Bacterial Contamination of Human Organ-cultured Corneas

PURPOSE.The aim of this study was to detect and identify bacterial contaminants in human corneoscleral tissue after organ culture storage. METHODS.Seventy-two corneoscleral rims and corneal buttons trephined from organ cultured corneoscleral discs using aseptic technique and 45 organ-cultured donor corneoscleral rims postpenetrating keratoplasty) were subjected to a mechanical extraction technique using a Stomacher laboratory blender. As a control, 28 of the corneoscleral rims and buttons were halved; one half of each corneoscleral rim and button was decontaminated in formalin for 48 hours before thorough washing in balanced salt solution. Corneal specimens, culture medium, and transport (5% dextran) medium were cultured in brain–heart infusion broth at 37°C for 5 days. Bacterial isolates were identified after culture of turbid enrichment broth. RESULTS.Bacterial contamination was demonstrated in 29% (21 of 72) of the corneoscleral rims and 15% (11 of 72) of the corneal buttons that were trephined aseptically from corneoscleral discs and in 29% (13 of 45) of postkeratoplasty corneoscleral rims. Bacterial contaminants were not isolated from controls. Isolated microorganisms included coagulase-negative Staphylococcus, Staphylococcus aureus, Streptococcus viridans, Pseudomonas sp, and Bacillus sp. A correlation was not demonstrated between contamination and cause of death, death to enucleation, death to culture time, or time in culture. Postkeratoplasty endophthalmitis was not evident in the patients who had received corneal buttons from those corneoscleral discs that had contaminated corneoscleral rims. CONCLUSIONS.Bacterial contamination exists in corneoscleral tissue after organ culture storage. The difference in distribution of bacteria and percentage of contamination between the peripheral and central corneas causes us to question the value of routine postpenetrating keratoplasty corneoscleral rim cultures.