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Analyse protéomique et propriétés de ré-épithélialisation des membranes amniotiques humaines après lyophilisation en vue d’une greffe de la surface oculaire

The graft of human amniotic membrane contributes to the healing of corneal perforating ulcers and so to save a large number of eyes suffering of severe chemical burns. This biological material is used for the treatment of ocular surface diseases because of its capacity to reduce inflammation and promote a quicker wound healing. For clinical use, the amniotic membrane is separated from the amnion. Subsequently, this membrane is stored by cryopreservation at -80 ° C, which potentially complicates the delivery of membranes. Consequently, the Eye bank of the University Hospital wishes to develop freeze-drying of MAH to facilitate its use and distribution. 

For this study, human placentas were obtained from healthy women after cesarean. The human amniotic membranes are divided into two parts, one part that has been lyophilized and the other frozen. The proteomic study is conducted by comparing lyophilized to cryopreserved human amniotic membrane by targeting growth factors and also a global proteomic analyze is done. Comparatives histological and cell culture analysis were also done.

For in vivo analysis, a lesion in 18 rabbits on a single cornea will be done mechanically. 6 of them received a lyophilized amniotic membrane graft, 6 frozen membrane and 6 did not receive a transplant. The wound healing process was observed for one month. 

Targeting analysis for proteins of Interest in the re-epithelialization process and results of histologic analysis protein do not show significant difference between the lyophilized portion and the frozen portion. The culture of epithelial cells on lyophilized membranes is comparable to that cultured on frozen membranes. These results are also confirmed on an in vivo model (rabbit). 

In vitro histological and proteomic analysis showed no significant difference between frozen and lyophilized membranes. The in vivo analysis does not show significant difference in the process of wound healing. These results will be confirmed by confirmed by structural analysis in electron microscopy. 

If in vivo analysis tends to confirm the results obtained in vitro, these membranes could be considered to be used clinically and also to be sent to countries to respond to humanitarian needs in African countries (Cameroon).