Extracellular Vesicles from Mesenchymal Stem Cells as Novel Treatments for Musculoskeletal Diseases

[EN] Mesenchymal stem/stromal cells (MSCs) represent a promising therapy for musculoskeletal diseases. There is compelling evidence indicating that MSC effects are mainly mediated by paracrine mechanisms and in particular by the secretion of extracellular vesicles (EVs). Many studies have thus suggested that EVs may be an alternative to cell therapy with MSCs in tissue repair. In this review, we summarize the current understanding of MSC EVs actions in preclinical studies of (1) immune regulation and rheumatoid arthritis, (2) bone repair and bone diseases, (3) cartilage repair and osteoarthritis, (4) intervertebral disk degeneration and (5) skeletal muscle and tendon repair. We also discuss the mechanisms underlying these actions and the perspectives of MSC EVs-based strategies for future treatments of musculoskeletal disorders.This work has been funded by grant SAF2017-85806-R (Ministerio de Ciencia, Innovación y Universidades, Spain, FEDER.Alcaraz Tormo, MJ.; Compañ, Á.; Guillem Salazar, MI. (2019). Extracellular Vesicles from Mesenchymal Stem Cells as Novel Treatments for Musculoskeletal Diseases. Cells. 9(1):1-21. https://doi.org/10.3390/cells9010098S12191Musculoskeletal Conditions https://www.who. int/news-room/fact-sheets/detail/musculoskeletal-conditionsHofer, H. R., & Tuan, R. S. (2016). Secreted trophic factors of mesenchymal stem cells support neurovascular and musculoskeletal therapies. Stem Cell Research & Therapy, 7(1). doi:10.1186/s13287-016-0394-0Wang, L., Wang, L., Cong, X., Liu, G., Zhou, J., Bai, B., … Liu, Y. (2013). Human Umbilical Cord Mesenchymal Stem Cell Therapy for Patients with Active Rheumatoid Arthritis: Safety and Efficacy. Stem Cells and Development, 22(24), 3192-3202. doi:10.1089/scd.2013.0023Franceschetti, T., & De Bari, C. (2017). The potential role of adult stem cells in the management of the rheumatic diseases. Therapeutic Advances in Musculoskeletal Disease, 9(7), 165-179. doi:10.1177/1759720x17704639Freitag, J., Bates, D., Boyd, R., Shah, K., Barnard, A., Huguenin, L., & Tenen, A. (2016). Mesenchymal stem cell therapy in the treatment of osteoarthritis: reparative pathways, safety and efficacy – a review. BMC Musculoskeletal Disorders, 17(1). doi:10.1186/s12891-016-1085-9Vega, A., Martín-Ferrero, M. A., Del Canto, F., Alberca, M., García, V., Munar, A., … García-Sancho, J. (2015). Treatment of Knee Osteoarthritis With Allogeneic Bone Marrow Mesenchymal Stem Cells. Transplantation, 99(8), 1681-1690. doi:10.1097/tp.0000000000000678Cui, G.-H., Wang, Y. Y., Li, C.-J., Shi, C.-H., & Wang, W.-S. (2016). Efficacy of mesenchymal stem cells in treating patients with osteoarthritis of the knee: A meta-analysis. Experimental and Therapeutic Medicine, 12(5), 3390-3400. doi:10.3892/etm.2016.3791Iaquinta, M. R., Mazzoni, E., Bononi, I., Rotondo, J. C., Mazziotta, C., Montesi, M., … Martini, F. (2019). Adult Stem Cells for Bone Regeneration and Repair. Frontiers in Cell and Developmental Biology, 7. doi:10.3389/fcell.2019.00268Marolt Presen, D., Traweger, A., Gimona, M., & Redl, H. (2019). Mesenchymal Stromal Cell-Based Bone Regeneration Therapies: From Cell Transplantation and Tissue Engineering to Therapeutic Secretomes and Extracellular Vesicles. Frontiers in Bioengineering and Biotechnology, 7. doi:10.3389/fbioe.2019.00352Jo, C. H., Chai, J. W., Jeong, E. C., Oh, S., & Yoon, K. S. (2020). Intratendinous Injection of Mesenchymal Stem Cells for the Treatment of Rotator Cuff Disease: A 2-Year Follow-Up Study. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 36(4), 971-980. doi:10.1016/j.arthro.2019.11.120Klimczak, A., Kozlowska, U., & Kurpisz, M. (2018). Muscle Stem/Progenitor Cells and Mesenchymal Stem Cells of Bone Marrow Origin for Skeletal Muscle Regeneration in Muscular Dystrophies. Archivum Immunologiae et Therapiae Experimentalis, 66(5), 341-354. doi:10.1007/s00005-018-0509-7Ferreira, J. R., Teixeira, G. Q., Santos, S. G., Barbosa, M. A., Almeida-Porada, G., & Gonçalves, R. M. (2018). Mesenchymal Stromal Cell Secretome: Influencing Therapeutic Potential by Cellular Pre-conditioning. Frontiers in Immunology, 9. doi:10.3389/fimmu.2018.02837Aggarwal, S., & Pittenger, M. F. (2005). Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood, 105(4), 1815-1822. doi:10.1182/blood-2004-04-1559Ren, G., Zhang, L., Zhao, X., Xu, G., Zhang, Y., Roberts, A. I., … Shi, Y. (2008). Mesenchymal Stem Cell-Mediated Immunosuppression Occurs via Concerted Action of Chemokines and Nitric Oxide. Cell Stem Cell, 2(2), 141-150. doi:10.1016/j.stem.2007.11.014Chabannes, D., Hill, M., Merieau, E., Rossignol, J., Brion, R., Soulillou, J. P., … Cuturi, M. C. (2007). A role for heme oxygenase-1 in the immunosuppressive effect of adult rat and human mesenchymal stem cells. Blood, 110(10), 3691-3694. doi:10.1182/blood-2007-02-075481Bernardo, M. E., & Fibbe, W. E. (2013). Mesenchymal Stromal Cells: Sensors and Switchers of Inflammation. Cell Stem Cell, 13(4), 392-402. doi:10.1016/j.stem.2013.09.006Selmani, Z., Naji, A., Zidi, I., Favier, B., Gaiffe, E., Obert, L., … Deschaseaux, F. (2008). Human Leukocyte Antigen-G5 Secretion by Human Mesenchymal Stem Cells Is Required to Suppress T Lymphocyte and Natural Killer Function and to Induce CD4+CD25highFOXP3+Regulatory T Cells. Stem Cells, 26(1), 212-222. doi:10.1634/stemcells.2007-0554Di Nicola, M., Carlo-Stella, C., Magni, M., Milanesi, M., Longoni, P. D., Matteucci, P., … Gianni, A. M. (2002). Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood, 99(10), 3838-3843. doi:10.1182/blood.v99.10.3838Gunawardena, T. N. A., Rahman, M. T., Abdullah, B. J. J., & Abu Kasim, N. H. (2019). Conditioned media derived from mesenchymal stem cell cultures: The next generation for regenerative medicine. Journal of Tissue Engineering and Regenerative Medicine, 13(4), 569-586. doi:10.1002/term.2806Arslan, F., Lai, R. C., Smeets, M. B., Akeroyd, L., Choo, A., Aguor, E. N. E., … de Kleijn, D. P. (2013). Mesenchymal stem cell-derived exosomes increase ATP levels, decrease oxidative stress and activate PI3K/Akt pathway to enhance myocardial viability and prevent adverse remodeling after myocardial ischemia/reperfusion injury. Stem Cell Research, 10(3), 301-312. doi:10.1016/j.scr.2013.01.002Tian, T., Wang, Y., Wang, H., Zhu, Z., & Xiao, Z. (2010). Visualizing of the cellular uptake and intracellular trafficking of exosomes by live-cell microscopy. Journal of Cellular Biochemistry, 111(2), 488-496. doi:10.1002/jcb.22733Feng, D., Zhao, W.-L., Ye, Y.-Y., Bai, X.-C., Liu, R.-Q., Chang, L.-F., … Sui, S.-F. (2010). Cellular Internalization of Exosomes Occurs Through Phagocytosis. Traffic, 11(5), 675-687. doi:10.1111/j.1600-0854.2010.01041.xXu, J., Wang, Y., Hsu, C.-Y., Gao, Y., Meyers, C. A., Chang, L., … James, A. W. (2019). Human perivascular stem cell-derived extracellular vesicles mediate bone repair. eLife, 8. doi:10.7554/elife.48191Morrison, T. J., Jackson, M. V., Cunningham, E. K., Kissenpfennig, A., McAuley, D. F., O’Kane, C. M., & Krasnodembskaya, A. D. (2017). Mesenchymal Stromal Cells Modulate Macrophages in Clinically Relevant Lung Injury Models by Extracellular Vesicle Mitochondrial Transfer. American Journal of Respiratory and Critical Care Medicine, 196(10), 1275-1286. doi:10.1164/rccm.201701-0170ocLener, T., Gimona, M., Aigner, L., Börger, V., Buzas, E., Camussi, G., … Portillo, H. A. del. (2015). Applying extracellular vesicles based therapeutics in clinical trials – an ISEV position paper. Journal of Extracellular Vesicles, 4(1), 30087. doi:10.3402/jev.v4.30087Raposo, G., & Stoorvogel, W. (2013). Extracellular vesicles: Exosomes, microvesicles, and friends. Journal of Cell Biology, 200(4), 373-383. doi:10.1083/jcb.201211138Qiu, G., Zheng, G., Ge, M., Wang, J., Huang, R., Shu, Q., & Xu, J. (2018). Mesenchymal stem cell-derived extracellular vesicles affect disease outcomes via transfer of microRNAs. Stem Cell Research & Therapy, 9(1). doi:10.1186/s13287-018-1069-9Van Niel, G., D’Angelo, G., & Raposo, G. (2018). Shedding light on the cell biology of extracellular vesicles. Nature Reviews Molecular Cell Biology, 19(4), 213-228. doi:10.1038/nrm.2017.125Lai, R. C., Tan, S. S., Yeo, R. W. Y., Choo, A. B. H., Reiner, A. T., Su, Y., … Lim, S. K. (2016). MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA. Journal of Extracellular Vesicles, 5(1), 29828. doi:10.3402/jev.v5.29828Théry, C., Witwer, K. W., Aikawa, E., Alcaraz, M. J., Anderson, J. D., Andriantsitohaina, R., … Atkin-Smith, G. K. (2018). Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. Journal of Extracellular Vesicles, 7(1), 1535750. doi:10.1080/20013078.2018.1535750Tofiño-Vian, M., Guillén, M. I., & Alcaraz, M. J. (2018). Extracellular vesicles: A new therapeutic strategy for joint conditions. Biochemical Pharmacology, 153, 134-146. doi:10.1016/j.bcp.2018.02.004Wong, D. E., Banyard, D. A., Santos, P. J. F., Sayadi, L. R., Evans, G. R. D., & Widgerow, A. D. (2019). Adipose-derived stem cell extracellular vesicles: A systematic review✰. Journal of Plastic, Reconstructive & Aesthetic Surgery, 72(7), 1207-1218. doi:10.1016/j.bjps.2019.03.008Zhou, Y., Xu, H., Xu, W., Wang, B., Wu, H., Tao, Y., … Qian, H. (2013). Exosomes released by human umbilical cord mesenchymal stem cells protect against cisplatin-induced renal oxidative stress and apoptosis in vivo and in vitro. Stem Cell Research & Therapy, 4(2), 34. doi:10.1186/scrt194De Jong, O. G., Van Balkom, B. W. M., Schiffelers, R. M., Bouten, C. V. C., & Verhaar, M. C. (2014). Extracellular Vesicles: Potential Roles in Regenerative Medicine. Frontiers in Immunology, 5. doi:10.3389/fimmu.2014.00608Robbins, P. D., & Morelli, A. E. (2014). Regulation of immune responses by extracellular vesicles. Nature Reviews Immunology, 14(3), 195-208. doi:10.1038/nri3622Burrello, J., Monticone, S., Gai, C., Gomez, Y., Kholia, S., & Camussi, G. (2016). Stem Cell-Derived Extracellular Vesicles and Immune-Modulation. Frontiers in Cell and Developmental Biology, 4. doi:10.3389/fcell.2016.00083Siegel, G., Schäfer, R., & Dazzi, F. (2009). The Immunosuppressive Properties of Mesenchymal Stem Cells. Transplantation, 87(Supplement), S45-S49. doi:10.1097/tp.0b013e3181a285b0Fierabracci, A., Del Fattore, A., Luciano, R., Muraca, M., Teti, A., & Muraca, M. (2015). Recent Advances in Mesenchymal Stem Cell Immunomodulation: The Role of Microvesicles. Cell Transplantation, 24(2), 133-149. doi:10.3727/096368913x675728Mokarizadeh, A., Delirezh, N., Morshedi, A., Mosayebi, G., Farshid, A.-A., & Mardani, K. (2012). Microvesicles derived from mesenchymal stem cells: Potent organelles for induction of tolerogenic signaling. Immunology Letters, 147(1-2), 47-54. doi:10.1016/j.imlet.2012.06.001Conforti, A., Scarsella, M., Starc, N., Giorda, E., Biagini, S., Proia, A., … Bernardo, M. E. (2014). Microvescicles Derived from Mesenchymal Stromal Cells Are Not as Effective as Their Cellular Counterpart in the Ability to Modulate Immune Responses In Vitro. Stem Cells and Development, 23(21), 2591-2599. doi:10.1089/scd.2014.0091Carreras-Planella, L., Monguió-Tortajada, M., Borràs, F. E., & Franquesa, M. (2019). Immunomodulatory Effect of MSC on B Cells Is Independent of Secreted Extracellular Vesicles. Frontiers in Immunology, 10. doi:10.3389/fimmu.2019.01288Chen, W., Huang, Y., Han, J., Yu, L., Li, Y., Lu, Z., … Xiao, Y. (2016). Immunomodulatory effects of mesenchymal stromal cells-derived exosome. Immunologic Research, 64(4), 831-840. doi:10.1007/s12026-016-8798-6Harting, M. T., Srivastava, A. K., Zhaorigetu, S., Bair, H., Prabhakara, K. S., Toledano Furman, N. E., … Olson, S. D. (2017). Inflammation-Stimulated Mesenchymal Stromal Cell-Derived Extracellular Vesicles Attenuate Inflammation. STEM CELLS, 36(1), 79-90. doi:10.1002/stem.2730Reis, M., Mavin, E., Nicholson, L., Green, K., Dickinson, A. M., & Wang, X. (2018). Mesenchymal Stromal Cell-Derived Extracellular Vesicles Attenuate Dendritic Cell Maturation and Function. Frontiers in Immunology, 9. doi:10.3389/fimmu.2018.02538Ji, L., Bao, L., Gu, Z., Zhou, Q., Liang, Y., Zheng, Y., … Feng, X. (2019). Comparison of immunomodulatory properties of exosomes derived from bone marrow mesenchymal stem cells and dental pulp stem cells. Immunologic Research, 67(4-5), 432-442. doi:10.1007/s12026-019-09088-6Blazquez, R., Sanchez-Margallo, F. M., de la Rosa, O., Dalemans, W., Ã lvarez, V., Tarazona, R., & Casado, J. G. (2014). Immunomodulatory Potential of Human Adipose Mesenchymal Stem Cells Derived Exosomes on in vitro Stimulated T Cells. Frontiers in Immunology, 5. doi:10.3389/fimmu.2014.00556Zhang, B., Yin, Y., Lai, R. C., Tan, S. S., Choo, A. B. H., & Lim, S. K. (2014). Mesenchymal Stem Cells Secrete Immunologically Active Exosomes. Stem Cells and Development, 23(11), 1233-1244. doi:10.1089/scd.2013.0479Zhang, B., Yeo, R. W. Y., Lai, R. C., Sim, E. W. K., Chin, K. C., & Lim, S. K. (2018). Mesenchymal stromal cell exosome–enhanced regulatory T-cell production through an antigen-presenting cell–mediated pathway. Cytotherapy, 20(5), 687-696. doi:10.1016/j.jcyt.2018.02.372TOH, W. S., ZHANG, B., LAI, R. C., & LIM, S. K. (2018). Immune regulatory targets of mesenchymal stromal cell exosomes/small extracellular vesicles in tissue regeneration. Cytotherapy, 20(12), 1419-1426. doi:10.1016/j.jcyt.2018.09.008Budoni, M., Fierabracci, A., Luciano, R., Petrini, S., Di Ciommo, V., & Muraca, M. (2013). The Immunosuppressive Effect of Mesenchymal Stromal Cells on B Lymphocytes is Mediated by Membrane Vesicles. Cell Transplantation, 22(2), 369-379. doi:10.3727/096368911x582769bDi Trapani, M., Bassi, G., Midolo, M., Gatti, A., Takam Kamga, P., Cassaro, A., … Krampera, M. (2016). Differential and transferable modulatory effects of mesenchymal stromal cell-derived extracellular vesicles on T, B and NK cell functions. Scientific Reports, 6(1). doi:10.1038/srep24120Henao Agudelo, J. S., Braga, T. T., Amano, M. T., Cenedeze, M. A., Cavinato, R. A., Peixoto-Santos, A. R., … Camara, N. O. S. (2017). Mesenchymal Stromal Cell-Derived Microvesicles Regulate an Internal Pro-Inflammatory Program in Activated Macrophages. Frontiers in Immunology, 8. doi:10.3389/fimmu.2017.00881Lo Sicco, C., Reverberi, D., Balbi, C., Ulivi, V., Principi, E., Pascucci, L., … Tasso, R. (2017). Mesenchymal Stem Cell-Derived Extracellular Vesicles as Mediators of Anti-Inflammatory Effects: Endorsement of Macrophage Polarization. STEM CELLS Translational Medicine, 6(3), 1018-1028. doi:10.1002/sctm.16-0363Ti, D., Hao, H., Tong, C., Liu, J., Dong, L., Zheng, J., … Han, W. (2015). LPS-preconditioned mesenchymal stromal cells modify macrophage polarization for resolution of chronic inflammation via exosome-shuttled let-7b. Journal of Translational Medicine, 13(1). doi:10.1186/s12967-015-0642-6Firestein, G. S. (2003). Evolving concepts of rheumatoid arthritis. Nature, 423(6937), 356-361. doi:10.1038/nature01661Goronzy, J. J., & Weyand, C. M. (2009). Developments in the scientific understanding of rheumatoid arthritis. Arthritis Research & Therapy, 11(5), 249. doi:10.1186/ar2758Casado, J. G., Blázquez, R., Vela, F. J., Álvarez, V., Tarazona, R., & Sánchez-Margallo, F. M. (2017). Mesenchymal Stem Cell-Derived Exosomes: Immunomodulatory Evaluation in an Antigen-Induced Synovitis Porcine Model. Frontiers in Veterinary Science, 4. doi:10.3389/fvets.2017.00039Cosenza, S., Toupet, K., Maumus, M., Luz-Crawford, P., Blanc-Brude, O., Jorgensen, C., & Noël, D. (2018). Mesenchymal stem cells-derived exosomes are more immunosuppressive than microparticles in inflammatory arthritis. Theranostics, 8(5), 1399-1410. doi:10.7150/thno.21072Yang, Y., Hutchinson, P., & Morand, E. F. (1999). Inhibitory effect of annexin I on synovial inflammation in rat adjuvant arthritis. Arthritis & Rheumatism, 42(7), 1538-1544. doi:10.1002/1529-0131(199907)42:73.0.co;2-3Headland, S. E., Jones, H. R., Norling, L. V., Kim, A., Souza, P. R., Corsiero, E., … Perretti, M. (2015). Neutrophil-derived microvesicles enter cartilage and protect the joint in inflammatory arthritis. Science Translational Medicine, 7(315), 315ra190-315ra190. doi:10.1126/scitranslmed.aac5608Tofiño-Vian, M., Guillén, M. I., Pérez del Caz, M. D., Silvestre, A., & Alcaraz, M. J. (2018). Microvesicles from Human Adipose Tissue-Derived Mesenchymal Stem Cells as a New Protective Strategy in Osteoarthritic Chondrocytes. Cellular Physiology and Biochemistry, 47(1), 11-25. doi:10.1159/000489739Ando, Y., Matsubara, K., Ishikawa, J., Fujio, M., Shohara, R., Hibi, H., … Yamamoto, A. (2014). Stem cell-conditioned medium accelerates distraction osteogenesis through multiple regenerative mechanisms. Bone, 61, 82-90. doi:10.1016/j.bone.2013.12.029Lu, Z., Chen, Y., Dunstan, C., Roohani-Esfahani, S., & Zreiqat, H. (2017). Priming Adipose Stem Cells with Tumor Necrosis Factor-Alpha Preconditioning Potentiates Their Exosome Efficacy for Bone Regeneration. Tissue Engineering Part A, 23(21-22), 1212-1220. doi:10.1089/ten.tea.2016.0548Qin, Y., Wang, L., Gao, Z., Chen, G., & Zhang, C. (2016). Bone marrow stromal/stem cell-derived extracellular vesicles regulate osteoblast activity and differentiation in vitro and promote bone regeneration in vivo. Scientific Reports, 6(1). doi:10.1038/srep21961Wang, X., Omar, O., Vazirisani, F., Thomsen, P., & Ekström, K. (2018). Mesenchymal stem cell-derived exosomes have altered microRNA profiles and induce osteogenic differentiation depending on the stage of differentiation. PLOS ONE, 13(2), e0193059. doi:10.1371/journal.pone.0193059Shirley, D., Marsh, D., Jordan, G., McQuaid, S., & Li, G. (2005). Systemic recruitment of osteoblastic cells in fracture healing. Journal of Orthopaedic Research, 23(5), 1013-1021. doi:10.1016/j.orthres.2005.01.013Lee, D. Y., Cho, T.-J., Kim, J. A., Lee, H. R., Yoo, W. J., Chung, C. Y., & Choi, I. H. (2008). Mobilization of endothelial progenitor cells in fracture healing and distraction osteogenesis. Bone, 42(5), 932-941. doi:10.1016/j.bone.2008.01.007Furuta, T., Miyaki, S., Ishitobi, H., Ogura, T., Kato, Y., Kamei, N., … Ochi, M. (2016). Mesenchymal Stem Cell-Derived Exosomes Promote Fracture Healing in a Mouse Model. STEM CELLS Translational Medicine, 5(12), 1620-1630. doi:10.5966/sctm.2015-0285Li, W., Liu, Y., Zhang, P., Tang, Y., Zhou, M., Jiang, W., … Zhou, Y. (2018). Tissue-Engineered Bone Immobilized with Human Adipose Stem Cells-Derived Exosomes Promotes Bone Regeneration. ACS Applied Materials & Interfaces, 10(6), 5240-5254. doi:10.1021/acsami.7b17620Hirschi, K. K., Li, S., & Roy, K. (2014). Induced Pluripotent Stem Cells for Regenerative Medicine. Annual Review of Biomedical Engineering, 16(1), 277-294. doi:10.1146/annurev-bioeng-071813-105108Sabapathy, V., & Kumar, S. (2016). hi PSC ‐derived iMSC s: NextGen MSC s as an advanced therapeutically active cell resource for regenerative medicine. Journal of Cellular and Molecular Medicine, 20(8), 1571-1588. doi:10.1111/jcmm.12839Zhang, J., Liu, X., Li, H., Chen, C., Hu, B., Niu, X., … Wang, Y. (2016). Exosomes/tricalcium phosphate combination scaffolds can enhance bone regeneration by activating the PI3K/Akt signaling pathway. Stem Cell Research & Therapy, 7(1). doi:10.1186/s13287-016-0391-3Narayanan, R., Huang, C.-C., & Ravindran, S. (2016). Hijacking the Cellular Mail: Exosome Mediated Differentiation of Mesenchymal Stem Cells. Stem Cells International, 2016, 1-11. doi:10.1155/2016/3808674Zhu, Y., Jia, Y., Wang, Y., Xu, J., & Chai, Y. (2019). Impaired Bone Regenerative Effect of Exosomes Derived from Bone Marrow Mesenchymal Stem Cells in Type 1 Diabetes. STEM CELLS Translational Medicine, 8(6), 593-605. doi:10.1002/sctm.18-0199Ferreira, E., & Porter, R. M. (2018). Harnessing extracellular vesicles to direct endochondral repair of large bone defects. Bone & Joint Research, 7(4), 263-273. doi:10.1302/2046-3758.74.bjr-2018-0006Moya, A., Paquet, J., Deschepper, M., Larochette, N., Oudina, K., Denoeud, C., … Petite, H. (2018). Human Mesenchymal Stem Cell Failure to Adapt to Glucose Shortage and Rapidly Use Intracellular Energy Reserves Through Glycolysis Explains Poor Cell Survival After Implantation. STEM CELLS, 36(3), 363-376. doi:10.1002/stem.2763Moya, A., Larochette, N., Paquet, J., Deschepper, M., Bensidhoum, M., Izzo, V., … Logeart-Avramoglou, D. (2016). Quiescence Preconditioned Human Multipotent Stromal Cells Adopt a Metabolic Profile Favorable for Enhanced Survival under Ischemia. STEM CELLS, 35(1), 181-196. doi:10.1002/stem.2493Potier, E., Ferreira, E., Andriamanalijaona, R., Pujol, J.-P., Oudina, K., Logeart-Avramoglou, D., & Petite, H. (2007). Hypoxia affects mesenchymal stromal cell osteogenic differentiation and angiogenic factor expression. Bone, 40(4), 1078-1087. doi:10.1016/j.bone.2006.11.024Jia, Y., Zhu, Y., Qiu, S., Xu, J., & Chai, Y. (2019). Exosomes secreted by endothelial progenitor cells accelerate bone regeneration during distraction osteogenesis by stimulating angiogenesis. Stem Cell Research & Therapy, 10(1). doi:10.1186/s13287-018-1115-7Zhang, Y., Hao, Z., Wang, P., Xia, Y., Wu, J., Xia, D., … Xu, S. (2019). Exosomes from human umbilical cord mesenchymal stem cells enhance fracture healing through HIF‐1α‐mediated promotion of angiogenesis in a rat model of stabilized fracture. Cell Proliferation, 52(2), e12570. doi:10.1111/cpr.12570Qi, X., Zhang, J., Yuan, H., Xu, Z., Li, Q., Niu, X., … Li, X. (2016). Exosomes Secreted by Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells Repair Critical-Sized Bone De

Extracellular vesicles: A new therapeutic strategy for joint conditions

[EN] Extracellular vesicles (EVs) are attracting increasing interest since they might represent a more convenient therapeutic tool with respect to their cells of origin. In the last years much time and effort have been expended to determine the biological properties of EVs from mesenchymal stem cells (MSCs) and other sources. The immunoregulatory, anti-inflammatory and regenerative properties of MSC EVs have been demonstrated in in vitro studies and animal models of rheumatoid arthritis or osteoarthritis. This cell-free approach has been proposed as a possible better alternative to MSC therapy in autoimmune conditions and tissue regeneration. In addition, EVs show great potential as biomarkers of disease or delivery systems for active molecules. The standardization of isolation and characterization methods is a key step for the development of EV research. A better understanding of EV mechanisms of action and efficacy is required to establish the potential therapeutic applications of this new approach in joint conditions.This work has been funded by grants SAF2013-48724R (MINECO,FEDER, Spain) and PROMETEOII/2014/071 (Generalitat Valenciana, Spain).Tofiño-Vian, M.; Guillen Salazar, MI.; Alcaraz Tormo, MJ. (2018). Extracellular vesicles: A new therapeutic strategy for joint conditions. Biochemical Pharmacology. 153:134-146. https://doi.org/10.1016/j.bcp.2018.02.004S13414615

Nrf2 as a therapeutic target for rheumatic diseases

[EN] Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a master regulator of cellular protective processes. Rheumatic diseases are chronic conditions characterized by inflammation, pain, tissue damage and limitations in function. Main examples are rheumatoid arthritis, systemic lupus erythematosus, osteoarthritis and osteoporosis. Their high prevalence constitutes a major health problem with an important social and economic impact. A wide range of evidence indicates that Nrf2 may control different mechanisms involved in the physiopathology of rheumatic conditions. Therefore, the appropriate expression and balance of Nrf2 is necessary for regulation of oxidative stress, inflammation, immune responses, and cartilage and bone metabolism. Numerous studies have demonstrated that Nrf2 deficiency aggravates the disease in experimental models while Nrf2 activation results in immunoregulatory and anti-inflammatory effects. These reports reinforce the increasing interest in the pharmacologic regulation of Nrf2 and its potential applications. Nevertheless, a majority of Nrf2 inducers are electrophilic molecules which may present off-target effects. In recent years, novel strategies have been sought to modulate the Nrf2 pathway which has emerged as a therapeutic target in rheumatic conditions.This work has been funded by grant SAF2017-85806-R (MINECO, FEDER, Spain).Ferrandiz Manglano, ML.; Nacher-Juan, J.; Alcaraz Tormo, MJ. (2018). Nrf2 as a therapeutic target for rheumatic diseases. Biochemical Pharmacology. 152:338-346. https://doi.org/10.1016/j.bcp.2018.04.010S33834615

Myeloid Heme Oxygenase-1 Regulates the Acute Inflammatory Response to Zymosan in the Mouse Air Pouch

[EN] Heme oxygenase-1 (HO-1) is induced by many stimuli to modulate the activation and function of different cell types during innate immune responses. Although HO-1 has shown anti-inflammatory effects in different systems, there are few data on the contribution of myeloid HO-1 and its role in inflammatory processes is not well understood. To address this point, we have used HO-1(M-KO) mice with myeloid-restricted deletion of HO-1 to specifically investigate its influence on the acute inflammatory response to zymosan in vivo. In the mouse air pouch model, we have shown an exacerbated inflammation in HO-1(M-KO) mice with increased neutrophil infiltration accompanied by high levels of inflammatory mediators such as interleukin-1 beta, tumor necrosis factor-a, and prostaglandin E-2. The expression of the degradative enzyme matrix metalloproteinase-3 (MMP-3) was also enhanced. In addition, we observed higher levels of serum MMP-3 in HO-1(M-KO) mice compared with control mice, suggesting the presence of systemic inflammation. Altogether, these findings demonstrate that myeloid HO-1 plays an anti-inflammatory role in the acute response to zymosan in vivo and suggest the interest of this target to regulate inflammatory processes.This work has been funded by grants SAF2010-22048, SAF2013-4874R (MINECO, FEDER), PROMETEO/2010/047, and PROMETEOII/2014/071 (Generalitat Valenciana).Brines, R.; Catalán, L.; Alcaraz Tormo, MJ.; Ferrandiz Manglano, ML. (2018). Myeloid Heme Oxygenase-1 Regulates the Acute Inflammatory Response to Zymosan in the Mouse Air Pouch. Oxidative Medicine and Cellular Longevity. 2018. https://doi.org/10.1155/2018/5053091S2018Grochot-Przeczek, A., Dulak, J., & Jozkowicz, A. (2011). Haem oxygenase-1: non-canonical roles in physiology and pathology. Clinical Science, 122(3), 93-103. doi:10.1042/cs20110147Busserolles, J., Megías, J., Terencio, M. C., & Alcaraz, M. J. (2006). Heme oxygenase-1 inhibits apoptosis in Caco-2 cells via activation of Akt pathway. The International Journal of Biochemistry & Cell Biology, 38(9), 1510-1517. doi:10.1016/j.biocel.2006.03.013Devesa, I., Ferrándiz, M. L., Terencio, M. C., Joosten, L. A. B., van den Berg, W. B., & Alcaraz, M. J. (2005). Influence of heme oxygenase 1 modulation on the progression of murine collagen-induced arthritis. Arthritis & Rheumatism, 52(10), 3230-3238. doi:10.1002/art.21356García-Arnandis, I., Guillén, M. I., Castejón, M. A., Gomar, F., & Alcaraz, M. J. (2010). Haem oxygenase-1 down-regulates high mobility group box 1 and matrix metalloproteinases in osteoarthritic synoviocytes. Rheumatology, 49(5), 854-861. doi:10.1093/rheumatology/kep463Clérigues, V., Guillén, M. I., Castejón, M. A., Gomar, F., Mirabet, V., & Alcaraz, M. J. (2012). Heme oxygenase-1 mediates protective effects on inflammatory, catabolic and senescence responses induced by interleukin-1β in osteoarthritic osteoblasts. Biochemical Pharmacology, 83(3), 395-405. doi:10.1016/j.bcp.2011.11.024Gozzelino, R., Jeney, V., & Soares, M. P. (2010). Mechanisms of Cell Protection by Heme Oxygenase-1. Annual Review of Pharmacology and Toxicology, 50(1), 323-354. doi:10.1146/annurev.pharmtox.010909.105600Naito, Y., Takagi, T., & Higashimura, Y. (2014). Heme oxygenase-1 and anti-inflammatory M2 macrophages. Archives of Biochemistry and Biophysics, 564, 83-88. doi:10.1016/j.abb.2014.09.005Orozco, L. D., Kapturczak, M. H., Barajas, B., Wang, X., Weinstein, M. M., Wong, J., … Araujo, J. A. (2007). Heme Oxygenase-1 Expression in Macrophages Plays a Beneficial Role in Atherosclerosis. Circulation Research, 100(12), 1703-1711. doi:10.1161/circresaha.107.151720Perrella, M., & Yet, S.-F. (2003). Role of Heme Oxygenase-1 in Cardiovascular Function. Current Pharmaceutical Design, 9(30), 2479-2487. doi:10.2174/1381612033453776Abraham, N. G., Li, M., Vanella, L., Peterson, S. J., Ikehara, S., & Asprinio, D. (2008). Bone marrow stem cell transplant into intra-bone cavity prevents type 2 diabetes: Role of heme oxygenase-adiponectin. Journal of Autoimmunity, 30(3), 128-135. doi:10.1016/j.jaut.2007.12.005Naito, Y., Takagi, T., & Yoshikawa, T. (2004). Heme oxygenase-1: a new therapeutic target for inflammatory bowel disease. Alimentary Pharmacology & Therapeutics, 20, 177-184. doi:10.1111/j.1365-2036.2004.01992.xBrines, R., Maicas, N., Ferrándiz, M. L., Loboda, A., Jozkowicz, A., Dulak, J., & Alcaraz, M. J. (2012). Heme Oxygenase-1 Regulates the Progression of K/BxN Serum Transfer Arthritis. PLoS ONE, 7(12), e52435. doi:10.1371/journal.pone.0052435Poss, K. D., & Tonegawa, S. (1997). Heme oxygenase 1 is required for mammalian iron reutilization. Proceedings of the National Academy of Sciences, 94(20), 10919-10924. doi:10.1073/pnas.94.20.10919Yachie, A., Niida, Y., Wada, T., Igarashi, N., Kaneda, H., Toma, T., … Koizumi, S. (1999). Oxidative stress causes enhanced endothelial cell injury in human heme oxygenase-1 deficiency. Journal of Clinical Investigation, 103(1), 129-135. doi:10.1172/jci4165Radhakrishnan, N., Yadav, S. P., Sachdeva, A., Pruthi, P. K., Sawhney, S., Piplani, T., … Yachie, A. (2011). Human Heme Oxygenase-1 Deficiency Presenting With Hemolysis, Nephritis, and Asplenia. Journal of Pediatric Hematology/Oncology, 33(1), 74-78. doi:10.1097/mph.0b013e3181fd2aaeTzima, S., Victoratos, P., Kranidioti, K., Alexiou, M., & Kollias, G. (2009). Myeloid heme oxygenase–1 regulates innate immunity and autoimmunity by modulating IFN-β production. Journal of Experimental Medicine, 206(5), 1167-1179. doi:10.1084/jem.20081582Rossi, M., Thierry, A., Delbauve, S., Preyat, N., Soares, M. P., Roumeguère, T., … Hougardy, J.-M. (2017). Specific expression of heme oxygenase-1 by myeloid cells modulates renal ischemia-reperfusion injury. Scientific Reports, 7(1). doi:10.1038/s41598-017-00220-wJais, A., Einwallner, E., Sharif, O., Gossens, K., Lu, T. T.-H., Soyal, S. M., … Esterbauer, H. (2014). Heme Oxygenase-1 Drives Metaflammation and Insulin Resistance in Mouse and Man. Cell, 158(1), 25-40. doi:10.1016/j.cell.2014.04.043Konrad, F. M., Knausberg, U., Höne, R., Ngamsri, K.-C., & Reutershan, J. (2015). Tissue heme oxygenase-1 exerts anti-inflammatory effects on LPS-induced pulmonary inflammation. Mucosal Immunology, 9(1), 98-111. doi:10.1038/mi.2015.39Underhill, D. M. (2003). Macrophage recognition of zymosan particles. Journal of Endotoxin Research, 9(3), 176-180. doi:10.1177/09680519030090030601MORONEY, M.-A., ALCARAZ, M. J., FORDER, R. A., CAREY, F., & HOULT, J. R. S. (1988). Selectivity of Neutrophil 5-Lipoxygenase and Cyclo-oxygenase Inhibition by an Anti-inflammatory Flavonoid Glycoside and Related Aglycone Flavonoids. Journal of Pharmacy and Pharmacology, 40(11), 787-792. doi:10.1111/j.2042-7158.1988.tb05173.xClausen, B. E., Burkhardt, C., Reith, W., Renkawitz, R., & Förster, I. (1999). Transgenic Research, 8(4), 265-277. doi:10.1023/a:1008942828960Posadas, I., Terencio, M. C., Guillén, I., Ferrándiz, M. L., Coloma, J., Payá, M., & Alcaraz, M. J. (2000). Co-regulation between cyclo-oxygenase-2 and inducible nitric oxide synthase expression in the time-course of murine inflammation. Naunyn-Schmiedeberg’s Archives of Pharmacology, 361(1), 98-106. doi:10.1007/s002109900150Vicente, A. M., Guillén, M. I., Habib, A., & Alcaraz, M. J. (2003). Beneficial Effects of Heme Oxygenase-1 Up-Regulation in the Development of Experimental Inflammation Induced by Zymosan. Journal of Pharmacology and Experimental Therapeutics, 307(3), 1030-1037. doi:10.1124/jpet.103.057992Galli, S. J., Borregaard, N., & Wynn, T. A. (2011). Phenotypic and functional plasticity of cells of innate immunity: macrophages, mast cells and neutrophils. Nature Immunology, 12(11), 1035-1044. doi:10.1038/ni.2109Chung, S. W., Liu, X., Macias, A. A., Baron, R. M., & Perrella, M. A. (2008). Heme oxygenase-1–derived carbon monoxide enhances the host defense response to microbial sepsis in mice. Journal of Clinical Investigation, 118(1), 239-247. doi:10.1172/jci32730Chiang, N., Shinohara, M., Dalli, J., Mirakaj, V., Kibi, M., Choi, A. M. K., & Serhan, C. N. (2013). Inhaled Carbon Monoxide Accelerates Resolution of Inflammation via Unique Proresolving Mediator–Heme Oxygenase-1 Circuits. The Journal of Immunology, 190(12), 6378-6388. doi:10.4049/jimmunol.1202969Murphy, G., Cockett, M. I., Stephens, P. E., Smith, B. J., & Docherty, A. J. P. (1987). Stromelysin is an activator of procollagenase. A study with natural and recombinant enzymes. Biochemical Journal, 248(1), 265-268. doi:10.1042/bj2480265Steenport, M., Khan, K. M. F., Du, B., Barnhard, S. E., Dannenberg, A. J., & Falcone, D. J. (2009). Matrix Metalloproteinase (MMP)-1 and MMP-3 Induce Macrophage MMP-9: Evidence for the Role of TNF-α and Cyclooxygenase-2. The Journal of Immunology, 183(12), 8119-8127. doi:10.4049/jimmunol.0901925Blom, A. B., van Lent, P. L., Libregts, S., Holthuysen, A. E., van der Kraan, P. M., van Rooijen, N., & van den Berg, W. B. (2006). Crucial role of macrophages in matrix metalloproteinase–mediated cartilage destruction during experimental osteoarthritis : Involvement of matrix metalloproteinase 3. Arthritis & Rheumatism, 56(1), 147-157. doi:10.1002/art.22337Houseman, M., Potter, C., Marshall, N., Lakey, R., Cawston, T., Griffiths, I., … Isaacs, J. D. (2012). Baseline serum MMP-3 levels in patients with Rheumatoid Arthritis are still independently predictive of radiographic progression in a longitudinal observational cohort at 8 years follow up. Arthritis Research & Therapy, 14(1), R30. doi:10.1186/ar3734Green, M. J. (2003). Serum MMP-3 and MMP-1 and progression of joint damage in early rheumatoid arthritis. Rheumatology, 42(1), 83-88. doi:10.1093/rheumatology/keg03

Microvesicles from Human Adipose Tissue-Derived Mesenchymal Stem Cells as a New Protective Strategy in Osteoarthritic Chondrocytes

[EN] Background/Aims: Chronic inflammation contributes to cartilage degeneration during the progression of osteoarthritis (OA). Adipose tissue-derived mesenchymal stem cells (ADMSC) show great potential to treat inflammatory and degradative processes in OA and have demonstrated paracrine effects in chondrocytes. In the present work, we have isolated and characterized the extracellular vesicles from human AD-MSC to investigate their role in the chondroprotective actions of these cells. Methods: AD-MSC were isolated by collagenase treatment from adipose tissue from healthy individuals subjected to abdominal lipectomy surgery. Microvesicles and exosomes were obtained from conditioned medium by filtration and differential centrifugation. Chondrocytes from OA patients were used in primary culture and stimulated with 10 ng/ml interleukin(IL)-1 beta in the presence or absence of AD-MSC microvesicles, exosomes or conditioned medium. Protein expression was investigated by ELISA and immunofluorescence, transcription factor-DNA binding by ELISA, gene expression by real-time PCR, prostaglandin E-2 (PGE(2)) by radioimmunoassay, and matrix metalloproteinase (MMP) activity and nitric oxide (NO) production by fluorometry. Results: In OA chondrocytes stimulated with IL-1 beta, microvesicles and exosomes reduced the production of inflammatory mediators tumor necrosis factor-alpha, IL-6, PGE(2) and NO. The downregulation of cyclooxygenase-2 and microsomal prostaglandin E synthase-1 would lead to the decreased PGE(2) production while the effect on NO could depend on the reduction of inducible nitric oxide synthase expression. Treatment of OA chondrocytes with extracellular vesicles also decreased the release of MMP activity and MMP-13 expression whereas the production of the anti-inflammatory cytokine IL-10 and the expression of collagen II were significantly enhanced. The reduction of inflammatory and catabolic mediators could be the consequence of a lower activation of nuclear factor-kappa B and activator protein-1. The upregulation of annexin A1 specially in MV may contribute to the anti-inflammatory and chondroprotective effects of AD-MSC. Conclusions: Our data support the interest of AD-MSC extracellular vesicles to develop new therapeutic approaches in joint conditions. (C) 2018 The Author(s) Published by S. Karger AG, BaselThis work was supported by grants SAF2013-4874R (MINECO, FEDER) and PROMETEOII/2014/071 (Generalitat Valenciana), Spain.Tofiño-Vian, M.; Guillen Salazar, MI.; Perez Del Caz, M.; Silvestre, A.; Alcaraz Tormo, MJ. (2018). Microvesicles from Human Adipose Tissue-Derived Mesenchymal Stem Cells as a New Protective Strategy in Osteoarthritic Chondrocytes. Cellular Physiology and Biochemistry. 47(1):11-25. https://doi.org/10.1159/000489739S112547

Targeting inflammasome by the inhibition of caspase-1 activity using capped mesoporous silica nanoparticles

[EN] Acute inflammation is a protective response of the body to harmful stimuli, such as pathogens or damaged cells. However, dysregulated inflammation can cause secondary damage and could thus contribute to the pathophysiology of many diseases. Inflammasomes, the macromolecular complexes responsible for caspase-1 activation, have emerged as key regulators of immune and inflammatory responses. Therefore, modulation of inflammasome activity has become an important therapeutic approach. Here we describe the design of a smart nanodevice that takes advantage of the passive targeting of nanoparticles to macrophages and enhances the therapeutic effect of caspase-1 inhibitor VX-765 in vivo. The functional hybrid systems consisted of MCM-41-based nanoparticles loaded with anti-inflammatory drug VX-765 (S2-P) and capped with poly-L-lysine, which acts as a molecular gate. S2-P activity has been evaluated in cellular and in vivo models of inflammation. The results indicated the potential advantage of using nanodevices to treat inflammatory diseases. (C) 2017 Elsevier B.V. All rights reserved.The authors wish to express their gratitude to the Spanish government (Projects MAT2015-64139-C4-1-R and SAF2014-52614-R (MINECO/FEDER)) and the Generalitat Valencia (Projects PROMETEOII/2014/061 and PROMETEOII/2014/047) for support. A.G-F. is grateful to the Spanish government for an FPU grant.García-Fernández, A.; García-Laínez, G.; Ferrandiz Manglano, ML.; Aznar, E.; Sancenón Galarza, F.; Alcaraz, MJ.; Murguía, JR.... (2017). Targeting inflammasome by the inhibition of caspase-1 activity using capped mesoporous silica nanoparticles. Journal of Controlled Release. 248:60-70. https://doi.org/10.1016/j.jconrel.2017.01.002S607024

Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts

[EN] Osteoarthritis (OA) affects all articular tissues leading to pain and disability. The dysregulation of bone metabolism may contribute to the progression of this condition. Adipose-derived mesenchymal stem cells (ASC) are attractive candidates in the search of novel strategies for OA treatment and exert anti-inflammatory and cytoprotective effects on cartilage. Chronic inflammation in OA is a relevant factor in the development of cellular senescence and joint degradation. In this study, we extend our previous observations of ASC paracrine effects to study the influence of conditioned medium and extracellular vesicles from ASC on senescence induced by inflammatory stress in OA osteoblasts. Our results in cells stimulated with interleukin- (IL-) 1 beta indicate that conditioned medium, microvesicles, and exosomes from ASC downregulate senescence-associated beta-galactosidase activity and the accumulation of gamma H2AX foci. In addition, they reduced the production of inflammatory mediators, with the highest effect on IL6 and prostaglandin E-2. The control of mitochondrial membrane alterations and oxidative stress may provide a mechanism for the protective effects of ASC in OA osteoblasts. We have also shown that microvesicles and exosomes mediate the paracrine effects of ASC. Our study suggests that correction of abnormal osteoblast metabolism by ASC products may contribute to their protective effects.This work has been funded by Grants SAF2013-48724-R (MINECO/FEDER) and PROMETEOII/2014/071 (Generalitat Valenciana).Tofiño, M.; Guillen Salazar, MI.; Perez Del Caz, M.; Castejon, M.; Alcaraz Tormo, MJ. (2017). Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts. Oxidative Medicine and Cellular Longevity. https://doi.org/10.1155/2017/7197598

Contemporary use of cefazolin for MSSA infective endocarditis: analysis of a national prospective cohort

Objectives: This study aimed to assess the real use of cefazolin for methicillin-susceptible Staphylococcus aureus (MSSA) infective endocarditis (IE) in the Spanish National Endocarditis Database (GAMES) and to compare it with antistaphylococcal penicillin (ASP). Methods: Prospective cohort study with retrospective analysis of a cohort of MSSA IE treated with cloxacillin and/or cefazolin. Outcomes assessed were relapse; intra-hospital, overall, and endocarditis-related mortality; and adverse events. Risk of renal toxicity with each treatment was evaluated separately. Results: We included 631 IE episodes caused by MSSA treated with cloxacillin and/or cefazolin. Antibiotic treatment was cloxacillin, cefazolin, or both in 537 (85%), 57 (9%), and 37 (6%) episodes, respectively. Patients treated with cefazolin had significantly higher rates of comorbidities (median Charlson Index 7, P <0.01) and previous renal failure (57.9%, P <0.01). Patients treated with cloxacillin presented higher rates of septic shock (25%, P = 0.033) and new-onset or worsening renal failure (47.3%, P = 0.024) with significantly higher rates of in-hospital mortality (38.5%, P = 0.017). One-year IE-related mortality and rate of relapses were similar between treatment groups. None of the treatments were identified as risk or protective factors. Conclusion: Our results suggest that cefazolin is a valuable option for the treatment of MSSA IE, without differences in 1-year mortality or relapses compared with cloxacillin, and might be considered equally effective