Comparison between Chondrogenic Markers of Differentiated Chondrocytes from Adipose Derived Stem Cells and Articular Chondrocytes In Vitro

Document Type : Original Article

Authors

1 Department of Anatomical Sciences and Molecular Biology, Medical School, Esfahan University of Medical Sciences, Esfahan, Islamic Republic of Iran

2 Department of Immunology, Medical School, Esfahan University of Medical Sciences, Esfahan, Esfahan, Islamic Republic of Iran

3 Amin Hospital, Medical School, Esfahan University of Medical Sciences, Esfahan, Esfahan, Islamic Republic of Iran

4 Department of Immunology, Medical School, Esfahan University of Medical Sciences, Esfahan, Islamic Republic of Iran

Abstract

 

Objective(s):
Osteoarthritis is one of the most common diseases in middle-aged population in the world. Cartilage tissue engineering (TE) has been presented as an effort to introduce the best combination of cells, biomaterial scaffolds and stimulating growth factors to produce a cartilage tissue similar to the natural articular cartilage. In this study, the chondrogenic potential of adipose derived stem cells (ADSCs) was compared with natural articular chondrocytes cultured in alginate scaffold.
 
Materials and Methods:
Human ADSCs were obtained from subcutaneous adipose tissue and human articular chondrocytes from non-weight bearing areas of knee joints. Cells were seeded in 1.5% alginate and cultured in chondrogenic media for three weeks with and without TGFβ3. The genes expression of types II and X collagens was assessed by Real Time PCR and the amount of aggrecan (AGC) and type I collagen measured by ELISA and the content of glycosaminoglycan evaluated by GAG assay.
Results:
Our findings showed that type II collagen, GAG and AGC were expressed, in differentiated ADSCs. Meanwhile, they produced a lesser amount of types II and X collagens but more AGC, GAG and type I collagen in comparison with natural chondrocytes (NCs).
Conclusion:
Further attempt should be carried out to optimize achieving type II collagen in DCs, as much as, natural articular chondrocytes and decline of the production of type I collagen in order to provide efficient hyaline cartilage after chondrogenic induction, prior to the usage of harvested tissues in clinical trials.

Keywords

References

1. Csaki C, Matis U, Mobasheri A, Ye H, Shakibaei M. Chondrogenesis, osteogenesis and adipogenesis of canine mesenchymal stem cells: a biochemical, morphological and ultrastructural study. Histochem Cell Biol 2007; 128: 507–520.
2. Buckwalter JA, Mankin HJ. Articular cartilage: degeneration and osteoarthritis, repair, regeneration and transplantation. Instr Course Lect 1998; 47:487-504.
3. Mano JF, Reis RL. Osteochondral defects: present situation and tissue engineering approaches. J Tissue Eng Regen Med 2007; 1: 261-273.
4. HunzikerEB. Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. Osteoarthritis Cartilage 2002; 10: 432-463.
5. Browne JE, Branch TP. Surgical alternatives for treatment of articular cartilage lesions. J Am Acad Orthop Surg 2000; 8:180-189.
6. Langer R, VacantiJP. Tissue engineering. Science 1993; 260: 920-926.
Mardani M et al Differentiated and Native Chondrocytes in Alginate
Iran J Basic Med Sci, Vol. 16, No. 6, Jun 2013
 
770
7. Guilak F, Butler DL, Goldstein SA. Functional tissue engineering: the role of biomechanics in articular cartilage repair. Clin Orthop Relat Res 2001; 391: 295-305.
8. Song L, Baksh D, Tuan RS. Mesenchymal stem cell-based cartilage tissue engineering: cells, scaffold and biology. Cytotherapy 2004; 6: 596-601.
9. Chaikof EL, Matthew H, Kohn J, Mikos AG, Prestwich GD, Yip CM. Biomaterials and scaffolds in reparative medicine. Ann N Y Acad Sci 2002; 961:96–105.
10. Naughton GK. From lab bench to market: critical issues in tissue engineering. Ann N Y Acad Sci 2002; 961:372–385.
11. Hilfiker A, Kasper C, Hass R, Haverich A. Mesanchymal stem cells and progeniator cells in connective tissue engineering and regenerative medicine: is there a future for transplantation? Langenbecks Arch Surg 2011; 396: 489-497.
12. Adkisson HD 4th, Martin JA, Amendola RL, Milliman C, Mauch KA, Katwal AB, et al. The potential of`human allogeneic juvenile chondrocytes for restoration of articular cartilage. Am J Sports Med 2010; 38:1324-1333.
13. Mahmoudifar N, Doran PM. Extent of cell differentiation and capacity for cartilage synthesis in human adult adipose-derived stem cells: comparison with fetal chondrocytes. Biotechnol Bioeng 2010; 107:393-401.
14. Keeney M, Lai JH, Yang F. Recent progress in cartilage tissue engineering. Curr Opin Biotechnol 2011; 22:734-740.
15. Gimble J, Guilak F. Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy 2003; 5:362-369.
16. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001; 7: 211-228.
17. Erickson GR, Gimble JM, Franklin DM, Rice HE, Awad H, Guilak F. Chondrogenic potential of adipose tissue-derived stromal cells in vitro and in vivo. Biochem Biophys Res Commun 2002; 290: 763-769.
18. Hashemi Beni B, Razavi S, Esfandiari E, Karbasi S, Mardani M, Nasresfahani M. Induction of chondrogenic differentiation of human adipose-derived stem cells with TGF-ß3 in pellet culture system. Iran J Basic Med Sci 2008; 11:10-17.
19. Hashemi beni B, Razavi S, Esfandiari E, Karbasi S, Mardani M, Farzaneh Sadeghi. The effect of BMP-6 growth factor on differentiation of adipose-derived stem cells into chondrocyte in pellet culture. JIMS 2009; 27:613-644.
20. Estes BT, Wu AW, Guilak F. Potent induction of chondrocytic differentiation of human adipose-derived adult stem cells by bone morphogenetic protein 6. Arthritis Rheum 2006; 54: 1222-1232.
21. Diekman BO, Rowland CR, Lennon DP, Caplan AI, GuilakF. Chondrogenesis of adult stem cells from adipose tissue and bone marrow: induction by growth factors and cartilage-derived matrix. Tissue Eng part A 2010;16:523-533.
22. Guilak F, Estes BT, Diekman BO, Moutos FT, Gimble JM. 2010 Nicolas Andry Award: multipotent adult stem cells from adipose tissue for musculoskeletal tissue engineering. Clin Orthop Relat Res 2010; 468:2530-2540.
23. Hunziker EB. Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. Osteoarthritis Cartilage 2002; 10: 432-463.
24. Freed LE, Marquis JC, Nohria A, Emmanual J, Mikos AG, Langer R. Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. J Biomed Mater Res 1993; 27:11–23.
25. Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials 1999; 20:45–53.
26. Fragonas E, Valente M, Pozzi-Mucelli M, Toffanin R, Rizzo R, Silvestri F, et al. Articular cartilage repair in rabbits by using suspensions of allogenic chondrocytes in alginate. Biomaterials 2000; 21:795–801.
27. Chang SC, Rowley JA, Tobias G, Genes NG, Roy AK, Mooney DJ, et al. Injection molding of chondrocyte/alginate constructs in the shape of facial implants. J Biomed Mater Res 2001; 55: 503–511.
28.
Awad HA, Wickham MQ, Leddy HA, Gimble JM, Guilak F. Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. Biomaterials 2004; 25: 3211–3222.
29.
Leddy HA, Awad HA, Guilak F. Molecular diffusion in tissue-engineered cartilage constructs: effects of scaffold material, time, and culture conditions. J Biomed Mater Res B Appl Biomater 2004; 70: 397–406.
30. Barry F, Boynton RE, Liu B, Murphy JM. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation- dependent gene expression of matrix components. Exp Cell Res 2001; 268:189-200.
 
31. Pelttari K, Steck E, Richter W. The use of mesnchymal stem cells for chondrogenesis. Injury 2008; 39: 58–65.
 
32. Gleghorn JP, Dines ME, Cox M, Ayala P, Bonassar LJ. Comparison of chondrogenesis by MSCs and chondrocytes in alginate: evaluation of biochemical composition and mechanical properties (Abstract). Trans Orthop Res Soc 2005.Poster #1782.
 
33. Mauck RL, Yuan X, Tuan RS. Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture. Osteoarthritis Cartilage 2006; 14: 179-189. 34. Jakobsen RB, Shahdadfar A, Reinholt FP, Brinchmann JE .Chondrogenesis in a hyaluronic acid scaffold: comparison between chondrocytes and MSC from bone marrow and adipose tissue. Knee Surg Sports Traumatol Arthrosc 2010; 18:1407–1416. 35. Seda Tigli RS, Ghosh S, Laha MM, Shevde NK, Daheron L, Gimble J, et al. Comparative chondrogenesis of human cell sources in 3D scaffolds. J Tissue Eng Regen Med 2009; 3:348-360.
 
36. Ruettger A, Neumann S, Wiederanders B, Huber R. Comparison of different methods for preparation and characterization of total RNA from cartilage samples to uncover osteoarthritis in vivo . BMC Res Notes 2010; 3:7.
 
37. Erickson GR, Gimble JM, Franklin DM, Rice HE, Awad H, Guilak F. Chondrogenic Potential of Adipose Tissue-Derived Stromal Cells in vitro and in vivo. Biochem Biophys Res Commun 2002; 290: 763–769. 38. Aguiar DJ, Knudson W, Knudson CB. Internalization of the hyaluronan receptor CD44 by chondrocytes. Exp Cell Res 1999; 252: 292–302.
 
39. Shen G.The role of type X collagen in facilitating and regulating endochondral ossification of articular cartilage. Orthod Craniofac Res 2005; 8: 11-17. 40. Wang Y, Kim UJ, Blasioli DJ, Kim HJ, Kaplan DL. In
Differentiated and Native Chondrocytes in Alginate Mardani M et al
Iran J Basic Med Sci, Vol. 16, No. 6, Jun 2013
 
771
vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells. Biomaterials 2005; 26: 7082–7094.
41. Johnstone B, Hering TM, Caplan AI, Goldberg VM, Yoo JU. In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 1998; 238: 265-272.
42. Bernstein P, Dong M, Corbeil D, Gelinsky M, Gunter KP, Fickert S. Pellet culture elicits superior chondrogenic redifferentiation than alginate-based systems. Biotechnol Prog 2009; 25:1146-1152.
43. Mwale F, Stachura D, Roughley P, Antoniou J. Limitations of using aggrecan and type X collagen as markers of chondrogenesis in mesenchymal stem cell differentiation. J Orthop Res 2006; 24:1791-1798.
44. Sekiya I, Colter DC, Prockop DJ. BMP-6 enhances chondrogenesis in a subpopulation of human marrow stromal cells. Biochem Biophys Res Commun 2001; 284: 411–418.
45. Zhang Z, McCaffery JM, Spencer RG, Francomano CA. Hyaline cartilage engineered by chondrocytes in pellet culture: histological, immunohistochemical and ultrastructural analysis in comparison with cartilage explants. J Anat 2004; 205: 229–237.
46. Riesle J, Hollander AP, Langer R, Freed LE, Vunjak-Novakovic G. Collagen in tissue-engineered cartilage: types, structure, and crosslinks. J Cell Biochem 1998; 71: 313–327.
47. Tallheden T, Karlsson C, Brunner A, Van Der Lee J, Hagg R, Tommasini R, et al. Gene expression during redifferentiation of human articular chondrocytes. Osteoarthritis Cartilage 2004; 12: 525–535.
48. Cheng NC, Estes BT, Awad HA, Guilak F. Chondrogenic differentiation of adipose-derived adult stem cells by a porous scaffold derived from native articular cartilage extracellular matrix. Tissue Eng Part A 2009;15: 231–241.
49. Kim HJ, ImGI. Chondrogenic differentiation of adipose tissue-derived mesenchymal stem cells: greater doses of growth factor are necessary. J Orthop Res 2009; 27:612–619.
50. Karlsson C, Brantsing C, Svensson T, Bribsy H, Asp J, Tallheden T, et al. Differentiation of human mesenchymal stem cells and articular chondrocytes: analysis of chondrogenic potential and expression pattern of differentiation-related transcription factors. J Orthop Res 2007; 25: 152-163.
51. Yang IH, Kim SH, Kim YH, Sun HJ, Kim SJ, Lee JW. Comparison of phenotypic characterization between "alginate bead" and "pellet" culture system as chondrogenic differentiation models for human mesenchymal stem cells.Yonsei Med J 2004; 45:891-900.
52. Park H, Temenoff JS, Tabata Y, Caplan AI, Mikos AG. Injectable biodegradable hydrogel composites for rabbit marrow mesenchymal stem cell and growth factor delivery for cartilage tissue engineering. Biomaterials 2007; 28: 3217-3227.
53. Park H, Temenoff JS, Holland TA, Tabata Y, Mikos AG. Delivery of TGF-Beta1and chondrocytes via injectable, biodegradable hydrogels for cartilage tissue engineering applications. Biomaterials 2005; 26: 7095-7103.
54. Grimaud E, Heymann D, Redini F. Recent advances in TGF-beta effects on chondrocyte metabolism. Potential therapeutic roles of TGF-beta in cartilage disorders. Cytokine Growth Factor Rev 2002;13:241-257.
55. Redini F, Galera P, Mauviel A, Loyau G, Pujol P. Transforming growth factor β stimulates collagen and glycosaminoglycan biosynthesis in cultural rabbit articular chondrocytes. FEBS Lett 1998; 234:172-176.
56. Skantze KA, brinckerhoff CE, Collier JP. Use of agarose culture to measure the effect of transforming growth factor beta and epidermal growth factor on rabbit articular chondrocytes. Cancer Res 1985; 45:4416-4421.
57. Becker JC, Beckbauer M, Domschke W, Herbst H, Pohle T. Fibrin glue, healing of gastric mucosal injury, and expression of growth factors: results from a human in vivo study. Gastrointest Endosc 2005; 61: 560–567.
58. Mackay AM, Beck SC, Murphy JM, Barry FP, Chichester CO, PittengerMF. Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng 1998; 4: 415–428.
59. Majumdar MK, Banks V, Peluso DP, Morris EA. Isolation, characterization, and chondrogenic potential of human bone marrow-derived multipotential stromal cells. J Cell Physiol 2000; 185: 98-106.
60. Zhang X, Ziran N, Goater JJ, Schwarz EM, Puzas JE, Rosier RN, et al. Primary murine limb bud mesenchymal cells in long-term culture complete chondrocyte differentiation: TGF-beta delays hypertrophy and PGE2 Inhibits terminal differentiation. Bone 2004; 34:809−817.
61. Williams CG, Kim TK, Taboas A, Malik A, Manson P, Elisseeff J. In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in photopolymerizing hydrogel. Tissue Eng 2003; 9:679-688.

Files

Share

How to cite

Statistics