Abstract
Human amniotic membrane (HAM) has biological properties which are useful for wound healing. HAM is notably one of the therapeutic alternatives for venous leg ulcer care. Indeed, a prospective clinical study has demonstrated that cryopreserved HAM transplantation for leg ulcer is feasible, safe and has beneficial effects: 80 % of the patients had a significant clinical response. Nevertheless, at the end of the 3-month follow-up period, only 20 % of the ulcers were totally closed. The aim of this work was to create and characterize a model of epidermized HAM. The method of HAM desepithelialization was validated by histology, immunohistochemistry and scanning electron microscopy. Then, de-epithelialized HAM was seeded with primary keratinocytes. After 21 days of culture, 15 at the air–liquid interface, the model obtained was analyzed histologically and by immunohistochemistry. The amniotic basement membrane was preserved during enzymatic desepithelialization of HAM. Primary keratinocytes proliferated on HAM: the model obtained showed involucrin expression and had a good basement membrane. As re-epithelialization is an important step for ulcer closure, a model of epidermized HAM could be used to speed up the healing of such wounds.
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Acknowledgments
This work was funded by the “Altération Cutanée” Grant 2007 from Coloplast Foundation (Paris, France) and by the “Aide-jeunes” from Oséo-Anvar (Paris, France). The authors want to thank Anne-Claude Gabiot, Sophie Robin and Delphine Binda for technical and scientific assistance. The authors also want to thank the Department of Histology of the University of Franche-Comté and particularly Claude Collard and Annie La Roche for processing the cryosections. The authors finally want to thank the AICT unit of the French Blood Establishment of Bourgogne Franche-Comté (EFS-BFC), especially Nathalie Pothier, Joëlle Pizzuto and Fabienne Pouthier.
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Tauzin, H., Rolin, G., Viennet, C. et al. A skin substitute based on human amniotic membrane. Cell Tissue Bank 15, 257–265 (2014). https://doi.org/10.1007/s10561-014-9427-z
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DOI: https://doi.org/10.1007/s10561-014-9427-z