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dc.contributor.authorFranck, Debra
dc.contributor.authorGil, Eun Seok
dc.contributor.authorAdam, Rosalyn Mare
dc.contributor.authorKaplan, David L.
dc.contributor.authorChung, Yeun Goo
dc.contributor.authorEstrada, Carlos R.
dc.contributor.authorMauney, Joshua Robert
dc.date.accessioned2013-06-03T16:30:01Z
dc.date.issued2013
dc.identifier.citationFranck, Debra, Eun Seok Gil, Rosalyn Mare Adam, David L. Kaplan, Yeun Goo Chung, Carlos R. Estrada, and Joshua Robert Mauney. 2013. Evaluation of silk biomaterials in combination with extracellular matrix coatings for bladder tissue engineering with primary and pluripotent cells. PLoS ONE 8(2): e56237.en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:10708043
dc.description.abstractSilk-based biomaterials in combination with extracellular matrix (ECM) coatings were assessed as templates for cell-seeded bladder tissue engineering approaches. Two structurally diverse groups of silk scaffolds were produced by a gel spinning process and consisted of either smooth, compact multi-laminates (Group 1) or rough, porous lamellar-like sheets (Group 2). Scaffolds alone or coated with collagen types I or IV or fibronectin were assessed independently for their ability to support attachment, proliferation, and differentiation of primary cell lines including human bladder smooth muscle cells (SMC) and urothelial cells as well as pluripotent cell populations, such as murine embryonic stem cells (ESC) and induced pluripotent stem (iPS) cells. AlamarBlue evaluations revealed that fibronectin-coated Group 2 scaffolds promoted the highest degree of primary SMC and urothelial cell attachment in comparison to uncoated Group 2 controls and all Group 1 scaffold variants. Real time RT-PCR and immunohistochemical (IHC) analyses demonstrated that both fibronectin-coated silk groups were permissive for SMC contractile differentiation as determined by significant upregulation of α-actin and SM22α mRNA and protein expression levels following TGFβ1 stimulation. Prominent expression of epithelial differentiation markers, cytokeratins, was observed in urothelial cells cultured on both control and fibronectin-coated groups following IHC analysis. Evaluation of silk matrices for ESC and iPS cell attachment by alamarBlue showed that fibronectin-coated Group 2 scaffolds promoted the highest levels in comparison to all other scaffold formulations. In addition, real time RT-PCR and IHC analyses showed that fibronectin-coated Group 2 scaffolds facilitated ESC and iPS cell differentiation toward both urothelial and smooth muscle lineages in response to all trans retinoic acid as assessed by induction of uroplakin and contractile gene and protein expression. These results demonstrate that silk scaffolds support primary and pluripotent cell responses pertinent to bladder tissue engineering and that scaffold morphology and fibronectin coatings influence these processes.en_US
dc.language.isoen_USen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofdoi:10.1371/journal.pone.0056237en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3567020/pdf/en_US
dash.licenseLAA
dc.subjectDNA transcriptionen_US
dc.subjectagricultureen_US
dc.subjectcropsen_US
dc.subjectfibersen_US
dc.subjectsilken_US
dc.subjectbiologyen_US
dc.subjectbiochemistryen_US
dc.subjectcytochemistryen_US
dc.subjectextracellular matrixen_US
dc.subjectbiotechnologyen_US
dc.subjecttissue engineeringen_US
dc.subjectdevelopmental biologyen_US
dc.subjectcell differentiationen_US
dc.subjectgeneticsen_US
dc.subjectgene expressionen_US
dc.subjectprotein translationen_US
dc.subjectmolecular cell biologyen_US
dc.subjectcellular typesen_US
dc.subjectstem cellsen_US
dc.subjectembryonic stem cellsen_US
dc.subjectinduced pluripotent stem cellsen_US
dc.subjectmuscle cellsen_US
dc.subjectmaterials scienceen_US
dc.subjectbiomaterialsen_US
dc.subjectmedicineen_US
dc.subjecturologyen_US
dc.subjectureteric disordersen_US
dc.subjectbladder disordersen_US
dc.titleEvaluation of Silk Biomaterials in Combination with Extracellular Matrix Coatings for Bladder Tissue Engineering with Primary and Pluripotent Cellsen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalPLoS ONEen_US
dash.depositing.authorAdam, Rosalyn Mare
dc.date.available2013-06-03T16:30:01Z
dc.identifier.doi10.1371/journal.pone.0056237*
dash.contributor.affiliatedMauney, Joshua
dash.contributor.affiliatedEstrada, Carlos
dash.contributor.affiliatedChung, Yeun Goo
dash.contributor.affiliatedAdam, Rosalyn


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