Comparison of a Suggested Model of Fibrosis in Human Dermal Fibroblastsby Serum from Systemic Sclerosis Patients with TransformingGrowth Factor β Induced in vitro Model

Document Type : Original Article

Authors
Elnaz Dadashzadeh 1 , 2
Marie Saghaeian Jazi 3 , 4
Nafiseh Abdolahi 5
Saeed Mohammadi 1 , 6
Mohsen Saeidi 1 , 7
1 Stem Cell Research center, Golestan University of Medical Sciences, Gorgan, Iran.
2 Department of Medical Immunology, Golestan University of Medical Sciences, Gorgan, Iran.
3 Stem Cell Research center, Golestan University of Medical Sciences, Gorgan, Iran
4 Metabolic Disorders Research Center, Golestan University of Medical Sciences, Gorgan, Iran
5 Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran
6 Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran
7 Department of Medical Immunology, Golestan University of Medical Sciences, Gorgan, Iran
10.22088/IJMCM.BUMS.11.1.31
Abstract
Systemic sclerosis (SSc) is a chronic autoimmune disease, featuring fibrosis in multiple organs. The
serum from SSc patients contain inflammatory mediators, contributing to SSc pathogenesis and could
be used to develop cell culture models. Here, we compared the fibrotic effects of serum samples from
SSc patients with TGFβ1 on human dermal fibroblasts (HDFs). HDF cells were cultured in four
different culture media supplementations; 10% SSc serum, 10% healthy human serum, 10% fetal
bovine serum or 10% FBS supplemented with 10 ng/Ml human TGFβ. The collagen content in cell
layers was measured by spectrophotometric Picro
-Sirius red staining
. The mRNA expression of
α
-
SMA, COL I and III, TGFβ1
, arginase and E
-Cadherin genes were determined by real time RT
-PCR
.
TGF
-β1 levels in cell culture supernatants were measured using ELISA. Cell layer collagen content
was significantly increased following TGF
-β1 treatment, compared with FBS group and SSc serum
treatment in comparison with healthy controls. Although not statistically significant, the mRNA
expression of α
-SMA, COLI and III, TGFβ1, and arginase increased upon TGF
-β1 treatment in
comparison with FBS group, and in SSc serum treatment group in comparison with healthy controls. E-Cadherin decreased following TGF-β1 treatment and SSc serum treatment in comparison with their
counterparts. TGF
-β1 levels increased in cell culture supernatants of HDF cells exposed to TGF
-β1
and SSc serum. An in vitro model of SSc serum
-induced fibrosis using human HDF cells was evaluated
in comparison to the TGF
-β1 fibrosis induced model and data suggested that it may be used in
documenting the role of pro
-fibrotic factors in serum or plasma from SSc patients.
Keywords
Fibrosis, human dermal fibroblast, systemic sclerosis, autoimmune disease, TGF-β