Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis
Synovitis of the knee synovium is proven to be a precursor of knee osteoarthritis (OA), leading to a radiologically advanced stage of the disease. This study was conducted to elucidate the expression pattern of different inflammatory factors—NF-kB, iNOS, and MMP-9 in a subpopulation of synovial cell...
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MDPI AG
2021-06-01
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| Series: | International Journal of Molecular Sciences |
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| author | Marko Ostojic Ante Zevrnja Katarina Vukojevic Violeta Soljic |
| author_facet | Marko Ostojic Ante Zevrnja Katarina Vukojevic Violeta Soljic |
| author_sort | Marko Ostojic |
| collection | DOAJ |
| description | Synovitis of the knee synovium is proven to be a precursor of knee osteoarthritis (OA), leading to a radiologically advanced stage of the disease. This study was conducted to elucidate the expression pattern of different inflammatory factors—NF-kB, iNOS, and MMP-9 in a subpopulation of synovial cells. Thirty synovial membrane intra-operative biopsies of patients (ten controls, ten with early OA, and ten with advanced OA, according to the Kellgren–Lawrence radiological score) were immunohistochemically stained for NF-kB, iNOS, and MMP9, and for different cell markers for macrophages, fibroblasts, leukocytes, lymphocytes, blood vessel endothelial cells, and blood vessel smooth muscle cells. The total number of CD68+/NF-kB+ cells/mm<sup>2</sup> in the intima of early OA patients (median = 2359) was significantly higher compared to the total number of vimentin+/Nf-kB+ cells/mm<sup>2</sup> (median = 1321) and LCA+/NF-kB+ cells/mm<sup>2</sup> (median = 64) (<i>p</i> < 0.001 and <i>p</i> < 0.0001, respectively). The total number of LCA+/NF-kB+ cells/mm<sup>2</sup> in the subintima of advanced OA patients (median = 2123) was significantly higher compared to the total number of vimentin+/NF-kB+ cells/mm<sup>2</sup> (median = 14) and CD68+/NF-kB+ cells/mm<sup>2</sup> (median = 29) (<i>p</i> < 0.0001). The total number of CD68+/iNOS+ cells/mm<sup>2</sup> in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/iNOS+ cells/mm<sup>2</sup> and LCA+/iNOS+ cells/mm<sup>2</sup> (<i>p</i> < 0.0001 and <i>p</i> < 0.001, respectively). The total number of CD68+/MMP-9+ cells/mm<sup>2</sup> in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/MMP-9+ cells/mm<sup>2</sup> and CD5+/MMP-9+ cells/mm<sup>2</sup> (<i>p</i> < 0.0001). Macrophages may have a leading role in OA progression through the NF-kB production of inflammatory factors (iNOS and MMP-9) in the intima, except in advanced OA, where leukocytes could have a dominant role through NF-kB production in subintima. The blocking of macrophageal and leukocyte NF-kB expression is a possible therapeutic target as a disease modifying drug. |
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| last_indexed | 2024-03-10T10:21:22Z |
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| spelling | doaj.art-51fc21535bc7460aa3a12fc6803726052023-11-22T00:24:27ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-06-012212646110.3390/ijms22126461Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee OsteoarthritisMarko Ostojic0Ante Zevrnja1Katarina Vukojevic2Violeta Soljic3Department of Orthopaedics and Traumatology, University Hospital Mostar, 88 000 Mostar, Bosnia and HerzegovinaPublic Health Centre, First Responders Unit, 21 000 Split, CroatiaDepartment of Anatomy, Histology and Embryology, School of Medicine, University of Split, Soltanska 2, 21 000 Split, CroatiaDepartment of Histology and Embryology, School of Medicine, University of Mostar, Kralja Petra Kresimira IV, 88 000 Mostar, Bosnia and HerzegovinaSynovitis of the knee synovium is proven to be a precursor of knee osteoarthritis (OA), leading to a radiologically advanced stage of the disease. This study was conducted to elucidate the expression pattern of different inflammatory factors—NF-kB, iNOS, and MMP-9 in a subpopulation of synovial cells. Thirty synovial membrane intra-operative biopsies of patients (ten controls, ten with early OA, and ten with advanced OA, according to the Kellgren–Lawrence radiological score) were immunohistochemically stained for NF-kB, iNOS, and MMP9, and for different cell markers for macrophages, fibroblasts, leukocytes, lymphocytes, blood vessel endothelial cells, and blood vessel smooth muscle cells. The total number of CD68+/NF-kB+ cells/mm<sup>2</sup> in the intima of early OA patients (median = 2359) was significantly higher compared to the total number of vimentin+/Nf-kB+ cells/mm<sup>2</sup> (median = 1321) and LCA+/NF-kB+ cells/mm<sup>2</sup> (median = 64) (<i>p</i> < 0.001 and <i>p</i> < 0.0001, respectively). The total number of LCA+/NF-kB+ cells/mm<sup>2</sup> in the subintima of advanced OA patients (median = 2123) was significantly higher compared to the total number of vimentin+/NF-kB+ cells/mm<sup>2</sup> (median = 14) and CD68+/NF-kB+ cells/mm<sup>2</sup> (median = 29) (<i>p</i> < 0.0001). The total number of CD68+/iNOS+ cells/mm<sup>2</sup> in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/iNOS+ cells/mm<sup>2</sup> and LCA+/iNOS+ cells/mm<sup>2</sup> (<i>p</i> < 0.0001 and <i>p</i> < 0.001, respectively). The total number of CD68+/MMP-9+ cells/mm<sup>2</sup> in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/MMP-9+ cells/mm<sup>2</sup> and CD5+/MMP-9+ cells/mm<sup>2</sup> (<i>p</i> < 0.0001). Macrophages may have a leading role in OA progression through the NF-kB production of inflammatory factors (iNOS and MMP-9) in the intima, except in advanced OA, where leukocytes could have a dominant role through NF-kB production in subintima. The blocking of macrophageal and leukocyte NF-kB expression is a possible therapeutic target as a disease modifying drug.https://www.mdpi.com/1422-0067/22/12/6461osteoarthritissynovitisinducible nitric oxide synthasenuclear factor kappa Bmacrophage |
| spellingShingle | Marko Ostojic Ante Zevrnja Katarina Vukojevic Violeta Soljic Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis International Journal of Molecular Sciences osteoarthritis synovitis inducible nitric oxide synthase nuclear factor kappa B macrophage |
| title | Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis |
| title_full | Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis |
| title_fullStr | Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis |
| title_full_unstemmed | Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis |
| title_short | Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis |
| title_sort | immunofluorescence analysis of nf kb and inos expression in different cell populations during early and advanced knee osteoarthritis |
| topic | osteoarthritis synovitis inducible nitric oxide synthase nuclear factor kappa B macrophage |
| url | https://www.mdpi.com/1422-0067/22/12/6461 |
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