Activation of endogenous latent transforming growth factor β1 with the ascorbic acid-ferric chloride system for osteoarthritis treatment and osteochondral repair

Abstract

The management of osteoarthritis (OA) and osteochondral defects faces challenges due to heightened catabolic activity from pro-inflammatory mediators and a lack of reparative cells. Transforming growth factor β1 (TGFβ1) plays a crucial role in cartilage maintenance and cellular recruitment, making it a promising therapeutic target. However, the high cost and unpredictable effects of exogenous TGFβ1 limit its clinical application. Notably, TGFβ1 is primarily found in a latent form within joint tissues, especially during early injury stages. This study proposes that activating endogenous TGFβ1 may serve as an effective treatment strategy. We demonstrate that an ascorbic acid (AA) and ferric chloride (AA/Fe) Fenton reaction system can activate latent TGFβ1 in knee joint tissues. Treatment with AA/Fe-activated synovial fluid protects chondrocytes from interleukin1β-induced damage and enhances chemotactic responses in joint tissues. Intra-articular AA/Fe injections in rats significantly phosphorylated SMAD2/3 and reduced cartilage degradation. Additionally, we developed poly(lactic-co-glycolic acid) microspheres for sustained AA/Fe release within a thermosensitive or photocrosslinkable hydrogel, showing high biocompatibility. These formulations effectively prevented cartilage degeneration and promoted osteochondral repair. Our findings confirm that AA/Fe reliably activates endogenous TGFβ1, providing a novel cell-free and cost-efficient treatment approach for OA and osteochondral defects.

Keywords

ascorbic acid / cartilage / Fenton / osteoarthritis / osteochondral defects

References

[1] M. P. Murphy, L. S. Koepke, M. T. Lopez, X. Tong, T. H. Ambrosi, G. S. Gulati, O. Marecic, Y. Wang, R. C. Ransom, M. Y. Hoover, H. Steininger, L. Zhao, M. P. Walkiewicz, N. Quarto, B. Levi, D. C. Wan, I. L. Weissman, S. B. Goodman, F. Yang, M. T. Longaker, C. K. F. Chan, Nat. Med. 2022, 26, 1583.

[2] L. Fu, L. Li, Q. Bian, B. Xue, J. Jin, J. Li, Y. Cao, Q. Jiang, H. Li, Nature 2023, 618, 740.

[3] L. S. Lohmander, P. M. Englund, L. L. Dahl, E. M. Roos, Am. J. Sports Med. 2007, 35, 1756.

[4] K. P. Spindler, R. W. Wright, N. Engl. J. Med. 2008, 359, 2135.

[5] X. Liu, X. Jin, P. X. Ma, Nat. Mater. 2011, 10, 398.

[6] Y. Liu, G. Dzidotor, T. T. Le, T. Vinikoor, K. Morgan, E. J. Curry, R. Das, A. McClinton, E. Eisenberg, L. N. Apuzzo, K. T. M. Tran, P. Prasad, T. J. Flanagan, S. W. Lee, H. M. Kan, M. T. Chorsi, K. W. H. Lo, C. T. Laurencin, T. D. Nguyen, Sci. Transl. Med. 2022, 14, eabi7282.

[7] R. Shkhyan, B. Van Handel, J. Bogdanov, S. Lee, Y. Yu, M. Scheinberg, N. W. Banks, S. Limfat, A. Chernostrik, C. E. Franciozi, M. P. Alam, V. John, L. Wu, G. B. Ferguson, A. Nsair, F. A. Petrigliano, C. T. Vangsness, K. Vadivel, P. Bajaj, L. Wang, N. Q. Liu, D. Evseenko, Ann. Rheum. Dis. 2018, 77, 760.

[8] K. Futrega, P. G. Robey, T. J. Klein, R. W. Crawford, M. R. Doran, Commun. Biol. 2021, 4, 29.

[9] E. N. Blaney Davidson, E. L. Vitters, P. M. van der Kraan, W. B. van den Berg, Ann. Rheum. Dis. 2006, 65, 1414.

[10] G. Wang, S. Chen, Z. Xie, S. Shen, W. Xu, W. Chen, X. Li, Y. Wu, L. Li, B. Liu, X. Ding, A. Qin, S. Fan, Ann. Rheum. Dis. 2020, 79, 1111.

[11] D. Wu, J. Guo, B. Qi, H. Xiao, Endocr. J. 2021, 68, 1347.

[12] J. Massagué, D. Sheppard, Cell 2023, 186, 4007.

[13] E. Anitua, M. Sánchez, M. de la Fuente, J. Azofra, M. Zalduendo, J. J. Aguirre, I. Andía, Int. J. Rheumatol. 2009, 2009, 747432.

[14] J. F. Schlaak, I. Pfers, K. H. Meyer Zum Büschenfelde, E. Märker-Hermann, Clin. Exp. Rheumatol. 1996, 14, 155.

[15] S. Zielinski, K. Bartels, K. Cebulski, C. Kühne, J. Kekow, Adv. Exp. Med. Biol. 2000, 477, 477.

[16] M. B. Albro, A. D. Cigan, R. J. Nims, K. J. Yeroushalmi, S. R. Oungoulian, C. T. Hung, G. A. Ateshian, Osteoarthr. Cartil. 2012, 20, 1374.

[17] S. Wang, Y. Niu, P. Jia, Z. Liao, W. Guo, R. C. Chaves, K. H. Tran-Ba, L. He, H. Bai, S. Sia, L. J. Kaufman, X. Wang, Y. Zhou, Y. Dong, J. J. Mao, Bioact. Mater. 2021, 15, 316.

[18] Y. Niu, Z. Yang, Y. Yang, X. Wang, P. Zhang, L. Lv, S. Wang, Y. Liu, Y. Liu, Y. Zhou, NPJ Regen. Med. 2023, 8, 56.

[19] O. Tatti, P. Vehviläinen, K. Lehti, J. Keski-Oja, Exp. Cell Res. 2008, 314, 2501.

[20] Y. Yao, C. Hu, Q. Song, Y. Li, X. Da, Y. Yu, H. Li, I. M. Clark, Q. Chen, Q. K. Wang, Cardiovasc. Res. 2020, 116, 956.

[21] M. H. Barcellos-Hoff, Cancer Res. 1993, 53, 3880.

[22] M. F. Jobling, J. D. Mott, M. T. Finnegan, V. Jurukovski, A. C. Erickson, P. J. Walian, S. E. Taylor, S. Ledbetter, C. M. Lawrence, D. B. Rifkin, M. H. Barcellos-Hoff, Radiat. Res. 2006, 166, 839.

[23] W. Diaodiao, T. Miaotian, R. Pengcheng, T. Zhuang, Z. Gang, L. Yubo, S. Yuyang, Y. Peng, S. Wenqiang, Y. Qi, J. Orthop. Translat. 2025, 53, 99.

[24] Q. Song, D. Wang, H. Li, Z. Wang, S. Sun, Z. Wang, Y. Liu, S. Lin, G. Li, S. Zhang, P. Zhang, Bioact. Mater. 2023, 32, 304.

[25] F. Danhier, E. Ansorena, J. M. Silva, R. Coco, A. Le Breton, V. Préat, J. Controlled Release 2012, 161, 505.

[26] J. Paik, S. T. Duggan, S. J. Keam, Drugs 2019, 79, 455.

[27] W. Pan, B. Wu, C. Nie, T. Luo, Z. Song, J. Lv, Y. Tan, C. Liu, M. Zhong, T. Liao, Z. Wang, G. Yi, L. Zhang, X. Liu, B. Li, J. Chen, L. Zheng, ACS Nano 2023, 17, 11253.

[28] H. Ma, C. He, Y. Cheng, D. Li, Y. Gong, J. Liu, H. Tian, X. Chen, Biomaterials 2014, 35, 8723.

[29] T. Ma, G. Xu, T. Gao, G. Zhao, G. Huang, J. Shi, J. Chen, J. Song, J. Xia, X. Ma, ACS Appl. Mater. Interfaces 2024, 16, 21383.

[30] G. Liu, Q. Guo, C. Liu, J. Bai, H. Wang, J. Li, D. Liu, Q. Yu, J. Shi, C. Liu, C. Zhu, B. Li, H. Zhang, Acta Biomater. 2023, 169, 317.

[31] C. Liu, Q. Yu, Z. Yuan, Q. Guo, X. Liao, F. Han, T. Feng, G. Liu, R. Zhao, Z. Zhu, H. Mao, C. Zhu, B. Li, Bioact. Mater. 2022, 25, 445.

[32] H. Qian, T. Lei, L. Hua, Y. Zhang, D. Wang, J. Nan, W. Liu, Y. Sun, Y. Hu, P. Lei, Bioact. Mater. 2023, 24, 450.

[33] A. Samuni, J. Aronovitch, D. Godinger, M. Chevion, G. Czapski, Eur. J. Biochem. 1983, 137, 119.

[34] K. Hill-Mündel, D. Nohr, Biochem. Biophys. Res. Commun. 2021, 546, 90.

[35] Y. Yang, C. Zhang, Y. Song, Y. Li, P. Li, M. Huang, F. Meng, M. Zhang, Nucleic Acids Res. 2022, 50, 9632.

[36] H. P. Lee, L. Gu, D. J. Mooney, M. E. Levenston, O. Chaudhuri, Nat. Mater. 2017, 16, 1243.

[37] Y. Qin, X. B. Hu, W. T. Fan, J. Yan, S. B. Cheng, Y. L. Liu, W. H. Huang, Adv. Sci. 2021, 8, 2003738.

[38] A. Plaas, J. Velasco, D. J. Gorski, J. Li, A. Cole, K. Christopherson, J. D. Sandy, Osteoarthr. Cartil. 2011, 19, 1081.

[39] K. W. Finnson, W. L. Parker, Y. Chi, C. D. Hoemann, M. B. Goldring, J. Antoniou, A. Philip, Osteoarthr. Cartil. 2010, 18, 1518.

[40] E. G. J. Ripmeester, J. S. J. J. Steijns, K. A. P. Wijnands, R. H. M. J. Stassen, V. Pitelka, L. C. W. Peeters, A. Cremers, N. M. S. A. Astryde, A. Chabronova, D. A. M. Surtel, P. J. Emans, G. G. H. van den Akker, B. van Rietbergen, L. W. van Rhijn, M. M. J. Caron, T. J. M. Welting, Cartilage 2024, 19, 19476035241233659.

[41] C. B. Hamilton, M. A. Pest, V. Pitelka, A. Ratneswaran, F. Beier, B. M. Chesworth, Osteoarthr. Cartil. 2015, 23, 1178.

[42] E. Anitua, M. Sánchez, M. de la Fuente, J. Azofra, M. Zalduendo, J. J. Aguirre, I. Andía, Int. J. Rheumatol. 2009, 2009, 747432.

[43] J. F. Schlaak, I. Pfers, K. H. Meyer Zum Büschenfelde, E. Märker-Hermann, Clin. Exp. Rheumatol. 1996, 14, 155.

[44] L. J. Moilanen, M. Hämäläinen, E. Nummenmaa, P. Ilmarinen, K. Vuolteenaho, R. M. Nieminen, L. Lehtimäki, E. Moilanen, Osteoarthr. Cartil. 2015, 23, 2017.

[45] K. Higa, N. Kitamura, K. Goto, T. Kurokawa, J. P. Gong, F. Kanaya, K. Yasuda, BMC Musculoskelet. Disord. 2017, 18, 210.

[46] C. Frippiat, Q. M. Chen, S. Zdanov, J. P. Magalhaes, J. Remacle, O. Toussaint, J. Biol. Chem. 2001, 276, 2531.

[47] R. A. Clark, S. J. Klebanoff, A. B. Einstein, A. Fefer, Blood 1975, 45, 161.

[48] T. Osaki, T. Homma, Y. Soma, S. Miyata, Y. Matsuda, J. Fujii, Redox Biol. 2025, 85, 103749.

[49] I. Jarocka-Karpowicz, A. Stasiewicz, E. Olchowik-Grabarek, S. Sękowski, A. Kacprowska, E. Skrzydlewska, Free Radical Biol. Med. 2025, 228, 251.

[50] D. F. Remst, E. N. Blaney Davidson, E. L. Vitters, R. A. Bank, W. B. van den Berg, P. M. van der Kraan, Cell Tissue Res. 2014, 355, 163.

[51] S. L. Wang, Y. L. Yu, C. L. Tang, F. Z. Lv, Exp. Ther. Med. 2013, 6, 1522.

[52] F. Lebrin, M. J. Goumans, L. Jonker, R. L. Carvalho, G. Valdimarsdottir, M. Thorikay, C. Mummery, H. M. Arthur, P. ten Dijke, EMBO J. 2004, 23, 4018.

[53] E. N. Blaney Davidson, D. F. Remst, E. L. Vitters, H. M. van Beuningen, A. B. Blom, M. J. Goumans, W. B. van den Berg, P. M. van der Kraan, J. Immunol. 2009, 182, 7937.

[54] Q. Hong, H. Kim, G. Y. Cai, X. M. Chen, J. C. He, K. Lee, Int. J. Biol. Sci. 2025, 21, 1649.

[55] H. Liu, Y. Qi, L. Wang, Y. Zhang, H. Chen, Y. Zhao, H. Min, G. Nie, H. Liang, ACS Nano 2025, 19, 25645.

[56] A. Butreddy, R. P. Gaddam, N. Kommineni, N. Dudhipala, C. Voshavar, Int. J. Mol. Sci. 2021, 22, 8884.

[57] W. Zhou, Y. Zhang, S. Meng, C. Xing, M. Ma, Z. Liu, C. Yang, T. Kong, Small Methods 2021, 5, 2000920.

[58] S. Yuan, H. Li, Z. Xu, Z. Yue, S. Chen, Q. Fu, Y. Chen, J. Controlled Release 2025, 384, 113920.

[59] J. Liu, J. Liu, S. Liu, P. Xiao, C. Du, J. Zhan, Z. Chen, L. Chen, K. Li, W. Huang, Y. Lei, Biomaterials 2025, 318, 123195.

[60] J. Liang, X. Huang, K. Qin, H. Wei, J. Yang, B. Liu, Z. Fan, Adv. Mater. 2025, 37, 2415417.

[61] Y. Li, H. Zhang, Y. Jiang, J. Yang, D. Cai, X. Bai, Interdiscip. Med. 2024, 2, e20230055.

[62] S. Han, H. Yang, X. Ni, Y. Deng, Z. Li, X. Xing, M. Du, Int. J. Biol. Macromol. 2023, 253, 126721.

[63] H. Qian, T. Lei, L. Hua, Y. Zhang, D. Wang, J. Nan, W. Liu, Y. Sun, Y. Hu, P. Lei, Bioact. Mater. 2023, 24, 450.

[64] L. Peterson, T. Minas, M. Brittberg, A. Nilsson, E. Sjögren-Jansson, A. Lindahl, Clin. Orthop. Relat. Res. 2000, 374, 212.

[65] P. Mainil-Varlet, T. Aigner, M. Brittberg, P. Bullough, A. Hollander, E. Hunziker, R. Kandel, S. Nehrer, K. Pritzker, S. Roberts, E. Stauffer, International Cartilage Repair Society, J. Bone. Joint. Surg. Am. 2023, 85, 45.

[66] S. W. O'Driscoll, F. W. Keeley, R. B. Salter, J. Bone Jt. Surg. 1986, 68, 1017.

RIGHTS & PERMISSIONS

2025 The Author(s). Interdisciplinary Medicine published by Wiley-VCH GmbH on behalf of Nanfang Hospital, Southern Medical University.