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Paracrine Effects of Adipose-Derived Cellular Therapies in an in Vitro Fibrogenesis Model of Human Vocal Fold Scarring

  • Mélanie Velier
    Affiliations
    Aix Marseille Univ, Institut National de la Santé Et de la Recherche Medicale, Institut National de la Recherche Agronomique, Centre recherche en CardioVasculaire et Nutrition, Marseille, France

    Cell Therapy Department, Hôpital de la Conception, Marseille, France
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  • Alexia Mattei
    Correspondence
    Address correspondence and reprint requests to Alexia Mattei, Service ORL et chirurgie cervico-faciale du Pr Michel, CHU Conception, 147 boulevard Baille, 13005 Marseille.
    Affiliations
    APHM, La Conception University Hospital, Department of Oto-Rhino-Laryngology and Head and Neck Surgery, Marseille, France

    Aix Marseille University, Centre National de la Recherche Scientifique, Laboratoire Parole et Language, Aix-en-Provence, France
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  • Stéphanie Simoncini
    Affiliations
    Aix Marseille Univ, Institut National de la Santé Et de la Recherche Medicale, Institut National de la Recherche Agronomique, Centre recherche en CardioVasculaire et Nutrition, Marseille, France
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  • Jérémy Magalon
    Affiliations
    Aix Marseille Univ, Institut National de la Santé Et de la Recherche Medicale, Institut National de la Recherche Agronomique, Centre recherche en CardioVasculaire et Nutrition, Marseille, France

    Cell Therapy Department, Hôpital de la Conception, Marseille, France
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  • Laurent Giraudo
    Affiliations
    Cell Therapy Department, Hôpital de la Conception, Marseille, France
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  • Laurent Arnaud
    Affiliations
    Cell Therapy Department, Hôpital de la Conception, Marseille, France
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  • Antoine Giovanni
    Affiliations
    APHM, La Conception University Hospital, Department of Oto-Rhino-Laryngology and Head and Neck Surgery, Marseille, France

    Aix Marseille University, Centre National de la Recherche Scientifique, Laboratoire Parole et Language, Aix-en-Provence, France
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  • Francoise Dignat-George
    Affiliations
    Aix Marseille Univ, Institut National de la Santé Et de la Recherche Medicale, Institut National de la Recherche Agronomique, Centre recherche en CardioVasculaire et Nutrition, Marseille, France

    Cell Therapy Department, Hôpital de la Conception, Marseille, France
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  • Florence Sabatier
    Affiliations
    Aix Marseille Univ, Institut National de la Santé Et de la Recherche Medicale, Institut National de la Recherche Agronomique, Centre recherche en CardioVasculaire et Nutrition, Marseille, France

    Cell Therapy Department, Hôpital de la Conception, Marseille, France
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  • Markus Gugatschka
    Affiliations
    Division of Phoniatrics, ENT University Hospital Graz, Medical University of Graz, Graz, Austria
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  • Tanja Grossmann
    Affiliations
    Division of Phoniatrics, ENT University Hospital Graz, Medical University of Graz, Graz, Austria
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      SUMMARY

      Objectives/Hypothesis

      Vocal folds (VF) scarring leads to severe dysphonia which negatively impacts daily life of patients. Current therapeutic options are limited due in large part to the high complexity of the micro-structure of the VF. Innovative therapies derived from adipose tissue such as stromal vascular fraction (SVF) or adipose derived stromal/ stem cells (ASC) are currently being evaluated in this indication and paracrine anti-fibrotic effects are considered as predominant mechanisms.

      Methods

      The paracrine anti-fibrotic effects of SVF and ASC from healthy donors were tested in an innovative in vitro fibrogenesis model employing human VF fiboblasts (hVFF) and the principles of macromolecular crowding (MMC). Biosynthesis of collogen and alpha-smooth-muscle actin (αSMA) expression in hVFF were quantified after five days of indirect coculture with ASC or SVF using silver stain, western blot and RT-qPCR analysis.

      Results

      Fibrogenesis was promoted by addition of transforming growth factor beta 1 (TGFβ1) combined with MMC characterized by an enhanced deposition of fibrillar collagens and the acquisition of a myofibroblast phenotype (overexpression of αSMA). Adipose-derived therapies led to a reduction in the αSMA expression and the collagen content was lower in hVFF co-cultivated with SVF.

      Conclusions

      ASC and SVF promoted significant prevention of fibrosis in an in vitro fibrogenesis model through paracrine mechanisms, supporting further development of adipose-derived cellular therapies in VF scarring.

      Key Words

      Abbreviations:

      VF (vocal fold), ECM (extracellular matrix), αSMA (alpha-smooth-muscle actin), ACTA2 (actin alpha 2), bFGF (basic fibroblasts growth factor), HGF (hepatocyte growth factor), MSC (mesenchymal stromal cells), SVF (stromal vascular fraction), ASC (adipose derived stromal cells), hVFF (human vocal fold fibroblasts), MMC (macromolecular crowding), TGF-β1 (transforming growth factor beta 1), DMEM (Dulbecco´s modified Eagle´s medium), FCS (fetal calf serum), SM (standard medium), GMP (good manufacturing practices), RT (reverse transcription), RT-qPCR (reverse transcription quantitative polymerase chain reaction), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), SDS-PAGE (sodium dodecylsulphate-polyacrylamide gel electrophoresis), VEGF (vascular endothelial growth factor), COL1A1 (Collagen 1 alpha 1)
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