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Artificial Polymeric Scaffolds as Extracellular Matrix Substitutes for Autologous Conjunctival Goblet Cell Expansion
Harvard Medical Sch, MA USA; Shanxi Medical University, Peoples R China.
Harvard Medical Sch, MA USA; Aarhus University Hospital NBG, Denmark.
Harvard Medical Sch, MA USA.
Tufts University, MA 02155 USA.
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2016 (English)In: Investigative Ophthalmology and Visual Science, ISSN 0146-0404, E-ISSN 1552-5783, Vol. 57, no 14, 6134-6146 p.Article in journal (Refereed) Published
Abstract [en]

PURPOSE. We fabricated and investigated polymeric scaffolds that can substitute for the conjunctival extracellular matrix to provide a substrate for autologous expansion of human conjunctival goblet cells in culture. METHODS. We fabricated two hydrogels and two silk films: (1) recombinant human collagen (RHC) hydrogel, (2) recombinant human collagen 2-methacryloylxyethyl phosphorylcholine (RHC-MPC) hydrogel, (3) arginine-glycine-aspartic acid (RGD) modified silk, and (4) poly-D-lysine (PDL) coated silk, and four electrospun scaffolds: (1) collagen, (2) poly(acrylic acid) (PAA), (3) poly(caprolactone) (PCL), and (4) poly(vinyl alcohol) (PVA). Coverslips and polyethylene terephthalate (PET) were used for comparison. Human conjunctival explants were cultured on scaffolds for 9 to 15 days. Cell viability, outgrowth area, and the percentage of cells expressing markers for stratified squamous epithelial cells (cytokeratin 4) and goblet cells (cytokeratin 7) were determined. RESULTS. Most of cells grown on all scaffolds were viable except for PCL in which only 3.6 +/- 2.2% of the cells were viable. No cells attached to PVA scaffold. The outgrowth was greatest on PDL-silk and PET. Outgrowth was smallest on PCL. All cells were CK7-positive on RHCMPC while 84.7 +/- 6.9% of cells expressed CK7 on PDL-silk. For PCL, 87.10 +/- 3.17% of cells were CK7-positive compared to PET where 67.10 +/- 12.08% of cells were CK7-positive cells. CONCLUSIONS. Biopolymer substrates in the form of hydrogels and silk films provided for better adherence, proliferation, and differentiation than the electrospun scaffolds and could be used for conjunctival goblet cell expansion for eventual transplantation once undifferentiated and stratified squamous cells are included. Useful polymer scaffold design characteristics have emerged from this study.

Place, publisher, year, edition, pages
ASSOC RESEARCH VISION OPHTHALMOLOGY INC , 2016. Vol. 57, no 14, 6134-6146 p.
Keyword [en]
polymeric scaffolds; goblet cells; cell growth
National Category
Neurosciences
Identifiers
URN: urn:nbn:se:liu:diva-134621DOI: 10.1167/iovs.16-20081ISI: 000392380000035PubMedID: 27832279OAI: oai:DiVA.org:liu-134621DiVA: diva2:1075938
Note

Funding Agencies|United States Department of Defense [W81XWH-07/P00001/CLIN 002]; NIH [RO1EY01947, RO1 EY020856, P30 EY003790]; Danish Society for Eye Health; Velux Foundation; Danish Eye Bank; Aarhus University; Danish Society of Ophthalmology; Synoptic Foundation; A.P. Moeller Foundation; Danish Eye Research Foundation; Case Western Reserve University Institute for Advanced Materials; Swedish Research Council

Available from: 2017-02-21 Created: 2017-02-21 Last updated: 2018-01-13

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