Deneysel Diz Osteoartrit Modeli Geliştirilen Sıçanlarda Serisinin Etkinliğinin TGF-beta/Smad Yolağı Üzerinden İncelenmesi

Öz

AMAÇ: Bu çalışmada; Monosodyum iyoKOAsetat (MIA) ile diz Osteoartrit (KOA) modeli oluşturulan sıçanlarda serisinin terapotik etkinliğinin incelenmesi ve bu etkinliğinin TGF-β/Smad yolağı üzerinden değerlendirilmesi amaçlanmıştır. MATERYAL VE METOD: Sıçanlarda KOA modeli diz eklemine MIA enjekte edilerek oluşturulmuş ve rastgele 3 gruba ayrılmıştır (1. grup (kontrol), 2. grup (KOA kontrol), 3. grup (KOA+serisin)). Sıçanlara, serisin 1, 7, 14 ve 21. günlerde (100 µL, 0,8 g/kg/mL) intratendinöz olarak uygulanmıştır. Sıçanlar 21 günün sonunda sakrifiye edilerek elde edilen serum örneklerinde TGF-β1, Smad2, CTGF, seviyeleri ELİSA yöntemi ile belirlenmiştir. Ayrıca diz eklem örneklerinde Hemotoksilen eozin boyası ile histopatolojik, TGF-β1 ve Smad2/3 antikorları ile immünohistokimyasal değerlendirmeleri gerçekleştirilmiştir. BULGULAR: Serum TGF-β1 ve CTGF düzeylerinde 2. grupda, 1. gruba göre anlamlı olarak anlamlı artış tespit edilmiş (P <0.05), tedavi verilen 3. grupta ise istatiksel olarak anlamlı azalma görülmüştür (P<0.05). Serum Smad2 düzeylerinde gruplar arasında anlamlı fark saptanmamıştır. Histopatolojik olarak 2. grupda subkondral kemik dokusu ve kıkırdak dejenerasyonu ve derin çatlaklar görülmüştür. 3. grup ise kondrosit hücrelerindeki dejenerasyonun azaldığı kıkırdak dokusunun kalınlığının arttığı ve kıkırdak matriksin normale yakın olduğu izlenmiştir. İmmünohistokimyasal olarak 2. grupda TGF-β1 ve Smad ekspresyonlarında artışlar görülmüştür. Bununla birlikte, 3. grup 2. gruba kıyasla bu ekspresyonlarda azalmalar sergilenmiştir. TARTIŞMA: Serisin sıçanlarda deneysel KOA modelinde TGF-β1/Smad yolağı üzerinden potansiyel etkinlik göstermektedir. Sonuç olarak, serisin ileri klinik çalışmalarla desteklenmesiyle KOA tedavisi için umut verici bir terapötik ajan olabilir.

Anahtar Kelimeler

• Serisin
• Diz osteoartrit
• TGF-β1/Smad yolağı

Investigation of The Efficacy of Sericin in Experimental Knee Osteoarthritis Model in Rats through the TGF-Beta/Smad Pathway

Öz

OBJECTIVE: This study investigates the therapeutic efficacy of sericin in rats with knee osteoarthritis (KOA) induced with monosodium iodoacetate (MIA), focusing on evaluating its effectiveness through the TGF-β/Smad pathway. MATERIALS AND METHODS: The KOA model was established by injecting MIA into the knee joint and the rats were randomly divided into three groups:group 1 (control), group 2 (KOA control), and group 3 (KOA+sericin). Sericin was administered intratendinously to rats on days 1,7,14, and 21 (50 µL,0.8 g/kg/mL). After 21 days, the rats were sacrificed, and serum samples were analyzed using the ELISA method to measure TGF-β1, Smad2, and CTGF levels. Additionally, knee joint samples underwent histopathological evaluations with hematoxylin-eosin staining and immunohistochemical assessment using TGF-β1 and Smad2/3 antibodies. RESULTS: Serum TGF-β1 and CTGF levels were significantly increased in group 2 vs. group 1 (P<0.05). A statistically significant decrease was observed in group 3 (P<0.05). Serum Smad2 levels were not significantly different between groups. Histopathologically, group 2 showed a narrowing of the joint space, degeneration of the cartilage, and deep fissures. On the other hand, group 3 showed marked joint space expansion, articular chondral regeneration, and immunostaining patterns similar to group 1. Immunohistochemically, group 2 exhibited a significant increase in TGF-β1 and Smad expression, whereas group 3 significantly decreased these expressions than group 2(P<0.01). DISCUSSION: Sericin demonstrates potential efficacy in the experimental KOA model in rats through the TGF-β1/Smad pathway. Consequently, sericin may emerge as a promising therapeutic agent for the treatment of KOA with further support from advanced clinical trials.

Anahtar Kelimeler

• Sericin
• Knee osteoarthritis
• TGF-β1/Smad pathway

Kaynakça

1. 1. Pulsatelli L, Addimanda O, Brusi V, Pavloska B, Meliconi R. New findings in osteoarthritis pathogenesis: therapeutic implications. Therapeutic advances in chronic disease. 2013;4(1):23-43.
2. 2. Cui A, Li H, Wang D, Zhong J, Chen Y, Lu H. Global, regional prevalence, incidence and risk factors of knee osteoarthritis in population-based studies. EClinicalMedicine. 2020;29:100587.
3. 3. Altay MA, Ertürk C, Bilge A, Yaptı M, Levent A, Aksoy N. Evaluation of prolidase activity and oxidative status in patients with knee osteoarthritis: relationships with radiographic severity and clinical parameters. Rheumatology international. 2015;35(10):1725-1731.
4. 4. Xia B, Chen D, Zhang J, Hu S, Jin H, Tong P. Osteoarthritis pathogenesis: a review of molecular mechanisms. Calcified tissue international. 2014;95(6):495-505.
5. 5. Lou Y, Song F, Kang Y, Xu Y. Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis. J Inflamm Res. 2023;16:5613-5628. doi:10.2147/jir.S419186
6. 6. Goldring MB, Otero M. Inflammation in osteoarthritis. Current opinion in rheumatology. 2011;23(5):471.
7. 7. Lee H, Choi H-S, Park Y, et al. Effects of deer bone extract on the expression of pro-inflammatory cytokine and cartilage-related genes in monosodium iodoacetate-induced osteoarthritic rats. Bioscience, Biotechnology, and Biochemistry. 2014;78(10):1703-1709.
8. 8. López-Reyes A, Medina-Luna D, Santamaría-Olmedo M, et al. Soluble inflammatory mediators of synoviocytes stimulated by monosodium urate crystals induce the production of oxidative stress, pain, and inflammation mediators in chondrocytes : Secretome of synoviocytes induces chondrocyte damage. Clin Rheumatol. Aug 2021;40(8):3265-3271. doi:10.1007/s10067-021-05676-w

9. 9. Liang Y, Shen L, Ni W, et al. CircGNB1 drives osteoarthritis pathogenesis by inducing oxidative stress in chondrocytes. Clin Transl Med. Aug 2023;13(8):e1358. doi:10.1002/ctm2.1358
10. 10. Boon MR, van der Horst G, van der Pluijm G, Tamsma JT, Smit JW, Rensen PC. Bone morphogenetic protein 7: a broad-spectrum growth factor with multiple target therapeutic potency. Cytokine & growth factor reviews. 2011;22(4):221-229.
11. 11. Lilja-Maula L, Syrjä P, Laurila H, et al. Comparative study of transforming growth factor-β signalling and regulatory molecules in human and canine idiopathic pulmonary fibrosis. Journal of comparative pathology. 2014;150(4):399-407.
12. 12. Miao MZ, Su QP, Cui Y, et al. Redox-active endosomes mediate α5β1 integrin signaling and promote chondrocyte matrix metalloproteinase production in osteoarthritis. Sci Signal. Oct 31 2023;16(809):eadf8299. doi:10.1126/scisignal.adf8299
13. 13. Pottie P, Presle N, Terlain B, Netter P, Mainard D, Berenbaum F. Obesity and osteoarthritis: more complex than predicted! Ann Rheum Dis. Nov 2006;65(11):1403-5. doi:10.1136/ard.2006.061994
14. 14. Sakao K, Takahashi KA, Arai Y, et al. Asporin and transforming growth factor-β gene expression in osteoblasts from subchondral bone and osteophytes in osteoarthritis. Journal of Orthopaedic Science. 2009;14(6):738-747.
15. 15. Wang N, Zhang S. Up-regulating MiR-146 inhibits osteoarthritis in rats through suppressing TGF-β/smad signaling pathway. Panminerva Med. Jan 24 2020;doi:10.23736/s0031-0808.19.03822-9
16. 16. Atala A, Kasper FK, Mikos AG. Engineering complex tissues. Science translational medicine. 2012;4(160):160rv12-160rv12.
17. 17. Aramwit P, Luplertlop N, Kanjanapruthipong T, Ampawong S. Effect of urea-extracted sericin on melanogenesis: potential applications in post-inflammatory hyperpigmentation. Biological research. 2018;51:1-13.
18. 18. Zhang X, Tsukada M, Morikawa H, Aojima K, Zhang G, Miura M. Production of silk sericin/silk fibroin blend nanofibers. Nanoscale research letters. 2011;6:1-8.
19. 19. Panilaitis B, Altman GH, Chen J, Jin H-J, Karageorgiou V, Kaplan DL. Macrophage responses to silk. Biomaterials. 2003;24(18):3079-3085.
20. 20. Gholipourmalekabadi M, Khosravimelal S, Nokhbedehghan Z, et al. Modulation of hypertrophic scar formation using amniotic membrane/electrospun silk fibroin bilayer membrane in a rabbit ear model. ACS Biomaterials Science & Engineering. 2019;5(3):1487-1496.
21. 21. Mumtaz S, Ali S, Qureshi MZ, Muhammad A, Manan A, Akbar Mughal T. Antioxidant and anti-aging role of silk sericin in D-galactose induced mice model. Saudi J Biol Sci. Dec 2023;30(12):103872. doi:10.1016/j.sjbs.2023.103872
22. 22. Dash R, Acharya C, Bindu P, Kundu S. Antioxidant potential of silk protein sericin against hydrogen peroxide-induced oxidative stress in skin fibroblasts. BMB reports. 2008;41(3):236-241.
23. 23. Qi C, Liu J, Jin Y, et al. Photo-crosslinkable, injectable sericin hydrogel as 3D biomimetic extracellular matrix for minimally invasive repairing cartilage. Biomaterials. 2018;163:89-104.
24. 24. Yamada EF, Bobinski F, Martins DF, Palandi J, Folmer V, da Silva MD. Photobiomodulation therapy in knee osteoarthritis reduces oxidative stress and inflammatory cytokines in rats. Journal of Biophotonics. 2020;13(1):e201900204.
25. 25. Kader S, Jabbari E. Material Properties and Cell Compatibility of Photo-Crosslinked Sericin Urethane Methacryloyl Hydrogel. Gels. 2022;8(9):543.
26. 26. Dokumacioglu E, İskender H, Yenice G, et al. Effects of astaxanthin on biochemical and histopathological parameters related to oxidative stress on testes of rats on high fructose regime. Andrologia. 2018;50(7):e13042.
27. 27. Kim D-W, Jo Y-Y, Garagiola U, et al. Increased level of vascular endothelial growth factors by 4-hexylresorcinol is mediated by transforming growth factor-β1 and accelerates capillary regeneration in the burns in diabetic animals. International journal of molecular sciences. 2020;21(10):3473.
28. 28. Zhang J, Su L, Liu Z, et al. A responsive hydrogel modulates innate immune cascade fibrosis to promote ocular surface reconstruction after chemical injury. Journal of Controlled Release. 2024;365:1124-1138.
29. 29. Scognamiglio F, Travan A, Donati I, Borgogna M, Marsich E. A hydrogel system based on a lactose-modified chitosan for viscosupplementation in osteoarthritis. Carbohydrate polymers. 2020;248:116787.
30. 30. Jianwei H, Cao W, Azeem I, Shao Z. Epigenetics of osteoarthritis: Histones and TGF-β1. Clinica Chimica Acta. 2020;510:593-598.
31. 31. Jann J, Gascon S, Roux S, Faucheux N. Influence of the TGF-β superfamily on osteoclasts/osteoblasts balance in physiological and pathological bone conditions. International journal of molecular sciences. 2020;21(20):7597.
32. 32. van der Kraan PM, Goumans M-J, Blaney Davidson E, Ten Dijke P. Age-dependent alteration of TGF-β signalling in osteoarthritis. Cell and tissue research. 2012;347(1):257-265.
33. 33. Cherifi C, Monteagudo S, Lories RJ. Promising targets for therapy of osteoarthritis: a review on the Wnt and TGF-β signalling pathways. Therapeutic advances in musculoskeletal disease. 2021;13:1759720X211006959.
34. 34. Van Beuningen H, Glansbeek H, Van Der Kraan P, Van Den Berg W. Osteoarthritis-like changes in the murine knee joint resulting from intra-articular transforming growth factor-β injections. Osteoarthritis and Cartilage. 2000;8(1):25-33.
35. 35. van Beuningen HM, Glansbeek HL, van der Kraan PM, van den Berg WB. Differential effects of local application of BMP-2 or TGF-β1 on both articular cartilage composition and osteophyte formation. Osteoarthritis and Cartilage. 1998;6(5):306-317.
36. 36. Millward‐Sadler S, Wright M, Davies L, Nuki G, Salter D. Mechanotransduction via integrins and interleukin‐4 results in altered aggrecan and matrix metalloproteinase 3 gene expression in normal, but not osteoarthritic, human articular chondrocytes. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 2000;43(9):2091-2099.
37. 37. Madej W, Van Caam A, Davidson EB, Hannink G, Buma P, van der Kraan P. Ageing is associated with reduction of mechanically-induced activation of Smad2/3P signaling in articular cartilage. Osteoarthritis and Cartilage. 2016;24(1):146-157.
38. 38. Song CJ, Fu XM, Li J, Chen ZH. [Effects of sericine on TGF-beta1/Smad3 signal pathway of diabetic mephropathy rats kidney]. Zhongguo Ying Yong Sheng Li Xue Za Zhi. Feb 2011;27(1):102-5.
39. 39. Sayin D, Gundogdu G, Kilic-Erkek O, Gundogdu K, Coban HS, Abban-Mete G. Silk protein sericin: a promising therapy for Achilles tendinopathy-evidence from an experimental rat model. Clin Rheumatol. Dec 2023;42(12):3361-3373. doi:10.1007/s10067-023-06767-6