Assessment of the use of cryopreserved x freeze-dried amniotic membrane (AM) for reconstruction of ocular surface in rabbit model

Assessment of the use of cryopreserved x freeze-dried amniotic membrane (AM) for reconstruction of ocular surface in rabbit model

Author Libera, Rodrigo Doyle Autor UNIFESP Google Scholar
Melo, Gustavo Barreto de Google Scholar
Lima, Acacio de Souza Google Scholar
Haapalainen, Edna Freymuller Autor UNIFESP Google Scholar
Cristovam, Priscila Google Scholar
Pereira Gomes, Jose Alvaro Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Abstract Purpose: To determine the efficacy of freeze-dried amniotic membrane (AM) for reconstruction of the ocular surface in rabbit eyes. Methods: the sterilized, freeze-dried amniotic membrane (lyophilized or FD-AM) is a preservative method that uses the drying by freezing process to maintain the AM well preserved for a long time even at room temperature. This paper is an experimental animal interventional study. One eye of each of 15 male New Zealand rabbits (1.5-3.0 kg) had the central cornea marked with a 6.0 mm trephine. the marked area was deepithelialized with a No. 15 blade. the denuded corneal surface was covered as follows: Group 1: cryopreserved AM (n=6); Group 2: freeze-dried AM (n=6); and Group 3: not covered (control group, n=3). the AM in group 1 and 2 and the periphery of the denuded area in group 3 were secured with continuous 10-0 nylon sutures. the clinical evaluation was made by a blinded observer and graded on a four-point scale (1=minimal, 4=marked) for conjunctival and ciliary hyperemia, eyelid edema, corneal neovascularization, corneal opacity and reepithelialization on postoperative (PO) days 1, 7 and 30. After PO day 30, the rabbits were euthanized and their corneas were sent for histopathological and ultrastructural analysis to evaluate tissue inflammation, reepithelialization, and basement membrane integrity. Results: Two eyes in group 2 had a corneal infection and were excluded from the analysis. No statistically significant differences among the three groups were found (p>0.05) regarding the clinical evaluation on 1(st), 7(th) and 30(th) PO days. On transmission electron microscopy, the basement membrane in lyophilized and control groups was more continuous and homogeneous than in the glycerol group. Conclusions: the freeze-drying method seems to be a good option to preserve human amniotic membrane to be used in ocular surface reconstruction. This preservative method reduces the preservation costs and may enhance the use of AM, facilitating its storage and transport.
Keywords Amnion/transplantation
Freeze-drying
Cryopreservation
Tissue preservation/methods
Rabbits
Language English
Date 2008-09-01
Published in Arquivos Brasileiros de Oftalmologia. São Paulo: Consel Brasil Oftalmologia, v. 71, n. 5, p. 669-673, 2008.
ISSN 0004-2749 (Sherpa/Romeo, impact factor)
Publisher Consel Brasil Oftalmologia
Extent 669-673
Origin http://dx.doi.org/10.1590/S0004-27492008000500011
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000273039800011
SciELO ID S0004-27492008000500011 (statistics in SciELO)
URI http://repositorio.unifesp.br/handle/11600/30868

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