A smart magnetic hydrogel containing exosome promotes osteogenic commitment of human adipose-derived mesenchymal stem cells

Document Type : Original Article

Authors

1 Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

3 Medical Nanotechnology and Tissue Engineering Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

4 Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

5 Department of Health and Medical Sciences, University of Antwerp, Antwerp, Belgium

Abstract

Objective(s): Exosomes, as nano-sized extracellular vehicles acting as cell-to-cell communicators, are novel promising therapeutics in the area of bone tissue engineering. Moreover, magnetic nanoparticles, whose integration with other appropriate components is viewed as an intriguing approach to strengthen bone tissue engineering efficacy. We investigated the effect of magnetic enriched with exosomes on osteogenic differentiation.
Materials and Methods: Exosomes were isolated from human adipose-derived mesenchymal stem cells by Exo-spin™ kit (MSC-EX). Alginate (Alg) scaffold containing 1% (w/w) cobalt ferrite nanoparticles (CoFe2O4) was produced. MSC-EX were gently loaded onto Alg and Alg-cobalt ferrite (Alg-CF) scaffolds yielding Alg-EX and Alg-CF-EX scaffolds. The effects of MSC-Ex and magnetic hydrogel composite under an external static magnetic field (SMF) on proliferation and differentiation of MSCs were evaluated by alkaline phosphatase (ALP) activity measurement, alizarin red staining, and energy dispersive X-ray (EDX) analysis.
Results: Our results showed that Alg and Alg-CF scaffolds were not only cytotoxic but also supported AdMSCs proliferation. MSC-EX loading of the scaffolds enhanced AdMSCs proliferation significantly. According to the results, Alg-CF-EX scaffolds under magnetic stimulation exhibited the most potent effect on osteogenic differentiation of cultured AdMSCs as evidenced by higher ALP activity and mineralization.
Conclusion: We provided evidence that the combination of Alg hydrogel, CFNPs, and MSC-EX resulted in the construction of a bone tissue-engineering scaffold that highly supports the osteogenic commitment of MSCs.

Keywords


1. Kamolz L-P, Keck M, Kasper C. Wharton’s jelly mesenchymal stem cells promote wound healing and tissue regeneration. Stem Cell Res Ther 2014; 5: 1-2.
2. Guan J, Zhang J, Zhu Z, Niu X, Guo S, Wang Y, et al. Bone morphogenetic protein 2 gene transduction enhances the osteogenic potential of human urine-derived stem cells. Stem Cell Res Ther 2015; 6: 1-11.
3. Li W, Liu Y, Zhang P, Tang Y, Zhou M, Jiang W, et al. Tissue-engineered bone immobilized with human adipose stem cells-derived exosomes promotes bone regeneration. ACS Appl Mater Interfaces 2018; 10: 5240-5254.
4. Liang X, Ding Y, Zhang Y, Tse H-F, Lian Q. Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant 2014; 23: 1045-1059.
5. Hu G-w, Li Q, Niu X, Hu B, Liu J, Zhou S-m, et al. Exosomes secreted by human-induced pluripotent stem cell-derived mesenchymal stem cells attenuate limb ischemia by promoting angiogenesis in mice. Stem Cell Res Ther 2015; 6: 1-15.
6. Zhang J, Liu X, Li H, Chen C, Hu B, Niu X, et al. Exosomes/tricalcium phosphate combination scaffolds can enhance bone regeneration by activating the PI3K/Akt signaling pathway. Stem Cell Res Ther 2016; 7: 1-14.
7. Burger D, Viñas JL, Akbari S, Dehak H, Knoll W, Gutsol A, et al. Human endothelial colony-forming cells protect against acute kidney injury: role of exosomes. Am J Pathol 2015; 185: 2309-2323.
8. Doeppner TR, Herz J, Görgens A, Schlechter J, Ludwig A-K, Radtke S, et al. Extracellular vesicles improve post‐stroke neuroregeneration and prevent postischemic immunosuppression. Stem Cells Transl Med 2015; 4: 1131-1143.
9. Wang L, Wang J, Zhou X, Sun J, Zhu B, Duan C, et al. A new self-healing hydrogel containing hucMSC-derived exosomes promotes bone regeneration. Front Bioeng Biotechnol 2020; 8: 564731.
10. Hernandez-González AC, Tellez-Jurado L, Rodríguez-Lorenzo LM. Alginate hydrogels for bone tissue engineering, from injectables to bioprinting: A review. Carbohydr Polym 2020; 229: 115514.
11. Nasri‐Nasrabadi B, Kaynak A, Heidarian P, Komeily‐Nia Z, Mehrasa M, Salehi H, et al. Sodium alginate/magnesium oxide nanocomposite scaffolds for bone tissue engineering. Polym Adv Technol  2018; 29: 2553-2559.
12. Jain TK, Richey J, Strand M, Leslie-Pelecky DL, Flask CA, Labhasetwar V. Magnetic nanoparticles with dual functional properties: drug delivery and magnetic resonance imaging. Biomater 2008; 29: 4012-4021.
13. Corr SA, Byrne SJ, Tekoriute R, Meledandri CJ, Brougham DF, Lynch M, et al. Linear assemblies of magnetic nanoparticles as MRI contrast agents. J Am Chem Soc 2008; 130: 4214-4215.
14. Lee J-H, Jang J-t, Choi J-s, Moon SH, Noh S-h, Kim J-w, et al. Exchange-coupled magnetic nanoparticles for efficient heat induction. Nat Nanotechnol 2011; 6: 418-422.
15. Xia Y, Sun J, Zhao L, Zhang F, Liang X-J, Guo Y, et al. Magnetic field and nano-scaffolds with stem cells to enhance bone regeneration. Biomater 2018; 183: 151-170.
16. Srinivasan SY, Paknikar KM, Bodas D, Gajbhiye V. Applications of cobalt ferrite nanoparticles in biomedical nanotechnology. Nanomedicine (Lond) 2018; 13: 1221-1238.
17. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254.
18. Gandolfi MG, Gardin C, Zamparini F, Ferroni L, Esposti MD, Parchi G, et al. Mineral-doped poly (L-lactide) acid scaffolds enriched with exosomes improve osteogenic commitment of human adipose-derived mesenchymal stem cells. Nanomaterials (Basel) 2020; 10: 432-451.
19. Tschuschke M, Kocherova I, Bryja A, Mozdziak P, Angelova Volponi A, Janowicz K, et al. Inclusion biogenesis, methods of isolation and clinical application of human cellular exosomes. J Clin Med     2020; 9: 436.
20. Wang Y, Wang X, Shi J, Zhu R, Zhang J, Zhang Z, et al. A biomimetic silk fibroin/sodium alginate composite scaffold for soft tissue engineering. Sci Rep 2016; 6: 1-13.
21. Lee H-j, Ahn S-H, Kim GH. Three-dimensional collagen/alginate hybrid scaffolds functionalized with a drug delivery system (DDS) for bone tissue regeneration. Chem Mater 2012; 24: 881-891.
22. Zhao L, Zhang H, Xing Y, Song S, Yu S, Shi W, et al. Studies on the magnetism of cobalt ferrite nanocrystals synthesized by hydrothermal method. J Solid State Chem 2008; 181: 245-252.
23. Georgopanos P, Schneider GA, Dreyer A, Handge UA, Filiz V, Feld A, et al. Exceptionally strong, stiff and hard hybrid material based on an elastomer and isotropically shaped ceramic nanoparticles. Sci Rep 2017; 7: 1-9.
24. Mangal R, Srivastava S, Archer LA. Phase stability and dynamics of entangled polymer–nanoparticle composites. Nat Commun 2015; 6: 1-9.
25. Luckanagul J, Lee LA, Nguyen QL, Sitasuwan P, Yang X, Shazly T, et al. Porous alginate hydrogel functionalized with virus as three-dimensional scaffolds for bone differentiation. Biomacromolecules 2012; 13: 3949-3958.
26. Filippi M, Dasen B, Guerrero J, Garello F, Isu G, Born G, et al. Magnetic nanocomposite hydrogels and static magnetic field stimulate the osteoblastic and vasculogenic profile of adipose-derived cells. Biomaterials 2019; 223: 119468.
27. Zhang J, Zhao S, Zhu M, Zhu Y, Zhang Y, Liu Z, et al. 3D-printed magnetic Fe 3 O 4/MBG/PCL composite scaffolds with multifunctionality of bone regeneration, local anticancer drug delivery and hyperthermia. J Mater Chem B 2014; 2: 7583-7595.
28. Wang Q, Chen B, Cao M, Sun J, Wu H, Zhao P, et al. Response of MAPK pathway to iron oxide nanoparticles in vitro treatment promotes osteogenic differentiation of hBMSCs. Biomaterials 2016; 86: 11-20.
29. Hu S, Zhou Y, Zhao Y, Xu Y, Zhang F, Gu N, et al. Enhanced bone regeneration and visual monitoring via superparamagnetic iron oxide nanoparticle scaffold in rats J Tissue Eng Regen Med 2018; 12: 2085-2098.
30. Kim EC, Leesungbok R, Lee SW, Lee HW, Park SH, Mah SJ, et al. Effects of moderate intensity static magnetic fields on human bone marrow‐derived mesenchymal stem cells. Bioelectromagnetics 2015; 36: 267-276.
31. Huang J, Wang D, Chen J, Liu W, Duan L, You W, et al. Osteogenic differentiation of bone marrow mesenchymal stem cells by magnetic nanoparticle composite scaffolds under a pulsed electromagnetic field. Saudi Pharm J 2017; 25: 575-579.
32. Rosen AD. Mechanism of action of moderate-intensity static magnetic fields on biological systems. Cell Biochem Biophys 2003; 39: 163-173.
33. Kotani H, Iwasaka M, Ueno S, Curtis A. Magnetic orientation of collagen and bone mixture. J Appl Phys 2000; 87: 6191-6193.
34. Petecchia L, Sbrana F, Utzeri R, Vercellino M, Usai C, Visai L, et al. Electro-magnetic field promotes osteogenic differentiation of BM-hMSCs through a selective action on Ca 2+-related mechanisms. Sci Rep 2015; 5: 1-13.
35. Qin Y, Wang L, Gao Z, Chen G, Zhang C. Bone marrow stromal/stem cell-derived extracellular vesicles regulate osteoblast activity and differentiation in vitro and promote bone regeneration in vivo. Sci Rep 2016; 6: 1-11.
36. Furuta T, Miyaki S, Ishitobi H, Ogura T, Kato Y, Kamei N, et al. Mesenchymal stem cell‐derived exosomes promote fracture healing in a mouse model. Stem Cells Transl Med 2016; 5: 1620-30.
37. Camussi G, Deregibus MC, Bruno S, Cantaluppi V, Biancone L. Exosomes/microvesicles as a mechanism of cell-to-cell communication. Kidney Int 2010; 78: 838-848.
38. Baker N, Sohn J, Tuan RS. Promotion of human mesenchymal stem cell osteogenesis by PI3-kinase/Akt signaling, and the influence of caveolin-1/cholesterol homeostasis. Stem Cell Res Ther 2015; 6: 1-11.
39. Zhang L, Jiao G, Ren S, Zhang X, Li C, Wu W, et al. Exosomes from bone marrow mesenchymal stem cells enhance fracture healing through the promotion of osteogenesis and angiogenesis in a rat model of nonunion. Stem Cell Res Ther 2020; 11: 1-15.