Mehrafarin Fesharaki

Study of Human Chondrocyte Redifferntiation Capacity in Three-Dimensional Hydrogel Culture

Article 3, Volume 11, Issue 3, October 2008, Page 152-158  PDF (1.18 MB)

Document Type: Original Article

DOI: 10.22038/IJBMS.2008.5225

Authors

Ebrahim Esfandiary  ¹; Mehdi Shakibaei²; Noshin Amirpour¹; Mehrafarin Fesharaki³; Mohammad Hossein Nasr-esfahani^4,5; Fariba Moulavi⁴; Farahnaz Moulavi⁵; Khalilollah Nazem⁶; Masoud Hatef Dehghani⁷; Constanze Csaki²; Shahnaz Razavi¹

¹Department of Anatomical Science, Isfahan University of Medical Sciences, Isfahan, Iran

²Musculoskeletal Research Group, Institute of Anatomy, Ludwig-Maximilian-University Munich, D-80336 Munich, Germany

³Department of Physiology, Isfahan University of Medical Sciences, Isfahan, Iran

⁴Department of Stem Cells, Cell Science Research Center, Royan Institute, Isfahan Research Campus, ACERC, Isfahan, Iran

⁵Isfahan Fertility and Infertility Center, Isfahan, Iran.

⁶Department of Orthopedics, Isfahan University of Medical Sciences, Isfahan, Iran

⁷Orthopedics ward, Sadoghi Hospital, Isfahan, Iran

Abstract

Objective(s)
Articular cartilage tissue defects cannot be repaired by the proliferation of resident chondrocytes. Autologous chondrocyte transplantation (ACT) is a relatively new therapeutic approach to cover full thickness articular cartilage defects by in vitrogrown chondrocytes from the joint of a patient. Therefore, we investigated the redifferentiation capability of human chondrocytes maintained in alginate culture.
Materials and Methods
The cartilage specimens obtained from 50 patients who underwent total knee and hip operations at the teaching hospital of Isfahan University of Medical Sciences, Isfahan Iran. Isolated primary chondrocytes were first grown in monolayer cultures for 1 to 6 passages (each passage lasting about 3 days). At each passage, monolayer cells seeded in alginate culture and investigated morphologically and immuno-cytologically for expression of cartilage-specific markers (collagen type II and cartilage-specific proteoglycans).
Results
The chondrocytes from monolayer passages P1 to P4 introduced in alginate cultures regained a chondrocyte phenotype. Cells were interconnected by typical gap junctions and after few days, they produced a cartilage-specific extracellular matrix (collagen type II and cartilage-specific proteoglycans). In contrast, cells from monolayer passages P5 and P6 did not redifferentiate to chondrocytes in the alginate cultures.
Conclusion
Chondrocyte culture was established for the first time in Iran. The alginate culture conditions promote the redifferentiation of dedifferentiated chondrocytes that have still a chondrogenic potential. This procedure opens up a promising approach to produce sufficient numbers of differentiated chondrocytes for ACT. Indeed, in some patients the harvested cells were used immediately and successfully for transplantation.

Keywords

Alginate; Autologous; Chondrocytes; Collagen Type II; Proteoglycan; Transplantation

References

Kuettner KE, Memoli VA, Pauli BU, Wrobel NC, Thonar EJ, Daniel JC. Synthesis of cartilage matrix by mammalian chondrocytes in vitro. II. Maintenance of collagen and proteoglycan phenotype. J Cell Biol 1982; 93:751-757.

 

2.            Shakibaei M. Integrin expression on epiphyseal mouse chondrocytes in monolayer culture. Histol Histopathol 1995; 10:339-349.

 

3. Shakibaei M, de Souza P. Differentiation of mesenchymal limb bud cells to chondrocytes in alginate beads. Cell Biol Int 1997; 21:75-86.

 

4. Lin Z, Fitzgerald JB, Xu J, Willers C, Wood D, Grodzinsky AJ, et al. Gene expression profiles of human chondrocytes during passaged monolayer cultivation. J Orthop Res 2008; 26:1230-1237.

 

5. Watt FM.  Effect of seeding density on stability of the differentiated phenotype of pig articular chondrocytes in culture. J Cell Sci 1988; 89:373-378.

 

6. Shakibaei M, de Souza P, Merker H-J . Integrin expression and collagen type II implicated in maintenance of chondrocyte shape in monolayer culture: an immunomorphological study. Cell Biol Int 1997; 21:115-125.

 

7. Martin I, Vunjak-Novakovic G, Yang J, Langer R, Freed LE. Mammalian chondrocytes expanded in the presence of fibroblast growth factor 2 maintain the ability to differentiate and regenerate three-dimensional cartilaginous tissue. Exp Cell Res 1999; 253:681-688.

 

8. Schulze-Tanzil G, de Souza P, Villegas Castrejon H, John T, Merker HJ, Scheid A, et al. Redifferentiation of dedifferentiated human chondrocytes in high-density cultures. Tissue Cell Res 2002; 308:371-379.

 

9. Schulze-Tanzil G, Mobasheri A, De Souza P, John T, Shakibaei M. Loss of chondrogenic potential in dedifferentiated chondrocytes correlates with deficient Shc-Erk interaction and apoptosis. Osteoarthritis Cartilage 2004; 12:448-458.

 

10. Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L. Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med 1994; 331:889-895.

 

11. Marlovits S, Zeller P, Singer P, Resinger C, Vecsei V. Cartilage repair: generations of autologous chondrocyte transplantation. Eur J Radiol 2006; 57:24-31.

 

12. Barbero A, Ploegert S, Heberer M, Martin I. Plasticity of clonal populations of dedifferentiated adult human articular chondrocytes. Arthritis Rheum 2003; 48:1315-1325.

 

13. Lin YJ, Yen CN, Hu YC, Wu YC, Liao CJ, Chu IM. Chondrocytes culture in three-dimensional porous alginate scaffolds enhanced cell proliferation, matrix synthesis and gene expression. J Biomed Mater Res A 2008.  (In press).

 

14. Shakibaei M, Csaki C, Rahmanzadeh M, Putz R. Interaction between human chondrocytes and extracellular matrix in vitro: a contribution to autologous chondrocyte transplantation. Orthopade 2008; 37(5):440-7.

 

15. Grimshaw MJ, Mason RM. Bovine articular chondrocyte function in vitro depends upon oxygen tension. Osteoarthritis Cartilage 2000; 8:386-392.

 

16. Hauselmann HJ, Aydelotte MB, Schumacher BL, Kuettner KE, Gitelis SH, Thonar EJ. Synthesis and turnover of proteoglycans by human and bovine adult articular chondrocytes cultured in alginate beads. Matrix 1992; 12:116-129.

 

17. Hauselmann HJ, Fernandes RJ, Mok SS, Schmid TM, Block JA, Aydelotte MB, et al. Phenotypic stability of bovine articular chondrocytes after long-term culture in alginate beads. J Cell Sci 1994; 107:17-27.

 

18. Petit B, Masuda K, D'Souza AL, Otten L, Pietryla D, Hartmann DJ, et al. Characterization of cross linked collagens synthesized by mature articular chondrocytes cultured in alginate beads: comparison of two distinct matrix compartments. Exp Cell Res 1996; 225:151-161.

 

19. Hwang NS, Varghese S, Elisseeff J. Cartilage tissue engineering: Directed differentiation of embryonic stem cells in three-dimensional hydrogel culture. Methods Mol Biol 2007; 407:351-373.

 

20. Wang Y, de Isla N, Huselstein C, Wang B, Netter P, Stoltz JF, et al. Effect of alginate culture and mechanical stimulation on cartilaginous matrix synthesis of rat dedifferentiated chondrocytes. Biomed Mater Eng 2008; 18:S47-54.

 

21. Shakibaei M, John T, De Souza P, Rahmanzadeh R, Merker H-J. Signal transduction by 1-integrin receptors in human chondrocytes in vitro: collaboration with the insulin-like growth factor-I receptor. Biochem J 1999; 342:615-623.

 

22. Scott JE, Dorling J. Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie 1965; 5:221-233.

 

23.  Pitzke P, Bujía J, Wilmes E, Hammer C. Expression of ICAM-1 on isolated human nasal, auricular and costal chondrocytes. Acta Otolaryngol 1994; 114: 81-86.

 

24. Hunter W. Of the structure and diseases of articulating cartilages. Clin Orthop Relat Res. 1995; 317: 3-6.

 

25. Minas T, Nehrer S. Current concepts in the treatment of articular cartilage defects. Orthopedics. 1997; 20:525-538.

 

26. Bentley G, Minas T. Treating joint damage in young people. BMJ 2000; 320: 1585-1588.

 

27. Peterson L, Minas T, Brittberg M, Nilsson A, Sjogren-Jansson E, Lindahl A. Two- to 9-year outcome after autologous chondrocyte transplantation of the knee. Clin Orthop Relat Res 2000;  212-234.

 

28.Grundmann K, Zimmermann B, Barrach HJ, Merker H-J. Behaviour of epiphyseal mouse chondrocyte populations in monolayer culture. morphological and immunochemical studies. Virchows Arch A Pathol Anat Histol 1980; 389:167-187.

 

29.Van der Kraan PM, Buma P, Van Kuppevelt T, Van den Berg WB. Interaction of chondrocytes, extracellular matrix and growth factors: relevance for articular cartilage tissue engineering. Osteoarthritis Cartilage 2002; 10:631-637.

 

30. Darling EM, Athanasiou KA. Rapid phenotypic changes in passaged articular chondrocyte subpopulations. J Orthop Res 2005; 23:425-432.

 

31. Loty S, Foll C, Forest N, Sautier J-M. Association of enhanced expression of gap junctions with in vitro chondrogenic differentiation of rat nasal septal cartilage-released cells following their dedifferentiation and redifferentiation. Arch Oral Biol, 2000; 45:843-856.

 

32. Shakibaei M, Schulze-Tanzil G, de Souza P, John T, Rahmanzadeh M, Rahmanzadeh R, et al. Inhibition of mitogen activated protein kinase kinase induces apoptosis of human chondrocytes. J Biol Chem 2001; 276: 13289-13294.

 

33. Buschmann MD, Gluzband YA, Grodzinsky AJ, Kimura JH, Hunziker EB.  Chondrocytes in agarose culture synthesize a mechanically functional extracellular matrix. J Orthop Res 1992; 10:745-758.

 

34. Hauselmann HJ, Masuda K, Hunziker EB, Neidhart M, Mok SS, Michel BA, et al. Adult human chondrocytes cultured in alginate form a matrix similar to native human articular cartilage. Am J Physiol 1996; 271: C742-C752.

 

35. Atala A, Cima LG, Kim W, Paige KT, Vacanti JP, Retik AB, et al. Injectable alginate seeded with chondrocytes as a potential treatment for vesicoureteral reflux. J Urol 1993; 150:745-747.

 

36. Soon-Shiong P, Heintz RE, Merideth N, Yao QX, Yao Z, Zheng T, et al. Insulin independence in a type 1 diabetic patient after encapsulated islet transplantation. Lancet 1994; 343:950-951.

 

37. Freed LE, Vunjak-Novakovic G, Biron RJ, Eagles DB, Lesnoy DC, Barlow SK, et al. Biodegradable polymer scaffolds for tissue engineering. Biotechnology (N Y) 1994; 12:689-693.

 

 

 

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