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dc.contributor.authorGhartey-Tagoe, Esi B. (Esi Baawah)en_US
dc.date.accessioned2005-03-03T21:45:54Z
dc.date.available2005-03-03T21:45:54Z
dc.date.issued2004-01-09en_US
dc.identifier.urihttp://hdl.handle.net/1853/5130
dc.description.abstractThis study was conducted to determine if electroporation could deliver membrane-impermeant molecules intracellularly to intact, physiologically competent monolayers that mimic the intestinal epithelium. The long-term effects of electroporation on these monolayers were studied to determine the kinetics with which monolayers recover barrier function. The ability of electroporation to introduce biologically active molecules, e.g., plasmid DNA and siRNA, into these monolayers, to either express a protein of interest or modify cellular function, was also studied. Results showed that intracellular uptake of calcein, a small tracer molecule, and bovine serum albumin, a globular protein, occurred uniformly throughout the monolayers and increased as a function of voltage, pulse length, and pulse number. There was no significant difference in uptake resulting from single and multiple pulses of the same total exposure time. Barrier function recovery depended on the electroporation conditions applied, with some monolayers recovering normal physiologic function within a day. Electroporation also increased the permeability of the monolayers to calcein and BSA, possibly through a combination of increased paracellular and transmonolayer transport. When compared to cationic lipid transfection (lipofection), transfection of intestinal epithelial monolayers with reporter plasmids by electroporation was more efficient in situations where high concentrations of DNA, and as a result, higher levels of expression were needed. Although uptake of DNA was high after electroporation and increased with increasing amounts of DNA, overall expreseion efficiency was still low (~3%). Electroporation-mediated transfection of intestinal epithelial monolayers with a plasmid that expressed inflammation inhibitor protein, IκВα was not always successful, probably because of low levels of protein expression. Introduction of the much smaller siRNA molecules into the monolayers by electroporation, on the other hand, was very successful at inhibiting the production of the nuclear envelope proteins lamin A and lamin C. The results of these experiments demonstrated that electroporation can introduce a wide variety of molecules intracellularly into model intestinal epithelia. These results should be useful to identify optimal electroporation conditions for transporting drugs, proteins, and genes into intestinal and, possibly, other epithelia for local drug and gene therapy, as well as for development of improved models of intestinal epithelium.en_US
dc.format.extent5093179 bytes
dc.format.mimetypeapplication/pdf
dc.language.isoen_US
dc.publisherGeorgia Institute of Technologyen_US
dc.subjectGene deliveryen_US
dc.subjectMonolayer
dc.subjectIntestinal epithelium
dc.subjectDrug delivery
dc.subjectElectroporation
dc.titleElectroporation-Mediated Delivery Of Macromolecules To Intestinal Epithelial Modelsen_US
dc.typeDissertationen_US
dc.description.degreePh.D.en_US
dc.contributor.departmentBiomedical Engineeringen_US
dc.description.advisorCommittee Chair: Prausnitz, Mark; Committee Co-Chair: Neish, Andrew; Committee Member: Babensee, Julia; Committee Member: Cargill, Robert; Committee Member: Hess, Dennis; Committee Member: Sambanis, Athanassiosen_US


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