resultados El tratamiento con rodamina B produjo una inhibición parcial de la adherencia de ambas especies fúngicas a queratinocitos, pero no a fibroblastos. La digestión selectiva del heparán sulfato produjo un aumento de la unión de Malassezia a los queratinocitos y de ambas especies a los fibroblastos. La digestión del condroitín sulfato redujo la unión de C. albicans en los queratinocitos, pero favoreció la unión de la forma filamentada de esta levadura en los fibroblastos. Conclusiones Los GAG de superficie celular de queratinocitos parecen estar implicados en la adherencia de Candida y Malasezzia a la superficie celular. En los fibroblastos, por el contrario, su eliminación favorece la adherencia, sugiriendo la implicación de otro tipo de receptores (AU)

Background and objective Superficial mycoses are some of the most common diseases worldwide. The usual culprits — yeasts belonging to the genera Malassezia and Candida — are commensal species in the skin that can cause opportunistic infections. We aimed to determine whether these yeasts use glycosaminoglycans (GAGs) as adhesion receptors to mediate binding to epithelial cells. Material and methods In keratinocyte and dermal fibroblast cultures, we used rhodamine B and genistein to inhibit GAG synthesis to study the role these molecules play in the adhesion of Candida albicans (C. albicans) and Malassezia species to cells. We also analyzed GAG involvement by means of enzyme digestion, using specific lyases. Results Rhodamine B partially inhibited the adhesion of both fungi to keratinocytes but not to fibroblasts. Selective digestion of heparan sulfate enhanced the binding of Malassezia species to keratinocytes and of both fungi to fibroblasts. Chondroitin sulfate digestion decreased C. albicans adhesion to keratinocytes, but increased the adhesion of the filamentous forms of this species to fibroblasts. Conclusions Cell surface GAGs appear to play a role in the adhesion of C albicans and Malasezzia species to keratinocytes. In contrast, their adhesion to fibroblasts appears to be enhanced by GAG inhibition, suggesting that some other type of receptor is the mediator (AU)

Neutral endopeptidase depletion decreases colon cancer cell proliferation and TGF-B1 synthesis in indirect co-cultures with normal colon fibroblasts

Purpose Although recent studies have suggested that neutral endopeptidase (NEP) is implicated in the regulation of colon cancer (CC) cell growth and metastasis, the influence of the tumor microenvironment on this role of NEP has not been investigated so far. Normal colon fibroblasts (NCFs) constitute a component of the stroma surrounding a tumor in an early stage of its development. NCFs can influence transformed cells via different paracrine factors, including TGF-β1. This in vitro study was undertaken to evaluate the role of NEP in CC promotion in conditions of indirect co-culture of CC cells (LS180 and SW620) with NCFs (CCD-18Co) or their conditioned medium (CM-18Co). Methods We examined cell proliferation (with the BrdU assay) and invasiveness (using BME-coated inserts, 8 µm) of NEP-expressing, NEP-silenced (siRNA), and NEP-inhibited (with thiorphan, i.e. a NEP specific inhibitor) CC cells cultured alone or co-cultured with CCD-18Co or with their conditioned medium. The Western blot and ELISA methods were used to assess the level of TGF-β1. Results The results showed that the co-culture of the NEP-depleted CC cells with NCFs or their conditioned medium resulted in a significant decrease in cell proliferation in comparison with the proliferative potential of NEP-silenced/inhibited CC cells cultivated alone. In contrast, the NEP depletion did not influence the invasiveness of CC cells in the co-cultures. The co-culture of CC cells with CCD-18Co or CM-C18Co resulted in increased synthesis of TGF-β1, while the NEP downregulation decreased the synthesis of TGF-β1 in CC cells and abolished the stimulatory effect of the co-cultures on TGF-β1 production. Conclusions The results suggest that the expression of NEP by colon cancer cells is essential for their proliferation and TGF-β1 synthesis during paracrine interactions with NCFs (AU)

Budesonide up-regulates vitamin D receptor expression in human bronchial fibroblasts and enhances the inhibitory effect of calcitriol on airway remodeling

Introduction and objectives: Transforming growth factor β1 (TGFβ1) and dysregulated microRNA-21 (miR-21) expression is associated with TGFβ/Smad signaling pathway activation and fibrosis. While calcitriol has been shown to improve airway remodeling in asthmatic mice, its mechanism remains unknown. In this study, the effect of calcitriol on the TGFβ/Smad signaling pathway and miR-21 expression in human bronchial fibroblasts was investigated to explore the mechanism of action of calcitriol and the inhaled glucocorticoid, budesonide, in airway remodeling. Materials and methods: Human bronchial fibroblasts were pretreated with budesonide, calcitriol, or budesonide plus calcitriol, and stimulated with TGFβ1 for 48h. Quantitative real-time PCR was used to determine the expression of miR-21. Western blot was used to determine airway remodeling-related proteins, TGFβ/Smad signaling pathway-related proteins, glucocorticoid receptor, and vitamin D receptor (VDR) expression. Results: Both budesonide and calcitriol down-regulated miR-21 expression in human bronchial fibroblasts, up-regulated Smad7 expression, and inhibited the expression of airway remodeling-related proteins. Both budesonide and calcitriol up-regulated the low expression of VDR induced by TGFβ1 in human bronchial fibroblasts. The expression of VDR in the combined treatment group (budesonide plus calcitriol) was significantly higher than that in the calcitriol treatment group. The expression of collagen type I in the combined treatment group was significantly lower than that in the calcitriol treatment group. Conclusions: Calcitriol can up-regulate the expression of VDR in human bronchial fibroblasts and exert an anti-airway remodeling effect. Budesonide can up-regulate the expression of VDR in human bronchial fibroblasts and enhance the inhibitory effect of calcitriol on airway remodeling

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Commentaries on the publication entitled: "Structure and distribution of an unrecognized interstitium in human tissues" by Benias et al. (2018)

The claim made in this publication of the existence of a hitherto unknown interstitial space is based on studies with sample-based confocal laser endo-microscopy (pCLM). Due to postings on various web portals (New Cellvizio, EurekAlert, Google Scholar,...) the alleged discovery has found great resonance. Nevertheless, there are several critical issues in this publication, the most important being that this is not the discovery of an "unrecognized" interstitium as it has, in fact, been known for a long time

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Análisis de degradación de stents metálicos en modelo de cultivo "in vitro" con células epiteliales respiratorias y fibroblastos / Degradation analysis for metal stents in an in vitro cell culture model with respiratory epithelial cells and fibroblasts

Objetivo: investigar la posible degradación de stents metálicos tras co-cultivo con células respiratorias in vitro. Método: durante 21 días se han co-cultivado con la línea CRL-4011 (epitelial) y MRC-5 (fibroblastos) tres tipos de stents: Wallstent(R) (aleación de cobalto-cromo-níquel y molibdeno), Zilver PTX(R) y Zilver Flex(R) (nitinol, aleación de níquel-titanio, con y sin liberación de paclitaxel, respectivamente). Las mismas células sin stent sirvieron como control. Los sobrenadantes de los cultivos se recogieron los días 3, 9, 15 y 21, se alicuotaron y almacenaron a -800 C. Mediante espectrometría de masas (ICP/ MS) se investigaron los niveles de titanio, cromo, níquel, cobalto y molibdeno en los sobrenadantes, y también se han analizado los niveles de esos mismos elementos en el medio de cultivo original (antes de añadirlo a los cultivos celulares). Resultados: en todos los experimentos se encontraron mayores niveles de elementos metálicos en los sobrenadantes recogidos en el tercer día de cultivo, tanto de células epiteliales como de fibroblastos, con diferencias estadísticamente significativas (p<0,002). Los sobrenadantes de los cultivos de células epiteliales con Wallstent mostraron los niveles más altos de níquel y cobalto respecto a los controles (p = 0,001), y los niveles de titanio fueron más altos en los cultivos de Zilver Flex y PTX, constituidos por una aleación de níquel y titanio (p <0,001). Conclusiones: hemos detectado una rápida liberación en el sobrenadante de todos los cultivos de los elementos constitutivos de los tres stents que incluimos en los experimentos, y con niveles muy superiores a los cultivos controles

Objective: To investigate the possible degradation of metal stents after co-culture with respiratory cells in vitro. Methods: Three types of stents were co-cultured with the CRL-4011 line (epithelial) and the MRC-5 line (fibroblasts): Wallstent(R) (cobalt-chromium-nickelmolybdenum alloy), Zilver PTX(R) and Zilver Flex(R) (nitinol, nickel-titanium alloy, with and without paclitaxel release, respectively). The same stentless cells served as the control group. Culture supernatants were collected on days 3, 9, 15 and 21, aliquoted and stored at -80 ºC. The levels of titanium, chromium, nickel, cobalt and molybdenum in the supernatants were studied using inductively coupled plasma mass spectrometry (ICP-MS), and the levels of the same elements were also analyzed in the original culture medium (before adding them to the cell cultures). Results: In all experiments, higher levels of metal elements were found in the supernatants collected on the third day of culture, for both epithelial cells and fibroblasts, with statistically significant differences (p < 0.002). The supernatants of epithelial cell cultures with Wallstent showed the highest levels of nickel and cobalt in comparison to controls (p = 0.001), and titanium levels were higher in Zilver Flex and Zilver PTX cultures, consisting of a nickeltitanium alloy (p < 0.001). Conclusion: We have detected a rapid release in the supernatant of all the cultures of the constituent elements of the three stents that we included in the experiments, and with levels much higher than those of the control cultures

Importancia de la investigación sobre el estroma tumoral en el cáncer de mama / Importance of investigating tumour stroma in breast cancer

A pesar de los avances conseguidos en el tratamiento del cáncer de mama, todavía sigue siendo una causa importante de mortalidad en las mujeres. Por tanto, resulta necesario plantear nuevos enfoques de la fisiopatología de la enfermedad que contribuyan a realizar una mejor evaluación pronóstica, y a la mejora de las estrategias terapéuticas. Para ello, deberíamos considerar que el cáncer no es solo una transformación maligna de las células epiteliales y su progresión meramente autónoma; sino que, hoy en día, existen datos que apoyan el concepto del cáncer como un ecosistema basado en una sociología de diferentes tipos celulares, con sus interacciones complejas. Entre los diversos tipos de células que conforman el estroma tumoral, y que tienen un papel relevante en la progresión del cáncer de mama, se encuentran los fibroblastos asociados al cáncer, las células inflamatorias y las células endoteliales. Existen diferentes factores moleculares expresados por esas células que se asocian con el desarrollo de metástasis, tales como las metaloproteasas de matriz y sus inhibidores tisulares, citoquinas o receptores tipo toll. En base a la expresión de todos ellos, aquí proponemos 2 fenotipos de estroma del cáncer de mama con influencias marcadamente diferentes sobre el pronóstico de las pacientes. También analizamos los mecanismos involucrados en la interrelación tumor-estroma que pueden llevarnos a mejorar las estrategias terapéuticas en el cáncer de mama

Despite advances in the treatment of breast cancer, it remains an important cause of mortality in women. Therefore, it is necessary to propose new approaches to the pathophysiology of the disease that could help to improve prognostic evaluation and therapeutic strategies. To do this, we should consider that cancer is not only a malignant transformation of the epithelial cells or their purely autonomous growth; nowadays, there are data that support the concept of cancer as a system based on a sociology of different cell types, with complex interactions. Among the various types of cells that make up the tumour stroma and which play an important role in the progression of breast cancer are cancer-associated fibroblasts, inflammatory cells and endothelial cells. Several molecular factors expressed by these cells are associated with the development of metastases, such as matrix metalloproteases and their tissue inhibitors, cytokines or toll-like receptors. Based on the expression of all of these factors, here we propose two types of stroma from breast cancer that display markedly different influences on patient prognosis. We also analyse mechanisms involved in the tumour-stroma interrrelationship that could help to improve therapeutic strategies in breast cancer

Influencia del suero fetal bovino en el cultivo de fibroblastos gingivales / Influence of bovine fetal serum in the culture of gingival fibroblasts

Introducción: Los fibroblastos gingivales (FGs) son células responsables del mantenimiento de la homeostasis, cicatrización y tolerancia inmunitaria del tejido conjuntivo gingival. Por sus características fisiológicas, los FGs pueden ser un candidato en la terapia celular. Sin embargo, la conservación de su fenotipo en cultivo celulares requiere de condiciones estrictamente definidas. Una de ellas es la concentración de Suero Fetal Bovino (SFB). Nuestro objetivo es evaluar el efecto concentración-dependiente de la suplementación de SFB en el medio de cultivo sobre el comportamiento, crecimiento, proliferación y supervivencia de los fibroblastos gingivales humanos. Materiales y métodos: FGs fueron cultivados con DMEM (Dulbecco's modification of Eagle médium) y concentraciones de 0%, 0.5% y 10% de SFB durante tres semanas. Análisis morfológico, de proliferación e inmunohistoquímicos fueron llevados a cabo en el presente trabajo. Se llevó a cabo un análisis factorial de varianza (ANOVA) por medio el software R Project. Los resultados fueron considerados significativos con un valor de p<0,05. Resultados y Discusión: Los FGs cultivados con 10% de SFB alcanzaron un desarrollo morfológico más notorio y en menor tiempo comparados con los FGs cultivados a 0.5% y 0% de SFB. La efectividad de la concentración del SFB 0,5% fue mucho más alta en relación a la concentración del SFB 10%. La inmunodetención de la actina, vimentina y fibronectina fueron más notorias en los FGs tratados con 10% y 0.5% de SFB. Este estudio concluye que los FGs humanos presentan una mejor capacidad de supervivencia, desarrollo y proliferación cuando son cultivados en presencia de SFB

Introduction: Gingival fibroblasts (FGs) are cells responsible for the maintenance of homeostasis, healing and immune tolerance of the gingival connective tissue. Due to their physiological characteristics, FGs can be a candidate in cell therapy. However, the conservation of their phenotype in cell culture requires strictly defined conditions. One of them is the concentration of Bovine Fetal Serum (FBS). Our aim is to evaluate the concentration-dependent effect of SFB supplementation in the culture medium on the behavior, growth, proliferation and survival of human gingival fibroblasts. Materials and methods: FGs were cultured with DMEM (Dulbecco's modification of Eagle medium) and concentrations of 0%, 0.5% and 10% of SFB for three weeks. Morphological, proliferation and immunohistochemical analyzes were carried out in the present work. A factorial analysis of variance (ANOVA) was carried out using the R Project software. The results were considered significant with a value of p <0.05. Results and discussion: The FGs grown with 10% FBS reached a more noticeable morphological development and in a shorter time compared to the FGs grown at 0.5% and 0% FBS. The effectiveness of the concentration of the SFB 0.5% was much higher in relation to the concentration of the SFB 10%. The immunodetention of actin, vimentin and fibronectin were more evident in the FGs treated with 10% and 0.5% FBS. This study concludes that human FGs have a better capacity for survival, development and proliferation when they are grown in the presence of FBS

Estudio en fibroblastos humanos del sistema angiotensina en la fisiopatología de la fibrosis pulmonar idiopática / The study of angiotensin system human fibroblasts in the physiopathology of idiopathic pulmonary fibrosis

En el tejido pulmonar de modelos murinos, la angiotensina II induce la proliferación de fibroblastos, su diferenciación a miofibroblastos y la producción de procolágeno tras su unión al receptor I de la angiotensina. Hemos estudiado el comportamiento de fibroblastos pulmonares humanos procedentes de una línea celular comercial tras la estimulación con TGF-β1. Hemos observado que estos fibroblastos, cuando son estimulados, aumentan la expresión de bFGF, colágeno y α-SMA. Tras el bloqueo del receptor de Angiotensina II con Losartan a una concentración de 10 µM y la estimulación con TGF- β1, se produce una disminución, tanto de los niveles de bFGF como de la concentración de colágeno, sin que llegue a alcanzar la significación estadística con respecto a las células no tratadas. En cuanto a la expresión de α-SMA como marcador de transformación a miofibroblastos, no había diferencias entre las células tratadas con TGF-β1 y TGF-β1 más losartán

In murine model lung tissue, angiotensin II induces the proliferation of fibroblasts, their distinction from myofibroblasts and procollagen production after its binding with the type 1 receptor. We have studied the behavior of human lung fibroblasts from a commercial cell line after stimulation with TGF-β1. We observed that those fibroblasts, when stimulated, increased bFGF, collagen and α-SMA expression. After blocking the angiotensin II receptor with losartan at a concentration of 10 µM and stimulation with TGF- β1, there was a decrease in both bFGF levels and collagen concentration, without reaching statistical significance with regard to untreated cells. With regard to α-SMA expression as an indicator of transformation to myofibroblasts, there were no differences between cells treated with TGF-β1 and TGF-β1 with losartan

The effect of therapeutic ultrasound on fibroblast cells in vitro: the systematic review / Efecto del ultrasonido terapéutico en células fibroblásticas in vitro: revisión sistemática

Introduction: Therapeutic ultrasound is one of the most used physical resources in the area of physiotherapy for the treatment of injuries. However, the multiplicity of dosimetry used in clinical practice points to its indiscriminate use for pathologies that surround skeletal muscle and expresses the limitation of the available literature on the ideal dosimetric standardization to the tissue restoration, mechanism of action and its real effects on the treatment in question. Objective: The objective of this study was to promote a systematic review about the different effects and the dosimetric parameters of therapeutic ultrasonic irradiation on the process of restoration of fibroblast cells in vitro. Methods: To select the articles, three electronic data banks were consulted, with publication from January 2000 to September 2016. The studies were tracked by three freestanding reviewers, according to inclusion and exclusion criteria. Results: 669 articles were selected and after the application of inclusion and exclusion criteria, 647 were excluded. Among the exclusions reasons there are: the utilization of another physical method, exclusive focus on another type of cell line, other experimental models or the use of another language, reaching at the end 22 studies directed to qualitative analysis. Conclusion: The results of this study showed that the scientific basis is not enough to stablish real effects and dosimetric parameters of therapeutic ultrasonic on the process of restoration of fibroblast cells in vitro, due to the lack of generalization and conflict of found results

Introducción: El ultrasonido terapéutico es uno de los recursos físicos más utilizados en el área de fisioterapia para el tratamiento de lesiones. Sin embargo, la gran cantidad de dosimetrías utilizadas en la práctica clínica muestra su uso indiscriminado para patologías que circundan el músculo esquelético y además expresa la limitación de la literatura sobre la estandarización dosimétrica ideal para la restauración del tejido, mecanismo de acción y sus efectos reales sobre el tratamiento en cuestión. Objetivos: El objetivo de este estudio fue realizar una revisión sistemática sobre los diferentes efectos y parámetros dosimétricos de la irradiación ultrasónica terapéutica en el proceso de reparación de células fibroblásticas in vitro. Material y método: Para la selección de los artículos fueron consultadas tres bases de datos para buscar publicaciones entre enero de 2000 y septiembre de 2016. La búsqueda de trabajos se realizó por tres revisores independientes, conforme a los criterios de inclusión y exclusión. Resultados: Se seleccionaron 669 artículos y tras la aplicación de los criterios de inclusión, se excluyeron 647 estudios. Entre los motivos de exclusión están la utilización de otro medio físico, enfoque exclusivo de otro tipo de línea celular, otros modelos experimentales o el uso de otro idioma, quedando 22 estudios para el análisis cualitativo. Conclusión: Los hallazgos de este estudio mostraron que la base científica todavía es insuficiente para el establecimiento de los efectos reales y parámetros dosimétricos de la irradiación ultrasónica terapéutica en el proceso de reparación de células fibroblásticas in vitro, por la falta de generalización y conflicto de los resultados encontrados

Mercury exposure induces proinflammatory enzymes in vascular fibroblasts / La exposición a mercurio induce la expresión de enzimas pro-inflamatorias en fibroblastos vasculares

Introduction: Previous studies show that mercury exposure increases cardiovascular risk, although the underlying cellular mechanisms have still not been fully studied. The aim of this project is to study, in vascular fibroblasts (VF), the effect of HgCl2 exposure on the expression of enzymes involved in the synthesis of prostanoids and reactive oxygen species (ROS). These molecules have been shown to participate in the inflammatory response associated with cardiovascular diseases. Material and methods: Adventitial VF cultures of Sprague-Dawley rat aortas, shown to be α-actin negative by immunofluorescence, were exposed to HgCl2 (0.05-5μg/mL) for 48h. mRNA and protein levels of cyclooxygenase-2 (COX-2), microsomal prostaglandin E synthase 1 (mPGES-1), thromboxane A2 synthase (TXAS), NADPH oxidase 1 (NOX-1), and 4 (NOX-4) where analyzed using qRT-PCR and western blot, respectively. NOX activity was determined by chemiluminescence. Results: HgCl2 exposure increased COX-2, mPGES-1, TXAS, and NOX-1 expression and NOX activity, and decreased NOX-4 expression. The increase in NOX-1 and COX-2 expression was abolished by the treatment with inhibitors of COX-2 (10μM celecoxib) and NOX (300μM apocynin, 0.5μM ML-171). Conclusions: 1) HgCl2 increases the expression of pro-inflammatory enzymes involved in ROS and prostanoid synthesis in VF. 2) There is a reciprocal regulation between COX-2 and NOX-1 pathways. 3) These effects can contribute to explain the increase in cardiovascular risk associated to mercury (AU)

Introducción: Estudios previos muestran que la exposición a mercurio aumenta el riesgo cardiovascular, sin embargo, los mecanismos celulares subyacentes no han sido esclarecidos completamente. Nuestro objetivo es estudiar el efecto de la exposición a HgCl2 sobre la expresión de enzimas involucradas en la síntesis de prostanoides y especies reactivas de oxígeno (ROS) en fibroblastos vasculares (FV). Se ha demostrado la participación de estas moléculas en la respuesta inflamatoria asociada a enfermedades cardiovasculares. Métodos: FV de la adventicia de aorta de ratas Sprague-Dawley, caracterizados por inmunofluorescencia como negativos para α-actina, fueron estimulados con HgCl2 (0,05-5μg/ml) durante 48 horas. Se analizaron los niveles de ciclooxigenasa-2 (COX-2), prostaglandina E sintasa 1 microsomal (mPGES-1), tromboxano A2 sintasa (TXAS), NADPH oxidasa 1 (NOX-1) y 4 (NOX-4) mediante qRT-PCR y western blot, respectivamente. La actividad de NOX se determinó mediante quimioluminiscencia. Resultados: La exposición a HgCl2 aumentó la expresión de COX-2, mPGES-1, TXAS y NOX-1, disminuyendo la expresión de NOX-4. El tratamiento con inhibidores de COX-2 (10μM celecoxib) y NOX (300μM apocynin, 0.5μM ML-171) abolió el aumento de la expresión de NOX-1 y COX-2, respectivamente. Conclusiones: 1) HgCl2 aumenta la expresión de enzimas proinflamatorias implicadas en la síntesis de ROS y prostanoides en FV. 2) Hay una regulación recíproca entre las vías de COX-2 y NOX-1. 3) Estos efectos pueden contribuir a explicar el aumento del riesgo cardiovascular asociado a la exposición al mercurio (AU)

Varón de 49 años con fiebre, malestar general y nódulos pulmonares / 49 Years-old man with fever, malaise and pulmonary nodules

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Metaloproteinasa 11, potencial marcador y diana molecular en cáncer de próstata avanzado y resistente a la castración. Estudio en cultivo de fibroblastos peritumorales / Metalloproteinase 11, potential marker and molecular target in advanced and castration-resistant prostate cancer. Culture study of peritumoral fibroblasts

Objetivo: Analizar el comportamiento de la metaloprotesa 11 (MMP11) en fibroblastos cultivados procedentes de tumores prostáticos humanos con diferentes características clinicopatológicas. Material y métodos: Para este estudio se analizaron muestras de biopsias de próstata obtenidas por vía transrectal de tumores con diferentes características, tratados o no con privación androgénica (PA). Tras la optimización del método de cultivo, se aislaron y cultivaron los fibroblastos para realizar el estudio (PCR) del ARNm de MMP11. Resultados: Se estudiaron finalmente 37 casos: 5 muestras de hiperplasia benigna de próstata, 14 casos con neoplasias localizadas (7 de alto riesgo según la clasificación de D’Amico), 5 con tumores con metástasis óseas y 13 tratados con PA, de los que 6 cumplían los requisitos para ser definidos como resistentes a la castración. En los tumores sin PA, la expresión de MMP11 fue significativamente mayor (p = 0,001) en los fibroblastos de tumores de grados más altos. Se encontró una correlación significativa (p = 0,001) entre PSA y expresión de MMP11 fibroblástica y un aumento significativo de la expresión de MMP11 en los tumores metastásicos. En los tumores con PA se objetivó una expresión significativamente mayor de MMP11 en pacientes resistentes a la castración que en los sensibles a esta (p = 0,003). Conclusión: En tumores de próstata avanzados o en fases de mayor agresividad tumoral, la producción de MMP11 por los fibroblastos resulta significativamente mayor que en tumores no metastásicos o en fases de sensibilidad a la PA

Objective: To analyze the expression of metalloprotein 11 (MMP11) in cultured fibroblasts obtained from human prostate tumors with different clinical and pathological characteristics. Material and methods: For this study we analyzed samples of transrectal prostate biopsies from tumors with different characteristics, treated with or whithout androgen deprivation (AD). After optimization of the culture method, fibroblasts were isolated and cultured to perform the study (PCR) of MMP11 mRNA. Results: Finally, 37 cases were studied: 5 samples of benign prostatic hyperplasia, 14 cases with localized neoplasms (7 high-risk according to the D’Amico classification), 5 with metastasic tumors (bone metastases), and 13 treated with AD therapy, of which 6 fulfilled the requirements to be defined as resistant to castration. In tumors without AD therapy, MMP11 expression was significantly higher (P= .001) in fibroblasts of higher grade tumors. A significant (P= .001) correlation was found between PSA and expression of MMP11 in fibroblast s and a significant increase of MMP11 expression in metastatic tumors. In tumors with AD therapy, a significantly greater expression of MMP11 was observed in resistant to castration patients than in those sensitive to castration (P= .003). Conclusion: In advanced prostate tumors or in stages of increased tumor aggressiveness, the production of MMP11 by fibroblasts is significantly greater than in non-metastatic tumors or in AD sensitive tumors