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. 2021 Sep 9;11:17978. doi: 10.1038/s41598-021-97445-7

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© The Author(s) 2021

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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Immunohistochemical analysis for the effective time of metformin administration for joint capsular fibrosis. Immunohistochemical staining for TGF-β1, ACTA2, and CCN2 in the posterior capsule. Microscopic images of specimens immunostained for TGF-β1, ACTA2, and CCN2 with DAB and counterstained with hematoxylin in the (Im+/Met 0–2 weeks) group (a–c); and (Im+/Met 2–4 weeks) group (d–f) (Scale bar: 50 μm). Percentage of TGF-β1-positive cells (g), ACTA2-positive cells (h), and CCN2-positive cells (i) to the total number of cells in the posterior joint capsule. Data are expressed as the mean ± standard errors. *p < 0.05, **p < 0.01; n = 5. TGF-β1 transforming growth factor beta 1, ACTA2 actin alpha 2, CCN2 cellular communication network factor 2, Non-Im mice with non-immobilized knees, Im+/Met− mice with immobilized knees but no metformin, Im+/Met 0–4 weeks mice with immobilized knees and metformin administered from 0 to 4 weeks, Im+/Met 0–2 weeks mice with immobilized knees and metformin administered from 0 to 2 weeks, Im+/Met 2–4 weeks mice with immobilized knees and metformin administered from 2 to 4 weeks, DAB 3,3′-diaminobenzidine.