title: Scaffold-free tissue engineering of functional corneal stromal tissue.
abstract: Blinding corneal scarring is predominately treated with allogeneic graft tissue; however, there is a worldwide shortage of donor tissue leaving millions in need of therapy. Human corneal stromal stem cells (CSSC) have been shown produce corneal tissue when cultured on nanofibre scaffolding, but this tissue cannot be readily separated from the scaffold. In this study, scaffold-free tissue engineering methods were used to generate biomimetic corneal stromal tissue constructs that can be transplanted in vivo without introducing the additional variables associated with exogenous scaffolding. CSSC were cultured on substrates with aligned microgrooves, which directed parallel cell alignment and matrix organization, similar to the organization of native corneal stromal lamella. CSSC produced sufficient matrix to allow manual separation of a tissue sheet from the grooved substrate. These constructs were cellular and collagenous tissue sheets, approximately 4 μm thick and contained extracellular matrix molecules typical of corneal tissue including collagen types I and V and keratocan. Similar to the native corneal stroma, the engineered corneal tissues contained long parallel collagen fibrils with uniform diameter. After being transplanted into mouse corneal stromal pockets, the engineered corneal stromal tissues became transparent, and the human CSSCs continued to express human corneal stromal matrix molecules. Both in vitro and in vivo, these scaffold-free engineered constructs emulated stromal lamellae of native corneal stromal tissues. Scaffold-free engineered corneal stromal constructs represent a novel, potentially autologous, cell-generated, biomaterial with the potential for treating corneal blindness. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Department of Ophthalmology, University of Pittsburgh, PA, USA.
authors.forename: Fatima N
authors.initials: FN
authors.lastname: Syed-Picard
authors.affiliation: Department of Ophthalmology, University of Pittsburgh, PA, USA.|McGowan Institute for Regenerative Medicine, Pittsburgh, PA, USA.
authors.forename: Yiqin
authors.initials: Y
authors.lastname: Du
authors.affiliation: Department of Ophthalmology, University of Pittsburgh, PA, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Hertsenberg
authors.affiliation: Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA.
authors.forename: Rachelle
authors.initials: R
authors.lastname: Palchesko
authors.affiliation: Department of Ophthalmology, University of Pittsburgh, PA, USA.
authors.forename: Martha L
authors.initials: ML
authors.lastname: Funderburgh
authors.affiliation: McGowan Institute for Regenerative Medicine, Pittsburgh, PA, USA.|Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA.|Department of Materials Science and Engineering, Carnegie Mellon University, Pittsburgh, PA, USA.
authors.forename: Adam W
authors.identifier: 0000-0003-3338-5456
authors.initials: AW
authors.lastname: Feinberg
authors.affiliation: Department of Ophthalmology, University of Pittsburgh, PA, USA.|McGowan Institute for Regenerative Medicine, Pittsburgh, PA, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Funderburgh
mesh_terms: D000818:Animals; D003319:Corneal Stroma / Q000502:physiology* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D020007:Implants, Experimental; D051379:Mice; D008810:Mice, Inbred C57BL; D013234:Stem Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
keywords: cell sheet; cornea; human cells; ocular; scaffold-free; stem cells; transplantation