1. Dermal fibroblasts have different extracellular matrix profiles induced by TGF-β, PDGF and IL-6 in a model for skin fibrosis
- Author
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Anne Sofie Siebuhr, Michael J. Davies, Sandie Bondesen, Clare L. Hawkins, Morten A. Karsdal, Anne-Christine Bay-Jensen, and Pernille Juhl
- Subjects
Calcium Phosphates ,0301 basic medicine ,Indoles ,LIVER ,INTERLEUKIN-6 ,Receptor, Transforming Growth Factor-beta Type I ,lcsh:Medicine ,ACTIVATION ,Extracellular matrix ,chemistry.chemical_compound ,Rheumatic diseases ,0302 clinical medicine ,Transforming Growth Factor beta ,Fibrosis ,VITRO ,lcsh:Science ,Skin ,Platelet-Derived Growth Factor ,Multidisciplinary ,biology ,Chemistry ,Dermis ,Extracellular Matrix ,Nintedanib ,Collagen ,Platelet-derived growth factor receptor ,Type I collagen ,Cell biology ,I COLLAGEN ,Article ,Transforming Growth Factor beta1 ,Dermal fibroblast ,03 medical and health sciences ,medicine ,Humans ,030203 arthritis & rheumatology ,Interleukin-6 ,GROWTH-FACTOR-BETA ,SYSTEMIC-SCLEROSIS ,lcsh:R ,Fibroblasts ,medicine.disease ,Molecular biology ,Actins ,Fibronectins ,Fibronectin ,030104 developmental biology ,biology.protein ,lcsh:Q ,Transforming growth factor - Abstract
Different stimulants might induce different extracellular matrix profiles. It is essential to gain an understanding and quantification of these changes to allow for focused anti-fibrotic drug development. This study investigated the expression of extracellular matrix by dermal fibroblast mimicking fibrotic skin diseases as SSc using clinically validated biomarkers. Primary healthy human dermal fibroblasts were grown in media containing FICOLL. The cells were stimulated with PDGF-AB, TGF-β1, or IL-6. Anti-fibrotic compounds (iALK-5, Nintedanib) were added together with growth factors. Biomarkers of collagen formation and degradation together with fibronectin were evaluated by ELISAs in the collected supernatant. Immunohistochemical staining was performed to visualize fibroblasts and proteins, while selected gene expression levels were examined through qPCR. TGF-β and PDGF, and to a lesser extent IL-6, increased the metabolic activity of the fibroblasts. TGF-β primarily increased type I collagen and fibronectin protein and gene expression together with αSMA. PDGF stimulation resulted in increased type III and VI collagen formation and gene expression. IL-6 decreased fibronectin levels. iALK5 could inhibit TGF-β induced fibrosis while nintedanib could halt fibrosis induced by TGF-β or PDGF. Tocilizumab could not inhibit fibrosis induced in this model. The extent and nature of fibrosis are dependent on the stimulant. The model has potential as a pre-clinical model as the fibroblasts fibrotic phenotype could be reversed by an ALK5 inhibitor and Nintedanib.
- Published
- 2020
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