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  • 1
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract Numerous methods are proposed to reduce the surface thrombogenicity of vascular prostheses, among them endothelialization of the lumen which has had clinical application since 1985. One of the problems is to collect enough cells to rapidly obtain a complete monolayer at the time of implantation. Thus, material improvements are necessary to enhance cell adhesion and spreading. A collaboration with the Bakoulev Institute (Moscow) gave us the opportunity to study the cytocompatibility of carbon coated materials (PAN and Vitlan®), polyester coated with albumin and/or synthetic polysaccharide. Studies were carried out with cultured human umbilical vein endothelial cells (HUVEC). Two steps are distinguished: indirect tests (medium added with materials extracts), then direct tests (cells directly seeded onto materials). Neither PAN, Vitlan® nor polyesters extracts have provoked a toxic effect on HUVEC. Concerning attachment on materials, a maximum of 60% of seeded cells is reached. Cells could proliferate and confluency is obtained between days 5 and 10 for the best materials. SEM corroborated these results. On polysaccharide-coated polyester (M. 11), HUVEC produced significant levels of vWF after thrombin stimulation (ELISA assay): vWF was functional (ristocetin cofactor activity). In conclusion, PAN and M. 11 gave encouraging results and further studies remain to be investigated.
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  • 2
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Cell culture models are becoming prevalent in the investigation of tissue responses to implant materials. Cellular attachment and cell adhesion studies can aid in the development of more effective orthopaedic and dental implants. Cell attachment was studied on extracellular matrix proteins (type I, IV collagen, peptide solubilized elastin (PSE), fibronectin laminin). Human osteoprogenitor cells responded differently to these collagenous and non-collagenous proteins. PSE and type I or type IV collagen are the most effective proteins in cellular attachment and cell spreading. Cell behaviour was measured in the presence of macroporous materials (Porites astreoïdes from the West Indies and a bovine hydroxyapatite ceramic ENDOBON®) and bioartificial connective matrices comprising hydroxyapatite, peptide solubilized elastin, collagen, fibronectin and chondroïtin-6-sulfate, components of the extracellular matrix (ECM). Human osteoprogenitor cells responded differently to the materials tested according to the content of components of ECM. About 40% of attached cells were obtained on the composite materials PSE, collagen, fibronectin and chondroïtin-6-sulfate, and about 10% on the macroporous materials, whatever their porosity and their chemical components. These results demonstrate a need for more effective surface treatment to promote cell attachment, cell spreading and cell growth.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of materials science 10 (1999), S. 807-810 
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract Endothelial cell (EC) seeding of small caliber vascular grafts prior to their implantation has proved to significantly improve long-term patency in humans. We have previously demonstrated that a monolayer of EC could be obtained on type I collagen-coated knitted ultrathin polyster grafts (InterVascular, La Ciotat, France). Thus, the aim of the present work was to understand the nature of cell adhesion mechanisms involved in the cell /biomaterial interface, using HemaCarotid® (InterVascular) patches made of type I collagen-coated knitted ultrathin polyster (type I collagen is used to coat patches to attain low permeability). By means of quantitative attachment tests, adhesion blocking assays, RT–PCR for the expression of β1 integrin mRNA, indirect immunofluorescence with antivinculin antibody, we were able to show that EC are able to adhere to such surfaces by the means (non-unique) of cell surface receptors of the β1 integrin group. However, the latter are probably downregulated at the cell/biomaterial interface.
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  • 4
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: It is well known that cuffs of endotracheal tubes can induce ischemic injuries on tracheal epithelium, as a result of mechanical hyperpressure caused by the cuff on the airway tissue. Whether or not material components are leached out and may provoke a direct toxic effect on the respiratory epithelium is much less clear. To study the cytocompatibility of such materials, we have developed an in vitro cell system using human tracheal epithelial cells, arising from trachea superficial biopsies. In culture, cells have been characterized by morphological and immunocytochemical criteria. Ultrastructural observations suggest that our culture conditions are permissive for the expression of both squamous and secretory phenotypes. We have assessed the cytocompatibility of a cuff towards epithelial cells, first, by an indirect test, and second by a direct test. By the indirect test, using material extracts, we did not find any toxic effect towards human airway epithelial cells of the cuff components. By a direct test, we found a slight cell lysis after a 24 h incubation. Our study shows that this human tracheal epithelial cell system is a useful and relevant model which could be used in a quality control procedure for testing the cytocompatibility of materials for endotracheal use.
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  • 5
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: With a view to elaborating a bioactive bone substitute, the association of an artificial extracellular matrix, basically constituted of elastin-solubilized peptides (ESP) and type I+III collagens, with different types of calcium phosphates, was investigated. This paper describes the selective adsorption of ESP on some calcium phosphate samples, and the further association of the adsorbed peptides with type I+III collagens. A preliminary study of the material cytotoxicity was carried out, investigating the behaviour of human osteoblast cells in contact with the yielded composite material.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Journal of materials science 5 (1994), S. 18-24 
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: General cytotoxicity was assayed for ceramic (Al2O3, ZrO2/Y2O3, AIN, B4C, BN, SiC, Si3N4, TiB4, TiC, TiN) diamond and graphite powders, using 3T3 Balb/c permanent cell lines. Neutral red test was carried out in order to establish cell viability. Further investigations were undertaken on human differentiated cells (human umbilical venous endothelial cells): cell behaviour (MTT assay, total cell protein content) and differentiation (immunofluorescence) were studied. In both cases, no cytotoxic effect has been noticed. All the impurities contained at low concentration in these powders do not seem to present any effect. The correlation which has been previously observed between cytotoxicity-cell culture response and blood haemolysis for polymers has not been established here for ceramic powders. We conclude that all the ceramic powders tested here and therefore the corresponding bulk ceramics or ceramic coatings do not induce any cytotoxic effect.
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  • 7
    ISSN: 1573-4838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Bioactive materials such as hydroxyapatite ceramics are known to show a stable interfacial bond with tissues. The objective of this study was to investigate the behaviour of human bone differentiated cells isolated from bone marrow on the surface of a hydroxyapatite ceramic obtained from bovine spongiosa. Scanning electron microscopy and DNA synthesis analysis assessed by [3H]-thymidine incorporation showed cell colonization of the whole material. Immunological studies using monoclonal antibodies to osteocalcin and osteonectin and cytochemical analysis of alkaline phosphatase activity indicated that these cells did not lose their osteoblastic phenotype after 28 days of culture. Furthermore, this study demonstrates the in vitro interface between the material and human cells, which is reporduced in vivo.
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  • 8
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 42 (1998), S. 96-102 
    ISSN: 0021-9304
    Keywords: coral ; calcite ; cell culture ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: The two crystalline forms of CaCO3, aragonite (from natural coral) and calcite (from natural limestone), have been used with success as bone graft substitutes. However, natural coral transformed into calcite by heating has never been tested. The objective of this work was to study the proliferation and alkaline phosphatase, osteonectin, and osteocalcin expression of human bone marrow cells cultured on CaCO3 crystallized both in the aragonite form (natural coral) and in the calcite form (natural coral modified by heating). The methods used to characterize calcite obtained from the coral were volumic porosimetry, scanning electron microscopy (SEM) and X-ray diffraction. Cell colonization of the material was assessed by SEM performed on days 1, 7, 20, and 30 and [3H]thymidine incorporation was performed on days 3, 7, 12, 18, 25, and 32. Phenotypic expression was assessed by using in situ cytochemistry (alkaline phosphatase), immunocytochemistry (osteonectin and osteocalcin), and hybridization (osteocalcin, β-actin, and alkaline phosphatase mRNA). Results showed the transformation of aragonite into calcite after heating, the conservation of macroporosity, and a modification of the surface. Calcite appeared to have a smoother and more uniform surface than aragonite crystals. As for [3H]thymidine there was an increase incorporation from days 3 to 18, a stabilization from days 18 to 25, and a decrease from days 25 to 32. After 20 days of culture, immunological studies using monoclonal antibodies to osteocalcin, osteonectin, cytochemical analysis of alkaline phosphatase activity, and in situ hybridization using osteocalcin, β-actin, and alkaline phosphatase cDNA indicated that the cells had not lost their osteoblastic phenotype. These experiments demonstrate that coral crystallized in the aragonite or calcite form present a similar degree of specific cytocompatibility. © 1998 John Wiley & Sons, Inc. J. Biomed Mater Res, 42, 96-102, 1998.
    Additional Material: 5 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 27 (1993), S. 1367-1381 
    ISSN: 0021-9304
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: There is a need for viable small diameter vascular grafts, the luminal surface of which could be seeded by endothelial cells (ECs) to prevent thrombosis. In order to select candidates for EC seeding before implantation, the in vitro cytocompatibility of three different Pellethanes® (polyetherurethanes) using human ECs was investigated. The methodology included two stages depending on either direct contact between cells and materials or contact between cells and material extracts, obtained under standardized conditions. By the latter method, we observed a cytotoxic effect on cell growth with 2363-55 D Pellethane extract at a 50% (v/v) concentration in the nutrient medium, likely provoked by leachables and correlated with the lowest levels of tPA, PAI1, and vWF antigens in the supernatants. By the former method, we studied EC attachment and growth. Morphology was studied by classical means and completed by scintigraphy and microautoradiography after 111Indium-labeling of the EC monolayer. Differentiation was determined by the release of vWF antigen and measurement of vWF activity (multimeric organization) after human thrombin stimulation. Despite an inhibition of proliferation for both 55 D and 75 D types (compared to the control), a functional monolayer of ECs was obtained on 75 D. Pellethane 75 D could be the best support for in vitro endothelialization. © 1993 John Wiley & Sons, Inc.
    Additional Material: 9 Ill.
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  • 10
    ISSN: 0021-9304
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Three different exoskeletons of coral species Porites astreoides (P), Montastrea annularis (M), and Dichocoenia stokesi (D) were implanted for 2-20 weeks in rabbits. At 2, 4, 8, or 20 weeks, the exoskeletons presented variations in their resorptions depending on the species. To understand the variations in the decreasing speed of the implants despite their similar chemical composition, a study of the surface and architecture of the coral was carried out using scanning electronic microscopy, porosity was evaluated, and growth and differentiation of osteogenic cells cultured in vitro were observed for more than 1 month. At the cellular level, the surface of the implants was identical. Three-dimensional structures of the implants were variable, but the porosity values [P = 42.7%, M = 40.7%, and D = 17.4%] could not completely account for the differences in the resorbing process of the species. Standard histologic studies performed at 2, 4, 8, and 20 weeks after implantation produced the same pattern with P or M, showing aspects of rapid resorption; however, with D there were images resembling those of a foreign-body reaction. It seems that when resorption is not quick enough, a foreign body reaction develops which further slows down the process. This work focuses on the importance of porosity when using coral as bone substitute. © 1995 John Wiley & Sons, Inc.
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