The Effects of Epithelial Viability on Stromal Keratocyte Apoptosis in Porcine Corneas Stored in Optisol-GS

Purpose: To investigate the effect of corneal epithelium on the viability of corneal stromal keratocytes in Optisol-GS. Methods: After sterilization, corneoscleral buttons were excised and stored in Optisol-GS for various time periods. Group 1 corneas (n = 40) underwent mechanical corneal epithelial debridement before storage while group 2 corneas (n = 40) were stored with intact epithelium. Changes in corneal thickness, keratocyte density, and keratocyte apoptosis were investigated immediately, at 4 hours, and on days 1, 2, 3, 5, 7, and 14 in the preservation medium. The differences between group 1 and 2 corneas were analyzed. Results: Corneal thickness increased significantly in the second week of preservation in both groups, though more substantially in group 1. Significant corneal epithelial apoptosis was noticed in the first week in group 2 corneas. Corneal stromal keratocyte density decreased with prolonged preservation time. DNA laddering was detected by ligation-mediated polymerase chain reaction throughout the experiment periods in both groups, but the increase of keratocyte apoptosis was more significant after 5 days of preservation, especially in group 1. Conclusions: Stromal keratocytes underwent apoptosis in Optisol-GS. The absence of corneal epithelium during preservation further increased the stromal keratocyte apoptosis.

[1]  D. Weiss,et al.  Fluorescence microscopy and three-dimensional imaging of the porcine corneal keratocyte network , 1997, Graefe's Archive for Clinical and Experimental Ophthalmology.

[2]  A. Behrens,et al.  Flap quality in single versus multiple use of the same blade in the Flapmaker microkeratome. , 2004, Journal of refractive surgery.

[3]  G. Wollensak,et al.  Corneal Endothelial Cytotoxicity of Riboflavin/UVA Treatment in vitro , 2003, Ophthalmic Research.

[4]  T. Seiler,et al.  Stress‐strain measurements of human and porcine corneas after riboflavin–ultraviolet‐A‐induced cross‐linking , 2003, Journal of cataract and refractive surgery.

[5]  B. Seitz,et al.  Evaluation of corneal flap dimensions and cut quality using the SKBM automated microkeratome , 2003, Journal of cataract and refractive surgery.

[6]  Koujiro Tohyama,et al.  Comparative observations on corneas, with special reference to bowman's layer and descemet's membrane in mammals and amphibians , 2002, Journal of morphology.

[7]  E. K. Kim,et al.  Effect of microkeratome suction duration on corneal flap thickness and incision angle. , 2002, Journal of refractive surgery.

[8]  M. Boulton,et al.  Apoptosis in the endothelium of human corneas for transplantation. , 2000, Investigative ophthalmology & visual science.

[9]  E. Orwin,et al.  In vitro culture characteristics of corneal epithelial, endothelial, and keratocyte cells in a native collagen matrix. , 2000, Tissue engineering.

[10]  M. Su,et al.  Benzalkonium chloride and gentamicin cause a leak in corneal epithelial cell membrane. , 2000, Experimental Eye Research.

[11]  Dieter G. Weiss,et al.  The Keratocyte Network of Human Cornea: A Three-Dimensional Study Using Confocal Laser Scanning Fluorescence Microscopy , 2000, Cornea.

[12]  R. Mohan,et al.  Stromal-epithelial interactions in the cornea , 1999, Progress in Retinal and Eye Research.

[13]  M. Reim,et al.  Metabolic changes of the human donor cornea during organ-culture. , 1999, Acta ophthalmologica Scandinavica.

[14]  W M Bourne,et al.  Automated quantification of keratocyte density by using confocal microscopy in vivo. , 1999, Investigative ophthalmology & visual science.

[15]  H. Edelhauser,et al.  Comparison of rabbit and human corneas stored in Optisol-GS: changes in stromal sodium. , 1998, Current eye research.

[16]  S. Srivannaboon,et al.  Normal Human Corneal Cell Populations Evaluated by In Vivo Scanning Slit Confocal Microscopy , 1998, Cornea.

[17]  M M Jablonski-Stiemke,et al.  Storage of human corneas in dextran and chondroitin sulfate-based corneal storage medium: changes in stromal free sodium. , 1998, Archives of ophthalmology.

[18]  J. Marshall,et al.  Keratocyte density and size in conscious humans by digital image analysis of confocal images , 1998, Eye.

[19]  A. Berghold,et al.  Loss of corneal Langerhans cells during storage in organ culture medium, Optisol and McCarey-Kaufman medium , 1998, Eye.

[20]  C. Cullander,et al.  Assessment of cornea viability by confocal laser scanning microscopy and MTT assay. , 1997, Cornea.

[21]  Jian-ping Gao,et al.  Apoptosis in the Rabbit Cornea After Photorefractive Keratectomy , 1997, Cornea.

[22]  A. Blaschke,et al.  Apoptotic DNA fragmentation is detected by a semi-quantitative ligation-mediated PCR of blunt DNA ends , 1997, Cell Death and Differentiation.

[23]  D. Easty,et al.  Factors influencing the suitability of organ-cultured corneas for transplantation. , 1997, Investigative ophthalmology & visual science.

[24]  H. Grossniklaus,et al.  The corneal epithelium after optisol-GS storage. , 1996, Cornea.

[25]  M. Lynn,et al.  Donor factors associated with epithelial defects after penetrating keratoplasty. , 1996, Cornea.

[26]  G. Wilson,et al.  Apoptosis in the corneal epithelium. , 1996, Investigative ophthalmology & visual science.

[27]  S. Wilson,et al.  Epithelial injury induces keratocyte apoptosis: hypothesized role for the interleukin-1 system in the modulation of corneal tissue organization and wound healing. , 1996, Experimental eye research.

[28]  T. Møller-Pedersen,et al.  A three-dimensional study of the human corneal keratocyte density. , 1995, Current Eye Research.

[29]  E. Cohen,et al.  Factors associated with epithelial defects after penetrating keratoplasty. , 1994, Ophthalmic surgery.

[30]  P. McDonnell,et al.  Keratocyte loss and repopulation of anterior corneal stroma after de-epithelialization. , 1994, Archives of ophthalmology.

[31]  N. Ehlers,et al.  Histopathological findings in failed human epikeratophakia lenticules , 1994, Acta ophthalmologica.

[32]  G. Clover,et al.  Keratocyte networks visualised in the living cornea using vital dyes. , 1993, Journal of cell science.

[33]  H. Edelhauser,et al.  Comparison of Corneal Preservation Media for Corneal Hydration and Stromal Proteoglycan Loss , 1992, Cornea.

[34]  R. Laing,et al.  Optisol corneal storage medium. , 1991, American journal of ophthalmology.

[35]  R. Beuerman,et al.  Optisol corneal storage medium. , 1992, Archives of ophthalmology.

[36]  R. Meyer,et al.  Corneal epithelium in penetrating keratoplasty. , 1980, American journal of ophthalmology.

[37]  J. Friend,et al.  Corneal epithelial changes during midterm storage. , 1976, Investigative ophthalmology.

[38]  J. Friend,et al.  Corneal epithelial preservation. , 1975, Archives of ophthalmology.

[39]  A. Silverstein,et al.  The survival and rejection of epithelium in experimental corneal transplants. , 1969, Investigative ophthalmology.