causes chronic pulmonary infections in patients with cystic fibrosis (CF). mucoid conversion, defined by overproduction of the exopolysaccharide alginate, correlates with accelerated decline in CF patient lung function. Recalcitrance of the mucoid phenotype to clearance by antibiotics and the immune response is well documented.
View Article and Find Full Text PDFCellular transformation is the first step in cancer development. Two features of cellular transformation are proliferation in reduced serum and loss of contact inhibition. Electronic Cell-Substrate Impedance Sensing (ECIS) measurements have been used to measure cellular proliferation, cytotoxicity, apoptosis, and attachment.
View Article and Find Full Text PDFNonsteroidal anti-inflammatory drugs (NSAIDs) are known to prevent colorectal tumorigenesis. Although antitumor effects of NSAIDs are mainly due to inhibition of cyclooxygenase activity, there is increasing evidence that cyclooxygenase-independent mechanisms may also play an important role. The early growth response-1 (EGR-1) gene is a member of the immediate-early gene family and has been identified as a tumor suppressor gene.
View Article and Find Full Text PDFMulticontrast microscopy techniques were used to comprehensively and dynamically map the cellular contact area adhering to a substrate. The natural fringe patterns observed with interference reflection contrast microscopy were used to map the dynamic fingerprint of a porcine pulmonary artery endothelial cell's ventral surface and to examine the focal and/or close contacts to the substrate when exposed to a toxic agent Cytochalasin D. In addition, differential interference contrast microscopy sequentially imaged the overall cellular morphological responses to the agent.
View Article and Find Full Text PDFThe surface micromechanical properties of 2-hydroxyethyl methacrylate (HEMA) and 2-methacryloxyethyl trimethyl ammonium chloride (MAETAC) copolymer hydrogels are probed using atomic force microscopy. HEMA-MAETAC polyelectrolyte hydrogels with increasing positive charge concentrations ranging from 0 to 400mM in increments of 40mM, are fabricated using different proportions of HEMA and MAETAC monomers. Increasing proportions of positively charged MAETAC monomers produce hydrogels with increasingly swollen states and correspondingly decreasing measures of stiffness, or Young's modulus.
View Article and Find Full Text PDFIndium tin oxide (ITO) biosensors are used to perform simultaneous optical and electrical measurements in order to examine the dynamic cellular attachment, spreading, and proliferation of endothelial cells (ECs) as well as cytotoxic effects when exposed to cytochalasin D. A detailed description of the fabrication of these sensors is provided and their superior optical characteristics are qualitatively shown using four different microscopic images. Differential interference contrast microscopy (DICM) images were acquired simultaneously with micro-impedance measurements as a function of frequency and time.
View Article and Find Full Text PDFThis study analyzes the cellular microelectrode voltage measurement errors produced by active and passive current regulation, and the propagation of these errors into cellular barrier function parameter estimates. The propagation of random and systematic errors into these parameters is accounted for within a Riemannian manifold framework consistent with information geometry. As a result, the full non-linearity of the model parameter state dependence, the instrumental noise distribution, and the systematic errors associated with the voltage to impedance conversion, are accounted for.
View Article and Find Full Text PDFThis study quantifies the dynamic attachment and spreading of porcine pulmonary artery endothelial cells (PPAECs) on optically thin, indium tin oxide (ITO) biosensors using simultaneous differential interference contrast microscopy (DICM) and electrical microimpedance spectroscopy. A lock-in amplifier circuit monitored the impedance of PPAECs cultivated on the transparent ITO bioelectrodes as a function of frequency between 10 Hz and 100 kHz and as a function of time, while DICM images were simultaneously acquired. A digital image processing algorithm quantified the cell-covered electrode area as a function of time.
View Article and Find Full Text PDFUsing equilibrium swelling and non-equilibrium membrane potential measurements, this study assesses the charge density in two representative series of polyelectrolyte hydrogels and examines the morphological and proliferative responses of endothelial cells as a function of the prepared charge offset. The neutral monomers 2-hydroxyethylmethacrylate (HEMA) and poly(ethylene glycol) dimethacrylate (n = 1,000) (PEGDMA) were copolymerized with either the acidic monomer 2-sulfoethyl methacrylate (SEMA) or the basic monomer methacryloxy ethyltrimethylammonium chloride (MAETAC) to make membranes with pregelation charge offset concentrations varying from 0 to +/-200 mM. A thermodynamic analysis of swelling and membrane potential measurements quantified the hydrogel charge density state following equilibration at different ion strengths.
View Article and Find Full Text PDFEndothelial cell adhesion and barrier function play a critical role in many biological and pathophysiological processes. The decomposition of endothelial cell adhesion and barrier function into cell-cell and cell-matrix components using frequency dependent cellular micro-impedance measurements has, therefore, received widespread application. Few if any studies, however, have examined the precision of these model parameters.
View Article and Find Full Text PDFThe use of an optically thin indium-tin-oxide (ITO) electrode is presented for an optoelectric biosensor simultaneously recording optical images and microimpedance to examine time-dependent cellular growth. The transmittance of a 100 nm thick ITO electrode layer is approximately the same as the transmittance of a clean glass substrate, whereas the industry-standard Au(47.5 nm)/Ti(2.
View Article and Find Full Text PDFFrequency dependent cellular micro-impedance estimates obtained from a gold two-electrode configuration using phase sensitive detection have become increasingly used to evaluate cellular barrier model parameters. The results of this study show that cellular barrier function parameter estimates optimized using measurements obtained from this biosensor are highly susceptible to both time dependent and systematic instrumental artifacts. Based on a power spectral analysis of experimentally measured microelectrode voltages, synchronous, 60 Hz, and white Gaussian noise were identified as the most significant time dependent instrumental artifacts.
View Article and Find Full Text PDFThis study describes the fabrication and performance of an endothelial cell compatible, optically thin, indium tin oxide (ITO) microimpedance biosensor. The biosensor was constructed by sputtering a thin insulating layer of silicon nitride (Si(3)N(4)) onto a 100 nm thick ITO layer. Indium tin oxide electrodes were formed by chemically etching 250 or 500 microm diameter holes through the Si(3)N(4) insulating layer.
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