Efficacy of Amphotericin B Against Fusarium and Aspergillus in Corneal Storage Medium.

PURPOSE: The incidence of postkeratoplasty fungal infection is increasing in the United States, and our most commonly used corneal storage medium, Optisol-GS, contains antibiotics but no antifungal agents. We previously demonstrated the efficacy of amphotericin B additives in eliminating Candida albicans contaminants in Optisol-GS. The purpose of this study was to determine whether amphotericin B would also be efficacious against Fusarium solani and Aspergillus fumigatus. METHODS: Vials of Optisol-GS were supplemented with 0.255 µg/mL of amphotericin B. Half of the vials were inoculated with F. solani and half with A. fumigatus. Positive control vials were inoculated with the fungi but no amphotericin B. The vials were refrigerated, sampled, and plated at different time points. The plates were then incubated at 36°C for 48 hr after which fungal colony counts were performed. RESULTS: There was an average reduction in the growth of F. solani in the amphotericin B-supplemented vials of 44% on day 2, 79% on day 7, and 80% on day 14 when compared with the positive control vials. There was an average reduction in the growth of A. fumigatus in the amphotericin B-supplemented vials of 40% on day 2 and 14% on day 7 when compared with the positive control vials. Both amphotericin B-supplemented and control vials grew less than 2 colonies of A. fumigatus on day 14. CONCLUSIONS: This study suggests that amphotericin B additives in Optisol-GS reduce the growth of F. solani and A. fumigatus.

Systemic Fungal Infections in Donors for Corneal Transplantation.

PURPOSE: To determine the prevalence of postkeratoplasty fungal infection when corneal tissue from donors with a recent medical history of oral thrush or respiratory, urine, wound, sputum, bronchial, tracheal, or throat culture positive for fungus is identified before recovery and after decontamination of the corneal tissue with 5% povidone-iodine flush to the donors' eyes during recovery. METHODS: This is a prospective analysis of corneas from 42 donors with a documented medical history of fungus or positive fungal culture, which were recovered between January 2010 and November 2010. Standard aseptic swab of the donors' corneas before and after application of 5% povidone-iodine solution was performed. Culture results were analyzed in relationship to the donors' medical history and potential posttransplantation infections. RESULTS: Eighty-four eyes from 42 patients were swabbed for cultures during the study period. Seven eyes (8.3%) were positive for fungal growth before treatment with 5% povidone-iodine, whereas there were no positive fungal cultures after treatment (P = 0.007). Fifty-four corneas from this study group were used for corneal transplantation. There were no cases of fungal infection in any postkeratoplasty eyes transplanted from this study group. CONCLUSIONS: In this small study, the overall prevalence of fungal infections after corneal transplantation using corneal donor tissue from donors with a fungal-positive medical history is low. Corneal fungal contamination in donors with a history of fungal infection or a positive fungal culture can be significantly reduced with a 5% povidone-iodine flush.

Validation of cooling effect of insulated containers for the shipment of corneal tissue and recommendations for transport.

PURPOSE: To determine the cooling effect of generic insulated shipping containers in ambient and high-temperature environments. METHODS: Twenty-seven shipping containers were packed with wet ice according to industry standards. The ice in each container was weighed. Ambient temperatures were recorded by data loggers affixed to the exterior. Internal temperatures were recorded by data loggers packed inside the containers, for as long as the data loggers remained at ≤8°C. The cooling effect, or minutes per gram of ice a data logger maintained a temperature of ≤8°C, was calculated using linear regression; 8 similar containers were subjected to elevated summer temperatures. RESULTS: Small, medium, and large containers held mean masses of wet ice of 685, 1929, and 4439 g, respectively. The linear regression equation for grams of ice to duration of time at ≤8°C was y = 0.1994x + 385.13 for small containers, y = 0.1854x + 1273.3 for medium, and y = 0.5892x + 1410.3 for large containers, resulting in a cooling effect of 25.1 hours for small, 58.9 hours for medium, and 85.7 hours for large containers at ambient temperature. The duration of cooling effect in the summer profile group was consistent with that of the ambient temperature group. CONCLUSIONS: All of the container sizes successfully maintained proper cooling when packed with the appropriate grams of wet ice for the needed time interval. This study validates current practice for the shipment of corneal tissue in inexpensive, generic containers that can maintain effective cooling for the duration required for local, national, and international shipment.

Preparation of corneal donor eyes comparing 1% versus 5% povidone-iodine.

PURPOSE: To compare the effect of 1% versus 5% polyvinylpyrrolidone-iodine (PVP-I) chemical preparation (prep) of the eye on the recovery of organisms from donor globes before in situ recovery of donor corneal tissue. METHODS: One hundred consecutive pairs of donor corneas (200 eyes) were randomized to receive either 1% or 5% PVP-I drops applied to the conjunctival cul-de-sac, which was left in place for 2 minutes. Limbal cultures were obtained before and after prepping of the eye. RESULTS: Twenty-five different species of organisms were recovered. Native flora of the eye included coagulase-negative staphylococci (62%), Corynebacterium species (27%), streptococcal species (9.5%), gram-negative bacilli (14.5%), Staphylococcus aureus (5%), anaerobes (10%), and yeast (2%). After PVP-I instillation of the donor eye, 74 isolates were recovered from the 1% P-I group and 76 isolates from the 5% PVP-I group. Cultures were sterile after PVP-I prep in 49 eyes and 47 eyes in the 1% PVP-I group and 5% PVP-I group, respectively. Microorganism colony forming units were similar among post-prep cultures from both PVP-I groups. The effect of the PVP-I prep on the number of negative cultures and on the reduction in the number of isolates was highly significant for both the 1% PVP-I group and the 5% PVP-I group when compared with the limbal cultures taken before PVP-I instillation. CONCLUSIONS: This study found that 1% and 5% PVP-I solutions are equally effective for chemical prep of the donor eye. Because PVP-I is known to be toxic to the corneal endothelium and corneal fibroblasts, this study suggests that 1% PVP-I should be the preferred disinfectant for the recovery of corneal donors.

Minimizing the risk of disease transmission during corneal tissue processing.

Corneal transplantation is undergoing significant change because the dysfunctional portion of the cornea may now be selectively transplanted. After recovery of corneoscleral tissue, further processing of such tissue as in microkeratome preparation of endothelial keratoplasty lenticules is defined as "open-container processing" by the Eye Bank Association of America. Airborne bacterial contamination during preparation of corneal tissue is a potential source of postoperative infection. This review addresses ways to minimize the risk of disease transmission as corneal tissue is processed for lamellar keratoplasty, endothelial keratoplasty, or femtosecond laser-assisted penetrating keratoplasty and to minimize risk to eye bank personnel or physicians preparing the tissue. Secondly, quality assurance measures are described that qualify the environment in which corneal tissue is being processed. We propose that the environment in which corneal tissue is being processed must be able to demonstrate acceptable levels of airborne microbial contamination annually as measured by settle plates to estimate airborne bacterial sedimentation. It is recommended that any environment where corneal tissue is prepared should meet the minimum standard of a conventional operating room which is <25 colony-forming unit per 90-mm settle plate per 1-hour exposure.

Recipient risk factors for graft failure in the cornea donor study.

PURPOSE: To identify recipient factors that may be related to risk of corneal graft failure. DESIGN: Multicenter, prospective, double-masked, controlled clinical trial. PARTICIPANTS: One thousand ninety subjects undergoing corneal transplantation for a moderate-risk condition (principally Fuchs' dystrophy or pseudophakic corneal edema). METHODS: Donor corneas were assigned using a random approach without respect to recipient factors, and surgeons were masked to information about the donor cornea, including donor age. Surgery and postoperative care were performed according to the surgeons' usual routines, and subjects were followed up for 5 years. Baseline factors were evaluated for their association with graft failure. MAIN OUTCOME MEASURES: Graft failure, defined as a regraft or a cloudy cornea that was sufficiently opaque to compromise vision for a minimum of 3 consecutive months. RESULTS: Preoperative diagnosis of pseudophakic or aphakic corneal edema increased graft failure risk approximately 4-fold compared with Fuchs' dystrophy (27% vs. 7%). Prior glaucoma surgery with preoperative glaucoma medication use substantially increased the graft failure rate. Factors not strongly associated with graft failure included age, gender, diabetes, smoking, and graft size. CONCLUSIONS: The risk of graft failure is significantly increased in eyes with pseudophakic or aphakic corneal edema compared with Fuchs' dystrophy, independent of lens status, and in eyes with a history of glaucoma. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

In situ ultrahigh-resolution optical coherence tomography characterization of eye bank corneal tissue processed for lamellar keratoplasty.

PURPOSE: To use optical coherence tomography (OCT) as a noninvasive tool to perform in situ characterization of eye bank corneal tissue processed for lamellar keratoplasty. METHODS: A custom-built ultrahigh-resolution OCT (UHR-OCT) was used to characterize donor corneal tissue that had been processed for lamellar keratoplasty. Twenty-seven donor corneas were analyzed. Four donor corneas were used as controls, whereas the rest were processed into donor corneal buttons for lamellar transplantation by using hand dissection, a microkeratome, or a femtosecond laser. UHR-OCT was also used to noninvasively characterize and monitor the viable corneal tissue immersed in storage medium over 3 weeks. RESULTS: The UHR-OCT captured high-resolution images of the donor corneal tissue in situ. This noninvasive technique showed the changes in donor corneal tissue morphology with time while in storage medium. The characteristics of the lamellar corneal tissue with each processing modality were clearly visible by UHR-OCT. The in situ characterization of the femtosecond laser-cut corneal tissue was noted to have more interface debris than shown by routine histology. The effects of the femtosecond laser microcavitation bubbles on the corneal tissue were well visualized at the edges of the lamellar flap while in storage medium. CONCLUSIONS: The results of our feasibility study show that UHR-OCT can provide superb, in situ microstructural characterization of eye bank corneal tissue noninvasively. The UHR-OCT interface findings and corneal endothelial disc thickness uniformity analysis are valuable information that may be used to optimize the modalities and parameters for lamellar tissue processing. The UHR-OCT is a powerful approach that will allow us to further evaluate the tissue response to different processing techniques for posterior lamellar keratoplasty. It may also provide information that can be used to correlate with postoperative clinical outcomes. UHR-OCT has the potential to become a routine part of tissue analysis for any eye bank or centers creating customized lamellar corneal tissue for transplantation.