Organ-cultured corneal grafts from septic donors: a retrospective study.

PURPOSE: To evaluate the quality of corneal grafts from donors, who have died from septic multi-organ failure and who are called septic donors in the following. METHODS: One hundred and eighty-two corneal grafts from septic donors were stored in organ culture for 10-14 days. Graft evaluation was performed according to the criteria of the European Eye Bank Association. Only donor corneas with cell density values above 2000 cells/mm(2) were transplanted. Ninety-one patients who received these transplanted corneas were examined retrospectively with special emphasis on endophthalmitis, graft failure and incidence of immune reactions. RESULTS: Ninety-one of 182 donor corneas (50%) from septic donors were discarded mainly due to endothelial damage (61; 67%). Only seven (8%) were discarded due to medium contamination. In contrast, 452 of 1261 donor corneas (36%) from non-septic donors during the same period were discarded, again mainly due to endothelial damage (264; 58%). In this group, 48 donor corneas (11%) were discarded due to medium contamination. No patient who had received a graft from a septic donor has experienced endophthalmitis. The rate of immune reactions and graft failure was in the same range when compared to a larger group who received grafts from non-septic donors. CONCLUSION: Our data reveal no contraindication against the use of corneal grafts derived from septic donors, critical graft assessment in organ culture provided.

Herpes simplex virus infection in the media of donor corneas during organ culture: frequency and consequences.

BACKGROUND AND PURPOSE: According to polymerase chain reaction (PCR) studies 2-38% of organ culture donor corneas may contain herpes simplex virus (HSV) DNA, but there are only 6 reported instances of proven virus replication in a corneoscleral disc. Moreover there are only 6 patients reported in whom primary graft failure and extensive post-operative epithelial defects were probably caused by a herpetic infection of the corneal graft. Recently we observed virus replication in a donor cornea with subsequent complete endothelial necrosis in our cornea bank. The aim of this study was to investigate the possible correlation between herpetic donor cornea infection and endothelial necrosis in organ culture. METHODS: To evaluate the frequency of HSV as a reason for endothelial necrosis in organ culture we tested the media of 199 donor corneas discarded due to an altered endothelium in the years 1997 to 1999 by PCR for HSV. As a negative control group we screened the media of 117 transplanted corneas using PCR. RESULTS: In the control group we had only negative PCR results, in contrast to the corneas with severe or complete endothelial necrosis where HSV DNA was detected in 12 media of the corneas of 9 donors. Virus could be cultivated out of 7 media. CONCLUSIONS: (1) HSV replication is a common cause of severe endothelial necrosis in organ culture corneas. (2) Replication of the virus during organ culture comes close to a virus cultivation using the corneoscleral disc as a cell culture. (3) We consider the danger of transplanting active HSV to be very small if critical assessment of the graft prior to surgery is carried out.

Testing of corneoscleral discs and their culture media of seropositive donors for hepatitis B and C virus genomes.

UNLABELLED: The prevalence of donors seropositive for hepatitis B virus (HBV) surface antigen (HBsAg) or hepatitis C virus (HCV) antibody (anti-HCV) in western countries is estimated to be 0.5%-1%. There have been only two cases, however, published so far, where hepatitis B was suspected to have been transmitted by penetrating keratoplasty [4]. Concerning HCV, no suspected transmission by keratoplasty has been reported so far. This is also true for the time before serological screening for infectious diseases became mandatory for corneal donors. In the Lions Cornea Bank North Rhine Westfalia, 4.7% (HBV) and 3.2% (HCV) respectively of the corneas of the years 1995 to 1999 were discarded due to a "non-negative serology". In about 50% of these cases the screening test (ELISA) generated no valid signal and, therefore, a "questionable positivity" was assumed. Since in Germany corneal graft shortage still is a limiting factor for penetrating keratoplasty, this study was to evaluate the detectability of HBV-DNA and HCV-RNA in the serum samples, organ culture media and corneas of donors tested seropositive for HBsAg or anti-HCV in an attempt to obtain information as to the potential infectivity of this donor material. In this study, 29 corneas of 17 donors seropositive by ELISA for HBsAg and 27 corneas of 14 donors seropositive by ELISA for anti-HCV were evaluated. The organ culture media and the sera were screened for the presence of HBV-DNA or HCV-RNA by PCR. The corneoscleral discs were divided into a central trephinate (7 mm) and the corneoscleral rim. Concerning HBV-DNA both tissues were examined separately by polymerase chain reaction (PCR). In the case of HCV-RNA, a further, more sensitive nucleotide amplification method (NAT), the transcription mediated amplification (TMA), was used to test media, central corneas and cornealscleral rims. The media were additionally tested by PCR. Viral nucleic acid was detected in the sera from 6 of 17 HBsAg positive donors and from 6 of 14 anti-HCV positive donors. Viral genomes could not be detected in the organ culture media nor in the central corneas or corneoscleral rims by PCR at a detection limit of 1000 and 100 copies/ml. Concerning HCV-RNA, two media were positive in the TMA with 50-100 copies/ml. CONCLUSION: according to our results, the risk of transmitting hepatitis B or C virus by penetrating keratoplasty appears to be low: although hepatitis C virus RNA could be detected in 2 media (from two donors) out of 27 with a low concentration of virus copies between 50-100/ml. It remains open whether such a low virus particle number may cause infection in the recipient.