Comparing the Effect of Silk Fibroin-Based Scaffolds on Differentiation of Rabbit Chondrocytes

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Biomaterials Research, School of Materials Engineering, Isfahan University of Technology AND Isfahan University of Medical Sciences, Isfahan, Iran

2 Professor, Department of Biomaterials Research, School of Materials Engineering, Isfahan University of Technology AND Dental Materials Research Center, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran

3 Associate Professor, Biosensor Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

4 Assistant Professor, National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

Abstract

Background: Osteoarthritis is a degenerative disease caused by damage or trauma to articular cartilage, leading to pain, brittleness, limitation of joint motions and swelling of the tissue. Cartilage damage is common in older people and also athletes. Chondrocytes dedifferentiate in monolayer culture. Tissue engineering involves the use of scaffold to maintain the differentiation of the cells. In this study, maintaining chondrogenic differentiation of the chondrocytes within the pure silk fibroin (SF) and silk fibroin- chondroitin sulfate- alginate (SF-CHS-SA) was compared.Methods: Pure SF and SF-CHS-SA scaffolds were prepared through lyophilization. The microstructures of the scaffolds were studied by scanning electron microscopy (SEM). Chondrocytes were isolated from the articular cartilage tissue of rabbit and were cultured within the prepared scaffolds for 14 days. The percentage of chondrocytes viability was measured using 3-(5,4-dimethythiazol-2-yl)-5,2-diphenyltetrazolium bromide (MTT) assay, using extract of the scaffolds. Glycosaminoglycan (GAG) secretion and gene expression of collagen II were studied using alcian blue staining and real time polymerase chain reaction (real time-PCR), respectively.Findings: SEM showed that the composite scaffold had higher interconnected pores and pure SF scaffold had mainly closed pores. Results of the MTT assay confirmed no cytotoxicity of the prepared scaffolds. GAG secretion and collagen II expression were significantly higher in the SF-CHS-SA scaffold than the pure SF scaffold (P < 0.05).Conclusion: The SF-CHS-SA scaffold is a much more suitable substrate for maintaining differentiation of the chondrocytes than pure SF scaffold or monolayer culture of chondrocytes.

Keywords

References

  1. Chung C, Burdick JA. Engineering cartilage tissue. Adv Drug Deliv Rev 2008; 60(2): 243-62.
  2. Yokoyama A, Sekiya I, Miyazaki K, Ichinose S, Hata Y, Muneta T. In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel. Cell Tissue Res 2005; 322(2): 289-98.
  3. Doostmohammadi A, Monshi A, Salehi R, Fathi MH, Seyedjafari E, Shafiee A, et al. Cytotoxicity evaluation of 63s bioactive glass and bone-derived hydroxyapatite particles using human bone-marrow stem cells. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2011; 155(4): 323-6.
  4. Karbasi S, Mirzadeh H, Orang F, Urban J. A comparison between cell viability of chondrocytes on a biodegradable polyester urethane scaffold and alginate beads in different oxygen tension and pH. Iran Polym J 2005; 14 (9): 823-30.
  5. Lanza R, Langer R, Vacanti J. Principles of tissue engineering. 4th ed. New York, NY: Academic Press; 2013. p. 1-15.
  6. Atala A, Lanza R. Methods of tissue engineering.1st ed. New York, NY: Academic Press; 2002. p. 1027-36.
  7. Nesic D, Whiteside R, Brittberg M, Wendt D, Martin I, Mainil-Varlet P. Cartilage tissue engineering for degenerative joint disease. Adv Drug Deliv Rev 2006; 58(2): 300-22.
  8. Bueno EM, Glowacki J. Biologic foundations for skeletal tissue engineering. 1st ed. San Rafael, CA: Morgan and Claypool Publishers; 2011. p. 30-38.
  9. 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(16): 3211-22.
  10. De FL, Grigolo B, Roseti L, Facchini A, Fini M, Giavaresi G, et al. Transplantation of chondrocytes seeded on collagen-based scaffold in cartilage defects in rabbits. J Biomed Mater Res A 2005; 75(3): 612-22.
  11. Eyrich D, Brandl F, Appel B, Wiese H, Maier G, Wenzel M, et al. Long-term stable fibrin gels for cartilage engineering. Biomaterials 2007; 28(1): 55-65.
  12. YANG CHUN. Enhanced physicochemical properties of collagen by using EDC/NHS-crosslinking. Bull Mater Sci 2012; 35(5): 913-8.
  13. Lv Q, Hu K, Feng Q, Cui F. Fibroin/collagen hybrid hydrogels with crosslinking method: preparation, properties, and cytocompatibility. J Biomed Mater Res A 2008; 84(1): 198-207.
  14. Wenk E, Merkle HP, Meinel L. Silk fibroin as a vehicle for drug delivery applications. J Control Release 2011; 150(2): 128-41.
  15. Vepari C, Kaplan DL. Silk as a Biomaterial. Prog Polym Sci 2007; 32(8-9): 991-1007.
  16. Wang Y, Kim HJ, Vunjak-Novakovic G, Kaplan DL. Stem cell-based tissue engineering with silk biomaterials. Biomaterials 2006; 27(36): 6064-82.
  17. Zhang X, Reagan MR, Kaplan DL. Electrospun silk biomaterial scaffolds for regenerative medicine. Adv Drug Deliv Rev 2009; 61(12): 988-1006.
  18. Asakura T, Kuzuhara A, Tabeta R, Saito H. Conformational characterization of Bombyx mori silk fibroin in the solid state by high-frequency carbon-13 cross polarization-magic angle spinning NMR, x-ray diffraction, and infrared spectroscopy. Macromolecules 1985; 18(10): 1841-5.
  19. Min BM, Jeong L, Lee KY, Park WH. Regenerated silk fibroin nanofibers: water vapor-induced structural changes and their effects on the behavior of normal human cells. Macromol Biosci 2006; 6(4): 285-92.
  20. Nazarov R, Jin HJ, Kaplan DL. Porous 3-D scaffolds from regenerated silk fibroin. Biomacromolecules 2004; 5(3): 718-26.
  21. Bhardwaj N, Chakraborty S, Kundu SC. Freeze-gelled silk fibroin protein scaffolds for potential applications in soft tissue engineering. Int J Biol Macromol 2011; 49(3): 260-7.
  22. Yan LP, Oliveira JM, Oliveira AL, Caridade SG, Mano JF, Reis RL. Macro/microporous silk fibroin scaffolds with potential for articular cartilage and meniscus tissue engineering applications. Acta Biomater 2012; 8(1): 289-301.
  23. Cao Y, Wang B. Biodegradation of silk biomaterials. Int J Mol Sci 2009; 10(4): 1514-24.
  24. Wang Y, Rudym DD, Walsh A, Abrahamsen L, Kim HJ, Kim HS, et al. In vivo degradation of three-dimensional silk fibroin scaffolds. Biomaterials 2008; 29(24-25): 3415-28.
  25. Hardy JG, Scheibel TR. Composite materials based on silk proteins. Progress in Polymer Science 2010; 35(9): 1093-115.
  26. Hu K, Cui F, Lv Q, Ma J, Feng Q, Xu L, et al. Preparation of fibroin/recombinant human-like collagen scaffold to promote fibroblasts compatibility. J Biomed Mater Res A 2008; 84(2): 483-90.
  27. Gobin AS, Butler CE, Mathur AB. Repair and regeneration of the abdominal wall musculofascial defect using silk fibroin-chitosan blend. Tissue Eng 2006; 12(12): 3383-94.
  28. Sionkowska A, Planecka A. Preparation and characterization of silk fibroin/chitosan composite sponges for tissue engineering. Journal of Molecular Liquids 2013; 178(0): 5-14.
  29. Garcia-Fuentes M, Meinel AJ, Hilbe M, Meinel L, Merkle HP. Silk fibroin/hyaluronan scaffolds for human mesenchymal stem cell culture in tissue engineering. Biomaterials 2009; 30(28): 5068-76.
  30. Yan SQ, Zhang Q, Wang JN, Li MZ. Characterization of silk fibroin/hyaluronic acid blend films cross-linked with EDC. Journal of Fiber Bioengineering and Informatics 2010; 3(2): 62-7.
  31. Ming J, Zuo B. A novel silk fibroin/sodium alginate hybrid scaffolds. Polym Eng Sci 2014; 54(1): 129-36.
  32. Shokrgozar MA, Bonakdar S, Dehghan MM, Emami SH, Montazeri L, Azari S, et al. Biological evaluation of polyvinyl alcohol hydrogel crosslinked by polyurethane chain for cartilage tissue engineering in rabbit model. J Mater Sci Mater Med 2013; 24(10): 2449-60.
Journal of Isfahan Medical School
Volume 32, Issue 286 - Serial Number 286
March and April 2014
Pages 740-751

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History

  • Receive Date: 08 April 2014
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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