Abstract
Amniotic membrane (AM) is currently collected from caesarean section placenta, processed into allograft pieces and cryopreserved by the Comprehensive Tissue Centre (CTC) in Edmonton, Alberta, Canada for regional and national ophthalmological and dermatological transplant applications. Anti-inflammatory, anti-immunogenic, and anti-fibrosis properties of AM allografts are important for post-transplant epithelialization. The purpose of this study was to evaluate quality of cryopreserved AM allografts. AM allograft sections were cultured for bacterial and fungal growth after recovery from donors and after antibiotic decontamination. Non-linear controlled-rate cooling was used to cryopreserve AM allografts (sizes: 3 cm2, 5 cm2, 10 cm2). Post-thaw AM viability was assessed by a fluorescent membrane integrity assay (ethidium bromide/Syto® 13) and confocal microscopy. Cell Tissue Organ Surveillance System (CTOSS) forms provided information on AM quality from surgical end-users. While 40.9% of AMs showed positive bacterial culture results upon recovery, with 55.6% of positive culture due to Propionibacterium sp, 100% of cultures were negative after the antibiotic decontamination (n = 22). CTOSS feedback indicated quality of 16.3% AM allografts was excellent, 61.2% satisfactory and 2.0% unsatisfactory (n = 49). Percent viabilities for cryopreserved AM allografts, including 3 cm2 (n = 8), 5 cm2 (n = 7) and 10 cm2 (n = 6) were 57.8 ± 11.0%, 42.7 ± 8.5%, and 47.7 ± 8.2%, respectively, with no statistically significant difference among the allograft sizes (p = 0.54). There was statistical significant difference between percent viability of “fresh” allografts (47.7 ± 13.6%) when compared to cryopreserved samples (p = 0.78). The results of this study provide important information on quality of AM allografts for transplant applications. The CTC takes great pride in providing safe allografts of high quality, through excellence in practice and leadership in research. Disclosure: Author was or is employed by University of Alberta.
Published Version
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