RESUMEN
BACKGROUND: Breast cancer is the most common cancer in women globally, and diagnosing it early and finding potential drug candidates against multi-drug resistant metastatic breast cancers provide the possibilities of better treatment and extending life. METHODS: The current study aimed to evaluate the synergistic anti-metastatic activity of Curcumin (Cur) and Paclitaxel (Pacli) individually, the combination of Curcumin-Paclitaxel (CP), and also in conjugation with gold nanoparticles (AuNP-Curcumin (Au-C), AuNP-Paclitaxel (Au-P), and AuNP-Curcumin-Paclitaxel (Au-CP)) in various in vitro and in vivo models. RESULTS: The results from combination treatments of CP and Au-CP demonstrated excellent synergistic cytotoxic effects in triple-negative breast cancer cell lines (MDA MB 231 and 4T1) in in vitro and in vivo mouse models. Detailed mechanistic studies were performed that reveal that the anti-cancer effects were associated with the downregulation of the expression of VEGF, CYCLIN-D1, and STAT-3 genes and upregulation of the apoptotic Caspase-9 gene. The group of mice that received CP combination therapy (with and without gold nanoparticles) showed a significant reduction in the size of tumor when compared to the Pacli alone treatment and control groups. CONCLUSIONS: Together, the results suggest that the delivery of gold conjugated Au-CP formulations may help in modulating the outcomes of chemotherapy. The present study is well supported with observations from cell-based assays, molecular and histopathological analyses.
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Carcinogénesis/efectos de los fármacos , Curcumina/farmacología , Oro/farmacología , Nanopartículas del Metal/administración & dosificación , Metástasis de la Neoplasia/tratamiento farmacológico , Paclitaxel/farmacología , Animales , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Línea Celular , Línea Celular Tumoral , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Femenino , Células HEK293 , Humanos , Ratones , Ratones Endogámicos BALB C , Neoplasias de la Mama Triple Negativas/tratamiento farmacológicoRESUMEN
Sepsis is an abnormal immune response to infection characterized by an overwhelming systemic inflammation and cell death. Non-apoptotic cell death pertaining to pyroptosis, necroptosis and autophagy contribute to sepsis pathogenesis apart from classical apoptotic cell death. The objective of the current study is to investigate the presence of molecular markers of relevance to apoptotic and non-apoptotic cell death in control healthy subjects and septic patient survivors. Sepsis survivors (N = 24) and healthy human volunteers (N = 16) [40 total subjects] were recruited into the study. Clinical intervention included antibiotic treatment regimen administered to patients upon clinical diagnosis of sepsis followed by blood draw 18-24 hr post-antibiotic dose. Serum samples analyzed by enzyme-linked immunosorbent assay (ELISA) and peripheral blood mononuclear cells (PBMCs) by flow cytometry analysis for identification of cell death markers. Cell death markers analyzed by ELISA and flow cytometry included caspase-1, caspase-3, MLKL, RIPK3, p62 and LC3B. Serum and peripheral blood mononuclear cells (PBMCs) of septic survivors and healthy controls analyzed for the presence of distinct cell death markers. Markers of relevance to apoptosis (caspase-3), pyroptosis (caspase-1), necroptosis (MLKL) and autophagy (p62 and LC3B) were compared between septic survivors and healthy controls. ELISA analysis suggested significant alterations in the serum levels of non-apoptotic cell death markers, caspase-1 and p62/SQSTM1, in septic survivors compared to healthy controls (p < 0.05). There was no significant difference in the serum levels of caspase-3 and MLKL between septic survivors and healthy control subjects (p> 0.05). Intracellular caspase-1 levels did not show any significant alterations between septic survivors and healthy control subjects (p > 0.05). Flow cytometry analysis suggested significant increase in the intracellular expression of caspase-3, MLKL and its associated kinase RIPK3, and p62/SQSTM1 (p < 0.05) in sepsis patient survivors when compared to healthy human subjects. The current observational study identified significantly elevated levels of non-apoptotic cell death markers in sepsis patients compared to healthy controls. Noteworthy observation is the significant modulation of non-apoptotic cell death markers in serum samples derived from septic survivors post-antibiotic administration compared to healthy control subjects. Preliminary results serve as a basis for further mechanistic investigations to elucidate the role of distinct cell death markers in the prediction of clinical outcomes in sepsis.
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Biomarcadores , Muerte Celular , Sepsis/metabolismo , Sobrevivientes , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Caspasas , Muerte Celular/genética , Muerte Celular/inmunología , Cuidados Críticos , Enfermedad Crítica , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunofenotipificación , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Sepsis/diagnóstico , Sepsis/etiología , Sepsis/terapiaRESUMEN
A scientific approach is presented describing the fabrication of nanoprobe (GloTrack) that can act as cardiac precursor label to segregate cells from cardiac/non cardiac origins and traced by magnetic resonance imaging (MRI). Signal regulatory protein alpha (SIRPA) and kinase domain receptor (KDR) recognizing antibodies, form a layer on super paramagnetic iron oxide nanoparticle - poly-ethylene glycol (SPION-PEG) complex, and bind to protein expressed on the surface of cardiac muscle cells. Physical attributes size, distribution, labelling efficiency, echocardiogram (ECG) changes and bio-distribution by MRI were analysed. The results indicate that GloTrack has an average size of 471â¯nm, exhibits negative potential and promotes labelling efficiency. The bio-distribution of GloTrack in in vivo experiments was traceable in 7T MRI showing high accumulation of GloTrack in cardiac muscles as compared to the liver and spleen. ECG data revealed that GloTrack segregated cardiac precursors has the potential benefit in treating heart failure, thereby paving way in the development of minimal cell manipulation with targeted cell delivery approaches.
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Sistemas de Liberación de Medicamentos , Nanopartículas Magnéticas de Óxido de Hierro , Imagen por Resonancia Magnética , Miocardio/citología , Células Madre/metabolismo , Animales , Anticuerpos Monoclonales , Separación Celular , Ecocardiografía , Inyecciones Intraperitoneales , Isoproterenol/administración & dosificación , Isoproterenol/efectos adversos , Hígado , Nanopartículas Magnéticas de Óxido de Hierro/química , Nanopartículas Magnéticas de Óxido de Hierro/ultraestructura , Ratones , Ratones Endogámicos C57BL , Micelas , Microscopía Confocal , Microscopía Electrónica de Transmisión , Infarto del Miocardio/inducido químicamente , Miocardio/metabolismo , Polietilenglicoles/química , Espectrometría Raman , BazoRESUMEN
The increasing demand for organs for transplantation necessitates the development of substitutes to meet the structural and physiological functions. Tissue decellularization and recellularization aids in retaining the three-dimensional integrity, biochemical composition, tissue ultra-structure, and mechanical behavior, which makes them functionally suitable for organ transplantation. Herein, we attempted to rebuild functional liver grafts in small animal model (Wistar rat) with a potential of translation. A soft approach was adopted using 0.1% SDS (Sodium Dodecyl Sulfate) for decellularization and primary hepatocytes were used as a potential cell source for recellularization. The decellularization process was evaluated and confirmed using histology, DNA content, ultra-structure analysis. The resultant scaffold was re-seeded with the rat hepatocytes and their biocompatibility was assessed by its metabolic functions and gene expression. The structural components of the Extracellular matrix (ECM) (Laminins, Collagen type I, Reticulins) were conserved and the liver cell-specific proteins like CK-18, alpha-fetoprotein, albumin were expressed in the recellularized scaffold. The functionality and metabolic activity of the repopulated scaffold were evident from the albumin and urea production. Expression of Cytokeratin-19 (CK-19), Glucose 6-Phosphatase (G6P), Albumin, Gamma Glutamyl Transferase (GGT) genes has distinctly confirmed the translational signals after the repopulation process. Our study clearly elucidates that the native extracellular matrix of rat liver can be utilized as a scaffold for effective recellularization for whole organ regeneration.