Absence of detectable SARS-CoV-2 replication in ex vivo cultured cornea and cornea-derived epithelial cells.

PURPOSE: To study the possibility of SARS-CoV-2 to infect human corneal cells and tissues under standard corneal culture conditions using explants of COVID-19 donors and primary cornea-derived epithelial cells. METHODS: Cornea isolated from deceased COVID-19 donors was cultured for 4 weeks, and SARS-CoV-2 replication was monitored by qRT-PCR. Furthermore, primary corneal epithelial cells from healthy donors were cultured ex vivo and infected with SARS-CoV-2 and human cytomegalovirus (HCMV) as a control. Infection status was assessed by western blotting and reporter gene expression using green fluorescent protein-expressing viral strains. ACE2 and TMPRSS2 receptor expression levels in cornea and epithelial cells were assessed by qRT-PCR. RESULTS: We did not detect SARS-CoV-2 replication in 10 corneas isolated from deceased COVID-19 patients and cultured for 4 weeks, indicating absence of infection under natural conditions. Furthermore, high-titer SARS-CoV-2 infection of ex vivo cultured cornea-derived epithelial cells did not result in productive virus replication. In contrast, the same cells were highly permissive for HCMV. This phenotype could potentially be explained by low ACE2 and TMPRSS2 transcriptional activity in cornea and cornea-derived epithelial cells. CONCLUSIONS: Our data suggest that cornea and limbal epithelial cells are refractory to productive SARS-CoV-2 infection. This could be due to the absence of robust receptor expression levels necessary for viral entry. This study adds further evidence to support the very low possibility of transmission of SARS-CoV-2 from an infected corneal transplant donor to a recipient in corneal organ cultures.

Factors influencing the contamination rate of human organ-cultured corneas.

PURPOSE: To assess the influence of donor, environment and storage factors on the contamination rate of organ-cultured corneas, to consider the microbiological species causing corneal contamination and to investigate the corresponding sensitivities. METHODS: Data from 1340 consecutive donor corneas were analysed retrospectively. Logistic regression analysis was used to assess the influence of different factors on the contamination rate of organ-cultured corneas for transplantation. RESULTS: The mean annual contamination rate was 1.8 ± 0.4% (range: 1.3-2.1%); 50% contaminations were of fungal origin with exclusively Candida species, and 50% contaminations were of bacterial origin with Staphylococcus species being predominant. The cause of donor death including infection and multiple organ dysfunction syndrome increased the risk of bacterial or fungal contamination during organ culture (p = 0.007 and p = 0.014, respectively). Differentiating between septic and aseptic donors showed an increased risk of contamination for septic donors (p = 0.0020). Mean monthly temperature including warmer months increased the risk of contamination significantly (p = 0.0031). Sex, donor age, death to enucleation, death to corneoscleral disc excision and storage time did not increase the risk of contamination significantly. CONCLUSION: The genesis of microbial contamination in organ-cultured donor corneas seems to be multifactorial. The main source of fungal or bacterial contamination could be resident species from the skin flora. The rate of microbial contamination in organ-cultured donor corneas seems to be dependent on the cause of donor death and mean monthly temperature.

Evaluation of Factors Limiting Corneal Donation.

BACKGROUND This study aimed to investigate factors limiting corneal donation at the University Hospital Tübingen. MATERIAL AND METHODS We retrospectively studied all hospital deaths from January 2012 to December 2015, considering each deceased patient as a potential corneal donor. During this period an ophthalmic resident managed corneal donor procurement on a full-time basis. Various factors limiting corneal donation were examined. RESULTS Among the 3412 deaths, 2937 (86.1%) displayed nonfulfillment of corneal donation. Consent for corneal donation was obtained in 475 cases (13.9%). The mean annual corneal donation rate was 13.9 donors per 100 deaths (range: 11.2-17.8). The leading causes of nonfulfillment of corneal donations were refusal to donate (49.8%, 1698 cases) and medical contraindications (23.6%, 805 cases). After next-of-kin interview of 2173 potential donors (109 potential donors were excluded because of logistical problems), willingness to participate in corneal donation was present in 475 cases (21.9%), whereas in 1698 cases (78.1%) corneal donation was refused. CONCLUSIONS Our study showed refusal to donate is the most important factor limiting corneal donation. It seems that increasing the knowledge of people about corneal donation through public education and media are necessary to address the corneal shortage.