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Published byPriscilla Whitehead Modified over 6 years ago
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Mice deficient in the NF-κB negative regulator, itch, develop severe osteoarthritis and reduced synovial lymphatic drainage due to m1 macrophages-induced lymphatic endothelial cell inflammation W. Wang, H. Xu, W. Sun, H. Wang, M. Zuscik, L. Xing Osteoarthritis and Cartilage Volume 24, Pages S31-S32 (April 2016) DOI: /j.joca Copyright © Terms and Conditions
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Fig. 1 Itch−/− mice have more severe OA and decreased synovial lymphatic drainage. 3.5 m-old Itch−/− and WT littermates were received MLI on right Knee and sham surgery on left knee. (A&B) Mice received ultrasound imaging of their knees before and 10 wks post-surgery and data were analyzed by Amira (A) Representative 2D and 3D images of with highlighted joint space (green outline). (B) Changes on joint space volume were calculated. (C&D) Lymphatic drainage of knee joints was assessed before and 10 weeks after MLI by NIR-ICG imaging. (C) Representative NIR- ICG lymphatic imaging showing retention of more ICG in OA joints of Itch−/− mice. (B) The ICG clearance was calculated. (E&F) Joint tissue sections were subjected to AB/OG-staining and histological analyses. Values are mean ± SD of 8-10 knee joints. *p<0.05 vs. WTOA samples. Osteoarthritis and Cartilage , S31-S32DOI: ( /j.joca ) Copyright © Terms and Conditions
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Fig. 2 Increased macrophages and lymphatic vessels in Itch−/− OA joints and Itch−/− macrophages stimulate LEC inflammation. 6-m-old Itch−/− mice and WT littermates were received MLI and sham surgery and sacrificed at 5 weeks post-MLI. Frozen sections of knee joints were subjected IHC with anti-PDPN for LECs (red) and anti-F/4/80 for macrophages (green). Imagines from whole slide digital microscopy showing enriched lymphatic vessels and macrophages in OA joints from Itch−/− mice (A) and some of F4-80+ macrophages (red arrows) are localized adjacent to PDPN+ LECs (yellow arrows)(B). Bone marrow (BM) cells from Itch−/− and WT littermates were cultured with M-CSF for 3 days to generate BM macrophages (BMM). (C) Itch−/− and WT BMMs were treated with IL-1. The expression of M1 cytokines were determined by qPCR. (D) CM (40%) from BMMs and M1s were added to mLEC cultured for 24 hrs, and expression of inflammatory genes was measured by qPCR. Values are mean ± SD of 3 wells. The fold changed was calculated by normalized with value from WT BMMs as 1. *p<0.05 vs WT M1s. Osteoarthritis and Cartilage , S31-S32DOI: ( /j.joca ) Copyright © Terms and Conditions
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