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. 2022 Jun 17;12:10275. doi: 10.1038/s41598-022-14491-5

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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NIN limits serum-stimulated growth of smooth muscle cells from circular smooth muscle layer of adult rat colon (CSMC) in vitro. (A–C) Representative phase contrast images of culture model of CSMC at passage 14 in vitro after plating, showing sub-confluent appearance (A), progressive growth to post-confluence with overlapping cell layers (B) and retraction in local areas at when maintained at high density (e.g., arrows) (C). Scale bars, 200 µM. (D) Growth assays showing similar outcomes after 48 h stimulation between fetal calf serum (FCS, 5%) and adult rat serum (RS, 2.5%), with p < 0.05 vs control, n = 5 cell lines, each with triplicate assays. (E) Nintedanib inhibits stimulation of growth of CSMC by FCS or RS with greater sensitivity than pirfenidone. p < 0.05 vs control for all data with nintedanib > 0.1 µM; p < 0.05 for pirfenidone ≥ 2 mM; n = 5 cell lines with triplicate assays. (F) Similar outcomes to (E) for inhibition of FCS-induced growth of fibroblast 3T3 cells. *p < 0.05 vs control. (G) Lack of effect of nintedanib on FCS-induced growth of rat intestinal epithelial cells (IEC-18).