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. 2020 Jul 2;10:10868. doi: 10.1038/s41598-020-67730-y

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© The Author(s) 2020

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Identification of chondrocyte cell types in OA and the potential upstream regulators of these phenotypes. a shows a similarity across patients and a predominance of preFibrochondrocytes (preFC), Fibrochondrocytes (FC), Regulatory chondrocytes (RegC), Reparative chondrocytes (RepC) and some preHypertrophic chondrocytes (preHTC) in diseased areas designated ‘D’ corresponding to the medial tibial plateau (MT); a also shows a predominance of Homeostatic chondrocytes (HomC), some PreHTC and Hypertrophic chondrocytes (HTC) in non-diseased areas designated ‘N’ corresponding to OLT or the lateral tibial plateau. Potential upstream cytokines (b) and growth factors (e) were identified based on all highly expressed genes from each cluster and scaled by activation z-score; asterisks indicate the upstream mediators that were expressed by > 1% of acquired chondrocytes. Nearly all cytokines (c) and growth factors (f) were expressed by synoviocytes: 22 of 31 cytokines and 14 of 30 growth factors were only expressed by synoviocytes and not by chondrocytes (the proportion of acquired synoviocytes expressing each cytokine and growth factor are shown in the pie charts). (d) Violin plots show expression levels of representative cytokines (TNF, IL6, IL1B and IL1A) and a growth factor (IGF1) across all cell types in OA synovia and the HLA-DRA⁺ cell subtypes. (g) Representative immunofluorescence staining of synovium and cartilage (intact and damaged regions of a single patient specimen) for IL6 and CD14. As expected, the damaged cartilage is hypercellular compared to intact cartilage due to the development of multicellular chondrons in damaged cartilage with osteoarthritis. IL6 was highly expressed by CD14⁺ synovial fibroblasts (SSF, SSF and iFIB), I- MΦ, and SMC, but not chondrocytes from either intact or damaged regions of cartilage.