Artificial Polymeric Scaffolds as Extracellular Matrix Substitutes for Autologous Conjunctival Goblet Cell Expansion.
OBJECTIVE
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 RHC-MPC 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.
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States 2Department of Ophthalmology, The Second Hospital of Shanxi Medical University, Taiyuan, China.
Min
M
He
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States 3Department of Ophthalmology, Aarhus University Hospital NBG, Aarhus, Denmark.
Thomas
T
Storr-Paulsen
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States.
Annie L
AL
Wang
Department of Biomedical Engineering, Tufts University, Medford, Massachusetts, United States.
Chiara E
CE
Ghezzi
Department of Biomedical Engineering, Tufts University, Medford, Massachusetts, United States.
Siran
S
Wang
Department of Macromolecular Science and Engineering, Case Western Reserve University, Cleveland, Ohio, United States.
Matthew
M
Fullana
Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States 7Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States.
Dimitrios
D
Karamichos
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States 8Department of Oral Biology, University of Oslo, Norway 9Department of Ophthalmology, Vestre Viken Hospital Trust, Drammen, Norway 10Faculty of Health Sciences, National Centre for Optics, Vision and Eye Care, University College of Southeast Norway, Norway.
Tor P
TP
Utheim
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States 8Department of Oral Biology, University of Oslo, Norway.
Rakibul
R
Islam
Department of Clinical and Experimental Medicine, Linkoping University, Linkoping, Sweden 12Swedish Medical Nanoscience Center, Department of Neurosciences, Karolinska Institutet, Stockholm, Sweden.
May
M
Griffith
Department of Clinical and Experimental Medicine, Linkoping University, Linkoping, Sweden 12Swedish Medical Nanoscience Center, Department of Neurosciences, Karolinska Institutet, Stockholm, Sweden.
M Mirazul
MM
Islam
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States.
Robin R
RR
Hodges
Department of Macromolecular Science and Engineering, Case Western Reserve University, Cleveland, Ohio, United States.
Gary E
GE
Wnek
Department of Biomedical Engineering, Tufts University, Medford, Massachusetts, United States.
David L
DL
Kaplan
Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States.
Darlene A
DA
Dartt
D000293:Adolescent; D000328:Adult; D000368:Aged; D001672:Biocompatible Materials; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology*; D005109:Extracellular Matrix / Q000737:chemistry*; D005260:Female; D020397:Goblet Cells / Q000166:cytology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D057139:Nanofibers; D011108:Polymers; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*; D055815:Young Adult
D001672:Biocompatible Materials; D011108:Polymers