Telomerized fibroblasts as a candidate 3D in vitro model of pathological hypertrophic scars

Shadrin VS, Kozhin PM, Shoshina OO, Luzgina NG, Rusanov AL
About authors

Orekhovich Research Institute of Biomedical Chemistry, Moscow, Russia

Correspondence should be addressed: Valerian S. Shadrin
Pogodinskaya, 10, str. 8, Moscow, 119121; moc.liamg@nirdahsnairelav

About paper

Funding: this research was supported by the Russian Ministry of Science and Higher Education and was conducted under the Federal Targeted Program on Research and Development in Priority Fields of Science and Technology for 2014–2020 (Agreement 05.604.21.0219, Project ID RFMEFI60419X0219).

Author contribution: Luzgina NG, Rusanov AL conceived the study and proposed its design; Shadrin VS, Kozhin PM, Shoshina OO, Luzgina NG, Rusanov AL analyzed the literature, analyzed and interpreted the experimental data and wrote the manuscript; Shadrin VS, Kozhin PM planned and conducted the experiment; Shadrin VS wrote the manuscript.

Received: 2020-08-28 Accepted: 2020-09-02 Published online: 2020-09-27

The search for the optimal cell model for studying the pathogenesis of pathological scars is a pressing challenge. This study aimed at evaluating the feasibility of using telomerized fibroblasts for the in vitro 3D modeling of pathological hypertrophic scars. NF and Fb-hTERT cells were cultured as monolayers and spheroids in the absence and in the presence of TGFβ1. The metabolic activity of the cultured cells was assessed using the MTT assay. Cell migration was estimated using the scratch assay. The expression of genes associated with fibrous scar tissue growth was measured by qRT-PCR. Fb-hTERT cells were more metabolically active than NF cells in the presence of TGFβ1 (for 1 ng/ml: 179 ± 12% vs. 135 ± 13% respectively; p < 0,05). Spheroids grown from Fb-hTERT cells were significantly larger than those derived from NF cells. In the presence of TGFβ1, the expression of proteins associated with extracellular matrix production (COL1A1, COL3A1, FN1) was lower in Fb-hTERT cells than in NF cells (more than 25, 20 and 2-fold, respectively; p < 0.05). Intact NF cells were more active in closing the scratch than Fb-hTERT cells: on day 2, the gap closure rate was 2.28 times higher in NF cells (p < 0.05). Exposure to TGFβ1 stimulated Fb-hTERT, unlike NF cells, to close the gap 2 times faster on day 2 (p < 0.05). Thus, telomerized fibroblasts have a few phenotypic traits observed in keloid fibroblasts; still there are some limitations that should be accounted for when using Fb-hTERT cells for the modeling of pathological hypertrophic scars.

Keywords: fibroblasts, in vitro model, hTERT, hypertrophic scar, keloid scar, TGFβ1