Porcine endothelial cells cocultured with smooth muscle cells became procoagulant in vitro.

dc.contributor.author

Pang, Zhengyu

dc.contributor.author

Niklason, Laura E

dc.contributor.author

Truskey, George A

dc.coverage.spatial

United States

dc.date.accessioned

2011-04-15T16:46:34Z

dc.date.issued

2010-06

dc.description.abstract

Endothelial cell (EC) seeding represents a promising approach to provide a nonthrombogenic surface on vascular grafts. In this study, we used a porcine EC/smooth muscle cell (SMC) coculture model that was previously developed to examine the efficacy of EC seeding. Expression of tissue factor (TF), a primary initiator in the coagulation cascade, and TF activity were used as indicators of thrombogenicity. Using immunostaining, primary cultures of porcine EC showed a low level of TF expression, but a highly heterogeneous distribution pattern with 14% of ECs expressing TF. Quiescent primary cultures of porcine SMCs displayed a high level of TF expression and a uniform pattern of staining. When we used a two-stage amidolytic assay, TF activity of ECs cultured alone was very low, whereas that of SMCs was high. ECs cocultured with SMCs initially showed low TF activity, but TF activity of cocultures increased significantly 7-8 days after EC seeding. The increased TF activity was not due to the activation of nuclear factor kappa-B on ECs and SMCs, as immunostaining for p65 indicated that nuclear factor kappa-B was localized in the cytoplasm in an inactive form in both ECs and SMCs. Rather, increased TF activity appeared to be due to the elevated reactive oxygen species levels and contraction of the coculture, thereby compromising the integrity of EC monolayer and exposing TF on SMCs. The incubation of cocultures with N-acetyl-cysteine (2 mM), an antioxidant, inhibited contraction, suggesting involvement of reactive oxygen species in regulating the contraction. The results obtained from this study provide useful information for understanding thrombosis in tissue-engineered vascular grafts.

dc.description.version

Version of Record

dc.identifier

https://www.ncbi.nlm.nih.gov/pubmed/20055662

dc.identifier.eissn

1937-335X

dc.identifier.uri

https://hdl.handle.net/10161/3327

dc.language

eng

dc.language.iso

en_US

dc.publisher

Mary Ann Liebert Inc

dc.relation.ispartof

Tissue Eng Part A

dc.relation.isversionof

10.1089/ten.TEA.2009.0448

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Tissue Engineering Part a

dc.subject

Animals

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Coculture Techniques

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Cytoplasm

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Endothelial Cells

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Myocytes, Smooth Muscle

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Reactive Oxygen Species

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Swine

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Thromboplastin

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Tissue Engineering

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Transcription Factor RelA

dc.title

Porcine endothelial cells cocultured with smooth muscle cells became procoagulant in vitro.

dc.type

Journal article

duke.date.pubdate

2010-6-0

duke.description.issue

6

duke.description.volume

16

pubs.author-url

https://www.ncbi.nlm.nih.gov/pubmed/20055662

pubs.begin-page

1835

pubs.end-page

1844

pubs.issue

6

pubs.organisational-group

Biomedical Engineering

pubs.organisational-group

Duke

pubs.organisational-group

Duke Science & Society

pubs.organisational-group

Initiatives

pubs.organisational-group

Institutes and Provost's Academic Units

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Pratt School of Engineering

pubs.publication-status

Published

pubs.volume

16

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