Inhibitory effect of low‑intensity pulsed ultrasound on the fibrosis of the infrapatellar fat pad through the regulation of HIF‑1α in a carrageenan‑induced knee osteoarthritis rat model

Kitagawa,Takashi; Kawahata,Hirohisa; Aoki,Motokuni; Kudo,Shintarou

doi:10.3892/br.2022.1562

Inhibitory effect of low‑intensity pulsed ultrasound on the fibrosis of the infrapatellar fat pad through the regulation of HIF‑1α in a carrageenan‑induced knee osteoarthritis rat model

Authors:
- Takashi Kitagawa
- Hirohisa Kawahata
- Motokuni Aoki
- Shintarou Kudo
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Affiliations: Graduate School of Health Sciences, Morinomiya University of Medical Sciences, Suminoe‑ku, Osaka 559‑8611, Japan
Published online on: August 9, 2022 https://doi.org/10.3892/br.2022.1562
Article Number: 79

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Abstract

Fibrotic changes in the infrapatellar fat pad (IFP) are involved in the pathogenesis of knee osteoarthritis (KOA). HIF‑1α is a transcription factor that is activated during hypoxia and is suggested to play a role in fibrosis in various organs. However, its participation in the fibrotic changes in IFP remains unclear. Therefore, we investigated the role of HIF‑1α in IFP fibrosis using a carrageenan‑induced KOA rat model and evaluated the potential of low‑intensity pulsed ultrasound (LIPUS) as a novel treatment for KOA. A rat model was prepared by intra‑articular injection of 0.5% carrageenan (50 µl) using 8‑week‑old male Wistar rats. Fibrosis of the IFP was evaluated histologically by hematoxylin and eosin and Sirius Red staining at 1 and 2 weeks after intra‑articular injection. The mRNA expression levels of HIF‑1α and fibrosis‑related molecules, CTGF and VEGF, were analyzed using reverse transcription‑quantitative PCR, and the DNA binding activity of HIF‑1α was assessed using a binding assay. In addition, the effect of irradiation with LIPUS on the fibrosis of IFP was verified. Histological studies demonstrated a significant increase in the fibrosis of IFP 1 and 2 weeks after intra‑articular injection of carrageenan, accompanied by overexpression of CTGF and VEGF, which was followed by upregulation of transcriptional activation of HIF‑1α. Moreover, intervention with LIPUS for 2 weeks after injection of carrageenan attenuated fibrosis of IFP, accompanied by a significant reduction in the transcriptional activation of HIF‑1α and decreased the gene expression levels of HIF‑1α, CTGF, and VEGF. The present study demonstrated that activation of HIF‑1α promoted fibrosis of IFP in carrageenan‑induced arthritis in rats and that intervention with LIPUS decreased the activity of HIF‑1α and inhibited fibrosis. These results suggest that LIPUS may serve as a novel approach for the treatment of KOA, through its modulation of HIF‑1α.

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1	Sellam J and Berenbaum F: The role of synovitis in pathophysiology and clinical symptoms of osteoarthritis. Nat Rev Rheumatol. 6:625–635. 2010.PubMed/NCBI View Article : Google Scholar
2	Dieppe PA and Lohmander LS: Pathogenesis and management of pain in osteoarthritis. Lancet. 365:965–973. 2005.PubMed/NCBI View Article : Google Scholar
3	Hunter DJ and Bierma-Zeinstra S: Osteoarthritis. Lancet. 393:1745–1759. 2019.PubMed/NCBI View Article : Google Scholar
4	Hill CL, Gale DG, Chaisson CE, Skinner K, Kazis L, Gale ME and Felson DT: Knee effusions, popliteal cysts, and synovial thickening: Association with knee pain in osteoarthritis. J Rheumatol. 28:1330–1337. 2001.PubMed/NCBI
5	Belluzzi E, Stocco E, Pozzuoli A, Granzotto M, Porzionato A, Vettor R, De Caro R, Ruggieri P, Ramonda R, Rossato M, et al: Contribution of infrapatellar fat pad and synovial membrane to knee osteoarthritis pain. Biomed Res Int. 2019(6390182)2019.PubMed/NCBI View Article : Google Scholar
6	Leese J and Davies DC: An investigation of the anatomy of the infrapatellar fat pad and its possible involvement in anterior pain syndrome: A cadaveric study. J Anat. 237:20–28. 2020.PubMed/NCBI View Article : Google Scholar
7	Macchi V, Picardi EEE, Fontanella CG, Porzionato A, Stecco C, Tortorella C, Favero M, Natali A and De Caro R: The characteristics of the lobular arrangement indicate the dynamic role played by the infrapatellar fat pad in knee kinematics. J Anat. 235:80–87. 2019.PubMed/NCBI View Article : Google Scholar
8	Macchi V, Stocco E, Stecco C, Belluzzi E, Favero M, Porzionato A and De Caro R: The infrapatellar fat pad and the synovial membrane: An anatomo-functional unit. J Anat. 233:146–154. 2018.PubMed/NCBI View Article : Google Scholar
9	Nakanishi S, Morimoto R, Kitano M, Kawanishi K, Tanaka A and Kudo S: Difference in movement between superficial and deep parts of the infrapatellar fat pad during knee extension. J Funct Morphol Kinesiol. 6(68)2021.PubMed/NCBI View Article : Google Scholar
10	Distel E, Cadoudal T, Durant S, Poignard A, Chevalier X and Benelli C: The infrapatellar fat pad in knee osteoarthritis: An important source of interleukin-6 and its soluble receptor. Arthritis Rheum. 60:3374–3377. 2009.PubMed/NCBI View Article : Google Scholar
11	Favero M, El-Hadi H, Belluzzi E, Granzotto M, Porzionato A, Sarasin G, Rambaldo A, Iacobellis C, Cigolotti A, Fontanella CG, et al: Infrapatellar fat pad features in osteoarthritis: A histopathological and molecular study. Rheumatology (Oxford). 56:1784–1793. 2017.PubMed/NCBI View Article : Google Scholar
12	Bastiaansen-Jenniskens YM, Wei W, Feijt C, Waarsing JH, Verhaar JA, Zuurmond AM, Hanemaaijer R, Stoop R and van Osch GJ: Stimulation of fibrotic processes by the infrapatellar fat pad in cultured synoviocytes from patients with osteoarthritis: A possible role for prostaglandin f2α. Arthritis Rheum. 65:2070–2080. 2013.PubMed/NCBI View Article : Google Scholar
13	Maculé F, Sastre S, Lasurt S, Sala P, Segur JM and Mallofré C: Hoffa's fat pad resection in total knee arthroplasty. Acta Orthop Belg. 71:714–717. 2005.PubMed/NCBI
14	Fontanella CG, Macchi V, Carniel EL, Frigo A, Porzionato A, Picardi EEE, Favero M, Ruggieri P, de Caro R and Natali AN: Biomechanical behavior of Hoffa's fat pad in healthy and osteoarthritic conditions: Histological and mechanical investigations. Australas Phys Eng Sci Med. 41:657–667. 2018.PubMed/NCBI View Article : Google Scholar
15	Henderson NC, Rieder F and Wynn TA: Fibrosis: From mechanisms to medicines. Nature. 587:555–566. 2020.PubMed/NCBI View Article : Google Scholar
16	Wynn TA and Ramalingam TR: Mechanisms of fibrosis: Therapeutic translation for fibrotic disease. Nat Med. 18:1028–1040. 2012.PubMed/NCBI View Article : Google Scholar
17	Wynn TA: Cellular and molecular mechanisms of fibrosis. J Pathol. 214:199–210. 2008.PubMed/NCBI View Article : Google Scholar
18	Sun K, Tordjman J, Clément K and Scherer PE: Fibrosis and adipose tissue dysfunction. Cell Metab. 18:470–477. 2013.PubMed/NCBI View Article : Google Scholar
19	Halberg N, Khan T, Trujillo ME, Wernstedt-Asterholm I, Attie AD, Sherwani S, Wang ZV, Landskroner-Eiger S, Dineen S, Magalang UJ, et al: Hypoxia-inducible factor 1alpha induces fibrosis and insulin resistance in white adipose tissue. Mol Cell Biol. 29:4467–4483. 2009.PubMed/NCBI View Article : Google Scholar
20	Qing L, Lei P, Liu H, Xie J, Wang L, Wen T and Hu Y: Expression of hypoxia-inducible factor-1α in synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritis. Exp Ther Med. 13:63–68. 2017.PubMed/NCBI View Article : Google Scholar
21	Sotobayashi D, Kawahata H, Anada N, Ogihara T, Morishita R and Aoki M: Therapeutic effect of intra-articular injection of ribbon-type decoy oligonucleotides for hypoxia inducible factor-1 on joint contracture in an immobilized knee animal model. J Gene Med. 18:180–192. 2016.PubMed/NCBI View Article : Google Scholar
22	Ohtomo S, Nangaku M, Izuhara Y, Takizawa S, de Strihou CV and Miyata T: Cobalt ameliorates renal injury in an obese, hypertensive type 2 diabetes rat model. Nephrol Dial Transplant. 23:1166–1172. 2008.PubMed/NCBI View Article : Google Scholar
23	Sotobayashi D and Kawahata H: Beneficial effect of low-intensity pulsed ultrasound on progression of joint contracture. J Judo Ther. 27:125–132. 2019.(In Japanese).
24	Itaya N, Yabe Y, Hagiwara Y, Kanazawa K, Koide M, Sekiguchi T, Yoshida S, Sogi Y, Yano T, Tsuchiya M, et al: Effects of low-intensity pulsed ultrasound for preventing joint stiffness in immobilized knee model in rats. Ultrasound Med Biol. 44:1244–1256. 2018.PubMed/NCBI View Article : Google Scholar
25	Hansra P, Moran EL, Fornasier VL and Bogoch ER: Carrageenan-induced arthritis in the rat. Inflammation. 24:141–155. 2000.PubMed/NCBI View Article : Google Scholar
26	Clockaerts S, Bastiaansen-Jenniskens YM, Runhaar J, Van Osch GJ, Van Offel JF, Verhaar JA, De Clerck LS and Somville J: The infrapatellar fat pad should be considered as an active osteoarthritic joint tissue: A narrative review. Osteoarthritis Cartilage. 18:876–882. 2010.PubMed/NCBI View Article : Google Scholar
27	Wang W, Liu Y, Yang C, Qi X, Li S, Liu C and Li X: Mesoporous bioactive glass combined with graphene oxide scaffolds for bone repair. Int J Biol Sci. 15:2156–2169. 2019.PubMed/NCBI View Article : Google Scholar
28	Zarella MD, Breen DE, Plagov A and Garcia FU: An optimized color transformation for the analysis of digital images of hematoxylin & eosin stained slides. J Pathol Inform. 6(33)2015.PubMed/NCBI View Article : Google Scholar
29	Aigner T, Cook JL, Gerwin N, Glasson SS, Laverty S, Little CB, McIlwraith W and Kraus VB: Histopathology atlas of animal model systems-overview of guiding principles. Osteoarthritis Cartilage. 18 (Suppl 3):S2–S6. 2010.PubMed/NCBI View Article : Google Scholar
30	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
31	Seol D, Choe H, Zheng H, Jang K, Ramakrishnan PS, Lim TH and Martin JA: Selection of reference genes for normalization of quantitative real-time PCR in organ culture of the rat and rabbit intervertebral disc. BMC Res Notes. 4(162)2011.PubMed/NCBI View Article : Google Scholar
32	Baumann B, Hayashida T, Liang X and Schnaper HW: Hypoxia-inducible factor-1α promotes glomerulosclerosis and regulates COL1A2 expression through interactions with Smad3. Kidney Int. 90:797–808. 2016.PubMed/NCBI View Article : Google Scholar
33	Forsythe JA, Jiang BH, Iyer NV, Agani F, Leung SW, Koos RD and Semenza GL: Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol. 16:4604–4613. 1996.PubMed/NCBI View Article : Google Scholar
34	Higgins DF, Biju MP, Akai Y, Wutz A, Johnson RS and Haase VH: Hypoxic induction of Ctgf is directly mediated by Hif-1. Am J Physiol Renal Physiol. 287:F1223–F1232. 2004.PubMed/NCBI View Article : Google Scholar
35	Yoon KH, Tak DH, Ko TS, Park SE, Nam J and Lee SH: Association of fibrosis in the infrapatellar fat pad and degenerative cartilage change of patellofemoral joint after anterior cruciate ligament reconstruction. Knee. 24:310–318. 2017.PubMed/NCBI View Article : Google Scholar
36	Han W, Aitken D, Zhu Z, Halliday A, Wang X, Antony B, Cicuttini F, Jones G and Ding C: Hypointense signals in the infrapatellar fat pad assessed by magnetic resonance imaging are associated with knee symptoms and structure in older adults: A cohort study. Arthritis Res Ther. 18(234)2016.PubMed/NCBI View Article : Google Scholar
37	Ding N, Wei B, Fu X, Wang C and Wu Y: Natural products that target the NLRP3 inflammasome to treat fibrosis. Front Pharmacol. 11(591393)2020.PubMed/NCBI View Article : Google Scholar
38	Takahashi I, Matsuzaki T, Kuroki H and Hoso M: Induction of osteoarthritis by injecting monosodium iodoacetate into the patellofemoral joint of an experimental rat model. PLoS One. 13(e0196625)2018.PubMed/NCBI View Article : Google Scholar
39	Inomata K, Tsuji K, Onuma H, Hoshino T, Udo M, Akiyama M, Nakagawa Y, Katagiri H, Miyatake K, Sekiya I, et al: Time course analyses of structural changes in the infrapatellar fat pad and synovial membrane during inflammation-induced persistent pain development in rat knee joint. BMC Musculoskelet Disord. 20(8)2019.PubMed/NCBI View Article : Google Scholar
40	Ekundi-Valentim E, Santos KT, Camargo EA, Denadai-Souza A, Teixeira SA, Zanoni CI, Grant AD, Wallace J, Muscará MN and Costa SK: Differing effects of exogenous and endogenous hydrogen sulphide in carrageenan-induced knee joint synovitis in the rat. Br J Pharmacol. 159:1463–1474. 2010.PubMed/NCBI View Article : Google Scholar
41	Ashraf S, Mapp PI, Shahtaheri SM and Walsh DA: Effects of carrageenan induced synovitis on joint damage and pain in a rat model of knee osteoarthritis. Osteoarthritis Cartilage. 26:1369–1378. 2018.PubMed/NCBI View Article : Google Scholar
42	Egners A, Erdem M and Cramer T: The response of macrophages and neutrophils to hypoxia in the context of cancer and other inflammatory diseases. Mediators Inflamm. 2016(2053646)2016.PubMed/NCBI View Article : Google Scholar
43	Semba H, Takeda N, Isagawa T, Sugiura Y, Honda K, Wake M, Miyazawa H, Yamaguchi Y, Miura M, Jenkins DM, et al: HIF-1α-PDK1 axis-induced active glycolysis plays an essential role in macrophage migratory capacity. Nat Commun. 7(11635)2016.PubMed/NCBI View Article : Google Scholar
44	Yabe Y, Hagiwara Y, Suda H, Ando A, Onoda Y, Tsuchiya M, Hatori K and Itoi E: Joint immobilization induced hypoxic and inflammatory conditions in rat knee joints. Connect Tissue Res. 54:210–217. 2013.PubMed/NCBI View Article : Google Scholar
45	Zhao K, Weng L, Xu T, Yang C, Zhang J, Ni G, Guo X, Tu J, Zhang D, Sun W and Kong X: Low-intensity pulsed ultrasound prevents prolonged hypoxia-induced cardiac fibrosis through HIF-1α/DNMT3a pathway via a TRAAK-dependent manner. Clin Exp Pharmacol Physiol. 48:1500–1514. 2021.PubMed/NCBI View Article : Google Scholar
46	Costa V, Carina V, Conigliaro A, Raimondi L, De Luca A, Bellavia D, Salamanna F, Setti S, Alessandro R, Fini M and Giavaresi G: miR-31-5p is a LIPUS-mechanosensitive MicroRNA that targets HIF-1α signaling and cytoskeletal proteins. Int J Mol Sci. 20(1569)2019.PubMed/NCBI View Article : Google Scholar
47	Yang PF, Li D, Zhang SM, Wu Q, Tang J, Huang LK, Liu W, Xu XD and Chen SR: Efficacy of ultrasound in the treatment of osteoarthritis of the knee. Orthop Surg. 3:181–187. 2011.PubMed/NCBI View Article : Google Scholar