Evaluation of Silk Biomaterials in Combination with Extracellular Matrix Coatings for Bladder Tissue Engineering with Primary and Pluripotent Cells

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Evaluation of Silk Biomaterials in Combination with Extracellular Matrix Coatings for Bladder Tissue Engineering with Primary and Pluripotent Cells

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dc.contributor.author Franck, Debra
dc.contributor.author Gil, Eun Seok
dc.contributor.author Adam, Rosalyn Mare
dc.contributor.author Kaplan, David L.
dc.contributor.author Chung, Yeun Goo
dc.contributor.author Estrada, Carlos R.
dc.contributor.author Mauney, Joshua Robert
dc.date.accessioned 2013-06-03T16:30:01Z
dc.date.issued 2013
dc.identifier.citation Franck, 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.issn 1932-6203 en_US
dc.identifier.uri http://nrs.harvard.edu/urn-3:HUL.InstRepos:10708043
dc.description.abstract Silk-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.iso en_US en_US
dc.publisher Public Library of Science en_US
dc.relation.isversionof doi:10.1371/journal.pone.0056237 en_US
dc.relation.hasversion http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3567020/pdf/ en_US
dash.license LAA
dc.subject DNA transcription en_US
dc.subject agriculture en_US
dc.subject crops en_US
dc.subject fibers en_US
dc.subject silk en_US
dc.subject biology en_US
dc.subject biochemistry en_US
dc.subject cytochemistry en_US
dc.subject extracellular matrix en_US
dc.subject biotechnology en_US
dc.subject tissue engineering en_US
dc.subject developmental biology en_US
dc.subject cell differentiation en_US
dc.subject genetics en_US
dc.subject gene expression en_US
dc.subject protein translation en_US
dc.subject molecular cell biology en_US
dc.subject cellular types en_US
dc.subject stem cells en_US
dc.subject embryonic stem cells en_US
dc.subject induced pluripotent stem cells en_US
dc.subject muscle cells en_US
dc.subject materials science en_US
dc.subject biomaterials en_US
dc.subject medicine en_US
dc.subject urology en_US
dc.subject ureteric disorders en_US
dc.subject bladder disorders en_US
dc.title Evaluation of Silk Biomaterials in Combination with Extracellular Matrix Coatings for Bladder Tissue Engineering with Primary and Pluripotent Cells en_US
dc.type Journal Article en_US
dc.description.version Version of Record en_US
dc.relation.journal PLoS ONE en_US
dash.depositing.author Adam, Rosalyn Mare
dc.date.available 2013-06-03T16:30:01Z

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