Study of chondrogenic potential of stem cells in co-culture with chondrons

Document Type : Original Article


1 Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

2 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

3 Department of Biochemistry and Laboratory Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

4 Department of Orthopedy, Tabriz University of Medical Sciences, Tabriz, Iran

5 Department of Immunolog, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

6 Umblical Cord Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran


Objective(s): Three-dimensional biomimetic scaffolds have widespread applications in biomedical tissue engineering due to similarity of their nanofibrous architecture to native extracellular matrix. Co-culture system has stimulatory effect on chondrogenesis of adult mesenchymal stem cells. This work presents a co-culture strategy using human articular chondrons and adipose-derived stem cells (ASCs) from infrapatellar fat pad (IPFP) for cartilage tissue production.
Materials and Methods: Isolated stem cells were characterized by flowcytometry. Electrospun and polycaprolactone (PCL) scaffolds (900 nm fiber diameter) was obtained from Bon Yakhteh (Tehran- Iran) and human infrapatellar fat pad-derived stem cells (IPFP-ASCs) were seeded on them. IPFP- ASCs on scaffolds were co-cultured with articular chondrons using transwell. After 21 day, chondrogenic differentiation of stem cell was evaluated by determining the genes expression of collagen2, aggrecan and Indian hedgehog using real- time RT-PCR.
Results: Genes expression of collagen2, aggrecan by IPFP-ASCs did not alter significantly in comparison with control group. Howevers, expression of Indian hedgehog decreased significantly compared to control group (P˂ 0.05).
Conclusion: These findings indicate that chondrons obtained from osteoarthritic articular cartilage did not stimulate chondrogenic differentiation of IPFP-ASCs in co-culture.


1.   Buckwalter J, Mankin H. Articular cartilage: degeneration and osteoarthritis, repair, regeneration, and transplantation. Instr Course lect 1997; 47:487-504.
2. Buckwalter JA, Mankin HJ, Grodzinsky AJ. Articular cartilage and osteoarthritis. Instr Course Lect 2005; 54:465.
3. Caplan AI. Tissue engineering designs for the future: new logics, old molecules. Tissue Eng 2000; 6:1-8.
4. Tuli R, Li W-J, Tuan RS. Current state of cartilage tissue engineering. Arthritis Res Ther  2003; 235-238.
5. Magazine E. From UMassWiki. Tissue Eng 1993; 60:920.
6. Tuan RS. Stemming cartilage degeneration: adult mesenchymal stem cells as a cell source for articular cartilage tissue engineering. Arthritis Rheum  2006; 54:3075-3078.
7. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284:143-147.
8. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002; 13:4279-4295.
9. Dragoo J, Samimi B, Zhu M, Hame S, Thomas B, Lieberman J, et al. Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads. J Bone Joint Surg Br  2003; 85:740-747.
10. Woodruff MA, Hutmacher DW. The return of a forgotten polymer—polycaprolactone in the 21st century. Prog Polym Sci  2010; 35:1217-1256.
11. Lee HU, Jeong YS, Jeong SY, Park SY, Bae JS, Kim HG, et al. Role of reactive gas in atmospheric plasma for cell attachment and proliferation on biocompatible poly ɛ-caprolactone film. Appl Surf Sci 2008; 254:5700-5705.
12. Yildirim ED, Pappas D, Güçeri S, Sun W. Enhanced cellular functions on polycaprolactone tissue scaffolds by O2 plasma surface modification. Plasma Proc Polym  2011; 8:256-267.
13. Wang YQ, Cai JY. Enhanced cell affinity of poly (L-lactic acid) modified by base hydrolysis: Wettability and surface roughness at nanometer scale. Curr Appl Phys  2007; 7:108-111.
14. Ng R, Zhang X, Liu N, Yang ST. Modifications of nonwoven polyethylene terephthalate fibrous matrices via NaOH hydrolysis: Effects on pore size, fiber diameter, cell seeding and proliferation. Proc Biochem 2009; 44:992-998.
15. Lee HU, Kang YH, Jeong SY, Koh K, Kim JP, Bae JS, et al. Long-term aging characteristics of atmospheric-plasma-treated poly (ɛ-caprolactone) films and fibres. Polym Degrad Stab    2011; 96:1204-1209.
16. Hendriks J, Riesle J, van Blitterswijk CA. Co-culture in cartilage tissue engineering. J Tissue Eng Regen Med  2007; 1:170-178.
17. Arana CJ, Diamandis EP, Kandel RA. Cartilage tissue enhances proteoglycan retention by nucleus pulposus cells in vitro. Arthritis Rheum  2010; 62:3395-3403.
18. Bonasia DE, Martin JA, Marmotti A, Amendola RL, Buckwalter JA, Rossi R, et al. Cocultures of adult and juvenile chondrocytes compared with adult and juvenile chondral fragments: in vitro matrix production. Am J Sports Med  2011; 39:2355-2361.
19. Szirmai J. The concept of the chondron as a biomechanical unit.  Biopolymere und Biomechanik von Bindegewebssystemen: Springer;1974.p. 87-91.
20. Poole CA, Matsuoka A, Schofield JR. Chondrons from articular cartilage. III. Morphologic changes in the cellular microenvironment of chondrons isolated from osteoarthritic cartilage. Arthritis Rheum 1991; 34:22-35.
21. Salmivirta K, Talts JF, Olsson M, Sasaki T, Timpl R, Ekblom P. Binding of mouse nidogen-2 to basement membrane components and cells and its expression in embryonic and adult tissues suggest complementary functions of the two nidogens. Exp Cell Res  2002; 279:188-201.
22. Poole CA, Honda T, Skinner SJ, Schofield JR, Hyde KF, Shinkai H. Chondrons from articular cartilage (II): analysis of the glycosaminoglycans in the cellular microenvironment of isolated canine chondrons. Connect Tissue Res 1990; 24:319-330.
23. Poole CA, Flint MH, Beaumont BW. Chondrons extracted from canine tibial cartilage: preliminary report on their isolation and structure. J Orthop Res 1988; 6:408-419.
24. Smith GN Jr, Hasty KA, Brandt KD. Type XI collagen is associated with the chondrocyte surface in suspension culture. Matrix 1989;9:186-192.
25. Rosenberg LC. Structure and function of dermatan sulfate proteoglycans in articular cartilage. In Articular Cartilage and Osteoarthritis 1992; 45-63. New York: Raven Press.
26. Van der Kraan P, Van den Berg W. Chondrocyte hypertrophy and osteoarthritis: role in initiation and progression of cartilage degeneration? Osteoarthritis  Cartilage 2012; 20:223-232.
27. Wei F, Zhou J, Wei X, Zhang J, Fleming BC, Terek R, et al. Activation of Indian hedgehog promotes chondrocyte hypertrophy and upregulation of MMP-13 in human osteoarthritic cartilage. Osteoarthritis  Cartilage  2012; 20:755-763.
28. Kabiri A, Esfandiari E, Hashemibeni B, Kazemi M, Mardani M, Esmaeili A. Effects of FGF-2 on human adipose tissue derived adult stem cells morphology and chondrogenesis enhancement in Transwell culture. Biochem Biophys Res Commun  2012; 424:234-238.
29. Lee GM, Poole CA, Kelley SS, Chang J, Caterson B. Isolated chondrons: a viable alternative for studies of chondrocyte metabolism in vitro. Osteoarthritis  Cartilage  1997; 5:261-274.
30. Heydarkhan-Hagvall S, Schenke-Layland K, Dhanasopon AP, Rofail F, Smith H, Wu BM, et al. Three-dimensional electrospun ECM-based hybrid scaffolds for cardiovascular tissue engineering. Biomaterials 2008; 29:2907-2914.
31. Li WJ, Jiang YJ, Tuan RS. Chondrocyte phenotype in engineered fibrous matrix is regulated by fiber size. Tissue Eng  2006; 12:1775-1785.
32. Ye K, Felimban R, Traianedes K, Moulton SE, Wallace GG, Chung J, et al. Chondrogenesis of infrapatellar fat pad derived adipose stem cells in 3D printed chitosan scaffold. PloS One 2014; 9:e99410.
33. Liu X, Sun H, Yan D, Zhang L, Lv X, Liu T, et al. In vivo ectopic chondrogenesis of BMSCs directed by mature chondrocytes. Biomaterials  2010; 31:9406-9414.
34. Zhang Z, Fan J, Becker K, Graff R, Lee G, Francomano C. Comparison of gene expression profile between human chondrons and chondrocytes: a cDNA microarray study. Osteoarthritis Cartilage  2006; 14:449-459.
35. Wang QG, Magnay JL, Nguyen B, Thomas CR, Zhang Z, El Haj AJ, et al. Gene expression profiles of dynamically compressed single chondrocytes and chondrons. Biochem Biophys Res Commun 2009; 379:738-742.
36. Shieh A, Athanasiou K. Dynamic compression of single cells. Osteoarthritis Cartilage  2007; 15:328-334.
37. Woo KM, Jun JH, Chen VJ, Seo J, Baek JH, Ryoo HM, et al. Nano-fibrous scaffolding promotes osteoblast differentiation and biomineralization. Biomaterials 2007; 28:335-343.
38. Tayalia P, Mooney DJ. Controlled growth factor delivery for tissue engineering. Adv Mater  2009; 21:3269-3285.
39. Aung A, Gupta G, Majid G, Varghese S. Osteoarthritic chondrocyte–secreted morphogens induce chondrogenic differentiation of human mesenchymal stem cells. Arthritis Rheum 2011; 63:148-158.
40. Kapoor M, Martel-Pelletier J, Lajeunesse D, Pelletier JP, Fahmi H. Role of proinflammatory cytokines in the pathophysiology of osteoarthritis. Nat Rev Rheumatol  2011; 7:33-42.
41. Goldring MB, Otero M. Inflammation in osteo-arthritis. Curr Opin Rheumatol  2011; 23:471.
42. Fermor B, Weinberg J, Pisetsky D, Misukonis M, Fink C, Guilak F. Induction of cyclooxygenase-2 by mechanical stress through a nitric oxide-regulated pathway. Osteoarthritis  Cartilage 2002; 10:792-798.
43. Estes B, Fermor B, Guilak F, editors. The influence of interleukin-1 and mechanical stimulation on human adipose derived adult stem cells undergoing chondrogenesis. Transactions of the Orthopaedic Research Society Annual Meeting  2004.
44. Majumdar MK, Wang E, Morris EA. BMP‐2 and BMP‐9 promotes chondrogenic differentiation of human multipotential mesenchymal cells and overcomes the inhibitory effect of IL‐1. J Cell Physiol  2001; 189:275-284.
45. Heldens GT, Blaney Davidson EN, Vitters EL, Schreurs BW, Piek E, van den Berg WB, et al. Catabolic factors and osteoarthritis-conditioned medium inhibit chondrogenesis of human mesenchymal stem cells. Tissue Eng Part A  2011; 18:45-54.
46. Buhrmann C, Mobasheri A, Matis U, Shakibaei M. Research article Curcumin mediated suppression of nuclear factor-κB promotes chondrogenic differentiation of mesenchymal stem cells in a high-density co-culture microenvironment  2010.
47. Felka T, Schafer R, Schewe B, Benz K, Aicher WK. Hypoxia reduces the inhibitory effect of IL-1beta on chondrogenic differentiation of FCS-free expanded MSC. Osteoarthritis Cartilage 2009; 17:1368-1376.