Dermal fibroblasts have different extracellular matrix profiles induced by TGF-β, PDGF and IL-6 in a model for skin fibrosis

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.

Biomarkers of extracellular matrix formation are associated with acute-on-chronic liver failure

“…This is not surprising as both PRO-C3 and PRO-C6 are biomarkers of collagen formation derived from fibroblasts and thus correlate directly to fibroblast activity. 24 Both markers are well known to be associated with increased severity of fibrosis and portal hypertension. 5 , 7 , 17 Interestingly, the plasma levels of the collagen type IV formation marker PRO-C4 were similar between patients with AD and ACLF.…”

Mechanotopography‐Driven Design of Dispersible Nanofiber‐Laden Hydrogel as a 3D Cell Culture Platform for Investigating Tissue Fibrosis

“…This cell type has been widely used to study fibrotic behavior. [ 61–63 ] The cells were cultured in Dulbecco's modified Eagle's medium supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Thermo Fisher). The fibroblasts were dispersed in a pre‐gel solution (1 × 10 6 cells mL –1 ) and photocrosslinked to encapsulate into a hydrogel.…”

A Human Cellular Model for Colorectal Anastomotic Repair: The Effect of Localization and Transforming Growth Factor-β1 Treatment on Collagen Deposition and Biomarkers

“…Specifically, TGF-β1 increased the synthesis of type I collagen measured by COL1A1 mRNA and the biomarker PINP but not type III collagen synthesis measured by COL3A1 mRNA or the biomarker PRO-C3. Treatment of normal human dermal fibroblasts with TGF-β1 also resulted in increased type I collagen but not type III secretion or deposition [ 50 ]. Although TGF-β1 increased PRO-C6 in the conditioned media, this was most likely due to concurrent increases in cell numbers, as TGF-β1 reduced COL6A1 mRNA levels in the CoReFs.…”

“…Although TGF-β1 increased PRO-C6 in the conditioned media, this was most likely due to concurrent increases in cell numbers, as TGF-β1 reduced COL6A1 mRNA levels in the CoReFs. Juhl et al [ 50 ] found that TGF-β1 downregulated COL6A1 in normal human dermal fibroblasts.…”