Mapping cell diversity and dynamics in inflammatory temporomandibular joint osteoarthritis with pain at single-cell resolution
Supawadee Jariyasakulroj, Yang Shu, Ziying Lin, Jingyi Chen, Qing Chang, Pao-Fen Ko, Jian-Fu Chen
Supawadee Jariyasakulroj, Yang Shu, Ziying Lin, Jingyi Chen, Qing Chang, Pao-Fen Ko, Jian-Fu Chen
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Research Article Cell biology Inflammation

Mapping cell diversity and dynamics in inflammatory temporomandibular joint osteoarthritis with pain at single-cell resolution

Abstract

Temporomandibular joint (TMJ) osteoarthritis with pain is a highly prevalent disorder affecting patients’ quality of life. A comprehensive understanding of cell type diversity and its dynamics in painful TMJ osteoarthritis (TMJOA) is lacking. Here, we utilized an inflammatory TMJOA mouse model via intra-articular injection of CFA. TMJOA mice exhibited cartilage remodeling, bone loss, synovitis, increased osteoarthritis (OA) score, and orofacial pain, recapitulating hallmark symptoms in patients. Single-cell transcriptomic profiling of the TMJ was performed in conjunction with mouse genetic labeling, tissue clearing, light sheet and confocal 3D imaging, multiplex RNAscope, and immunodetection. We visualized, reconstructed, and analyzed the distribution and density of nociceptive innervation of TMJ at single-axon levels. We systematically mapped the heterogeneity and anatomical position of blood endothelial cells, synovial fibroblasts, and immune cells, including Cx3cr1-positive barrier macrophages. Importantly, TMJOA mice exhibited enhanced neurovascular coupling, sublining fibroblast hyperplasia, inflammatory immune cell expansion, disrupted signaling-dependent cell-cell interaction, and a breakdown of the sandwich-like organization consisting of synovial barrier macrophages and fibroblasts. By utilizing a mouse model with combined TMJ pain history and OA, we reveal the cellular diversity, anatomical structure, and cell dynamics of the TMJ at single-cell resolution, which facilitate our understanding and potential targeting of TMJOA.

Authors

Supawadee Jariyasakulroj, Yang Shu, Ziying Lin, Jingyi Chen, Qing Chang, Pao-Fen Ko, Jian-Fu Chen

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Figure 4

scRNA-Seq analysis of all cells in the TMJ of control and CFA mice.

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scRNA-Seq analysis of all cells in the TMJ of control and CFA mice. (A) ...
(A) Schematic diagram of TMJ inoculation and experimental procedure of single-cell analysis. (B) Uniform manifold approximation and projection (UMAP) visualization of major cell types highlighted with different colors in TMJ. (C) Dot plot depicting selected markers enriched for each cell population within the TMJ. (D) UMAP plot for both control (red) and CFA (blue) with the major cell types. Red arrow indicates the reduced osteocyte population in CFA group, while black arrowheads represent the increased monocyte, macrophage, and osteoclast subsets in CFA group compared with control. The red arrowhead indicates an increased endothelial cell population in the CFA group compared with the control. (E) Proportion of each cell cluster in CFA group versus control. Red arrow indicates the reduced osteocyte population, and black arrowheads represent the increased macrophages in CFA group compared with control. The red arrowhead indicates an increased endothelial cell population in the CFA group compared with the control. (F) Feature plots showing the expression of Ccr2 (monocytes), C1qa (macrophages), and Acp5 (osteoclasts), which are distributed closely but are separated from each other in UMAP plot and are increased in CFA group compared with control. Normalized expression levels for each cell are color-coded and overlaid onto the UMAP plot. Myh11, myosin heavy chain 11; Acta2, α–smooth muscle actin; ND, not determined.