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Cell Uptake (cell + uptake)
Selected AbstractsFabrication of Galactosylated Polyethylenimine and Plasmid DNA Multilayers on poly (D,L -lactic acid) Films for in situ Targeted Gene Transfection,ADVANCED ENGINEERING MATERIALS, Issue 5 2009Yan Hu This study presents surface-mediated targeted in situ gene delivery from gene-tagged poly(D,L -lactic acid) (PDLLA) films, which were fabricated via a layer-by-layer (LbL) assembly technique with galactosylated polyethylenimine (GP) and plasmid DNA (pDNA, pSV-,-galactosidase). A linear growth of GP/pDNA multilayered films was observed. The pDNA was continuously released from multilayered films for over 32,h. The multilayered structure degraded and simultaneously formed GP/pDNA complexes in situ when exposing to a physiological environment. The pDNA was well protected by GP against DNase I digestion within formed GP/pDNA complexes. Our results demonstrated that GP contributes to receptor-mediated targeting for cell uptake and in situ gene transfection. The results reported here are potentially important for gene therapy, surface engineering of biomaterials, tissue engineering and implant technology. [source] Physico-chemical requirements for cellular uptake of pAntp peptideFEBS JOURNAL, Issue 5 2001Role of lipid-binding affinity The pAntp peptide, corresponding to the third helix of the Antennapedia homeodomain, is internalized by a receptor-independent process into eucaryotic cells. The precise mechanism of entry remains unclear but the interaction between the phospholipids of plasma membrane and pAntp is probably involved in the translocation process. In order to define the role of peptide,lipid interaction in this mechanism and the physico-chemical properties that are necessary for an efficient cellular uptake, we have carried out an Ala-Scan mapping. The peptides were labeled with a fluorescent group (7-nitrobenz-2-oxo-1,3-diazol-4-yl-; NBD) and their cell association was measured by flow cytometry. Furthermore, we determined the fraction of internalized peptide by using a dithionite treatment. Comparison between cell association and cell uptake suggests that the affinity of pAntp for the plasma membrane is required for the import process. To further investigate which are the physico-chemical requirements for phospholipid-binding of pAntp, we have determined the surface partition coefficient of peptides by titrating them with phospholipid vesicles having different compositions. In addition, we estimated by circular dichroism the conformation adopted by these peptides in a membrane-mimetic environment. We show that the phospholipid binding of pAntp depends on its helical amphipathicity, especially when the negative surface charge density of phospholipid vesicles is low. The cell uptake of pAntp, related to lipid-binding affinity, requires a minimal hydrophobicity and net charge. As pAntp does not seem to translocate through an artificial phospholipid bilayer, this might indicate that it could interact with other cell surface components or enters into cells by a nonelucidated biological mechanism. [source] Length-Dependent Uptake of DNA-Wrapped Single-Walled Carbon Nanotubes,ADVANCED MATERIALS, Issue 7 2007L. Becker A length threshold for cell uptake of DNA-wrapped single-walled carbon nanotubes (SWNTs) by human lung fibroblasts (IMR90) is identified. Competitive uptake experiments with well-defined and characterized length fractions show that SWNTs above the length threshold are excluded from the cell, whereas SWNTs labeled with Cy3-derivatized DNA below the threshold are able to access the cell interior, as shown in the fluorescence image and on the cover. [source] Transglutaminase 2 limits murine peritoneal acute gout-like inflammation by regulating macrophage clearance of apoptotic neutrophilsARTHRITIS & RHEUMATISM, Issue 10 2006David M. Rose Objective Monosodium urate monohydrate (MSU) crystals have remarkable inflammatory potential. However, gouty inflammation is spontaneously self-limited, an occurrence recognized since antiquity. Gouty synovitis is driven and sustained by neutrophil influx. Importantly, macrophage phagocytosis of apoptotic (but not necrotic) neutrophils is antiinflammatory. Therefore, we tested the hypothesis that efficient clearance of apoptotic neutrophils by macrophages is one of the factors that restrains the progression of gouty inflammation. Macrophage expression of transglutaminase 2 (TG2), a multifunctional protein with reciprocally regulated transamidation and purine nucleotide,binding activities, promotes apoptotic leukocyte uptake. In this study, we tested the specific role of macrophage TG2 expression in MSU crystal,induced inflammation. Methods We studied MSU crystal,induced peritonitis in TG2,/, and congenic TG2+/+ mice. We also studied the effects of TG2 on apoptotic cell uptake by cultured macrophages. Results TG2,/, mice demonstrated more progressive neutrophilic accumulation than did TG2+/+ mice, which was associated with delayed clearance of apoptotic neutrophils during MSU crystal,induced peritonitis. We observed defective phagocytosis of apoptotic leukocytes by TG2,/, peritoneal macrophages, which was corrected by soluble extracellular TG2. Transamidation catalytic activity of TG2 was not required to mediate macrophage uptake of apoptotic leukocytes. In contrast, the TG2 nucleotide binding site residue K173 was critical for this TG2 function. TG2 bound to GDP, ADP, or ATP (but not to GTP) rescued defective apoptotic leukocyte uptake by TG2,/, macrophages. Conclusion Enhancement of apoptotic neutrophil uptake by macrophage-derived TG2 restrains gout-like neutrophilic peritoneal inflammation. Differential binding of TG2 by purine nucleotides may contribute to clinical variability in the extent and duration of gouty inflammation. [source] Surface-Functionalized Ultrasmall Superparamagnetic Nanoparticles as Magnetic Delivery Vectors for CamptothecinCHEMMEDCHEM, Issue 6 2009Feride Cengelli Abstract Drug,nanoparticle conjugates: The anticancer drug camptothecin (CPT) was covalently linked at the surface of ultrasmall superparamagnetic iron oxide nanoparticles (USPIOs) via a linker, allowing drug release by cellular esterases. Nanoparticles were hierarchically built to achieve magnetically-enhanced drug delivery to human cancer cells and antiproliferative activity. The linking of therapeutic drugs to ultrasmall superparamagnetic iron oxide nanoparticles (USPIOs) allowing intracellular release of the active drug via cell-specific mechanisms would achieve tumor-selective magnetically-enhanced drug delivery. To validate this concept, we covalently attached the anticancer drug camptothecin (CPT) to biocompatible USPIOs (iron oxide core, 9,10,nm; hydrodynamic diameter, 52,nm) coated with polyvinylalcohol/polyvinylamine (PVA/aminoPVA). A bifunctional, end-differentiated dicarboxylic acid linker allowed the attachment of CPT to the aminoPVA as a biologically labile ester substrate for cellular esterases at one end, and as an amide at the other end. These CPT,USPIO conjugates exhibited antiproliferative activity in,vitro against human melanoma cells. The intracellular localization of CPT,USPIOs was confirmed by transmission electron microscopy (iron oxide core), suggesting localization in lipid vesicles, and by fluorescence microscopy (CPT). An external static magnetic field applied during exposure increased melanoma cell uptake of the CPT,USPIOs. [source] ,Danger' effect of low-density lipoprotein (LDL) and oxidized LDL on human immature dendritic cellsCLINICAL & EXPERIMENTAL IMMUNOLOGY, Issue 3 2007R. Zaguri Summary Dendritic cell (DC) maturation may accelerate autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis, and may contribute to accelerated atherosclerosis seen in these patients. The immune system responds to both exogenous and endogenous ,dangerous' signals that can induce dendritic cell maturation. We have found that autologous plasma contains danger signals that induce up-regulation of major histocompatibility complex (MHC) class II and co-stimulatory molecules in immature DCs (iDCs). The objective of this study was to determine whether low-density lipoprotein (LDL) and/or oxidized LDL (oxLDL) constitute danger signals, and to assess the effect of exposure to LDL and oxLDL following monocyte differentiation into iDCs in lipoprotein-deficient serum (LPDS). IDCs were generated in the presence of autologous plasma or LPDS. Expression of maturation and migration molecules was evaluated using flow cytometry, and morphology was assessed by light microscopy. Pro- or anti-apoptotic effect was determined using annexin V and propidium iodide binding. Phagocytosis of apoptotic cells was evaluated using autologous plasma or LPDS. LDL and oxLDL were clearly able to slightly up-regulate levels of HLA-DR and co-stimulatory molecule CD86. High oxLDL concentrations (50,100 µg/ml) were associated with expression of additional maturation molecules. Moreover, iDCs that were prepared in LPDS showed partial maturation following exposure to LDL and oxLDL, and improved tolerogenic apoptotic cell uptake. This study suggests that oxLDL, and to some extent LDL, are at least partly responsible for the iDC ,danger' response induced by autologous plasma. [source] | ||||||||||||||||