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Epithelial Cell Cultures (epithelial + cell_culture)
Selected AbstractsFunctionalized Poly(D,L -lactide) for Pulmonary Epithelial Cell CultureADVANCED ENGINEERING MATERIALS, Issue 4 2010Yuan-Min Lin Functional groups on a material surface affect the response of many cell types. As part of our strategy aimed at engineering lung tissue, we introduced functional groups into the surface of Poly(D,L -lactide) (PDLLA) films to improve its suitability for the culture of mature pulmonary epithelial cells (A549 line) using two different methods. The first method, aminolysis, can introduce primary amines into PDLLA films by transesterification using 1,15% of ethylenediamine in isopropanol. The second method, a branching modification, can generate amine-terminated or carboxylic acid-terminated tree-like branched architectures. All modified PDLLA surfaces exhibited lower water contact angles, i.e. are more hydrophilic than unmodified PDLLA. PDLLA treated with 15% ethylenediamine exhibited a rougher surface than the control, and PDLLA with branching modification had a droplet-like surface topography as visualized by atomic force microscopy (AFM). PDLLA treated with 15% ethylenediamine and branching modification with two and three generations enhanced the attachment of pulmonary epithelial cells measured using Hoechst dye. Immunostaining demonsatrated that amine-terminated branched architectures allowed for better focal adhesion point formation than the control 24,h after cell seeding. Furthermore, they also induced higher A549 cell populations and levels of activity after 4 days in culture measured using Hoechst dye and WST1 cell proliferation reagents, respectively. In contrast, carboxylic acid-terminated branching architectures were found to reduce the cell population size after 4 days. It was concluded that the concentration, type and distribution of surface functional groups can affect significantly the behavior of pulmonary epithelial cells growing on a PDLLA surface, and PDLLA film modified with two or three generations of amine-terminated branched architectures is a suitable 2D scaffold for the culture of of pulmonary epithelial cells. [source] Cultured epithelial cells response to phototherapy with low intensity laser,LASERS IN SURGERY AND MEDICINE, Issue 4 2007Fernanda P. Eduardo PhD Abstract Background and Objectives Little is known about the intracellular response of epithelial cells to phototherapy. The aim of this in vitro study was to analyze the effect of phototherapy with low-energy lasers with different wavelengths and powers on cultured epithelial cell growth under different nutritional conditions. Study Design/Materials and Methods Epithelial cell cultures (Vero cell line) grown in nutritional deficit in culture medium supplemented with 2% fetal bovine serum (FBS) were irradiated with low-energy laser from one to three times with a GaAlAs laser (660 nm) and InGaAlP (780 nm), 40 and 70 mW, respectively, with 3 or 5 J/cm2. Cell growth was indirectly assessed by measuring the cell mitochondrial activity. Results Nonirradiated cell cultures grown in nutritional regular medium supplemented with 10% FBS produced higher cell growth than all cultures grown in nutritional deficit irradiated or not. The overall cell growth of cultures grown under nutritionally deficit conditions was significantly improved especially when irradiated with 780 nm for three times. Conclusions Phototherapy with the laser parameters tested increases epithelial cell growth rate for cells stressed by growth under nutritionally deficient states. This cell growth improvement is directly proportional to the number of irradiations; however, was not enough to reach the full cell growth potential rate of Vero epithelial cell line observed when growing under nutritional regular condition. Lasers Surg. Med. 39: 365,372, 2007. © 2007 Wiley-Liss, Inc. [source] Alternative splicing of cyclooxygenase-1 gene: altered expression in leucocytes from patients with bronchial asthma and association with aspirin-induced 15-HETE releaseALLERGY, Issue 6 2007M. L. Kowalski Background:, Cyclooxygenase-1 (COX-1) is a key enzyme involved in generation of prostanoids, important mediators and modulators of asthmatic inflammation. In a subpopulation of aspirin-sensitive asthmatics (ASA) inhibition of COX-1 by nonsteroidal anti-inflammatory drugs results in activation of inflammatory cells and development of symptoms. Alternatively spliced variants of COX-1 lacking 111 bp from exon 9 were described previously but have never been identified in human leucocytes peripheral blood leucocytes (PBL) or upper airway epithelial cells. We aimed to assess the expression of spliced variants of COX-1 mRNA in PBLs from patients with asthma and in healthy subjects (HS) referring the expression to patients characteristics (including ASA-sensitivity) and to aspirin-triggered 15-hydroxyeicosatetraenoic acid (15-HETE) generation. Methods:, The study included 30 patients with ASA, 30 patients with aspirin-tolerant asthma (ATA) and 30 HS serving as controls. Nasal polyps for epithelial cell cultures were obtained from 10 patients with chronic rhinosinusitis. Expression of full length and spliced variants of COX-1 enzyme was detected by RT-PCR and presented as the ratio of full-length COX-1 to alternatively spliced COX-1 mRNA [COX-1 alternative splicing index (COX-1 AS index)]. Release of eicosanoids (PGE2 and 15-HETE) by PBLs was measured with enzyme immunoassay. Results:, In both PBLs and airway epithelial cells the expression of full-length product prevailed over spliced variants of COX-1 enzyme. Cyclooxygenase-1 AS index was significantly lower in asthmatics as compared to HS (1.96 ± 0.71 vs 2.41 ± 0.99, P < 0.05) indicating the relatively higher expression of the alternative transcript in asthmatic patients. Cyclooxygenase-1 AS index was not different between ASA and ATA groups (mean 1.90 ± 0.66 vs 2.02 ± 0.76, respectively, P = 0.39). There was no significant association between COX-1 AS index and mean daily dose of inhaled glucocorticosteroids or pulmonary function tests (FEV1, FVC) but in ASA group a weak correlation with daily dose of oral glucocorticosteroids was found (r = 0.39; P = 0.03). In ASA patients there was a significant positive correlation between the COX-1 AS index and the percentage of aspirin-triggered increase in 15-HETE generation (r = 0.51; P < 0.03). Conclusions:, Alternatively spliced variants of COX-1 mRNA are differently expressed in patients with bronchial asthma and may be associated with aspirin-triggered 15-HETE generation. [source] Interaction of cblA/adhesin-positive Burkholderia cepacia with squamous epitheliumCELLULAR MICROBIOLOGY, Issue 2 2002Umadevi Sajjan Summary A highly transmissible strain of Burkholderia cepacia from genomovar III carries the cable pilin gene, expresses the 22 kDa adhesin (cblA+ve/Adh +ve), binds to cytokeratin 13 (CK13) and is invasive. CK13 is expressed abundantly in the airway epithelia of cystic fibrosis (CF) patients. We have now investigated whether binding of cblA+ve/Adh +ve B. cepacia to CK13 potentiates bacterial invasion and epithelial damage using bronchial epithelial cell cultures differentiated into either squamous (CK13-enriched) or mucociliary (CK13-deficient) epithelia. Three different B. cepacia isolates (cblA+ve/Adh +ve, cblA+ve/Adh ,ve and cblA,ve/Adh ,ve) showed minimal binding to mucociliary cultures, and did not invade or cause cell damage. In contrast, the cblA+ve/Adh +ve isolate, but not others, bound to CK13-expressing cells in squamous cultures, caused cytotoxicity and stimulated IL-8 release within 2 h. By 24 h, this isolate invaded and migrated across the squamous culture, causing moderate to severe epithelial damage. A specific antiadhesin antibody, which blocked the initial binding of the cblA+ve/Adh +ve isolate to CK13, significantly inhibited all the pathologic effects. Transmission electron microscopy of squamous cultures incubated with the cblA+ve/Adh +ve isolate, revealed bacteria on the surface surrounded by filopodia by 2 h, and within the cells in membrane-bound vesicles by 24 h. Bacteria were also observed free in the cytoplasm, surrounded by intermediate filaments containing CK13. These findings suggest that binding of B. cepacia to CK13 is an important initial event and that it promotes bacterial invasion and epithelial damage. [source] | ||||||||||