Tissue Engineering and Regenerative Medicine

, Volume 10, Issue 4, pp 200–210 | Cite as

Silk gland fibroin from indian muga silkworm Antheraea assama as potential biomaterial

  • Subrata Kar
  • Sarmistha Talukdar
  • Shilpa Pal
  • Sunita Nayak
  • Pallavi Paranjape
  • S. C. Kundu
Original Article
First Online:
Received:
Revised:
Accepted:

Abstract

There is an increasing demand for new versatile biomaterials. Silk is reported by many researchers as an ideal biomaterial for different biomedical applications. Most of the studies are carried out on mulberry silk Bombyx mori, however silk from Indian non-mulberry silkworms are relatively unexplored. In this report we fabricate 2D matrices from the regenerated aqueous silk fibroin protein of the glands of non-mulberry Indian muga silkworm Antheraea assama (assamensis). Its biochemical, biophysical characteristics and its cytocompatibility for biomedical uses are evaluated. The properties of this muga gland fibroin are compared with silk gland fibroin from non-mulberry Indian tropical tasar silkworm Antheraea mylitta and with the fibroin from the cocoon of mulberry silkworm Bombyx mori. The gland fibroin of Antheraea assama is observed to consist of two polypeptides of approximately 250 kDa each linked by disulfide bond. Fourier transformed infrared spectroscopy and X-ray diffraction studies indicate random coil structure of dissolved fibroin solution. The alpha-helical structure in 2D films changed to beta-sheets upon ethanol treatment, imparting crystallinity and insolubility. The fibroin film is found to be the least hydrophilic, followed by B. mori and A. mylitta silk. Biocompatibility of the films from all three species is investigated through the cell attachment and spreading study of MG-63 human osteoblast-like cells. The cytocompatibility of non-mulberry fibroin matrices are comparable with that of standard tissue culture plates. The results indicate that the non-mulberry Indian muga silk gland fibroin is also suitable matrix as a natural biomaterial for tissue engineering applications.

Key words

muga silkworm fibroin film crystallinity biomaterial 
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    H Mori, M Tsukada, New silk protein: modification of silk protein by gene engineering for production of biomaterials, Rev Mol Biotech, 74, 95 (2000).CrossRefGoogle Scholar
  2. 2.
    C Vepari, DL Kaplan, Silk as a biomaterial, Prog Polym Sci, 32, 991 (2007).PubMedCrossRefGoogle Scholar
  3. 3.
    FG Omenetto, DL Kaplan, New opportunities for an ancient material, Science, 329, 528 (2010).PubMedCrossRefGoogle Scholar
  4. 4.
    DN Rockwood, RC Preda, T Y Ÿcel, et al., Materials fabrication from Bombyx mori silk fibroin, Nat Protoc, 6, 1612 (2011).PubMedCrossRefGoogle Scholar
  5. 5.
    SC Kundu, BC Dash, R Dash, et al., Natural protective glue protein, sericin bioengineered by silkworms: Potential for biomedical and biotechnological applications, Prog Polym Sci, 33, 998 (2008).CrossRefGoogle Scholar
  6. 6.
    B Mahendran, SK Ghosh, SC Kundu, Molecular phylogeny of silk-producing insects based on 16S ribosomal RNA and cytochrome oxidase subunit I genes, J Genet, 85, 31 (2006).PubMedCrossRefGoogle Scholar
  7. 7.
    YC Lin, M Ramadan, M Hronik-Tupaj, et al., Spatially controlled delivery of neurotrophic factors in silk fibroinĐbased nerve conduits for peripheral nerve repair, Ann Plas Surg, 67, 147 (2011).CrossRefGoogle Scholar
  8. 8.
    RM Reddy, GV Prasad, Silk-the prospective and compatible bio-material for advanced functional applications, Trends Appl Sci Res, 6, 89 (2011).CrossRefGoogle Scholar
  9. 9.
    MN Padamwar, AP Pawar, Silk sericin and its applications: a review, J Sci Ind Res India, 63, 323 (2004).Google Scholar
  10. 10.
    N Suzuki, A Fujimura, T Nagai, et al., Antioxidative activity of animal and vegetable dietary fibers, Biofactors, 21, 329 (2004).PubMedCrossRefGoogle Scholar
  11. 11.
    D Devi, NS Sarma, B Talukdar, et al., Study of the structure of degummed Antheraea assamensis (muga) silk fibre, J Text I, 102, 527 (2011).CrossRefGoogle Scholar
  12. 12.
    R Ahmed, A Kamra, SE Hasnain, Fibroin silk proteins from the nonmulberry silkworm Philosamia ricini are biochemically and immunologically distinct from those mulberry silkworm Bombyx mori, DNA Cell Biol, 23, 149, (2004).CrossRefGoogle Scholar
  13. 13.
    K Inouye, M Kurokawa, S Nishikawa, et al., Use of Bombyx mori silk fibroin as a substratum for cultivation of animal cells, J Biochem Biophys Meth, 37, 159 (1998).PubMedCrossRefGoogle Scholar
  14. 14.
    C Acharya, SK Ghosh, SC Kundu, Silk fibroin protein from mulberry and nonmulberry silkworms: cytotoxicity, biocompatibility and kinetics of L929 murine fibroblast adhesion, J Mater Sci Mater Med, 19, 2827 (2008).PubMedCrossRefGoogle Scholar
  15. 15.
    BB Mandal, SC Kundu, Non-bioengineered silk fibroin protein 3D scaffolds for potential biotechnological and tissue engineering applications, Macromol Biosci, 8, 807 (2008).PubMedCrossRefGoogle Scholar
  16. 16.
    S Talukdar, M Mandal, DW Hutmacher, et al., Engineered silk fibroin protein 3D matrices for in vitro tumor model, Biomaterials, 32, 2149 (2011).PubMedCrossRefGoogle Scholar
  17. 17.
    Y Wang, DD Rudym, A Walsh, et al., In vivo degradation of three-dimensional silk fibroin scaffolds, Biomaterials, 29, 3415 (2008).PubMedCrossRefGoogle Scholar
  18. 18.
    BB Mandal, SC Kundu, Osteogenic and adipogenic differentiation of rat bone marrow cells on non-mulberry and mulberry silk gland fibroin 3D scaffolds, Biomaterials, 30, 5019 (2009).PubMedCrossRefGoogle Scholar
  19. 19.
    N Kasoju, U Bora, Antheraea assama silk fibroin-based functional scaffold with enhanced blood compatibility for tissue engineering applications, Adv Eng Mater, 12, B139 (2010).CrossRefGoogle Scholar
  20. 20.
    A Datta, AK Ghosh, SC Kundu, Differential expression of the fibroin gene in developmental stages of silkworm, Antheraea mylitta (Saturniidae), Comp Biochem Physiol B, 129, 197 (2001).PubMedCrossRefGoogle Scholar
  21. 21.
    HY Kweon, IC Um, YH Park, Thermal behavior of regenerated Antheraea pernyi silk fibroin film treated with aqueous methanol, Polymer, 41, 7361 (2000).CrossRefGoogle Scholar
  22. 22.
    Z Zheng, Y Wei, S Yan, et al., Preparation of regenerated Antheraea yamamai silk fibroin film and controlled-molecular conformation changes by aqueous ethanol treatment, J Appl Polym Sc, 116, 461 (2010).CrossRefGoogle Scholar
  23. 23.
    CJ Wilson, RE Clegg, DI Leavesley, et al., Mediation of biomaterialĐcell interactions by adsorbed proteins: A Review, Tissue Eng, 11, 1 (2005).PubMedCrossRefGoogle Scholar
  24. 24.
    BB Mandal, SC Kundu, A novel method for dissolution and stabilisation of non mulberry silk gland protein fibroin using anionic surfactant sodium dodecyl sulphate, Biotechnol Bioeng, 99, 1482 (2008).PubMedCrossRefGoogle Scholar
  25. 25.
    S Talukdar, QT Nguyen, AC Chen, et al., Effect of initial cell seeding density on 3D-engineered silk fibroin scaffolds for articular cartilage tissue engineering, Biomaterials, 32, 8927 (2011).PubMedCrossRefGoogle Scholar
  26. 26.
    J Kundu, M Dewan, S Ghoshal, et al., Mulberry non-engineered silk gland protein vis-∧-vis silk cocoon protein engineered by silkworms as biomaterial matrices, J Mater Sci Mater Med, 19, 2679 (2008).PubMedCrossRefGoogle Scholar
  27. 27.
    N Kasoju, RR Bhonde, U Bora, Preparation and characterization of Antheraea assama silk fibroin based novel non-woven scaffold for tissue engineering applications, J Tissue Eng Regen Med, 7, 539 (2009).CrossRefGoogle Scholar
  28. 28.
    U Saxena, P Goswami, Silk mat as bio-matrix for the immobilization of cholesterol oxidase, Appl Biochem Biotechnol, 162, 1122 (2010).PubMedCrossRefGoogle Scholar
  29. 29.
    B Talukdar, M Saikia, PJ Handique, et al., Effect of organic solvents on tensile strength of muga silk produced by Antheraea assamensis, Int J Pure Appl Sci Technol, 7, 81 (2011).Google Scholar
  30. 30.
    S Dutta, B Talukdar, R Bharali, et al., Fabrication and characterization of biomaterial film from gland silk of muga and eri silkworms, Biopolymers, 99, 326 (2013).PubMedCrossRefGoogle Scholar
  31. 31.
    S Sofia, MB McCarthy, G Gronowicz, et al., Functionalized silk-based biomaterials for bone formation, J Biomed Mater Res, 54, 139 (2000).CrossRefGoogle Scholar
  32. 32.
    H Yamada, H Nakao, Y Takasu, et al., Preparation of undegraded native molecular fibroin solution from silkworm cocoons, Mater Sci Eng C, 14, 41 (2001).CrossRefGoogle Scholar
  33. 33.
    X Hu, DL Kaplan, P Cebe, Determining beta-sheet crystallinity in fibrous proteins by thermal analysis and Infrared spectroscopy, Macromolecules, 39, 6161 (2006).CrossRefGoogle Scholar
  34. 34.
    W Tao, M Li, C Zhao, Structure and properties of regenerated Antheraea pernyi silk fibroin in aqueous solution, Int J Biol Macromol, 40, 472 (2007).PubMedCrossRefGoogle Scholar
  35. 35.
    Q Lv, C Cao, Y Zhang, et al., The preparation of insoluble fibroin films induced by degummed fibroin or fibroin microspheres, J Mat Sci Mat Med, 15, 1193 (2004).Google Scholar
  36. 36.
    CL Du, J Jin, YC Li, et al., Novel silk fibroin/hydroxyapatite composite films: Structure and properties, Mater Sci Eng C, 29, 62 (2009).CrossRefGoogle Scholar
  37. 37.
    J Magoshi, S Nakamura, Studies on physical properties and structure of silk. Glass transition and crystallization of silk fibroin, J Appl Polym Sc, 19, 1013 (1975).CrossRefGoogle Scholar
  38. 38.
    D Cuvelier, M ThŽry, YS Chu, et al., The universal dynamics of cell spreading, Curr Biol, 17, 694 (2007).PubMedCrossRefGoogle Scholar
  39. 39.
    EA Cavalcanti-Adam, A Micoulet, J BlŸmmel, et al., Lateral spacing of integrin ligands influences cell spreading and focal adhesion assembly, Eur J Cell Biol, 85, 219 (2006).PubMedCrossRefGoogle Scholar
  40. 40.
    H Sezutsu, K Yukuhiro, Dynamic rearrangement within the Antheraea pernyi silk fibroin gene is associated with four types of repetitive units, J Mol Evol, 51, 329 (2000).PubMedGoogle Scholar
  41. 41.
    JS Hwang, JS Lee, TW Goo, et al., Cloning of the fibroin gene from the oak silkworm, Antheraea yamamai and its complete sequence, Biotechnol Lett, 23, 1321 (2001).CrossRefGoogle Scholar

Copyright information

© The Korean Tissue Engineering and Regenerative Medicine Society and Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Subrata Kar
    • 1
  • Sarmistha Talukdar
    • 1
  • Shilpa Pal
    • 1
  • Sunita Nayak
    • 1
  • Pallavi Paranjape
    • 1
  • S. C. Kundu
    • 1
  1. 1.Department of BiotechnologyIndian Institute of Technology KharagpurWest BengalIndia

Personalised recommendations