Debra L. Skelnik

Optisol Corneal Storage Medium

Optisol is an investigational, intermediate-term corneal storage medium containing chondroitin sulfate and dextran to enhance corneal dehydration during storage. We used scanning electron microscopy to grade endothelial cell morphologic characteristics in terms of cell shape, cell borders, cell swelling, and apical holes in pairs of corneas stored in Optisol and Dexsol. Optisolstored corneas showed significantly fewer morphologic changes after 14 days at 4 degrees C than did Dexsol-stored corneas. No significant differences were seen after 1 to 4 days at 26 degrees C. Temperature-reversal analysis showed no significant change in corneal thickness with warming after 2-week storage at 4 degrees C in either medium, although Optisol-stored corneas were significantly thinner than those stored in Dexsol at all times. The results of scanning electron microscopy suggest that preservation at refrigerator temperature for 2 weeks in Optisol is superior to preservation in Dexsol. Both media may be useful in preserving endothelial structure for limited periods at room temperature, which could provide a measure of safety in shipping or storage where refrigeration is unreliable.

Minnesota system corneal preservation.

The clinical and laboratory results with a modified Minnesota system of organ culture corneal preservation are presented. A refinement of our preservation technique using a closed system, as well as the addition of chondroitin sulphate to the medium is presented. Laboratory results show preservation of corneal endothelial integrity for at least 21 days with maintenance of normal corneal thickness. In addition, a 10-day quarantine system reduces the risk of donor contamination and secondary endophthalmitis. Preliminary results of the 34 degrees C and 4 degrees C closed Minnesota corneal preservation system using chondroitin sulphate show that it is safe and efficacious and allows intermediate to long-term maintenance of sterile thin tissue prior to corneal transplantation.

An In Vitro and Clinical Comparison of Corneal Storage with Chondroitin Sulfate Corneal Storage Medium with and without Dextran

The safety and efficacy of 1% dextran in Chondroitin Sulfate Corneal Storage Medium (CSM) in reducing corneal swelling after 4 degrees C storage was assessed in a corneal endothelial cell culture system. No difference was found in 3H-thymidine incorporation by cells incubated in either CSM-dextran medium or CSM medium alone. Subsequently, 21 pairs of corneas, stored in either CSM or CSM-dextran from 30 to 112 hours, were transplanted into 42 eyes of 42 patients, paired by diagnostic group and procedure. All CSM grafts and 19 of 21 CSM-dextran grafts were clear at 4 months with no primary donor failures in either group. Intraoperative corneal thickness was significantly greater in the CSM group (0.82 +/- 0.07 mm) than the CSM-dextran group (0.76 +/- 0.06 mm); however, the two groups did not differ thereafter. No differences in all endothelial morphometric parameters were noted between the two groups pre- and postoperatively. Average endothelial cell loss by 4 months was 13.0 +/- 16.4% for the CSM group and 16.4 +/- 15.5% for the CSM-dextran group. The addition of dextran to CSM medium results in significant intraoperative corneal thinning without adversely affecting endothelial DNA synthesis in vitro and endothelial survival in vivo.

Increased Endothelial Cell Loss After Transplantation of Corneas Preserved by a Modified Organ-culture Technique

Forty-seven donor corneas were preserved in McCarey-Kaufman (M-K) medium at 4 degrees C for 1 day, then in organ culture at 34 degrees C for approximately 1 month, then in M-K medium at 4 degrees C for an additional two days before transplantation. The central donor endothelium was examined by specular microscopy before and after organ culture and 2 months after keratoplasty. No significant change in central endothelial cell density occurred during organ culture. The 47 transplants were compared with 47 grafts preserved only in M-K medium at 4 degrees C for approximately 36 hours. All transplants were performed by the same surgeon over the same period, and the two groups contained similar types of surgical procedures. The organ-cultured grafts were thicker on the first post-operative day and took longer to epithelialize . Two months after keratoplasty all of the 94 grafts were clear and thin, but the mean central endothelial cell loss was 28% in the 47 organ-cultured transplants and 10% in the 47 transplants preserved only in M-K medium (P less than 0.0001). These results indicate that the endothelium of corneas preserved by organ culture at 34 degrees C and then placed in M-K medium at 4 degrees C for 2 days may be more susceptible to surgical trauma than those preserved only in M-K medium at 4 degrees C.

Neodymium:YAG laser interaction with Alcon IOGEL hydrogel intraocular lenses: An in vitro toxicity assay

ABSTRACT The objective of this study was to determine the potential toxicity generated by the interaction of the Nd:YAG laser and Alcon IOGEL intraocular hydrogel lens material. The IOGEL lens is composed of poly 2‐hydroxyethylmethacrylate, containing 38% water, previously shown to be highly biocompatible in a wide range of tissue culture and implantation experiments. In this study, intraocular lenses (IOLs) immersed in serum‐free cell culture medium were purposely exposed to exaggerated doses of laser energy to cause extensive damage. An IOLAB polymethylmethacrylate (PMMA) lens served as a control lens material. The resultant solutions were assayed for cytotoxicity in a bioassay system using fourth passage human corneal endothelial cells. No cytotoxicity was seen in the bioassay for the IOGEL hydrogel IOLs or the PMMA control IOL at any laser range/dosage tested over a 72‐hour incubation period. Hydrogel lenses exhibited decreasing yellowing with decreasing energy levels, and no lens discoloration was apparent at the lowest level of irradiation, 5 mJ/50 laser bursts; the PMMA control lens exhibited moderate yellowing at 15 mJ/50 bursts. Lens marking was moderate for all IOGEL IOLs; the PMMA lens marking was severe at the power level tested.

Safety and efficacy of 2% methylcellulose in cat and monkey cataract-implant surgery

We evaluated the safety and efficacy of 2% methycellulose as an adjunct for cataract extraction with implantation in cat and monkey models. When used intraoperatively, methylcellulose reduced the iridovitreal bulge during surgery. No significant increase in clinical inflammation occurred nor was there statistically significant intraocular pressure elevation at 24 hours, 7 days, or 90 days. In the cat model, the central corneal thickness increased at day seven in both control and methylcellulose eyes; this thickness persisted to 90 days. The endothelial cell loss decreased significantly at day 90 in methylcellulose eyes. In the monkey model, no statistically significant increase in corneal thickness occurred in control or methylcellulose eyes at day seven. The endothelial cell loss was greater than in the cat model in both control and methylcellulose eyes; there was no statistically significant difference between the two. Two percent methylcellulose was safe in both the cat and monkey models. It facilitated surgery in both models and reduced the endothelial cell loss in the cat eye.

Neodymium:YAG laser interaction with intraocular lenses: An in vitro toxicity assay

Use of the Nd:YAG laser is an effective technique to open an opacified posterior lens capsule. However, in the presence of a posterior chamber intraocular lens (IOL), precise focusing of the laser on the capsule is required to avoid pitting the lens optic. The question has been raised whether toxic products may result from laser damage to the IOL. We addressed this issue in the present study by exposing primary human corneal endothelial cell and human corneal organ cultures to solutions produced by purposefully hitting IOLs immersed in cell growth medium with a Nd:YAG laser. The lenses studied were lathe-cut polymethylmethacrylate (PMMA), injection-molded non-UV PMMA, injection-molded UV PMMA, and cast-molded UV PMMA. Samples of each material were irradiated in a holder containing 1 ml of cell culture medium using the following conditions: 5, 10, and 50 laser bursts at 10 millijoules (mJ), and 50 laser bursts at 5 mJ. The solutions were applied to the endothelial cell cultures (all materials) and to the corneal organ cultures (injection-molded non-UV lenses only). There was no toxicity in either assay for any of the materials studied.

pH of organ-culture-stored corneas

Abstract Changes in intracorneal and storage‐medium pH values of organ‐culture‐stored cat corneas were monitored over a 4‐week period. The intracorneal pH was determined using the phosphorus‐31 magnetic resonance spectroscopy (31P MRS) chemical shift of inorganic orthophosphate in conjunction with a standard pH titration curve. We incubated 32 adult cat corneas using two similar standard organ‐culture methods, one with chondroitin sulfate (method 1) and the other without (method 2). Time‐course data at 0, 1, 3 and 4 weeks of storage were used to calculate the rate of pH change. The intracorneal pH was not changed significantly for either organ‐culture method; however, the storage‐medium pH rate of change declined significantly for both methods (method 1, 0.15 pH units/week; method 2, 0.12 pH units/week). The difference between intracorneal and storage‐medium pH values over time increased at a rate of 0.12 and 0.11 pH units/week for method 1 and method 2, respectively. The declining storage‐medium pH in conjunction with the maintenance of intracorneal pH contributes to an increased metabolic demand on the cornea.