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)To read this article in full you will need to make a payment
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Article info
Publication history
Published online: June 13, 2022
Accepted:
May 17,
2022
Publication stage
In Press Corrected ProofFootnotes
This work was supported by PHC Amadeus Campus France 2017 (grant number FR05/2017).
Identification
Copyright
© 2022 The Voice Foundation. Published by Elsevier Inc. All rights reserved.
