Surface treatment to a biomaterial surface has been shown to modify and help cell growth. Our aim was to determine the best surface-modified system for the treatment of limbal stem cell deficiency (LSCD), which would facilitate expansion of autologous limbal epithelial cells, while maintaining cultivated epithelial cells in a less differentiated state. Commercially available contact lenses (CLs) were variously surface modified by plasma polymerization with ratios of acrylic acid to octadiene tested at 100% acrylic acid, 50:50% acrylic acid:octadiene, and 100% octadiene to produce high-, mid-, and no-acid. X-ray photoelectron spectroscopy was used to analyze the chemical composition of the plasma polymer deposited layer. Limbal explants cultured on high acid-modified CLs outgrew more cells. Immunofluorescence and RT2-PCR array results indicated that a higher acrylic acid content can also help maintain progenitor cells during ex vivo expansion of epithelial cells. This study provides the first evidence for the ability of high acid-modified CLs to preserve the stemness and to be used as substrates for the culture of limbal cells in the treatment of LSCD.
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References
1.
WhitcherJ.P., SrinivasanM., and UpadhyayM.P.Corneal blindness: a global perspective. Bull World Health Organ, 79, 214, 2001.
2.
O'CallaghanA.R., and DanielsJ.T.Concise review: limbal epithelial stem cell therapy: controversies and challenges. Stem Cells, 29, 1923, 2011.
3.
PellegriniG., et al.Long-term restoration of damaged corneal surfaces with autologous cultivated corneal epithelium. Lancet, 349, 990, 1997.
4.
TsaiR.J.F., and TsaiR.Y.N.Ex vivo expansion of corneal stem cells on amniotic membrane and their outcome. Eye Contact Lens, 36, 305, 2010.
5.
KolliS., et al.Successful clinical implementation of corneal epithelial stem cell therapy for treatment of unilateral limbal stem cell deficiency. Stem Cells, 28, 597, 2010.
6.
RamaP., et al.Limbal stem-cell therapy and long-term corneal regeneration. N Engl J Med, 363, 147, 2010.
7.
ChaiC., and LeongK.W.Biomaterials approach to expand and direct differentiation of stem cells. Mol Ther, 15, 467, 2007.
8.
SelvamS., ThomasP.B., and YiuS.C.Tissue engineering: current and future approaches to ocular surface reconstruction. Ocul Surf, 4, 120, 2006.
9.
HeY.G., et al.Experimental transplantation of cultured human limbal and amniotic epithelial cells onto the corneal surface. Cornea, 18, 570, 1999.
10.
HigaK., et al.Proliferation and differentiation of transplantable rabbit epithelial sheets engineered with or without an amniotic membrane carrier. Invest Ophthalmol Vis Sci, 48, 597, 2007.
11.
HanB., et al.A fibrin-based bioengineered ocular surface with human corneal epithelial stem cells. Cornea, 21, 505, 2002.
12.
LawrenceB.D., et al.Bioactive silk protein biomaterial systems for optical devices. Biomacromolecules, 9, 1214, 2008.
13.
ChirilaT.V., et al.Bombyx mori silk fibroin membranes as potential substrata for epithelial constructs used in the management of ocular surface disorders. Tissue Eng Part A, 14, 1203, 2008.
14.
AmbroseW.M., et al.Collagen vitrigel membranes for the in vitro reconstruction of separate corneal epithelial, stromal, and endothelial cell layers. J Biomed Mater Res B Appl Biomater, 90B, 818, 2009.
15.
Di GirolamoN., et al.Cultured human ocular surface epithelium on therapeutic contact lenses. Br J Ophthalmol, 91, 459, 2007.
16.
DeshpandeP., et al.Development of a surface-modified contact lens for the transfer of cultured limbal epithelial cells to the cornea for ocular surface diseases. Tissue Eng Part A, 15, 2889, 2009.
17.
ShorttA.J., et al.Characterization of the limbal epithelial stem cell niche: novel imaging techniques permit in vivo observation and targeted biopsy of limbal epithelial stem cells. Stem Cells, 25, 1402, 2007.
18.
EriksteinB.S., et al.Cellular stress induced by resazurin leads to autophagy and cell death via production of reactive oxygen species and mitochondrial impairment. J Cell Biochem, 111, 574, 2010.
19.
PerrotS., et al.Resazurin metabolism assay is a new sensitive alternative test in isolated pig cornea. Toxicol Sci, 72, 122, 2003.
20.
KosterM.I., et al.p63 is the molecular switch for initiation of an epithelial stratification program. Genes Dev, 18, 126, 2004.
21.
ParsaR., et al.Association of p63 with proliferative potential in normal and neoplastic human keratinocytes. J Invest Dermatol, 113, 1099, 1999.
22.
LevreroM., et al.The p53/p63/p73 family of transcription factors: overlapping and distinct functions. J Cell Sci, 113, 1661, 2000.
23.
WatanabeK., et al.Human limbal epithelium contains side population cells expressing the ATP-binding cassette transporter ABCG2. FEBS Lett, 565, 6, 2004.
24.
ChenZ., et al.Characterization of putative stem cell phenotype in human limbal epithelia. Stem Cells, 22, 355, 2004.
25.
Schlotzer-SchrehardtU., and KruseF.E.Identification and characterization of limbal stem cells. Exp Eye Res, 81, 247, 2005.
26.
Di GirolamoN., et al.Cultured human ocular surface epithelium on therapeutic contact lenses. Br J Ophthalmol, 91, 459, 2007.
27.
YangJ., BeiJ., and WangS.Enhanced cell affinity of poly (D,L-lactide) by combining plasma treatment with collagen anchorage. Biomaterials, 23, 2607, 2002.
28.
YangY., et al.Morphological behavior of osteoblast-like cells on surface-modified titanium in vitro. Biomaterials, 23, 1383, 2002.
29.
PellegriniG., et al.p63 identifies keratinocyte stem cells. Proc Natl Acad Sci U S A, 98, 3156, 2001.
30.
YangA., et al.p63 is essential for regenerative proliferation in limb, craniofacial and epithelial development. Nature, 398, 714, 1999.
31.
Di IorioE., et al.Isoforms of DeltaNp63 and the migration of ocular limbal cells in human corneal regeneration. Proc Natl Acad Sci U S A, 102, 9523, 2005.
32.
MoritaY., et al.Non-side-population hematopoietic stem cells in mouse bone marrow. Blood, 108, 2850, 2006.
33.
ZhouS., et al.The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med, 7, 1028, 2001.
34.
KawakitaT., et al.Greater growth potential of p63-positive epithelial cell clusters maintained in human limbal epithelial sheets. Invest Ophthalmol Vis Sci, 50, 4611, 2009.
NakatsuM.N., et al.Wnt/beta-catenin signaling regulates proliferation of human cornea epithelial stem/progenitor cells. Invest Ophthalmol Vis Sci, 52, 4734, 2011.
37.
VercauterenS.M., and SutherlandH.J.Constitutively active Notch4 promotes early human hematopoietic progenitor cell maintenance while inhibiting differentiation and causes lymphoid abnormalities in vivo. Blood, 104, 2315, 2004.
38.
YimE.K., et al.Nanopattern-induced changes in morphology and motility of smooth muscle cells. Biomaterials, 26, 5405, 2005.
39.
LinH.B., et al.Endothelial cell adhesion on polyurethanes containing covalently attached RGD-peptides. Biomaterials, 13, 905, 1992.
40.
ChengZ., and TeohS.H.Surface modification of ultra thin poly (epsilon-caprolactone) films using acrylic acid and collagen. Biomaterials, 25, 1991, 2004.
