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Exosomes derived from mesenchymal stem cells inhibit mitochondrial dysfunction-induced apoptosis of chondrocytes via p38, ERK, and Akt pathways

  • Hui QiEmail author
  • Dan-Ping Liu
  • Da-Wei Xiao
  • Da-Chuan Tian
  • Yong-Wei Su
  • Shao-Feng Jin
Article
  • 210 Downloads

Abstract

Osteoarthritis (OA) is the most common chronic joint disease worldwide. Chondrocyte, as the only resident cell type in cartilage, its apoptosis is of pathogenetic significance in OA. Mesenchymal stem cell (MSC)-based-therapy has been proved effective in OA in animals and clinical studies. Nowadays, the regenerative potential of MSC-based therapy is mostly attributed to its paracrine secretion, in which exosomes may play an important role. In the present study, we aimed to find out the significance of MSC-derived exosomes (MSC-Exos) on the viability of chondrocytes under normal and inflammatory conditions. Bone marrow MSCs (BMSCs) and chondrocytes from rabbits were cultured in vitro. BMSC-Exos were isolated by an ultracentrifugation method. Transmission electron microscopy and Western blot were used to identify exosomes. The internalization of BMSC-Exos into chondrocytes was observed by fluorescent microscope. The viability and apoptosis of chondrocytes induced by IL-1β were tested through MTT method, Hoechst33324 dying, and mitochondrial damage measurement. Phosphorylation of p38, ERK, and Akt were evaluated by Western blot. The results showed that BMSC-Exos were round-shaped. Co-culturing BMSC-Exos with chondrocytes could observe the uptake of BMSC-Exos by chondrocytes. The viability decreased, apoptosis occurred, and the mitochondrial membrane potential of chondrocytes changed a lot when IL-1β were given, but all the changes were almost abolished when BMSC-Exos was added. Furthermore, the phosphorylation of p38 and ERK were inhibited, and phosphorylation of Akt was promoted by BMSC-Exos compared with IL-1β group. The present study demonstrated that BMSC-Exos inhibited mitochondrial-induced apoptosis in response to IL-1β, and p38, ERK, and Akt pathways were involved. BMSC-Exo might represent a novel cell-free therapeutic approach for the treatment of OA.

Keywords

Exosome Mesenchymal stem cell Chondrocyte Apoptosis Mitochondrial dysfunction 

References

  1. Azmi AS, Bao B, Sarkar FH (2013) Exosomes in cancer development, metastasis and drug resistance: a comprehensive review. Cancer Metastasis Rev 32:623–642CrossRefGoogle Scholar
  2. Baglio SR, Pegtel DM, Baldini N (2012) Mesenchymal stem cell secreted vesicles provide novel opportunities in (stem) cell-free therapy. Front Physiol 3:359CrossRefGoogle Scholar
  3. Blanco FJ, Rego I, Ruiz-Romero C (2011) The role of mitochondria in osteoarthritis. Nat Rev Rheumatol 7:161–169CrossRefGoogle Scholar
  4. Bornes TD, Adesida AB, Jomha NM (2014) Mesenchymal stem cells in the treatment of traumatic articular cartilage defects: a comprehensive review. Arthritis Res Ther 16:432CrossRefGoogle Scholar
  5. Brittberg M, Peterson L, Sjögren-Jansson E, Tallheden T, Lindahl A (2003) Articular cartilage engineering with autologous chondrocyte transplantation. A review of recent developments. J Bone Joint Surg Am 85-A:109–115CrossRefGoogle Scholar
  6. Burger D, Viñas JL, Akbari S, Dehak H, Knoll W, Gutsol A, Carter A, Touyz RM, Allan DS, Burns KD (2015) Human endothelial colony-forming cells protect against acute kidney injury: role of exosomes. Am J Pathol 185:2309–2323CrossRefGoogle Scholar
  7. Chang L, Karin M (2001) Mammalian MAP kinase signalling cascades. Nature 410:37–40CrossRefGoogle Scholar
  8. Chen TS, Lai RC, Lee MM, Choo AB, Lee CN, Lim SK (2010) Mesenchymal stem cell secretes microparticles enriched in premicroRNAs. Nucl Acids Res 38:215–224CrossRefGoogle Scholar
  9. Chen Z, Yue SX, Zhou G, Greenfield EM, Murakami S (2015) ERK1 and ERK2 regulate chondrocyte terminal differentiation during endochondral bone formation. J Bone Miner Res 30:765–774CrossRefGoogle Scholar
  10. Cosenza S, Ruiz M, Toupet K, Jorgensen C, Noël D (2017) Mesenchymal stem cells derived exosomes and microparticles protect cartilage and bone from degradation in osteoarthritis. Sci Rep 7:16214CrossRefGoogle Scholar
  11. Cross M, Smith E, Hoy D, Nolte S, Ackerman I, Fransen M, Bridgett L, Williams S, Guillemin F, Hill CL, Laslett LL, Jones G, Cicuttini F, Osborne R, Vos T, Buchbinder R, Woolf A, March L (2014) The global burden of hip and knee osteoarthritis: estimates from the global burden of disease 2010 study. Ann Rheum Dis 73:1323–1330CrossRefGoogle Scholar
  12. Ferguson SW, Wang J, Lee CJ, Liu M, Neelamegham S, Canty JM, Nguyen J (2018) The microRNA regulatory landscape of MSC-derived exosomes: a systems view. Sci Rep 8:1419CrossRefGoogle Scholar
  13. Fresno Vara JA, Casado E, de Castro J, Cejas P, Belda-Iniesta C, González-Barón M (2004) PI3K/Aktsignalling pathway and cancer. Cancer Treat Rev 30:193–204CrossRefGoogle Scholar
  14. Goldring MB, Marcu KB (2009) Cartilage homeostasis in health and rheumatic diseases. Arthritis Res Ther 11:224CrossRefGoogle Scholar
  15. Hu GW, Li Q, Niu X, Hu B, Liu J, Zhou SM, Guo SC, Lang HL, Zhang CQ, Wang Y, Deng ZF (2015) Exosomes secreted by human-induced pluripotent stem cell-derived mesenchymal stem cells attenuate limb ischemia by promoting angiogenesis in mice. Stem Cell Res Ther 6:10CrossRefGoogle Scholar
  16. Hwang HS, Kim HA (2015) Chondrocyte apoptosis in the pathogenesis of osteoarthritis. Int J Mol Sci 16:26035–26054CrossRefGoogle Scholar
  17. Im GI (2016) Regeneration of articular cartilage using adipose stem cells. J Biomed Mater Res A 104:1830–1844CrossRefGoogle Scholar
  18. Im GI (2017) Clinical use of stem cells in orthopaedics. Eur Cell Mater 33:183–196CrossRefGoogle Scholar
  19. Lai RC, Arslan F, Lee MM, Sze NS, Choo A, Chen TS, Salto-Tellez M, Timmers L, Lee CN, El Oakley RM, Pasterkamp G, de Kleijn DP, Lim SK (2010) Exosome secreted by MSCs reduces myocardial ischemia/reperfusion injury. Stem Cell Res 4:214–222CrossRefGoogle Scholar
  20. Lai RC, Tan SS, Teh BJ, Sze SK, Arslan F, de Kleijn DP, Choo A, Lim SK (2012) Proteolytic potential of the MSC exosome proteome: implications for an exosome-mediated delivery of therapeutic proteasome. Int J Proteom 2012:971907CrossRefGoogle Scholar
  21. Li H, Liu D, Li C, Zhou S, Tian D, Xiao D, Zhang H, Gao F, Huang J (2017a) Exosomes secreted from mutant-HIF-1α-modified bone-marrow-derived mesenchymal stem cells attenuate early steroid-induced avascular necrosis of femoral head in rabbit. Cell Biol Int 41:1379–1390CrossRefGoogle Scholar
  22. Li J, Tan M, Xiang Q, Zhou Z, Yan H (2017b) Thrombin-activated platelet-derived exosomes regulate endothelial cell expression of ICAM-1 via microRNA-223 during the thrombosis-inflammation response. Thromb Res 154:96–105CrossRefGoogle Scholar
  23. Li J, Yu J, Zhang H, Wang B, Guo H, Bai J, Wang J, Dong Y, Zhao Y, Wang Y (2016) Exosomes-derived MiR-302b suppresses lung cancer cell proliferation and migration via TGFβRII inhibition. Cell Physiol Biochem 38:1715–1726CrossRefGoogle Scholar
  24. Liang X, Ding Y, Zhang Y, Tse HF, Lian Q (2014) Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant 23:1045–1059CrossRefGoogle Scholar
  25. Liu Y, Lin L, Zou R, Wen C, Wang Z, Lin F (2018a) MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis. Cell Cycle 17:2411–2422CrossRefGoogle Scholar
  26. Liu Y, Zou R, Wang Z, Wen C, Zhang F, Lin F (2018b) Exosomal KLF3-AS1 from hMSCs promoted cartilage repair and chondrocyte proliferation in osteoarthritis. Biochem J 475:3629–3638CrossRefGoogle Scholar
  27. Ma X, Wang J, Li J, Ma C, Chen S, Lei W, Yang Y, Liu S, Bihl J, Chen C (2018) Loading miR-210 in endothelial progenitor cells derived exosomes boosts their beneficial effects on hypoxia/reoxygeneation-injured human endothelial cells via protecting mitochondrial function. Cell Physiol Biochem 46:664–675CrossRefGoogle Scholar
  28. Maneiro E, Martín MA, de Andres MC, López-Armada MJ, Fernández-Sueiro JL, del Hoyo P, Galdo F, Arenas J, Blanco FJ (2003) Mitochondrial respiratory activity is altered in osteoarthritic human articular chondrocytes. Arthritis Rheum 48:700–708CrossRefGoogle Scholar
  29. Maniatopoulos C, Sodek J, Melcher AH (1988) Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats. Cell Tissue Res 254:317–330CrossRefGoogle Scholar
  30. Marcacci M, Berruto M, Brocchetta D, Delcogliano A, Ghinelli D, Gobbi A, Kon E, Pederzini L, Rosa D, Sacchetti GL, Stefani G, Zanasi S (2005) Articular cartilage engineering with Hyalograft C: 3-year clinical results. Clin Orthop Relat Res 435:96–105CrossRefGoogle Scholar
  31. Martini M, De Santis MC, Braccini L, Gulluni F, Hirsch E (2014) PI3K/AKT signaling pathway and cancer: an updated review. Ann Med 46:372–383CrossRefGoogle Scholar
  32. Mayan MD, Gago-Fuentes R, Carpintero-Fernandez P, Fernandez-Puente P, Filgueira-Fernandez P, Goyanes N, Valiunas V, Brink PR, Goldberg GS, Blanco FJ (2015) Articular chondrocyte network mediated by gap junctions: role in metabolic cartilage homeostasis. Ann Rheum Dis 74:275–284CrossRefGoogle Scholar
  33. Ochi M, Adachi N, Nobuto H, Yanada S, Ito Y, Agung M (2004) Articular cartilage repair using tissue engineering technique novel approach with minimally invasive procedure. Artif Organs 28:28–32CrossRefGoogle Scholar
  34. Phornphutkul C, Wu KY, Auyeung V, Chen Q, Gruppuso PA (2008) mTOR signaling contributes to chondrocyte differentiation. Dev Dyn 237:702–712CrossRefGoogle Scholar
  35. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147CrossRefGoogle Scholar
  36. Reza AM, Choi YJ, Yasuda H, Kim JH (2016) Human adipose mesenchymal stem cell-derived exosomal-miRNAs are critical factors for inducing anti-proliferation signalling to A2780 and SKOV-3 ovarian cancer cells. Sci Rep 6:38498CrossRefGoogle Scholar
  37. Roma-Rodrigues C, Fernandes AR, Baptista PV (2014) Exosome in tumour microenvironment: overview of the crosstalk between normal and cancer cells. Biomed Res Int 2014:179486CrossRefGoogle Scholar
  38. Shi B, Wang Y, Zhao R, Long X, Deng W, Wang Z (2018) Bone marrow mesenchymal stem cell-derived exosomal miR-21 protects C-kit+ cardiac stem cells from oxidative injury through the PTEN/PI3K/Aktaxis. PLoS One 13:e0191616CrossRefGoogle Scholar
  39. Sixt SU, Peters J (2010) Extracellular alveolar proteasome: possible role in lung injury and repair. Proc Am ThoracSoc 7:91–96CrossRefGoogle Scholar
  40. Sung DK, Chang YS, Kang S, Song HY, Park WS, Lee BH (2010) Comparative evaluation of hypoxic-ischemic brain injury by flow cytometric analysis of mitochondrial membrane potential with JC-1 in neonatal rats. J Neurosci Methods 193:232–238CrossRefGoogle Scholar
  41. Tao SC, Yuan T, Zhang YL, Yin WJ, Guo SC, Zhang CQ (2017) Exosomes derived from miR-140-5p-overexpressing human synovial mesenchymal stem cells enhance cartilage tissue regeneration and prevent osteoarthritis of the knee in a rat model. Theranostics 7:180–195CrossRefGoogle Scholar
  42. Tofiño-Vian M, Guillén MI, Pérez Del Caz MD, Silvestre A, Alcaraz MJ (2018) Microvesicles from human adipose tissue-derived mesenchymal stem cells as a new protective strategy in osteoarthritic chondrocytes. Cell Physiol Biochem 47:11–25CrossRefGoogle Scholar
  43. Tomasetti M, Lee W, Santarelli L, Neuzil J (2017) Exosome-derived microRNAs in cancer metabolism: possible implications in cancer diagnostics and therapy. Exp Mol Med 49:e285CrossRefGoogle Scholar
  44. Tomasetti M, Nocchi L, Staffolani S, Manzella N, Amati M, al GJ (2014) MicroRNA-126 suppresses mesothelioma malignancy by targeting IRS1 and interfering with the mitochondrial function. Antioxid Redox Signal 21:2109–2125CrossRefGoogle Scholar
  45. Vinatier C, Bouffi C, Merceron C, Gordeladze J, Brondello JM, Jorgensen C, Weiss P, Guicheux J, Noël D (2009) Cartilage tissue engineering: towards a biomaterial-assisted mesenchymal stem cell therapy. Curr Stem Cell Res Ther 4:318–329CrossRefGoogle Scholar
  46. Wang Y, Yu D, Liu Z, Zhou F, Dai J, Wu B, Zhou J, Heng BC, Zou XH, Ouyang H, Liu H (2017) Exosomes from embryonic mesenchymal stem cells alleviate osteoarthritis through balancing synthesis and degradation of cartilage extracellular matrix. Stem Cell Res Ther 8:189CrossRefGoogle Scholar
  47. Wei Y, Bai L (2016) Recent advances in the understanding of molecular mechanisms of cartilage degeneration, synovitis and subchondral bone changes in osteoarthritis. Connect Tissue Res 57:245–261CrossRefGoogle Scholar
  48. Wood DD, Ihrie EJ, Dinarello CA, Cohen PL (1983) Isolation of an interleukin-1-like factor from human joint effusions. Arthritis Rheum 26:975–983CrossRefGoogle Scholar
  49. Xin H, Li Y, Chopp M (2014) Exosomes/miRNAs as mediating cell-based therapy of stroke. Front Cell Neurosci 8:377CrossRefGoogle Scholar
  50. Zhu Y, Wang Y, Zhao B, Niu X, Hu B, Li Q, Zhang J, Ding J, Chen Y, Wang Y (2017) Comparison of exosomes secreted by induced pluripotent stem cell-derived mesenchymal stem cells and synovial membrane-derived mesenchymal stem cells for the treatment of osteoarthritis. Stem Cell Res Ther 8:64CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Hui Qi
    • 1
    Email author
  • Dan-Ping Liu
    • 2
  • Da-Wei Xiao
    • 2
  • Da-Chuan Tian
    • 2
  • Yong-Wei Su
    • 2
  • Shao-Feng Jin
    • 1
  1. 1.Beijing Research Institute of Traumatology and OrthopaedicsBeijing Jishuitan HospitalBeijingChina
  2. 2.Department of OrthopaedicsThe First Affiliated Hospital of Jinzhou Medical UniversityJinzhouChina

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