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Mapping of proteomic profile and effect of the spongy layer in the human amniotic membrane

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Abstract

The graft of human amniotic membrane (HAM) 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 HAM is denuded from its spongy layer, but this layer can be an important source of growth factors which promote re-epithelialization. The aim of our study is to provide a general view of protein expression of the HAM and the spongy layer and therefore to determine if the spongy layer and/or a specific part of HAM have a beneficial role in the process of wound healing in patients with corneal ulcers. For this study, human placentas were obtained from healthy women after vaginal delivery or caesarean section after signing the consent form. Mapping of protein expression is done by dividing the placenta in 2 equal parts, one with spongy layer and another without (conventional HAM). Each part is also divided in 3 zones depending on the distance from the umbilical cord. The proteomic analysis was done by ELISA, targeting growth factors (EGF, HGF, KGF, NGF and TGF-beta1) and pro inflammatory cytokine TNF-α in the HAM without spongy layer and in the spongy layer. In this study we observed significant difference in the total amount of protein extract between the different donors. We do not observe a significant difference in the growth factor level between the conventional HAM and the spongy layer. No variation was observed in the expression of HGF, KGF and NGF in different zone of HAM and neither between conventional HAM and spongy layer in each zone. (*p value < 0.05, **p value<0.01,***p value < 0.001). We do detect very low dose of TNF-α and no correlation with the amount of growth factors. In our study we demonstrated that keeping the spongy layer in conventional method of handling HAM can add more GF, and so probably have a positive affect the wound healing process. Variation in some growth factors expression has been observed between the placentas and therefore this may explain the variation in clinical results. No indicator for the selection of placentas with a higher rate of growth factor was found.

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Acknowledgements

Thanks to Charlotte QUETTEVILLE for the help she has provided in recruiting placentas.

Funding

This works was co-supported by European Union and Normandie Regional Council. Europe gets involved in Normandie with European Regional Development Fund (ERDF), (17B07001E).

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Authors and Affiliations

  1. Faculty of Medicine and Pharmacy, University of Rouen Normandy, Inserm U1234, Rouen, France

    Parvin Nazari Hashemi, Olivier Boyer & Marc Muraine

  2. Eye Bank of Normandy, Charles Nicolle Hospital, Rouen, France

    Fanny Chaventre & Marc Muraine

  3. Department of Ophthalmology, Charles Nicolle Hospital, Rouen University Hospital, Rouen, France

    Julie Gueudry & Marc Muraine

  4. Laboratory of Immunology and Biotherapy, Rouen University Hospital, Rouen, France

    Aurelie Bisson & Olivier Boyer

Authors
  1. Parvin Nazari Hashemi
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  2. Fanny Chaventre
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  3. Aurelie Bisson
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  4. Julie Gueudry
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  5. Olivier Boyer
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  6. Marc Muraine
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Correspondence to Parvin Nazari Hashemi.

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The authors do not have any conflict of interest.

Ethical approval

All procedures performed in this study were in accordance whit the ethical standard of French bioethics laws (N°AC-2013-1886), and the study complied with the tenets of the Declaration of Helsinki 1964 and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in this study.

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Nazari Hashemi, P., Chaventre, F., Bisson, A. et al. Mapping of proteomic profile and effect of the spongy layer in the human amniotic membrane. Cell Tissue Bank 21, 329–338 (2020). https://doi.org/10.1007/s10561-020-09821-8

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  • DOI: https://doi.org/10.1007/s10561-020-09821-8

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