Comparison of the Production of Extracellular Matrix in Nucleus Pulposus of Intervertebral Disc in Alginate and Chitosan-Gelatin Scaffolds

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Tissue Engineering, School of New Technologies, Tehran University of Medical Sciences, Tehran, Iran

2 Assistant Professor, Department of Anatomy and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Student of Medicine, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran

4 PhD Student, Department of Immunology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran

5 PhD Student, Department of Biostatistics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

Abstract

Background: Low back pain is an important disorder linked with degenerative changes in the intervertebral disc (IVD). Degradation of IVD is caused by decreased number of cells, reduced production and degradation of extracellular matrix of IVD tissue especially in nucleus pulposus (NP). Natural and synthetic scaffolds are used for regeneration of IVD in tissue engineering.  Aggrecan is an important proteoglycan in NP tissue of IVD. This study aimed to levels of aggrecan secreted by human NP cells of IVD in alginate and chitosan-gelatin scaffolds.Methods: Collagenase enzymatic hydrolysis was used to extract NP cells from NP tissue of patients with IVD hernia in Alzahra Hospital (Isfahan, Iran). Chitosan gel was mixed with gelatin gel and freeze dried to make the scaffold. An alginate scaffold was also prepared. Cellular suspension containing the extracted NP cells was transferred to each scaffold and cultured for 14 days. The levels of secreted aggrecan were investigated by enzyme-linked immunosorbent assay. A light microscope was used to assert the morphology of NP cells.Findings: Secretion of aggrecan had significant increases during the third to the 14th day. The increments were more considerable in alginate scaffolds. There were significant differences in secreted aggrecan between alginate and chitosan scaffolds on the seventh and 14th days. However, no such a significant difference was observed on the third day. The two scaffolds were significantly different in terms of the secretion of extracellular matrix by NP.Conclusion: Compared to the chitosan-gelatin scaffold, the alginate scaffold provided better conditions for aggrecan secretion in NP cells in vitro. The use of thus scaffold is suggested to culture NP cell in vivo.

Keywords

References

  1. Oegema TR, Jr. The role of disc cell heterogeneity in determining disc biochemistry: a speculation. Biochem Soc Trans 2002; 30(Pt 6): 839-44.
  2. Hunter CJ, Matyas JR, Duncan NA. The notochordal cell in the nucleus pulposus: a review in the context of tissue engineering. Tissue Eng 2003; 9(4): 667-77.
  3. Roberts S, Evans H, Trivedi J, Menage J. Histology and pathology of the human intervertebral disc. J Bone Joint Surg Am 2006; 88(Suppl 2): 10-4.
  4. Sobajima S, Vadala G, Shimer A, Kim JS, Gilbertson LG, Kang JD. Feasibility of a stem cell therapy for intervertebral disc degeneration. Spine J 2008; 8(6): 888-96.
  5. Sobajima S, Kim JS, Gilbertson LG, Kang JD. Gene therapy for degenerative disc disease. Gene Ther 2004; 11(4): 390-401.
  6. Meisel HJ, Siodla V, Ganey T, Minkus Y, Hutton WC, Alasevic OJ. Clinical experience in cell-based therapeutics: disc chondrocyte transplantation A treatment for degenerated or damaged intervertebral disc. Biomol Eng 2007; 24(1): 5-21.
  7. Nishimura K, Mochida J. Percutaneous reinsertion of the nucleus pulposus. An experimental study. Spine (Phila Pa 1976) 1998; 23(14): 1531-8.
  8. Watanabe T, Sakai D, Yamamoto Y, Iwashina T, Serigano K, Tamura F, et al. Human nucleus pulposus cells significantly enhanced biological properties in a coculture system with direct cell-to-cell contact with autologous mesenchymal stem cells. J Orthop Res 2010; 28(5): 623-30.
  9. Bronzino JD. The biomedical engineering handbook. 3rd ed. Boca Raton, FL: CRC/Taylor & Francis; 2006.
  10. Vunjak-Novakovic G, Freshney RI. Culture of cells for tissue engineering. 1st ed. Hoboken, NJ: John Wiley and Sons; 2006.
  11. Hutmacher DW. Scaffolds in tissue engineering bone and cartilage. Biomaterials 2000; 21(24): 2529-43.
  12. Guo JF, Jourdian GW, MacCallum DK. Culture and growth characteristics of chondrocytes encapsulated in alginate beads. Connect Tissue Res 1989; 19(2-4): 277-97.
  13. Stevens MM, Qanadilo HF, Langer R, Prasad S, V. A rapid-curing alginate gel system: utility in periosteum-derived cartilage tissue engineering. Biomaterials 2004; 25(5): 887-94.
  14. Leone G, Torricelli P, Chiumiento A, Facchini A, Barbucci R. Amidic alginate hydrogel for nucleus pulposus replacement. J Biomed Mater Res A 2008; 84(2): 391-401.
  15. Suh JK, Matthew HW. Application of chitosan-based polysaccharide biomaterials in cartilage tissue engineering: a review. Biomaterials 2000; 21(24): 2589-98.
  16. Khor E, Lim LY. Implantable applications of chitin and chitosan. Biomaterials 2003; 24(13): 2339-49.
  17. Lahiji A, Sohrabi A, Hungerford DS, Frondoza CG. Chitosan supports the expression of extracellular matrix proteins in human osteoblasts and chondrocytes. J Biomed Mater Res 2000; 51(4): 586-95.
  18. Elder SH, Nettles DL, Bumgardner JD. Synthesis and characterization of chitosan scaffolds for cartilage-tissue engineering. Methods Mol Biol 2004; 238: 41-8.
  19. Chenite A, Chaput C, Wang D, Combes C, Buschmann MD, Hoemann CD, et al. Novel injectable neutral solutions of chitosan form biodegradable gels in situ. Biomaterials 2000; 21(21): 2155-61.
  20. Lee JY, Nam SH, Im SY, Park YJ, Lee YM, Seol YJ, et al. Enhanced bone formation by controlled growth factor delivery from chitosan-based biomaterials. J Control Release 2002; 78(1-3): 187-97.
  21. Mao J, Zhao L, de Yao K, Shang Q, Yang G, Cao Y. Study of novel chitosan-gelatin artificial skin in vitro. J Biomed Mater Res A 2003; 64(2): 301-8.
  22. Roughley P, Hoemann C, DesRosiers E, Mwale F, Antoniou J, Alini M. The potential of chitosan-based gels containing intervertebral disc cells for nucleus pulposus supplementation. Biomaterials 2006; 27(3): 388-96.
  23. Dang JM, Sun DD, Shin-Ya Y, Sieber AN, Kostuik JP, Leong KW. Temperature-responsive hydroxybutyl chitosan for the culture of mesenchymal stem cells and intervertebral disk cells. Biomaterials 2006; 27(3): 406-18.
  24. Huang Y, Onyeri S, Siewe M, Moshfeghian A, Madihally SV. In vitro characterization of chitosan-gelatin scaffolds for tissue engineering. Biomaterials 2005; 26(36): 7616-27.
  25. Miranda SC, Silva GA, Hell RC, Martins MD, Alves JB, Goes AM. Three-dimensional culture of rat BMMSCs in a porous chitosan-gelatin scaffold: A promising association for bone tissue engineering in oral reconstruction. Arch Oral Biol 2011; 56(1): 1-15.
  26. Hong SR, Lee SJ, Shim JW, Choi YS, Lee YM, Song KW, et al. Study on gelatin-containing artificial skin IV: A comparative study on the effect of antibiotic and EGF on cell proliferation during epidermal healing. Biomaterials 2001; 22(20): 2777-83.
  27. Mao JS, Zhao LG, Yin YJ, Yao KD. Structure and properties of bilayer chitosan-gelatin scaffolds. Biomaterials 2003; 24(6): 1067-74.
  28. Thein-Han WW, Saikhun J, Pholpramoo C, Misra RD, Kitiyanant Y. Chitosan-gelatin scaffolds for tissue engineering: physico-chemical properties and biological response of buffalo embryonic stem cells and transfectant of GFP-buffalo embryonic stem cells. Acta Biomater 2009; 5(9): 3453-66.
  29. Sun LP, Wang S, Zhang ZW, Wang XY, Zhang QQ. Biological evaluation of collagen-chitosan scaffolds for dermis tissue engineering. Biomed Mater 2009; 4(5): 055008.
  30. Hashemibeni B, Razavi Sh, Esfandiary E, Karbasi S, Mardani M, Sadeghi F, et al. Effect of transforming growth factor-β3 and bone morphogenetic protein-6 growth factors on chondrogenic differentiation of adipose-derived stem cells in alginate scaffold. J Isfahan Med Sch 2010; 28(112): 607-20.
  31. Bertolo A, Mehr M, Aebli N, Baur M, Ferguson SJ, Stoyanov JV. Influence of different commercial scaffolds on the in vitro differentiation of human mesenchymal stem cells to nucleus pulposus-like cells. Eur Spine J 2012; 21(Suppl 6): S826-S838.
  32. Ghorbani M, Hashemibani B, Bahramian H, Karimi Z, Zamani S, Mirhosseini SA, et al. Recognition of cytokeratin 18 marker by flow cytometry of nucleus pulposus cells in human intervertebral disc and comparison of proliferation and morphology of these cells in chitosan-gelatin and alginate scaffolds. J Isfahan Med Sch 2012; 30(188): 633-48.
  33. Bahramian Renani H, Ghorbani M, Hashemibeni Beni B, Mirhosseini MM, Karimi Z, Zarkesh H, et al. Determination and comparison of specifics of nucleus pulposus cells of human intervertebral disc in alginate and chitosan-gelatin scaffolds. Adv Biomed Res 2012; 1: 81.
Journal of Isfahan Medical School
Volume 31, Issue 225 - Serial Number 225
January and February 2013
Pages 82-91

Files

History

  • Receive Date: 19 September 2012
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics