Abstract
Chondrocytes are the major functional elements of articular cartilage. Force has been demonstrated to influence the structure and function of articular cartilage and chondrocytes. Therefore, it is necessary to evaluate chondrocytes under different force conditions to gain deep insight into chondrocyte function. Six cartilage tissues from the distal tibia (referred to as the AT group) and five cartilage tissues from the trochlear surface of the talus (referred to as the ATa group) were obtained from 6 donors who had experienced fatal accidents. Single-cell RNA sequencing was used on these samples. A total of 149,816 cells were analyzed. Nine chondrocyte subsets were ultimately identified. Pseudotime analyses, enrichment analyses, cell–cell interaction studies, and single-cell regulatory network inference and clustering were performed for each cell type, and the differences between the AT and ATa groups were analyzed. Immunohistochemical staining was used to verify the existence of each chondrocyte subset and its distribution. The results suggested that reactive oxygen species related processes were active in the force-applied region, while tissue repair processes were common in the force-bearing region. Although the number of prehypertrophic chondrocytes was small, these chondrocytes seemed to play an important role in the ankle.
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Acknowledgements
We acknowledge all donors and authors contributed to this work
Funding
This work was supported by grants from National Natural Science Foundation of China (31872310), which supported the scRNA process and IHC methods.
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Contributions
Junjie Wang: analysis and interpretation of the data, drafting of the article, and critical revision of the article for important intellectual content. Zewen Sun: conception and design and critical revision of the article for important intellectual content. Chenghao Yu: provision of study materials or patients, administrative, technical, or logistic support. Haibo Zhao: administrative, technical, or logistic support. Mingyue Yan: administrative, technical, or logistic support. Shenjie Sun: provision of study materials or patients. Xu Han: provision of study materials or patients. Tingting Jiang: administrative, technical, or logistic support, collection and assembly of data. Tianrui Wang: conception and design, final approval of the article, provision of study materials or patients, administrative, technical, or logistic support. Tengbo Yu: conception and design, final approval of the article, provision of study materials or patients, obtaining of funding, administrative, technical, or logistic support. Yingze Zhang: conception and design, final approval of the article, provision of study materials or patients, obtaining of funding, administrative, technical, or logistic support.
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Supplementary information
Fig S1
Interaction possibility of each determined pathway among different cell types. The size of the circle represents the p value, while the color of the circle represents the interaction possibility. (PNG 1963 kb)
Fig S2
Specific TFs identified by SCENIC a to f From left to right, AUC histogram, scatter plot, ridge plots and violin plots, and parts of the motifs for the specific TFs in each cell type. a for FCs; b for HomCs; c to e for preHTCs, and f for SpCs) (PDF 4539 kb)
Fig S3
Heatmap of the differential expression levels for all scored TFs (PNG 481 kb)
Fig S4
PPI network for DEGs between the AT and ATa groups (Fig S4 for the AT group, Fig S5 for the ATa group). Hub genes are marked with red rectangles (PNG 2276 kb)
Fig S5
(PNG 1791 kb)
Table S1
Detailed DEGs of each cell type (XLSX 371 kb)
Table S2
Detailed results for GO enrichment analysis of each cell type (XLSX 147 kb)
Table S3
Detailed results for KEGG pathway enrichment analysis of each cell type (XLSX 68 kb)
Table S4
Gene targets for specific TFs of HomCs, FCs, RegCs and SpCs (XLSX 32 kb)
Table S5
DEGs between the AT and ATa groups, Genes with |log2FC| values greater than 0 were DEGs for the AT group, and genes with |log2FC| values less than 0 were DEGs for the ATa group (XLSX 226 kb)
Table S6
GO enrichment analysis for DEGs for which the |log2FC| was greater than 0.5 in each group (XLSX 31 kb)
Tables S7 and S8
GO enrichment analysis for DEGs with |log2FC| values greater than 0.25 in each cell type in the AT and ATa groups, respectively (XLSX 128 kb)
Table S8
(XLSX 106 kb)
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Wang, J., Sun, Z., Yu, C. et al. Single-cell RNA sequencing reveals differences between force application and bearing in ankle cartilage. Cell Biol Toxicol 39, 3235–3253 (2023). https://doi.org/10.1007/s10565-023-09829-2
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DOI: https://doi.org/10.1007/s10565-023-09829-2