Bacterial Contamination of Human Organ-Cultured Corneas

Zanetti, Elisa; Bruni, A.; Mucignat, Giorgio; Camposampiero, Davide; Frigo, Anna Chiara; Ponzin, Diego

doi:10.1097/01.ico.0000153099.96904.43

Cited by 33 publications

(22 citation statements)

References 17 publications

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“…It has been suggested that stromal oedema may allow bacteria to enter the donor cornea. 27 Finally, in the New Zealand National Eye Bank report, 16 the mean death-to-preservation time was 15.2 h and it did not differ between corneas that were suitable for transplantation and those that were unsuitable.…”

Section: Discussionmentioning

confidence: 99%

Influencing factors on the suitability of organ-cultured corneas

Gavrilov

Borderie

Laroche

et al. 2010

Eye

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Section: Discussionmentioning

confidence: 99%

Influencing factors on the suitability of organ-cultured corneas

Gavrilov

Borderie

Laroche

et al. 2010

Eye

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“…On one hand, some latent infections may be inhibited by the fresh antibiotic dose in the deswelling medium, but others may be boosted by the fresh nutrient supply or activated by the deswelling process itself [35]. On the other hand, it is known that Dextran 500 contained in the deswelling medium displays toxicity [5,17,23,25,36], and that the deswelling duration should be kept to the necessary minimum.…”

Section: Discussionmentioning

confidence: 99%

“…Zanetti et al used a combination of classic and automated test systems to assess their contamination rates [35]. In 1,694 corneas they saw 119 (7.02%) primary contaminations.…”

Section: Discussionmentioning

confidence: 99%

Detection of contamination during organ culture of the human cornea

Hermel

Salla

Hamsley

et al. 2009

Graefes Arch Clin Exp Ophthalmol

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“…[9][10][11][12][13][14][15] The Hamburg eye bank found 4.7% fungal and 2.6% bacterial contaminants in 4546 tested cultures. 9 In contrast, the Besançon group reported 1.1% fungal and 4.6% bacterial contamination in 1156 tested storage media.…”

Section: Discussionmentioning

confidence: 99%

Time Course of Antibiotic and Antifungal Concentrations in Corneal Organ Culture

Seiler

Tschopp

Zimmerli

et al. 2016

Cornea

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Purpose: Contamination with bacteria and/or fungi is a serious complication in organ-cultured corneas. Hence, antibiotic and antifungal agents are added to the culture medium. The concentration of different antimicrobial and antifungal additives to the media over time has so far not been investigated in detail and is the aim of this study.Methods: Nine human fresh corneoscleral discs were stored in corneal culture medium consisting of 2% fetal bovine serum and minimal essential medium. In addition, the culture medium contained 1200 mg/mL penicillin G, 25 mg/mL amphotericin B, 120 mg/mL streptomycin, and 100 mg/mL voriconazole. The concentration of amphotericin B used was 10 times higher than in clinical routine to facilitate its detection. The cultures were kept at 37°C for 28 days. At days 0, 7, 14, 21, and 28, samples of the culture medium were harvested for analysis of antimicrobial concentrations by liquid chromatography and electrospray ionization tandem mass spectrometry.Results: During corneal storage, the concentration of all antibiotics and antifungal agents declined significantly. By day 28, penicillin G was reduced to 14% of the original concentration. Amphotericin B and streptomycin retained approximately 60% of the original concentration to the end of the experiment and voriconazole maintained stable concentrations after an initial decline to approximately 80% at 7 days.Conclusions: Throughout the entire storage period, the concentrations of penicillin G, streptomycin, and voriconazole exceeded the minimum inhibitory concentrations of all common contaminants, obviating the need for a change of the medium for antimicrobial reasons. Based on the minimum inhibitory concentrations and our findings, the initial concentration of amphotericin B should be raised to 5 mg/mL. Warm preservation at 31 to 37°C is more commonly used in European eye banks: corneas can be stored up to 4 weeks in organ culture medium consisting of minimum essential medium with the addition of 2% fetal bovine serum.2-6 Antibiotics and antifungal agents are added to the organ culture media to kill contaminants and to avoid iatrogenic infections at transplantation. Despite this prophylaxis, severe complications like endophthalmitis can still occur. 7,8 Little is known about the decay characteristics and efficacy over time of antimicrobial agents kept at body temperature. Therefore, we studied the concentration of the most commonly used antimicrobial and antifungal additives over time under the standard conditions established at the local eye bank at the Department of Ophthalmology, University Hospital in Bern. METHODS Sample PreparationNine human corneoscleral discs unsuitable for transplantation were excised under sterile conditions from intact globes not later than 6 hours post mortem and stored each in a corneal culture medium consisting of 2% fetal bovine serum and minimal essential medium (Sigma Aldrich, St Louis, MO) following the standard protocol of the eye bank of the University Hospital of Bern. The culture medium contained 1...

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Bacterial Contamination of Human Organ-Cultured Corneas

Cited by 33 publications

References 17 publications

Influencing factors on the suitability of organ-cultured corneas

Influencing factors on the suitability of organ-cultured corneas

Detection of contamination during organ culture of the human cornea

Time Course of Antibiotic and Antifungal Concentrations in Corneal Organ Culture

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