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

dc.contributor.author Niklason, LE
dc.contributor.author Pang, Z
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.identifier https://www.ncbi.nlm.nih.gov/pubmed/20055662
dc.identifier.uri http://hdl.handle.net/10161/3327
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.language eng
dc.language.iso en_US
dc.relation.ispartof Tissue Eng Part A
dc.relation.isversionof 10.1089/ten.TEA.2009.0448
dc.subject Animals
dc.subject Coculture Techniques
dc.subject Cytoplasm
dc.subject Endothelial Cells
dc.subject Myocytes, Smooth Muscle
dc.subject Reactive Oxygen Species
dc.subject Swine
dc.subject Thromboplastin
dc.subject Tissue Engineering
dc.subject Transcription Factor RelA
dc.title Porcine endothelial cells cocultured with smooth muscle cells became procoagulant in vitro.
dc.type Journal article
dc.description.version Version of Record
duke.date.pubdate 2010-6-0
duke.description.issue 6
duke.description.volume 16
dc.relation.journal Tissue Engineering Part a
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
pubs.organisational-group Pratt School of Engineering
pubs.publication-status Published
pubs.volume 16
dc.identifier.eissn 1937-335X


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