Human Umbilical Cord Mesenchymal Stem Cells-Derived Small Extracellular Vesicles Can Be Considered as Cell-Free Therapeutics for Angiogenesis Promotion

Document Type : Original Article

Authors

1 Department of Hematology and Cell Therapy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Faculty of Pharmacy, Lorestan University of Medical Sciences, Khorramabad, Iran

Abstract

Objective:
Angiogenesis has critical roles in several physiological processes. Restoring angiogenesis in some
pathological conditions such as a few vascular diseases can be a therapeutic approach to controlling this issue.
Mesenchymal stem cells (MSCs) secrete specific intracellular products known as extracellular vesicles (EVs) with high
therapeutic potential which compared to their source cells, do not have the limitations of cell therapy. The angiogenic
effect of the human umbilical cord MSCs (hUCMSCs)-derived small EVs are evaluated in the present work. Aim of this
research is to show that hUCMSCs-derived small EVs cause differentiation of genes involved in angiogenesis like
FGFR-1, FGF, VEGF, and VEGFR-2.

Materials and Methods:
In this experimental study, MSCs were isolated from the human umbilical cord, and after
confirming their identities, their secreted EVs (including exosomes) were extracted by ultracentrifugation. The isolated
small EVs were characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), bicinchoninic
acid assay (BCA), and Western Blotting. Then, the human umbilical vein endothelial cells (HUVECs) were treated
with derived small EVs for 72 hours, and the expression of the angiogenic factors including FGFR-1, FGF, VEGF,
and VEGFR-2 was evaluated by quantitative real-time-polymerase chain reaction (qPCR). Angiogenesis was also
evaluated via a tube formation assay.

Results:
The results demonstrated that FGFR-1, FGF, VEGF, and VEGFR-2 could be elevated 2, 2, 3.5, and 2 times,
respectively, in EVs treated HUVECs, and derivative EVs can encourage tube formation in HUVECs.

Conclusion:
These findings imply that hUCMSCs-derived small EVs are valuable resources in promoting angiogenesis
and are very promising in cell-free therapy.

Keywords

References

  1. Soufi-zomorrod M, Hajifathali A, Kouhkan F, Mehdizadeh M, Rad SMAH, Soleimani M. MicroRNAs modulating angiogenesis: miR- 129-1 and miR-133 act as angio-miR in HUVECs. Tumor Biol. 2016; 37(7): 9527-9534.
  2. Komaki M, Numata Y, Morioka C, Honda I, Tooi M, Yokoyama N, et al. Exosomes of human placenta-derived mesenchymal stem cells stimulate angiogenesis. Stem Cell Res Ther. 2017; 8: 219.
  3. Bian X, Ma K, Zhang C, Fu X. Therapeutic angiogenesis using stem cell-derived extracellular vesicles: an emerging approach for treatment of ischemic diseases. Stem Cell Res Ther. 2019; 10: 158.
  4. Naito H, Iba T, Takakura N. Mechanisms of new blood-vessel formation and proliferative heterogeneity of endothelial cells. Int Immunol. 2020; 32(5): 295-305.
  5. Wang S, Zhu R, Li H, Li J, Han Q, Zhao RC. Mesenchymal stem cells and immune disorders: from basic science to clinical transition. Front Med. 2019; 13(2): 138-151.
  6. Keshtkar S, Azarpira N, Ghahremani MH. Mesenchymal stem cellderived extracellular vesicles: Novel frontiers in regenerative medicine. Stem Cell Res Ther. 2018; 9(1): 63.
  7. Pashoutan Sarvar D, Shamsasenjan K, Akbarzadehlaleh P. Mesenchymal stem cell-derived exosomes: new opportunity in cell-free therapy. Adv Pharm Bull. 2016; 6(3): 293-299.
  8. Brown C, McKee C, Bakshi S, Walker K, Hakman E, Halassy S, et al. Mesenchymal stem cells: Cell therapy and regeneration potential. J Tissue Eng Regen Med. 2019; 13(9): 1738-1755.
  9. Berebichez-Fridman R, Montero-Olvera PR. Sources and clinical applications of mesenchymal stem cells state-of-the-art review. Sultan Qaboos Univ Med J. 2018; 18(3): e264-e277.
  10. Lee M, Jeong SY, Ha J, Kim M, Jin HJ, Kwon SJ, et al. Low immunogenicity of allogeneic human umbilical cord blood-derived mesenchymal stem cells in vitro and in vivo. Biochem Biophys Res Commun. 2014; 446(4): 983-989.
  11. Rackov G, Garcia-Romero N, Esteban-Rubio S, Carrión-Navarro J, Belda-Iniesta C, Ayuso-Sacido A. Vesicle-mediated control of cell function: The role of extracellular matrix and microenvironment. Front Physiol. 2018; 9: 651.
  12. Park KS, Bandeira E, Shelke GV, Lässer C, Lötvall J. Enhancement of therapeutic potential of mesenchymal stem cell-derived extracellular vesicles. Stem Cell Res Ther. 2019; 10: 288.
  13. Campanella C, Caruso Bavisotto C, Logozzi M, Marino Gammazza A, Mizzoni D, Cappello F, et al. On the choice of the extracellular vesicles for therapeutic purposes. Int J Mol Sci. 2019; 20(2): 236.
  14. Ni YQ, Lin X, Zhan JK, Liu YS. Roles and functions of exosomal non-coding RNAs in vascular aging. Aging Dis. 2020; 11(1): 164- 178.
  15. Edgar JR. Q & A: What are exosomes, exactly? BMC Biol. 2016; 14: 46.
  16. Cheng L, Zhang K, Wu S, Cui M, Xu T. Focus on mesenchymal stem cell-derived exosomes: Opportunities and challenges in cellfree therapy. Stem Cells Int. 2017; 2017: 6305295.
  17. Nazimek K, Bryniarski K, Santocki M, Ptak W. Exosomes as mediators of intercellular communication: clinical implications. Pol Arch Med Wewn. 2015; 125(5): 370-380.
  18. Mushahary D, Spittler A, Kasper C, Weber V, Charwat V. Isolation, cultivation, and characterization of human mesenchymal stem cells. Cytom Part A. 2018; 93(1): 19-31.
  19. Riordan NH, Morales I, Fernández G, Allen N, Fearnot NE, Leckrone ME, et al. Clinical feasibility of umbilical cord tissue-derived mesenchymal stem cells in the treatment of multiple sclerosis. J Transl Med. 2018; 16(1): 57.
  20. Zhang K, Li Z. Molecular imaging of therapeutic effect of mesenchymal stem cell-derived exosomes for hindlimb ischemia treatment. Methods Mol Biol. 2020; 2150: 213-225.
  21. Zhang Y, Yan J, Li Z, Zheng J, Sun Q. Exosomes derived from human umbilical cord mesenchymal stem cells alleviate psoriasis-like skin inflammation. J Interferon Cytokine Res. 2022; 42(1): 8-18.
  22. Addi T, Poitevin S, McKay N, El Mecherfi KE, Kheroua O, Jourde- Chiche N, et al. Mechanisms of tissue factor induction by the uremic toxin indole-3 acetic acid through aryl hydrocarbon receptor/ nuclear factor-kappa B signaling pathway in human endothelial cells. Arch Toxicol. 2019; 93(1): 121-136.
  23. Kishore R, Khan M. More than tiny sacks: stem cell exosomes as cell-free modality for cardiac repair. Circ Res. 2016; 118(2): 330- 343.
  24. Fallah A, Sadeghinia A, Kahroba H, Samadi A, Heidari HR, Bradaran B, et al. Therapeutic targeting of angiogenesis molecular pathways in angiogenesis-dependent diseases. Biomed Pharmacother. 2019; 110: 775-785.
  25. Lukomska B, Stanaszek L, Zuba-Surma E, Legosz P, Sarzynska S, Drela K. Challenges and controversies in human mesenchymal stem cell therapy. Stem Cells Int. 2019; 2019: 9628536.
  26. Shang F, Yu Y, Liu S, Ming L, Zhang Y, Zhou Z, et al. Advancing application of mesenchymal stem cell-based bone tissue regeneration. Bioact Mater. 2021; 6(3): 666-683.
  27. Lee BC, Kang KS. Functional enhancement strategies for immunomodulation of mesenchymal stem cells and their therapeutic application. Stem Cell Res Ther. 2020; 11(1): 1-10.
  28. Teng X, Chen L, Chen W, Yang J, Yang Z, Shen Z. Mesenchymal stem cell-derived exosomes improve the microenvironment of infarcted myocardium contributing to angiogenesis and anti-inflammation. Cell Physiol Biochem. 2015; 37(6): 2415-2424.
  29. Álvaro-Afonso FJ, Sanz-Corbalán I, Lázaro-Martínez JL, Kakagia D, Papanas N. Adipose-derived mesenchymal stem cells in the treatment of diabetic foot ulcers: a review of preclinical and clinical studies. Angiology. 2020; 71(9): 853-863.
  30. Zhang C, Lin Y, Liu Q, He J, Xiang P, Wang D, et al. Growth differentiation factor 11 promotes differentiation of MSCs into endothelial- like cells for angiogenesis. J Cell Mol Med. 2020; 24(15): 8703- 8717.
  31. Wang Z, Zheng L, Lian C, Qi Y, Li W. Human umbilical cord-derived mesenchymal stem cells relieve hind limb ischemia by promoting angiogenesis in mice. Stem Cells Dev. 2019; 28(20): 1384-1397.
  32. Lai RC, Arslan F, Lee MM, Sze NS, Choo A, Chen TS, et al. Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. Stem Cell Res. 2010; 4(3): 214-222.
  33. Zhang J, Guan J, Niu X, Hu G, Guo S, Li Q, et al. Exosomes released from human induced pluripotent stem cells-derived MSCs facilitate cutaneous wound healing by promoting collagen synthesis and angiogenesis. J Transl Med. 2015; 13: 49.
  34. Tian T, Zhang HX, He CP, Fan S, Zhu YL, Qi C, et al. Surface functionalized exosomes as targeted drug delivery vehicles for cerebral ischemia therapy. Biomaterials. 2018; 150: 137-149.
  35. Zhao Y, Sun X, Cao W, Ma J, Sun L, Qian H, et al. Exosomes derived from human umbilical cord mesenchymal stem cells relieve acute myocardial ischemic injury. Stem Cells Int. 2015; 2015: 761643.
  36. Liang X, Zhang L, Wang S, Han Q, Zhao RC. Exosomes secreted by mesenchymal stem cells promote endothelial cell angiogenesis by transferring miR-125a. J Cell Sci. 2016; 129(11): 2182-2189.
  37. Huang JH, Xu Y, Yin XM, Lin FY. Exosomes derived from miR- 126-modified MSCs promote angiogenesis and neurogenesis and attenuate apoptosis after spinal cord injury in rats. Neuroscience. 2020; 424: 133-145.
  38. Raileanu VN, Whiteley J, Chow T, Kollara A, Mohamed A, Keating A, et al. Banking mesenchymal stromal cells from umbilical cord tissue: large sample size analysis reveals consistency between donors. Stem Cells Transl Med. 2019; 8(10): 1041-1054.
  39. Liu Y, Fang J, Zhang Q, Zhang X, Cao Y, Chen W, et al. Wnt10boverexpressing umbilical cord mesenchymal stem cells promote critical size rat calvarial defect healing by enhanced osteogenesis and VEGF-mediated angiogenesis. J Orthop Transl. 2020; 23: 29- 37.
  40. Karaman S, Leppänen VM, Alitalo K. Vascular endothelial growth factor signaling in development and disease. Development. 2018; 145(14): dev151019.
Cell Journal (Yakhteh)
Volume 24, Issue 11 - Serial Number 106
November 2022
Pages 689-696

Files

Cited by

Share

How to cite

Statistics