RESUMO
The entrapment of enzymes, drugs, cells or tissue fragments in alginates cross-linked with Ca(2+) or Ba(2+) has great potential in basic research, biotechnology and medicine. The swelling properties and, in turn, the mechanical stability are key factors in designing an optimally cross-linked hydrogel matrix. These parameters depend critically on the cross-linking process and seemingly minor modifications in manufacture have a large impact. Thus, sensitive and non-invasive tools are required to determine the spatial homogeneity and efficacy of the cross-linking process. Here, we show for alginate microcapsules (between 400 microm and 600 microm in diameter) that advanced (1)H NMR imaging, along with paramagnetic Cu(2+) as contrast agent, can be used to validate the cross-linking process. Two- and three-dimensional images and maps of the spin-lattice relaxation time T(1) of Ba(2+) cross-linked microcapsules exposed to external Cu(2+) yielded qualitative as well as quantitative information about the accumulation of Cu(2+) within and removal from microcapsules upon washing with Cu(2+) free saline solution. The use of Cu(2+) (having a slightly higher affinity constant to alginate than Ba(2+)) for gelling gave a complementary insight into the spatial homogeneity of the cross-linking process together with information about the mechanical stability of the microcapsules. The potential of this technique was demonstrated for alginates extracted from two different algal sources and cross-linked either externally by the conventional air-jet dropping method or internally by the "crystal gun" method.
Assuntos
Alginatos/química , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Alginatos/ultraestrutura , Bário , Fenômenos Biofísicos , Biofísica , Cápsulas , Meios de Contraste , Cobre , Reagentes de Ligações Cruzadas , Géis , Imageamento por Ressonância MagnéticaRESUMO
Cross-linked alginate microcapsules of sufficient mechanical strength can immunoisolate cells for the long-term treatment of hormone and other deficiency diseases in human beings. However, gelation of alginate by external Ba(2+) (or other divalent cations) produces non-homogeneous cross-linking of the polymeric mannuronic (M) and guluronic (G) acid chains. The stability of such microcapsules is rather limited. Here, we show that homogeneous cross-linking can be achieved by injecting BaCl(2) crystals into alginate droplets before they come into contact with external BaCl(2). The high effectiveness of this crystal gun method is demonstrated by confocal laser scanning microscopy and by advanced nuclear magnetic resonance imaging. Both techniques gave clear-cut evidence that homogeneous cross-linkage throughout the microcapsule is only obtained with simultaneous internal and external gelation. Atomic force microscopy showed a very smooth surface topography for microcapsules made by the crystal gun method, provided that excess Ba(2+) ions were removed immediately after gelation. In vitro experiments showed greatly suppressed swelling for crystal gun microcapsules. Even alginate extracted from Lessonia nigrescens (highly biocompatible) yielded microcapsules with long-term mechanical stability not hitherto possible. Encapsulation of rat islets, human monoclonal antibodies secreting hybridoma cells and murine mesenchymal stem cells transfected with cDNA encoding for bone morphogenetic protein (BMP-4) revealed that injection of BaCl(2) crystals has no adverse side effects on cell viability and function. However, the release of low-molecular weight factors (such as insulin) may be delayed when using alginate concentrations in the usual range.
Assuntos
Alginatos/química , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Espectroscopia de Ressonância Magnética/métodos , Microscopia de Força Atômica/métodos , Microscopia Confocal/métodos , Bário/química , Compostos de Bário/farmacologia , Proteína Morfogenética Óssea 4 , Proteínas Morfogenéticas Ósseas/química , Cátions , Linhagem Celular Tumoral , Sobrevivência Celular , Cloretos/farmacologia , Reagentes de Ligações Cruzadas/farmacologia , DNA Complementar/metabolismo , Humanos , Insulina/química , Íons , TransfecçãoRESUMO
Interactive television (ITV) is being used by increasing numbers of nursing schools and hospitals to provide educational access to geographically diverse learners and nurse employees. ITV-related research suggests that learner outcomes and satisfaction with such distance education courses are similar to those associated with the traditional classroom. The authors conducted an evaluation of a major nursing school's nursing telecourse program (NTP) to assess collaborative efforts with hospitals and regional campuses. The respondents, 15 site coordinators, were generally satisfied with the operational aspects of the NTP. Concerns raised by respondents included classroom space availability, operational costs, library resources, and communication equipment and processes. These stakeholders' perspectives were viewed as valuable in identifying problems and making improvements in the school's NTP.