Hydraulic conductivity of arterial endothelial cell and smooth muscle cell cocultures
Item
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Title
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Hydraulic conductivity of arterial endothelial cell and smooth muscle cell cocultures
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Identifier
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d_2009_2013:bd8a50cefa46:11558
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identifier
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12051
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Creator
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Mathura, Rishi A.,
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Contributor
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John M. Tarbell
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Date
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2012
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biomedical engineering
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Abstract
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This research investigates the hydraulic conductivity (Lp) of cocultured arterial endothelial cells (ECs) and smooth muscle cells (SMCs) exposed to pressure-driven transmural flow. Homotypic and heterotypic cellular interactions are explored through monoculturing and coculturing of bovine aortic ECs and SMCs. It is hypothesized that cocultured ECs and SMCs will have significantly different transport properties than ECs alone. This thesis is the first to examine Lp of arterial cocultures in the presence of physiological convective transmural flow.;Chapter 1 provides a general introduction to the arterial wall and reviews previous work on endothelial transport. Chapter 1 also provides a review of various EC-SMC coculture constructs and concludes with a description of specific aims that are proposed for the study of arterial coculture L p.;Chapter 2 reports on the Lp of EC-initiated cocultures that were constructed on porous membranes. Hydraulic conductivities of these cocultures were lower than the Lp of monocultured ECs. Monoculture resistances-in-series modeled coculture Lp values were also compared to actual coculture Lp values to distinguish the presence of heterotypic interactions in coculture. Immunofluorescence staining of VE-cadherin and morphometric analysis of these images revealed that ECs in the EC-initiated cocultures can have an elongated morphology compared to ECs in monoculture. Serum content in media was also shown to have a significant influence on EC Lp. EC-initiated cocultures on opposing sides of the porous membrane were used to mimic the EC-SMC arrangement in a normal vessel wall and produced one of the lowest Lp values that is reported in the literature.;Chapter 3 reports on the Lp of several SMC-initiated cocultures that were constructed on porous membranes. Hydraulic conductivities of these cocultures tended to be higher than the Lp of monocultured ECs. Monoculture resistances-in-series modeled coculture Lp values were also compared to actual coculture Lp values to distinguish the presence of heterotypic interactions in coculture. Immunofluorescence staining of VE-cadherin and morphometric analysis of these images revealed that ECs in the SMC-initiated cocultures can have a rounded morphology compared to ECs in monoculture. SMC-initiated cocultures that were configured on the luminal side of the porous membrane mimic the arrangement of ECs and SMCs in diseased states, such as intimal hyperplasia and atherogenesis, and produced a significantly higher Lp than ECs alone.;Chapter 4 reports on Lp of ECs in monoculture and coculture formats that have been exposed to transmural media flow. A transmural media flow apparatus was developed to interface with cultures on porous membranes. Endothelial cells in monoculture and shared-media cocultures with SMCs were exposed to 48 hours of transmural flow. Hydraulic conductivity of ECs that were exposed to transmural flow produced a higher Lp, which was not significant, compared to the Lp of ECs that were cultured in static media. Also, Lp of ECs derived from cocultures that were exposed to 48 hours of transmural flow was similar to the Lp of ECs from cocultures in static media. Arterial coculture configurations have produced a wide range of Lp values. Hydraulic conductivity in cocultures which mimic the EC-SMC arrangement in normal vasculature is lower than EC alone while the Lp in cocultures modeling the EC-SMC arrangement in diseased states can be higher than EC alone. These variations in L p are linked to the methods of coculturing that were used, and indicate the importance of serum content, inoculation order, culture time, and the proximity between ECs and SMCs. In general, EC-SMC coculturing produces more realistic arterial wall environments for the study of arterial transport.
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Type
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dissertation
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Source
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2009_2013.csv
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degree
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Ph.D.
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Program
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Engineering
