RESUMEN
Animal models of cirrhosis are of great interest to investigate the pathological process leading to the final stage of cirrhosis. The aim of this study was to analyze the different steps involved in the progressive development of cirrhosis using Fourier transform infrared spectral histology in 2 mouse models of cirrhosis, the STAM model of metabolic cirrhosis, and the carbon tetrachloride-induced cirrhosis model. Formalin-fixed, paraffin-embedded liver samples were obtained from 3 mice at 5 time points in each model to analyze the course of hepatic lesions up to the formation of cirrhosis. For each time point, adjacent 3-µm-thick liver sections were obtained for histologic stains and spectral histology. Fourier transform infrared acquisitions of liver sections were performed at projected pixel sizes of 25 µm × 25 µm and 6.25 µm × 6.25 µm. Spectral images were then preprocessed with an extended multiplicative signal correction and analyzed with common k-means clustering, including all stages in each model. In both models, the 2- and 4-class common k-means clustering in the 1000 to 1350 cm-1 range showed that spectral classes characterized by higher absorbance peaks of glycogen were predominant at baseline, then decreased markedly in early stages of hepatic damage, and almost disappeared in cirrhotic tissues. Concomitantly, spectral classes characterized by higher absorbance peaks of nucleic acids became progressively predominant during the course of hepatic lesions. These results were confirmed using k-means clustering on the peaks of interest identified for glycogen and nucleic acid content. Our study showed that the glycogen depletion previously described at the stage of cirrhosis is an early event in the pathological process, independently of the cause of cirrhosis. In addition, there was a progressive increase in the nucleic acid content, which may be linked to increased proliferation and polyploidy in response to cellular lesions.
Asunto(s)
Tetracloruro de Carbono , Ácidos Nucleicos , Ratones , Animales , Tetracloruro de Carbono/toxicidad , Análisis de Fourier , Estudios Longitudinales , Cirrosis Hepática/diagnóstico por imagen , Cirrosis Hepática/patología , Modelos Animales de Enfermedad , GlucógenoRESUMEN
Urea cycle disorders (UCD) are rare diseases that usually affect neonates or young children. During decompensations, hyperammonemia is neurotoxic, leading to severe symptoms and even coma and death if not treated rapidly. The aim was to describe a cohort of patients with adult onset of UCDs in a multicentric, retrospective and descriptive study of French adult patients with a diagnosis after 16 years of age of UCDs due to a deficiency in one of the 6 enzymes (arginase, ASL, ASS, CPS1, NAGS, OTC) or the two transporters (ORNT1 or citrin). Seventy-one patients were included (68% female, 32% male). The diagnosis was made in the context of (a) a metabolic decompensation (42%), (b) family history (55%), or (c) chronic symptoms (3%). The median age at diagnosis was 33 years (range 16-86). Eighty-nine percent of patients were diagnosed with OTC deficiency, 7% CPS1 deficiency, 3% HHH syndrome and 1% argininosuccinic aciduria. For those diagnosed during decompensations (including 23 OTC cases, mostly female), 89% required an admission in intensive care units. Seven deaths were attributed to UCD-6 decompensations and 1 epilepsy secondary to inaugural decompensation. This is the largest cohort of UCDs diagnosed in adulthood, which confirms the triad of neurological, gastrointestinal and psychiatric symptoms during hyperammonemic decompensations. We stress that females with OTC deficiency can be symptomatic. With 10% of deaths in this cohort, UCDs in adults remain a life-threatening condition. Physicians working in adult care must be aware of late-onset presentations given the implications for patients and their families.
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Trastornos Innatos del Ciclo de la Urea/diagnóstico , Adolescente , Adulto , Edad de Inicio , Anciano , Anciano de 80 o más Años , Aciduria Argininosuccínica/diagnóstico , Femenino , Francia , Humanos , Hiperamonemia/diagnóstico , Masculino , Persona de Mediana Edad , Ornitina/deficiencia , Enfermedad por Deficiencia de Ornitina Carbamoiltransferasa/diagnóstico , Estudios Retrospectivos , Factores Sexuales , Trastornos Innatos del Ciclo de la Urea/mortalidad , Adulto JovenRESUMEN
Background and study aims White bile is defined as a colorless fluid occasionally found in the biliary tract of patients with bile duct obstruction. Its significance is not clearly established. Our objective was to analyze the prognostic value of white bile in a series of patients with biliary obstruction due to biliary or pancreatic cancer. Patients and methods The study was conducted on a series of consecutive patients with malignant obstructive jaundice. They all underwent endoscopic retrograde cholangiopancreatography with collection of bile and biliary stent insertion. White bile was defined as bile duct fluid with bilirubin level <â20âµmol/L. Univariate and multivariate analyses were performed to identify variables associated with overall survival (OS). Results Seventy-three patients were included (32 pancreatic cancers, 41 bile duct cancers). Thirty-nine (53.4â%) had white bile. The mean bile duct bilirubin level in this group was 4.2â±â5.9âµmol/L vs 991â±â1039âµmol/L in patients with colored bile (Pâ<â0.0001). In the group of 54 patients not eligible for surgery, the multivariate analysis demonstrated an association between the presence of white bile and reduced OS (HR 2.3, 95â%CI 1.1-4.7; Pâ=â0.02). Other factors independently associated with OS were metastatic extension (HR 2.8, 95â%CI 1.4-5.7) and serum total bilirubin (HR 1.003, 95â%CI 1.001-1.006). There was a significant inverse correlation between serum and bile duct bilirubin levels (râ=â-0.43, Pâ=â0.0001). Conclusion White bile in patients with inoperable malignant biliary obstruction is an independent factor of poor survival.
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Alkaline phosphatase activity is a parameter included in biochemical liver test. These isoenzymes are of various cellular origin inducing physiological variations on age and sex. The alkaline phosphatase activity standardization as well as numerous international studies have made it possible to standardize the pediatric reference values. The hyperphosphatasemia etiologies are very well know but the hypophosphatasemia are hardly explored and can allow the diagnosis of pathologies including hypophosphatasia, a rare treatable disease.
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Fosfatasa Alcalina/sangre , Hipofosfatasia/sangre , Hipofosfatemia/sangre , Pediatría/normas , Fosfatasa Alcalina/análisis , Niño , Enfermedad/etiología , Humanos , Hiperfosfatemia/sangre , Hiperfosfatemia/diagnóstico , Hipofosfatasia/diagnóstico , Hipofosfatemia/diagnóstico , Pediatría/métodos , Valores de ReferenciaRESUMEN
Biochemical diagnosis of hereditary metabolic diseases requires the detection and simultaneous identification of a large number of compounds, hence the interest in metabolic profiles. Organic acid chromatography allows the identification of several hundred compounds and the quantification of the main molecules of interest. As part of the accreditation process for medical biology examinations according to standard NF EN ISO 15189, the group from the French society for inborn errors of metabolism (SFEIM) recommends an approach to accredit organic acid chromatography. Validation parameters and recommendations are discussed in this specific framework.
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Ácidos/orina , Cromatografía de Gases y Espectrometría de Masas/normas , Errores Innatos del Metabolismo/diagnóstico , Compuestos Orgánicos/orina , Urinálisis/normas , Acreditación , Ácidos/análisis , Adulto , Bioquímica/métodos , Bioquímica/normas , Niño , Servicios de Laboratorio Clínico/normas , Pruebas Diagnósticas de Rutina/métodos , Pruebas Diagnósticas de Rutina/normas , Femenino , Cromatografía de Gases y Espectrometría de Masas/métodos , Humanos , Recién Nacido , Compuestos Orgánicos/análisis , Fase Preanalítica/métodos , Fase Preanalítica/normas , Embarazo , Urinálisis/métodos , Toma de Muestras de Orina/métodos , Toma de Muestras de Orina/normas , Estudios de Validación como AsuntoRESUMEN
Biochemical diagnosis of hereditary metabolic diseases requires the detection and simultaneous identification of a large number of compounds, hence the interest in metabolic profiles. Amino acid chromatography allows the identification and quantification of more than forty compounds. As part of the accreditation process for medical biology examinations according to standard NF EN ISO 15189, the group from SFEIM recommends an approach to accredit amino acid chromatography. Validation parameters and recommendations are discussed in this specific framework.
Asunto(s)
Aminoácidos/análisis , Cromatografía/normas , Pruebas Diagnósticas de Rutina/normas , Errores Innatos del Metabolismo/diagnóstico , Acreditación/normas , Adulto , Aminoácidos/sangre , Aminoácidos/líquido cefalorraquídeo , Aminoácidos/orina , Amniocentesis/normas , Líquido Amniótico/química , Análisis Químico de la Sangre/métodos , Análisis Químico de la Sangre/normas , Recolección de Muestras de Sangre/normas , Niño , Cromatografía/métodos , Cromatografía Liquida/normas , Pruebas Diagnósticas de Rutina/métodos , Femenino , Humanos , Recién Nacido , Errores Innatos del Metabolismo/sangre , Errores Innatos del Metabolismo/líquido cefalorraquídeo , Errores Innatos del Metabolismo/orina , Tamizaje Neonatal/métodos , Tamizaje Neonatal/normas , Fase Preanalítica , Embarazo , Diagnóstico Prenatal/métodos , Diagnóstico Prenatal/normas , Espectrometría de Masas en Tándem/normas , Urinálisis/métodos , Urinálisis/normas , Toma de Muestras de Orina/normasRESUMEN
Biochemical diagnosis of hereditary metabolic diseases requires the detection and simultaneous identification of a large number of compounds, hence the interest in metabolic profiles. Acylcarnitine profile allows the identification and quantification of more than thirty compounds. As part of the accreditation process for medical biology examinations according to standard NF EN ISO 15189, the group from SFEIM recommends an approach to accredit acylcarnitine profile. Validation parameters and recommendations are discussed in this specific framework.
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Carnitina/análogos & derivados , Servicios de Laboratorio Clínico/normas , Pruebas Diagnósticas de Rutina/normas , Errores Innatos del Metabolismo/diagnóstico , Acreditación , Adulto , Amniocentesis/métodos , Amniocentesis/normas , Líquido Amniótico/química , Análisis Químico de la Sangre/métodos , Análisis Químico de la Sangre/normas , Recolección de Muestras de Sangre/métodos , Recolección de Muestras de Sangre/normas , Carnitina/análisis , Carnitina/sangre , Carnitina/orina , Niño , Cromatografía en Papel/normas , Femenino , Humanos , Recién Nacido , Masculino , Errores Innatos del Metabolismo/sangre , Errores Innatos del Metabolismo/orina , Tamizaje Neonatal/métodos , Tamizaje Neonatal/normas , Fase Preanalítica/métodos , Fase Preanalítica/normas , Embarazo , Diagnóstico Prenatal/métodos , Diagnóstico Prenatal/normas , Urinálisis/métodos , Urinálisis/normas , Toma de Muestras de Orina/métodos , Toma de Muestras de Orina/normasAsunto(s)
Biomarcadores/análisis , Técnicas y Procedimientos Diagnósticos/normas , Errores Innatos del Metabolismo/diagnóstico , Guías de Práctica Clínica como Asunto , Estudios de Validación como Asunto , Bioquímica/métodos , Bioquímica/normas , Biomarcadores/metabolismo , Técnicas de Laboratorio Clínico/métodos , Técnicas de Laboratorio Clínico/normas , Humanos , Espectrometría de Masas/métodos , Espectrometría de Masas/normas , Errores Innatos del Metabolismo/sangre , Errores Innatos del Metabolismo/genética , Guías de Práctica Clínica como Asunto/normas , Sociedades Científicas/organización & administración , Sociedades Científicas/normasRESUMEN
The evolution of cirrhosis is marked by quantitative and qualitative modifications of the fibrosis tissue and an increasing risk of complications such as hepatocellular carcinoma (HCC). Our purpose was to identify by FTIR imaging the spectral characteristics of hepatic fibrosis in cirrhotic patients with and without HCC. FTIR images were collected at projected pixel sizes of 25 and 2.7 µm from paraffinized hepatic tissues of five patients with uncomplicated cirrhosis and five cirrhotic patients with HCC and analyzed by k-means clustering. When compared to the adjacent histological section, the spectral clusters corresponding to hepatic fibrosis and regeneration nodules were easily identified. The fibrosis area estimated by FTIR imaging was correlated to that evaluated by digital image analysis of histological sections and was higher in patients with HCC compared to those without complications. Qualitative differences were also observed when fibrosis areas were specifically targeted at higher resolution. The partition in two clusters of the fibrosis tissue highlighted subtle differences in the spectral characteristics of the two groups of patients. These data show that the quantitative and qualitative changes of fibrosis tissue occurring during the course of cirrhosis are detectable by FTIR imaging, suggesting the possibility of subclassifying cirrhosis into different steps of severity.
Asunto(s)
Diagnóstico por Imagen , Cirrosis Hepática/diagnóstico por imagen , Cirrosis Hepática/patología , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/patología , Espectroscopía Infrarroja por Transformada de Fourier , Biopsia , Diagnóstico por Imagen/métodos , Humanos , Procesamiento de Imagen Asistido por Computador , Cirrosis Hepática/complicaciones , Neoplasias Hepáticas/etiología , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Carga TumoralRESUMEN
Fabry disease is a frequent lysosomal storage disorder secondary to the deficiency of alpha-galactosidase A enzyme. This X-linked genetic disease realizes progressive and systemic manifestations that affect both male and female. Fabry disease may present as "classical", as "late-onset" or "non-classical" forms. Symptoms and organ involvements of classical Fabry disease are acral pain crisis, cornea verticillata, hypertrophic cardiomyopathy, stroke and chronic kidney disease with proteinuria. Other common symptoms are often poorly recognized, such as gastrointestinal or ear involvements. In classical Fabry disease, symptoms first appear during childhood or during teenage years in males, but later in females. Patients with non-classical or late-onset Fabry disease have delayed manifestations or a single-organ involvement. Diagnosis is therefore difficult when classical organ involvements are missing, in paucisymptomatic patients or in late-onset forms. Recognition of Fabry disease is important because effective treatments are available. They have to be prescribed early. In male, diagnosis is made with alpha-galactosidase A enzyme activity dosage in leukocyte, that is very low or null in classical forms and under 30 percent in late-onset forms. Diagnosis is more challenging in females who may express normal residual enzyme activity. Other plasmatic biomarkers, such as lyso-globotriaosylceramide are interesting, especially in females. In this review, we aimed to summarize main clinical manifestations of Fabry disease to know when to evoke Fabry disease and propose a practical diagnosis algorithm to know how to diagnose.
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Enfermedad de Fabry/diagnóstico , Factores de Edad , Progresión de la Enfermedad , Enfermedad de Fabry/fisiopatología , Femenino , Humanos , Masculino , Factores SexualesRESUMEN
The metabolism of homocysteine is complex and involves many enzymes as well as vitamin-derived cofactors. Any dysregulation of this metabolism may lead to hyperhomocysteinemia (HHCy) which is responsible for many clinical disorders including thromboembolic events. HHCy may result from very different etiologies and is generally classified into three groups according to homocysteine concentrations: moderate (<30 µmol/L), intermediate (30-100 µmol/L) or major (>100 µmol/L). Major HHCy cases are generally due to monogenic defects of key enzymes involved in homocysteine metabolism, such as cystathionine-ß-synthase or 5,10-methylene-tetrahydrofolate reductase, or to any defect in vitamin B12 absorption, transport or metabolism. By contrast, moderate and intermediate HHCy tend to result from so-called "secondary" etiologies (e.g. tobacco, drugs, alcohol, vitamin deficiencies or pathological contexts). Here we describe the case of a patient with an unusually high plasma homocysteine concentration (1562 µmol/L) which was only explained by a combination of such secondary etiologies, among them chronic renal failure, hypothyroidism, the homozygous C677T MTHFR variant, a novel heterozygous variant of the MSR gene, and a vitamin deficiency. In addition, this patient exhibited a spectacular decline in homocysteine concentrations (returning to normal) after betaine and vitamin administration. In conclusion, this case highlights that major HHCy may also result from the combination of secondary etiologies, with vitamin deficiency as a triggering factor.
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Homocisteína/sangre , Hiperhomocisteinemia/etiología , Deficiencia de Vitamina B 12/sangre , Anciano , Betaína/administración & dosificación , Femenino , Homocistinuria/sangre , Homocistinuria/genética , Humanos , Hiperhomocisteinemia/sangre , Leucovorina/administración & dosificación , Metilenotetrahidrofolato Reductasa (NADPH2)/sangre , Metilenotetrahidrofolato Reductasa (NADPH2)/deficiencia , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Espasticidad Muscular/sangre , Espasticidad Muscular/genética , Trastornos Psicóticos/sangre , Trastornos Psicóticos/genética , Vitamina B 12/administración & dosificación , Complejo Vitamínico B/administración & dosificaciónRESUMEN
Recently, pre-analytical, analytical, and post-analytical issues have been addressed to implement biofluid FTIR spectroscopy as a novel diagnostic tool in the clinical setting. Although hemolysis, icterus, and hyperlipidemia are known to interfere with colorimetric and turbidimetric biochemical methods, there are no data on their impact on serum/plasma FTIR spectra. This study aimed at investigating the impact of hemoglobin, bilirubin, and triglycerides concentrations on plasma spectral analysis. Plasma samples with high concentrations of hemoglobin, conjugated bilirubin, or triglycerides were studied. To mimic the various concentrations observed in clinical setting, samples were diluted using normal plasma and analyzed using high-throughput FTIR spectroscopy. Hemolytic, icteric, and hyperlipidemic plasma spectra were compared with control plasma spectra. Unsupervised analysis of all spectra was performed using principal component analysis. The comparison between control and hemolytic plasmas did not show spectral differences in the range of hemoglobin concentrations observed in spurious or pathological hemolysis. By contrast, spectra from lipidemic plasmas had different spectral profiles compared with control plasma, exhibiting increased absorbance in lipid bands. Differences in the same spectral regions were observed in spectra from icteric plasma, which may be explained by the hyperlipidemia associated with cholestasis. PCA did not discriminate between control and hemolytic plasmas up to 1 g/L hemoglobin but confirmed the interference of bilirubin and triglycerides concentrations on spectral classification. Our results show that hemolysis does not have an impact on the plasma spectral profile except for high concentrations of hemoglobin rarely observed in clinical practice, whereas icterus and hyperlipidemia constitute significant confounding factors. Graphical abstract.
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Plasma/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Bilirrubina/sangre , Hemoglobinas/análisis , Hemólisis , Humanos , Hiperlipidemias/sangre , Ictericia/sangre , Triglicéridos/sangreRESUMEN
BACKGROUND: Atherosclerosis is an inflammatory process involving activation of monocytes recruited by various chemoattractant factors, among which lipoprotein(a) and its specific apolipoprotein apo(a). Lp(a) contains a specific apolipoprotein apo(a) which size is determined by a variable number of repeats of a specific structural domain, the kringle IV type 2 (IV-2). Lp(a) plasma concentration and apo(a) size is inversely correlated, and smaller apo(a) are major risk factors for coronary heart disease. DESIGN AND METHODS: The aim of this study was to evaluate the effect of recombinant apo(a) isoforms (containing 10, 18 or 34 kringles) on monocytes interacting with type I collagen. RESULTS: Apo(a) isoforms stimulated reactive oxygen species (ROS) and matrix metalloproteinase-9 (MMP-9) production by monocytes, and not modified monocytes adhesion on type I collagen. This effect was specific of apo(a) since no effect was observed in the presence of plasminogen and was inversely related to apo(a) size. The lysine analogue 6-aminohexanoic acid which blocks the lysine binding sites (LBS), and carboxypeptidase B (CpB) which cleaves carboxy-terminal lysine residues, abolished apo(a)-induced ROS and MMP-9 production, highlighting an effect mediated by apo(a) lysing-binding sites. CONCLUSIONS: These results indicate that activation of collagen-primed monocytes stimulated with apo(a) is a Kringle number-dependent effect and reinforce the hypothesis of a role for small size apo(a) isoforms in atherothrombosis.
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Apolipoproteínas A/farmacología , Colágeno Tipo I/farmacología , Monocitos/efectos de los fármacos , Proteínas Recombinantes/farmacología , Ácido Aminocaproico/farmacología , Animales , Apolipoproteínas A/biosíntesis , Apolipoproteínas A/química , Fibronectinas/farmacología , Células HEK293 , Humanos , Metaloproteinasa 9 de la Matriz/biosíntesis , Peso Molecular , Monocitos/citología , Monocitos/metabolismo , Plasminógeno/farmacología , Cultivo Primario de Células , Unión Proteica , Isoformas de Proteínas/biosíntesis , Isoformas de Proteínas/química , Isoformas de Proteínas/farmacología , Proteolisis , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/químicaRESUMEN
Infrared spectroscopy is a rapid, easy-to-operate, label-free and therefore cost-effective technique. Many studies performed on biofluids (eg, serum, plasma, urine, sputum, bile and cerebrospinal fluid) have demonstrated its promising application as a clinical diagnostic tool. Given all these characteristics, infrared spectroscopy appears to be an ideal candidate to be implemented into the clinics. However, before considering its translation, a clear effort is needed to standardise protocols for biofluid spectroscopic analysis. To reach this goal, careful investigations to identify and track errors that can occur during the pre-analytical phase is a crucial step. Here, we report for the first time, results of investigations into pre-analytical factors that can affect the quality of the spectral data acquired on serum and plasma, such as the impact of long-term freezing time storage of samples as well as the month-to-month reproducibility of the spectroscopic analysis. The spectral data discrimination has revealed to be majorly impacted by a residual water content variation in serum and plasma dried samples.
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Análisis Químico de la Sangre/métodos , Plasma/química , Suero/química , Espectrofotometría Infrarroja , Métodos Analíticos de la Preparación de la Muestra , Humanos , Humedad , Agua/químicaRESUMEN
The prevention of perinatal brain damage following preterm birth remains a public health priority. Melatonin has been shown to be a promising neuroprotectant in neonatal preclinical models of brain damage, but few studies have investigated melatonin secretion in newborns. We hypothesized that melatonin circulating levels would be lower in preterm compared to term infants. We conducted a prospective, longitudinal, multicenter study to assess melatonin, and 6-sulfatoxy-melatonin (aMT6s) concentrations, measured by radioimmunoassay. Among 209 neonates recruited, 110 were born before 34 gestational weeks (GW) and 99 born after 34 GW. Plasma melatonin concentrations, measured at birth and on Day 3 were below detectable levels (≤7 pg/mL) in 78% and 81%, respectively, of infants born before 34 GW compared to 57% and 34%, respectively, of infants born after 34 GW. The distribution of plasma melatonin concentrations was found to be correlated with gestational age at both time-points (p < 0.001). Median urine aMT6s concentrations were significantly lower in infants born before 34 GW, both on Day 1 (230 ng/L vs. 533 ng/L, p < 0.0001) and on Day 3 (197 ng/L vs. 359 ng/L, p < 0.0001). In conclusion, melatonin secretion appears very low in preterm infants, providing the rationale for testing supplemental melatonin as a neuroprotectant in clinical trials.
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Recien Nacido Prematuro/sangre , Melatonina/sangre , Madres , Biomarcadores , Encéfalo/embriología , Femenino , Humanos , Lactante , Recién Nacido , Melatonina/análogos & derivados , Neurogénesis , EmbarazoRESUMEN
Affecting more than 30% of the Western population, nonalcoholic fatty liver disease (NAFLD) is the most common liver disease and can lead to multiple complications, including nonalcoholic steatohepatitis (NASH), cancer, hypertension, and atherosclerosis. Insulin resistance and obesity are described as potential causes of NAFLD. However, we surmised that factors such as extracellular matrix remodeling of large blood vessels, skin, or lungs may also participate in the progression of liver diseases. We studied the effects of elastin-derived peptides (EDPs), biomarkers of aging, on NAFLD progression. We evaluated the consequences of EDP accumulation in mice and of elastin receptor complex (ERC) activation on lipid storage in hepatocytes, inflammation, and fibrosis development. The accumulation of EDPs induces hepatic lipogenesis (i.e., SREBP1c and ACC), inflammation (i.e., Kupffer cells, IL-1ß, and TGF-ß), and fibrosis (collagen and elastin expression). These effects are induced by inhibition of the LKB1-AMPK pathway by ERC activation. In addition, pharmacological inhibitors of EDPs demonstrate that this EDP-driven lipogenesis and fibrosis relies on engagement of the ERC. Our data reveal a major role of EDPs in the development of NASH, and they provide new clues for understanding the relationship between NAFLD and vascular aging.
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Diabetes Mellitus Tipo 2/complicaciones , Elastina/metabolismo , Regulación de la Expresión Génica , Hígado/metabolismo , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Fragmentos de Péptidos/metabolismo , Receptores de Superficie Celular/agonistas , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Índice de Masa Corporal , Células Cultivadas , Estudios de Cohortes , Dieta Alta en Grasa/efectos adversos , Progresión de la Enfermedad , Elastina/sangre , Elastina/genética , Matriz Extracelular/inmunología , Matriz Extracelular/metabolismo , Matriz Extracelular/patología , Femenino , Humanos , Lipogénesis , Hígado/inmunología , Hígado/patología , Masculino , Ratones Endogámicos C57BL , Ratones Mutantes , Enfermedad del Hígado Graso no Alcohólico/complicaciones , Enfermedad del Hígado Graso no Alcohólico/patología , Enfermedad del Hígado Graso no Alcohólico/fisiopatología , Obesidad Mórbida/complicaciones , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/genética , Prueba de Estudio Conceptual , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Transducción de SeñalRESUMEN
Maintenance of mesenchymal stem cells (MSCs) requires a tissue-specific microenvironment (i.e., niche), which is poorly represented by the typical plastic substrate used for two-dimensional growth of MSCs in a tissue culture flask. The objective of this study was to address the potential use of collagen-based medical devices (HEMOCOLLAGENE®, Saint-Maur-des-Fossés, France) as mimetic niche for MSCs with the ability to preserve human MSC stemness in vitro. With a chemical composition similar to type I collagen, HEMOCOLLAGENE® foam presented a porous and interconnected structure (>90%) and a relative low elastic modulus of around 60 kPa. Biological studies revealed an apparently inert microenvironment of HEMOCOLLAGENE® foam, where 80% of cultured human MSCs remained viable, adopted a flattened morphology, and maintained their undifferentiated state with basal secretory activity. Thus, three-dimensional HEMOCOLLAGENE® foams present an in vitro model that mimics the MSC niche with the capacity to support viable and quiescent MSCs within a low stiffness collagen I scaffold simulating Wharton's jelly. These results suggest that haemostatic foam may be a useful and versatile carrier for MSC transplantation for regenerative medicine applications.
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Microambiente Celular , Colágeno , Células Madre Mesenquimatosas , Preservación Biológica/métodos , Medicina Regenerativa/instrumentación , HumanosRESUMEN
In this study, we investigated the effect of [N-(5-chloro-2-hydroxyphenyl)-l-aspartato] chlorogallate (GS2), a new water soluble gallium complex, on cell invasion and on the expression and activity of matrix metalloproteinases (MMPs) in human metastatic HT-1080 fibrosarcoma and MDA-MB 231 breast carcinoma cells. The effect on cell invasion was studied using a modified Boyden chamber coated with a type-I collagen. We analyzed the effect of GS2 on MMP-2, MMP-9, and MMP-14 via zymography and enzymatic assay using high affinity fluorogenic substrates. The expression of MMP mRNA was analyzed via qRT-PCR. GS2 induced a decrease in cell invasion. A dose-dependent inhibition effect was observed on the activities of MMP-2, MMP-9, and MMP-14 with the IC50 values of 168, 82, and 20 µM, respectively. A decrease in the expression of MMP-14 mRNA was observed in both cell lines, whereas the expression of MMP-2 and MMP-9 mRNA was decreased only in the MDA-MB231 cells. Data obtained for the expression of MMP-14 mRNA were confirmed via Western blotting. In fact, MMP-14 expression was decreased in the presence of GS2. Overall, these data show that GS2 is a promising compound for anti-invasive and anticancer therapy.
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Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Complejos de Coordinación/farmacología , Fibrosarcoma/tratamiento farmacológico , Galio/farmacología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Metaloproteinasa 14 de la Matriz/metabolismo , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/patología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Femenino , Fibrosarcoma/enzimología , Fibrosarcoma/patología , Galio/química , Humanos , Metaloproteinasa 14 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/metabolismo , Invasividad Neoplásica , Células Tumorales CultivadasRESUMEN
[This corrects the article on p. 55 in vol. 7, PMID: 27014069.].
RESUMEN
Tumor cells are confronted to a type I collagen rich environment which regulates cell proliferation and invasion. Biological aging has been associated with structural changes of type I collagen. Here, we address the effect of collagen aging on cell proliferation in a three-dimensional context (3D).We provide evidence for an inhibitory effect of adult collagen, but not of the old one, on proliferation of human fibrosarcoma HT-1080 cells. This effect involves both the activation of the tyrosine kinase Discoidin Domain Receptor 2 (DDR2) and the tyrosine phosphatase SHP-2. DDR2 and SHP-2 were less activated in old collagen. DDR2 inhibition decreased SHP-2 phosphorylation in adult collagen and increased cell proliferation to a level similar to that observed in old collagen.In the presence of old collagen, a high level of JAK2 and ERK1/2 phosphorylation was observed while expression of the cell cycle negative regulator p21CIP1 was decreased. Inhibition of DDR2 kinase function also led to an increase in ERK1/2 phosphorylation and a decrease in p21CIP1 expression. Similar signaling profile was observed when DDR2 was inhibited in adult collagen. Altogether, these data suggest that biological collagen aging could increase tumor cell proliferation by reducingthe activation of the key matrix sensor DDR2.