Benefits of bone marrow-derived mesenchymal stem cells primed with estradiol in alleviating collagen-induced arthritis

Document Type : Original Article


Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran


Objective(s): To investigate the effects of the oestradiol (ES) pulsed bone marrow-derived mesenchymal stem cells (BM-MSC) to treat adjuvant-induced arthritis in Wistar rats.  
Materials and Methods: BM-MSCs were pulsed with ES (0, 10,100, and 1000 nM) for 24 hr. RA was induced by collagen and Freund’s Complete Adjuvant into the base of the tail of Wistar rats.
Results: The least effective concentration of ES that can promote potent anti-inflammatory properties in the MSC population is 100 nM. At this concentration, ES increases the inhibition of the polyclonal T lymphocyte proliferation, production of IDO, IL-10, Nitric oxide, and TGF-β, and expression of CXCR4 and CCR2 mRNA in the MSC population. Accordingly, the RA rats were treated with 2×106 MSCs or ES-pulsed MSCs (100 nM) on day 10 when all animals had developed signs of RA. ES-pulsed BM-MSCs reduced the severity of RA more profoundly than treatment with BM-MScs alone. The ability of ES-pulsed BM-MSCs to reduce symptoms and RA markers like CRP, RF, and nitric oxide was comparable to that of prednisolone. Prednisolone was more successful in reducing inflammatory cytokines than treatment with ES-pulsed BM-MSCs. ES-pulsed BM-MSCs were more successful in increasing anti-inflammatory cytokines than treatment with Prednisolone. The ability of ES-pulsed BM-MSCs to decrease the level of nitric oxide was comparable to that of prednisolone.
Conclusion: ES-pulsed BM-MSCs may be a helpful strategy in RA control.


1. Ahmed EA, Ahmed OM, Fahim HI, Mahdi EA, Ali TM, Elesawy BH, et al. Combinatory effects of bone marrow-derived mesenchymal stem cells and indomethacin on adjuvant-induced arthritis in wistar rats: roles of il-1β, il-4, nrf-2, and oxidative stress. Evid Based Complement Alternat Med 2021; 2021:8899143.
2. Zheng M, Jia H, Wang H, Liu L, He Z, Zhang Z, et al. Application of nanomaterials in the treatment of rheumatoid arthritis. RSC Adv 2021; 11:7129-7137.
3. Hajizadeh A, Abtahi Froushani SM, Tehrani AA, Azizi S, Bani Hashemi SR. Effects of naringenin on experimentally induced rheumatoid arthritis in wistar rats. Arch Razi Inst 2021; 76:903-912.
4. Etemadi S, Abtahi Froushani SM, Hashemi Asl SM, Mahmoudian A. Combined atorvastatin and pentoxifylline in ameliorating inflammation induced by complete Freund’s adjuvant. Inflammopharmacology 2022;30:935-944.
5. Roy HS, Singh R, Ghosh D. Recent advances in nanotherapeutic strategies that target nitric oxide pathway for preventing cartilage degeneration. Nitric Oxide 2021; 109-110:1-11.
6. George G, Shyni GL, Raghu KG. Current and novel therapeutic targets in the treatment of rheumatoid arthritis. Inflammopharmacology 2020; 28:1457-1476.
7. Rahmani-Kukia N, Abbasi A, Abtahi Froushani SM, Shahgaldi S, Mokarram P. The effects of 17 Beta-Estradiol primed mesenchymal stem cells on the biology of co-cultured neutrophil. Int Immunopharmacol 2020; 84:106602.
8. Chahal J, Gómez-Aristizábal A, Shestopaloff K, Bhatt S, Chaboureau A, Fazio A, et al. Bone marrow mesenchymal stromal cell treatment in patients with osteoarthritis results in overall improvement in pain and symptoms and reduces synovial inflammation. Stem Cells Transl Med 2019; 8:746-757.
9. Mirzamohammadi S, Aali E, Najafi R, Kamarul T, Mehrabani M, Aminzadeh A, et al. Effect of 17β-estradiol on mediators involved in mesenchymal stromal cell trafficking in cell therapy of diabetes. Cytotherapy 2015; 17:46-57.
10. Schrepfer S, Deuse T, Reichenspurner H, Fischbein MP, Robbins RC, Pelletier MP. Stem cell transplantation: the lung barrier. Transplant Proc 2007; 39:573-576.
11. Gao J, Dennis JE, Muzic RF, Lundberg M, Caplan AI. The dynamic in vivo distribution of bone marrow-derived mesenchymal stem cells after infusion. Cells Tissues Organs 2001; 169:12-20.
12. Maijenburg MW, Gilissen C, Melief SM, Kleijer M, Weijer K, Ten Brinke A, et al. Nuclear receptors Nur77 and Nurr1 modulate mesenchymal stromal cell migration. Stem Cells Dev 2012; 21:228-238.
13. Hofbauer LC, Heufelder AE. Role of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in bone cell biology. J Mol Med (Berl) 2001; 79:243-253.
14. Andersson A, Stubelius A, Karlsson MN, Engdahl C, Erlandsson M, Grahnemo L, et al. Estrogen regulates T helper 17 phenotype and localization in experimental autoimmune arthritis. Arthritis Res Ther 2015; 17:1-12.
15. Niada S, Giannasi C, Ferreira LM, Milani A, Arrigoni E, Brini AT. 17β-estradiol differently affects osteogenic differentiation of mesenchymal stem/stromal cells from adipose tissue and bone marrow. Differentiation 2016; 92:291-297.
16. Dai Z, Li Y, Quarles LD, Song T, Pan W, Zhou H, et al. Resveratrol enhances proliferation and osteoblastic differentiation in human mesenchymal stem cells via ER-dependent ERK1/2 activation. Phytomedicine 2007; 14:806-814.
17. Heidari Barchi Nezhad R, Asadi F, Mirzaei MR, Abtahi Froushani SM. Comparison of the effects of 17β- estradiol treated and untreated mesenchymal stem cells on ameliorating animal model of multiple sclerosis. Iran j Basic Med Sci 2018; 21:936-942.
18. Mansouri Motlagh B, Afzale Ahangaran N, Abtahi Froushani SM. Calcitriol modulates the effects of bone marrow-derived mesenchymal stem cells on macrophage functions. Iran j Basic Med Sci 2015; 18:672-676.
19. Jones EA, English A, Kinsey SE, Straszynski L, Emery P, Ponchel F, et al. Optimization of a flow cytometry-based protocol for detection and phenotypic characterization of multipotent mesenchymal stromal cells from human bone marrow. Cytometry B Clin Cytom 2006; 70:391-399.
20. Moases Ghaffary E, Abtahi Froushani SM. Immunomodulatory benefits of mesenchymal stem cells treated with Caffeine in adjuvant-induced arthritis. Life Sci 2020; 246:117420.
21. Brand DD, Latham KA, Rosloniec EF. Collagen-induced arthritis. Nature Protocols 2007; 2:1269-1275.
22. Bryan NS, Grisham MB. Methods to detect nitric oxide and its metabolites in biological samples. Free Radic Biol Med 2007; 43:645-657.
23. Pulli B, Ali M, Forghani R, Schob S, Hsieh KL, Wojtkiewicz G, et al. Measuring myeloperoxidase activity in biological samples. PLoS One 2013; 8:e67976.
24. Crisostomo PR, Wang M, Herring CM, Markel TA, Meldrum KK, Lillemoe KD, et al. Gender differences in injury induced mesenchymal stem cell apoptosis and VEGF, TNF, IL-6 expression: role of the 55 kDa TNF receptor (TNFR1). J Mol Cell Cardiol 2007; 42:142-149.
25. Zamani Mazdeh D, Mirshokraei P, Emami M, Mirshahi A, Karimi I. 17β-estradiol improves the efficacy of exploited autologous bone marrow-derived mesenchymal stem cells in non-union radial defect healing: A rabbit model. Res Vet Sci 2018; 118:11-18.
26. Golbahari S, Abtahi Froushani SM. Synergistic benefits of Nicotine and Thymol in alleviating experimental rheumatoid arthritis. Life Sci 2019; 239:117037.
27. Sung WY, Tsai WC. Rethink about the role of rheumatoid factor and anti-citrullinated protein antibody in rheumatoid arthritis. Rheumatol Immunol Res 2021; 2:19-25.
28. Yeo J, Lee YM, Lee J, Park D, Kim K, Kim J, et al. Nitric oxide-scavenging nanogel for treating rheumatoid arthritis. Nano Lett 2019; 19:6716-6724.
29. Khan AA, Alsahli MA, Rahmani AH. Myeloperoxidase as an active disease biomarker: Recent biochemical and pathological perspectives. Med Sci (Basel) 2018; 6:1-21.
30. Fernandes RM, da Silva NP, Sato EI. Increased myeloperoxidase plasma levels in rheumatoid arthritis. Rheumatol Int 2012; 32:1605-1609.
31. Sokal DM, McSloy A, Donegà M, Kirk J, Colas RA, Dolezalova N, et al. Splenic nerve neuromodulation reduces inflammation and promotes resolution in chronically implanted pigs. Front Immunol 2021; 12:649786.
32. Bazzazi H, Aghaei M, Memarian A, Asgarian-Omran H, Behnampour N, Yazdani Y. Th1-Th17 Ratio as a new insight in rheumatoid arthritis disease. Iran J Allergy Asthma Immunol 2018; 17:68-77.