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. 2021 Mar 1;10:e66532. doi: 10.7554/eLife.66532

Figure 6. PTH-induced osteoarthritis pain relief is inhibited by PTH type one receptor knockout on Nestin+ MSCs.

(A, B) 50% PWT at the LH and PAMWT at the left knee in sham-operated, PTH-treated DMM, and vehicle-treated DMM PTH type one receptor−/− (Pth1r−/−) and Pth1r+/+ mice at week 8 after sham or DMM surgery (n = 8/group). (C) Quantitative analysis of LH intensity, LH area, and LH swing speed compared with the RH in sham-operated, PTH-treated DMM, and vehicle-treated DMM Pth1r−/− and Pth1r+/+ mice at week 8 after sham or DMM surgery, based on catwalk analysis (n = 8/group). (D–F) Immunofluorescent and quantitative analysis of CGRP+ (green) sensory nerve fibers and EMUN+ vessels (red) in tibial subchondral bone of PTH-treated or vehicle-treated Pth1r−/− and Pth1r+/+ mice at week 8 after DMM surgery (n = 8/group). Scale bar: 50 μm. (G, H) Immunofluorescent and quantitative analysis of CGRP+ (green) sensory nerve fibers in synovium of PTH-treated or vehicle-treated Pth1r−/− and Pth1r+/+ mice at week 8 after DMM surgery (n = 8/group). Scale bar: 50 μm. (I) μCT images of the tibial subchondral bone medial compartment (coronal) at week 8 after DMM surgery. Arrowhead indicates the osteophyte. Scale bar: 500 μm. (J) Total volume measurement of osteophytes from the tibial plateau of sham-operated, PTH-treated DMM, and vehicle-treated DMM Pth1r−/− and Pth1r+/+ mice (n = 8/group). (K) Immunohistochemical quantification of COX2+ cells in the tibial subchondral bone of mice at week 4 after DMM surgery (n = 8/group). (L) Quantitative analysis of PGE2 in subchondral bone determined by enzyme-linked immunosorbent assay (ELISA) (n = 8/group). *p<0.05, **p<0.01. NS, no significant difference.

Figure 6—source data 1. Raw data of PAMWT, 50% PWT, catwalk analysis, CGRP staining, EMUN staining, CGRP staining in synovium, quantification of osteophyte volume, and COX2 staining, and quantification of level of PGE2.

Figure 6.

Figure 6—figure supplement 1. Expression levels of SP, P2X3 and PiEZO in nerve fibers were signigficantly decreased PTH1R knockout mice.

Figure 6—figure supplement 1.

(A–C) Immunofluorescence analysis of Substance P+ (SP+) (red), P2 × 3+ (red), and PIEZO2+ (red) sensory nerve fibers in tibial subchondral bone of sham-operated, PTH-treated DMM, and vehicle-treated Pth1r−/− and Pth1r+/+ mice at week 8 after DMM surgery. DAPI stains nuclei blue. Scale bar: 50 μm. (D–F) Quantitative analysis of the density of SP+, P2 × 3+, and PIEZO2+ sensory nerve fibers in tibial subchondral bone at week 8 after DMM surgery (n = 8/group).
Figure 6—figure supplement 1—source data 1. Raw data of SP staining, P2 × 3 staining, and PIEZO2 staining.