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2020, Number 2

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Biotecnol Apl 2020; 37 (2)

Novel contributions by Transmission Electron Immunomicroscopy on the intracellular trafficking of EGFR, PCNA, FOXO1A-P and LC3B and their biological response in clinical samples of diabetic foot ulcers treated with Heberprot-P®

Falcón CV, González BM, Acosta RN, Pentón AE, Guillén NG, Mendoza MY, Bringas PR, Pereira YK, García OA, Fernández MM, Garateix SR, Acosta MEF, González LNE, Rosales UCM, Berlanga AJ

Language: English
References: 31
Page: 2511-2514
PDF size: 599.61 Kb.


ABSTRACT

Diabetic foot ulcers (UPDs) is a diabetes complication and the leading cause of non-traumatic amputations. It was shown that Heberprot-P®, a therapeutic product based on epidermal growth factor (EGF), enhances the healing response or healing of chronic wounds (RCH) in UPDs. However, a better understanding of the cellular and molecular mechanisms of EGF receptor (EGFR) activation with its natural ligand in vivo, its intracellular traffic and the biological response to treatment was required. A study with electron immunomicroscopy was carried out for the detection and quantification of EGFR, its related molecules and autophagy vacuoles in fibroblast-like cells (FLCs) from UPDs. Samples were analyzed before and after treatment with Heberprot-P®. Before treatment, little EGFR and proliferating cell nuclear antigen (PCNA) were detected, predominating the transcriptional factor of phosphorylated hairpin head O1 (FOXO1A-P) in the cell nucleus and the induction of autophagy in FLCs from UPDs, possibly associated with the impairment of the functions of the FLCs in the RCH. Heberprot-P® induced an early increase in the immunodetection of EGFR and PCNA in the cell nucleus (15-60 minutes) and later in mitochondria (6 and 24 h). The nuclear functions of FOXO1A-P and autophagy decreased. EGFR and PCNA were shown associated with exosome-like structures and their accumulation in the extracellular matrix. All these were relevant for the restoration of FLCs functions, facilitating skin RCH. This was the first in vivo report demonstrating a prolonged biological effect (24 h) of Heberprot-P® infiltration in FLCs from UPDs, related to RCH, and supporting the current therapeutic protocol.


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