Biomaterials Research (생체재료학회지)

Korean Society for Biomaterials (한국생체재료학회)
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Hypoxic condition enhances chondrogenesis in synovium-derived mesenchymal stem cells

  • Bae, Hyun Cheol (Department of Orthopedic Surgery, Seoul National University Hospital) ;
  • Park, Hee Jung (Department of Orthopedic Surgery, Seoul National University Hospital) ;
  • Wang, Sun Young (Department of Orthopedic Surgery, Seoul National University Hospital) ;
  • Yang, Ha Ru (Department of Orthopedic Surgery, Seoul National University Hospital) ;
  • Lee, Myung Chul (Department of Orthopedic Surgery, Seoul National University Hospital) ;
  • Han, Hyuk-Soo (Department of Orthopedic Surgery, Seoul National University Hospital)
  • Received : 2018.06.04
  • Accepted : 2018.08.29
  • Published : 2018.12.31
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Abstract

Background: The chondrogenic differentiation of mesenchymal stem cells (MSCs) is regulated by many factors, including oxygen tensions, growth factors, and cytokines. Evidences have suggested that low oxygen tension seems to be an important regulatory factor in the proliferation and chondrogenic differentiation in various MSCs. Recent studies report that synovium-derived mesenchymal stem cells (SDSCs) are a potential source of stem cells for the repair of articular cartilage defects. But, the effect of low oxygen tension on the proliferation and chondrogenic differentiation in SDSCs has not characterized. In this study, we investigated the effects of hypoxia on proliferation and chondrogenesis in SDSCs. Method: SDSCs were isolated from patients with osteoarthritis at total knee replacement. To determine the effect of oxygen tension on proliferation and colony-forming characteristics of SDSCs, A colony-forming unit (CFU) assay and cell counting-based proliferation assay were performed under normoxic (21% oxygen) or hypoxic (5% oxygen). For in vitro chondrogenic differentiation, SDSCs were concentrated to form pellets and subjected to conditions appropriate for chondrogenic differentiation under normoxia and hypoxia, followed by the analysis for the expression of genes and proteins of chondrogenesis. qRT-PCR, histological assay, and glycosoaminoglycan assays were determined to assess chondrogenesis. Results: Low oxygen condition significantly increased proliferation and colony-forming characteristics of SDSCs compared to that of SDSCs under normoxic culture. Similar pellet size and weight were found for chondrogensis period under hypoxia and normoxia condition. The mRNA expression of types II collagen, aggrecan, and the transcription factor SOX9 was increased under hypoxia condition. Histological sections stained with Safranin-O demonstrated that hypoxic conditions had increased proteoglycan synthesis. Immunohistochemistry for types II collagen demonstrated that hypoxic culture of SDSCs increased type II collagen expression. In addition, GAG deposition was significantly higher in hypoxia compared with normoxia at 21 days of differentiation. Conclusion: These findings show that hypoxia condition has an important role in regulating the synthesis ECM matrix by SDSCs as they undergo chondrogenesis. This has important implications for cartilage tissue engineering applications of SDSCs.

Keywords

Acknowledgement

Supported by : Seoul National University Hospital

References

  1. Pastides P, Chimutengwende-Gordon M, Maffulli N, Khan W. Stem cell therapy for human cartilage defects: a systematic review. Osteoarthr Cartil. 2013;21:646-54. https://doi.org/10.1016/j.joca.2013.02.008
  2. Bornes TD, Adesida AB, Jomha NM. Mesenchymal stem cells in the treatment of traumatic articular cartilage defects: a comprehensive review. Arthritis Res Ther. 2014;16:432. https://doi.org/10.1186/s13075-014-0432-1
  3. Sakaguchi Y, Sekiya I, Yagishita K, Muneta T. Comparison of human stem cells derived from variousmesenchymal tissues: superiority of synovium as a cell source. Arthritis Rheum. 2005;52:2521. https://doi.org/10.1002/art.21212
  4. Kubosch EJ, Lang G, Furst D, Kubosch D, Izadpanah K, Rolauffs B, Sudkamp NP, Schmal H. The potential for synovium-derived stem cells in cartilage repair. Curr Stem Cell Res Ther. 2018;13:174-84. https://doi.org/10.2174/1574888X12666171002111026
  5. Yusuke O, Yo M, Mayu Y, Eriko GS, Nobuharu S, Takeshi M, Ichiro S, Chihiro A. Purified human synovium mesenchymal stem cells as a good resource for cartilage regeneration. PLoS One. 2015;10:e129096.
  6. Danisovica L, Varga I, Polakc S. Growth factors and chondrogenic differentiation of mesenchymal stem cells. Tissue Cell. 2012;44:69-73. https://doi.org/10.1016/j.tice.2011.11.005
  7. Rodrigo AS, Jean FW, Diego C, Arnold IC. Chondrogenic differentiation of mesenchymal stem cells: challenges and unfulfilled expectations. Tissue Eng Part B Rev. 2014;20:596-608. https://doi.org/10.1089/ten.teb.2013.0771
  8. Chiou SH, et al. Identification of CD133-positive radioresistant cells in atypical teratoid/rhabdoid tumor. PLoS One. 2008;3:e2090. https://doi.org/10.1371/journal.pone.0002090
  9. Das R, et al. The role of hypoxia in bone marrow-derived mesenchymal stem cells: considerations for regenerative medicine approaches. Tissue Eng Part B Rev. 2010;16:159. https://doi.org/10.1089/ten.teb.2009.0296
  10. Kim DS, et al. Effect of low oxygen tension on the biological characteristics of human bone marrow mesenchymal stem cells. Cell Stress Chaperones. 2016;21:1089-99. https://doi.org/10.1007/s12192-016-0733-1
  11. Paquet J, Deschepper M, Moya A, Logeart-Avramoglou D, Boisson-Vidal C, Petite H. Oxygen tension regulates human mesenchymal stem cell paracrine functions. Stem Cells Transl Med. 2015;4:809-21. https://doi.org/10.5966/sctm.2014-0180
  12. Roy S, Tripathy M, Mathur N, Jain A, Mukhopadhyay A. Hypoxia improves expansion potential of human cord blood-derived hematopoietic stem cells and marrow repopulation efficiency. Eur J Haematol. 2012;88:396-405. https://doi.org/10.1111/j.1600-0609.2012.01759.x
  13. Lafont JE. Lack of oxygen in articular cartilage: consequences for chondrocyte biology. Int J Exp Pathol. 2010;91:99-106. https://doi.org/10.1111/j.1365-2613.2010.00707.x
  14. Fermor B, Christensen SE, Youn I, Cernanec JM, Davies CM, Weinberg JB. Oxygen, nitric oxide and articular cartilage. Eur Cell Mater. 2007;13:56-65. https://doi.org/10.22203/eCM.v013a06
  15. Lafont JE, Talma S, Murphy CL. Hypoxia-inducible factor 2alpha is essential for hypoxic induction of the human articular chondrocyte phenotype. Arthritis Rheum. 2007;56:3297-306. https://doi.org/10.1002/art.22878
  16. Shang J, Liu H, Li J, Zhou Y, Fermor B, Gimble JM, Awad HA, Guilak F. Roles of hypoxia during the chondrogenic differentiation of mesenchymal stem cells. Curr Stem Cell Res Ther. 2014;9:141-7. https://doi.org/10.2174/1574888X09666131230142459
  17. Scherer K, Schunke M, Sellckau R, Hassenpflug J, Kurz B. The influence of oxygen and hydrostatic pressure on articular chondrocytes and adherent bone marrow cells in vitro. Biorheology. 2004;41:323-33.
  18. Lee S, Kim JH, Jo CH, Seong SC, Lee JC, Lee MC. Effect of serum and growth factors on chondrogenic differentiation of synoviumderived stromal cells. Tissue Eng Part A. 2009;15(11):3401-15. https://doi.org/10.1089/ten.tea.2008.0466
  19. Hunziker EB. Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. Osteoarthr Cartil. 2002;10:432. https://doi.org/10.1053/joca.2002.0801
  20. Lee JW, Kim YH, Kim SH, Han SH, Hahn SB. Chondrogenic differentiation of mesenchymal stem cells and its clinical applications. Yonsei Med J. 2004;45(Suppl):41-7. https://doi.org/10.3349/ymj.2004.45.Suppl.41
  21. Kadiyala S, Young RG, Thiede MA, Bruder SP. Culture expanded canine mesenchymal stem cells possess osteochondrogenic potential in vivo and in vitro. Cell Transplant. 1997;6(2):125-34. https://doi.org/10.1177/096368979700600206
  22. Meirelles Lda S, Nardi NB. Murine marrow-derived mesenchymal stem cell: isolation, in vitro expansion, and characterization. Br J Haematol. 2003;123(4):702-11. https://doi.org/10.1046/j.1365-2141.2003.04669.x
  23. Fan J, Varshney RR, Ren L, Cai D, Wang DA. Synovium-derived mesenchymal stem cells: a new cell sourcefor musculoskeletal regeneration. Tissue Eng Part B Rev. 2009;15:75.
  24. Archer CW, Dowthwaite GP, Francis-West P. Development of synovial joints. Birth Defects Res C Embryo Today. 2003;69:144. https://doi.org/10.1002/bdrc.10015
  25. Kim JH, Lee MC, Seong SC, Park KH, Lee S. Enhanced proliferation and chondrogenic differentiation of human synovium-derived stem cells expanded with basic fibroblast growth factor. Tissue Eng Part A. 2011;17:991. https://doi.org/10.1089/ten.tea.2010.0277
  26. Kanichai M, Ferguson D, Prendergast PJ, Campbell VA. Hypoxia promotes chondrogenesis in rat mesenchymal stem cells: a role for AKT and hypoxiainducible factor (HIF)-1alpha. J Cell Physiol. 2008;216(3):708-15. https://doi.org/10.1002/jcp.21446
  27. Khan WS, Adesida AB, Tew SR, Lowe ET, Hardingham TE. Bone marrowderived mesenchymal stem cells express the pericyte marker 3G5 in culture and show enhanced chondrogenesis in hypoxic conditions. J Orthop Res. 2010;28(6):834-40. https://doi.org/10.1002/jor.21043
  28. Khan WS, Adesida AB, Hardingham TE. Hypoxic conditions increase hypoxiainducible transcription factor $2{\alpha}$ and enhance chondrogenesis in stem cells from the infrapatellar fat pad of osteoarthritis patients. Arthritis Research & Therapy. 2007;9:R55. https://doi.org/10.1186/ar2211
  29. Baumgartner L, Arnhold S, Brixius K, Addicks K, Bloch W. Human mesenchymal stem cells: influence of oxygen pressure on proliferation and chondrogenic differentiation in fibrin glue in vitro. J Biomed Mater Res A. 2010;93(3):930-40.
  30. Werb Z, Chin JR. Extracellular matrix remodelling during morphogenesis. Ann N Y Acad Sci. 1998;857:110-8. https://doi.org/10.1111/j.1749-6632.1998.tb10111.x
  31. Hofbauer K, Gess B, Lohaus C, Meyer H, Katschinski D, Kurtz A. Oxygen tension regulates the expression of a group of procollagen hydroxylases. Eur J Biochem. 2003;270:4515-22. https://doi.org/10.1046/j.1432-1033.2003.03846.x
  32. Takahashi Y, Takahashi S, Shiga Y, Yoshimi T, Miura T. Hypoxic induction of prolyl 4-hydroxylase alpha (I) in cultured cells. J Biol Chem. 2000;275:14139-46. https://doi.org/10.1074/jbc.275.19.14139
  33. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the ${\Delta}{\Delta}CT$ method. Methods. 2001;25:402-8. https://doi.org/10.1006/meth.2001.1262
  34. Shang J, Liu H, Li J, Zhou Y. Roles of hypoxia during the Chondrogenic differentiation of mesenchymal stem cells. Methods. 2014;9:141-7.
  35. Han H-S, Lee S, Kim JH, Seong SC, Lee MC. Changes in Chondrogenic phenotype and gene expression profiles associated with the in vitro expansion of human synovium-derived cells. J Orthop Res. 2010;28:1283-91. https://doi.org/10.1002/jor.21129

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