Amniotic Membrane Modifies the Genetic Program Induced by TGFß, Stimulating Keratinocyte Proliferation and Migration in Chronic Wounds.

María Concepción López-Martínez,Francisco José Nicolás,Ander Izeta,Ángel Bernabé-García,Asunción Monfort,Dimitris Kletsas,Carmen Luisa Insausti,Anna Mrowiec,Gregorio Castellanos,Eva María García-Vizcaíno,Antonia Alcaraz,José María Moraleda

doi:10.1371/journal.pone.0135324

María Concepción López-Martínez, Francisco José Nicolás + Show 10 more

Open Access

https://doi.org/10.1371/journal.pone.0135324

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Abstract

BackgroundPost-traumatic large-surface or deep wounds often cannot progress to reepithelialisation because they become irresponsive in the inflammatory stage, so intervention is necessary to provide the final sealing epidermis. Previously we have shown that Amniotic Membrane (AM) induced a robust epithelialisation in deep traumatic wounds.Methods and FindingsTo better understand this phenomenon, we used keratinocytes to investigate the effect of AM on chronic wounds. Using keratinocytes, we saw that AM treatment is able to exert an attenuating effect upon Smad2 and Smad3 TGFß-induced phosphorylation while triggering the activation of several MAPK signalling pathways, including ERK and JNK1, 2. This also has a consequence for TGFß-induced regulation on cell cycle control key players CDK1A (p21) and CDK2B (p15). The study of a wider set of TGFß regulated genes showed that the effect of AM was not wide but very concrete for some genes. TGFß exerted a powerful cell cycle arrest; the presence of AM however prevented TGFß-induced cell cycle arrest. Moreover, AM induced a powerful cell migration response that correlates well with the expression of c-Jun protein at the border of the healing assay. Consistently, the treatment with AM of human chronic wounds induced a robust expression of c-Jun at the wound border.ConclusionsThe effect of AM on the modulation of TGFß responses in keratinocytes that favours proliferation together with AM-induced keratinocyte migration is the perfect match that allows chronic wounds to move on from their non-healing state and progress into epithelialization. Our results may explain why the application of AM on chronic wounds is able to promote epithelialisation.

Highlights

Wound healing is the body’s natural biological process for regenerating dermal and epidermal tissue, which involves a delicate balanced activity of inflammatory, vascular, connective tissue and epithelial cells [1].Acute wounds heal rapidly and proceed through the inflammatory, proliferation and remodelling phases of healing
Amniotic membrane attenuated the TGFß-induced phosphorylation of Smad2 and 3
When we looked into the JNK1 phosphorylation, the stimulation with Amniotic Membrane (AM) sustained its effect for 24 h and TGFß did not stimulate it further

Summary

Introduction

Wound healing is the body’s natural biological process for regenerating dermal and epidermal tissue, which involves a delicate balanced activity of inflammatory, vascular, connective tissue and epithelial cells [1].Acute wounds heal rapidly and proceed through the inflammatory, proliferation and remodelling phases of healing. Largesurface or deep wounds, with an important loss of soft tissues, often become senescent in the inflammatory or proliferation stages and cannot progress to re-epithelialisation [1, 2]. This failure in the re-epithelialisation process requires the need for intervention in order to provide the epithelial layer for the final sealing of the skin [1, 2]. Post-traumatic large-surface or deep wounds often cannot progress to reepithelialisation because they become irresponsive in the inflammatory stage, so intervention is necessary to provide the final sealing epidermis. We have shown that Amniotic Membrane (AM) induced a robust epithelialisation in deep traumatic wounds

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