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Fabrication of nanofibrous silkworm gland three-dimensional scaffold containing micro/nanoscale pores and study of its effects on adipose tissue-derived stem cell growth

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Abstract

An ideal tissue engineering scaffold requires a structure similar to that of the natural extracellular matrix. The structure of a scaffold is the most important parameter for determining the behavior and functions of cells. Therefore, the importance of designing a suitable scaffold has been greatly emphasized. Electrospinning has been extensively used to fabricate scaffolds for tissue engineering. In this study, we developed a nanofibrous scaffold made from silkworm gland (SG), which contains the silk protein fibroin and other proteins that have functions including antigenotoxicity and the promotion of cell growth. To investigate the physical and biological functions of SG nanofibrous (SGN) scaffold, we fabricated silk fibroin nanofibrous (SFN) scaffold as a control lacking the SG functional proteins. Field emission scanning electron microscope images revealed that the SGN three-dimensional scaffold had a homogenous pore distribution with a high porosity and interconnected pore walls. In biological tests, the SGN scaffold supported excellent cell viability and had a stronger antioxidant property compared to the SFN scaffold. In addition, the SGN scaffold efficiently supported the growth of adipose tissue-derived stem cells and their osteogenic and fibrocartilage differentiation. Therefore, this novel SGN three-dimensional scaffold shows high promise for use in tissue engineering.

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References

  1. Langer R, Vacanti J (1993) Tissue engineering. Science 260:920–926

    Article  Google Scholar 

  2. Berthiaume F, Maguire TJ, Yarmush ML (2011) Tissue engineering and regenerative medicine: history, progress, and challenges. Annu Rev Chem Biomol Eng 2:403–430

    Article  Google Scholar 

  3. Matthews JA, Wnek GE, Simpson DG, Bowlin GL (2002) Electrospinning of collagen nanofibers. Biomacromolecules 3:232–238

    Article  Google Scholar 

  4. Deville S, Saiz E, Nalla RK, Tomsia AP (2006) Freezing as a path to build complex composites. Science 311:515–518

    Article  Google Scholar 

  5. Deville S, Saiz E, Tomsia AP (2006) Freeze casting of hydroxyapatite scaffolds for bone tissue engineering. Biomaterials 27:5480–5489

    Article  Google Scholar 

  6. Nazarov R, Jin H-J, Kaplan DL (2004) Porous 3-D scaffolds from regenerated silk fibroin. Biomacromolecules 5:718–726

    Article  Google Scholar 

  7. Mandal BB, Kundu SC (2008) Non-bioengineered silk fibroin protein 3D scaffolds for potential biotechnological and tissue engineering applications. Macromol Biosci 8:807–818

    Article  Google Scholar 

  8. Altman GH, Diaz F, Jakuba C, Calabro T, Horan RL, Chen J, Lu H, Richmond J, Kaplan DL (2003) Silk-based biomaterials. Biomaterials 24:401–416

    Article  Google Scholar 

  9. Vacanti JP, Langer R (1999) Tissue engineering: the design and fabrication of living replacement devices for surgical reconstruction and transplantation. Lancet 354:S32–S34

    Article  Google Scholar 

  10. Kim B-S, Kim I-S (2011) Recent nanofiber technologies. Polym Rev 51:235–238

    Article  Google Scholar 

  11. Kim K-O, Akada Y, Kai W, Kim B-S, Kim I-S (2011) Cells attachment property of PVA hydrogel nanofibers incorporating hyaluronic acid for tissue engineering. J Biomater Nanobiotechnol 2:353

    Article  Google Scholar 

  12. Park J-C, Ito T, Kim K-O, Kim K-W, Kim B-S, Khil M-S, Kim H-Y, Kim I-S (2010) Electrospun poly (vinyl alcohol) nanofibers: effects of degree of hydrolysis and enhanced water stability. Polym J 42:273–276

    Article  Google Scholar 

  13. Wei K, Li Y, Kim KO, Nakagawa Y, Kim BS, Abe K, Chen GQ, Kim IS (2011) Fabrication of nano-hydroxyapatite on electrospun silk fibroin nanofiber and their effects in osteoblastic behavior. J Biomed Mater Res A 97:272–280

    Article  Google Scholar 

  14. Ma Z, Kotaki M, Inai R, Ramakrishna S (2005) Potential of nanofiber matrix as tissue-engineering scaffolds. Tissue Eng 11:101–109

    Article  Google Scholar 

  15. Boland ED, Telemeco TA, Simpson DG, Wnek GE, Bowlin GL (2004) Utilizing acid pretreatment and electrospinning to improve biocompatibility of poly (glycolic acid) for tissue engineering. J Biomed Mater Res B 71:144–152

    Article  Google Scholar 

  16. Yang F, Murugan R, Wang S, Ramakrishna S (2005) Electrospinning of nano/micro scale poly (L-lactic acid) aligned fibers and their potential in neural tissue engineering. Biomaterials 26:2603–2610

    Article  Google Scholar 

  17. Ki CS, Baek DH, Gang KD, Lee KH, Um IC, Park YH (2005) Characterization of gelatin nanofiber prepared from gelatin–formic acid solution. Polymer 46:5094–5102

    Article  Google Scholar 

  18. Ki CS, Kim JW, Hyun JH, Lee KH, Hattori M, Rah DK, Park YH (2007) Electrospun three-dimensional silk fibroin nanofibrous scaffold. J Appl Polym Sci 106:3922–3928

    Article  Google Scholar 

  19. Vepari C, Kaplan DL (2007) Silk as a biomaterial. Prog Polym Sci 32:991–1007

    Article  Google Scholar 

  20. Hwang JW, Lee HS, Kim H, Kim K-O, Choi Y-S (2012) Manufacture and characterization of silkworm gland hydrolysate. J Sericult Entomol Sci 50:76–81

    Google Scholar 

  21. Kim K-O, Lee Y, Hwang J-W, Kim H, Kim SM, Chang SW, Lee HS, Choi Y-S (2014) Wound healing properties of a 3-D scaffold comprising soluble silkworm gland hydrolysate and human collagen. Colloids Surf B 116:318–326

    Article  Google Scholar 

  22. Ishiyama M, Tominaga H, Shiga M, Sasamoto K, Ohkura Y, Ueno K (1996) A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol Pharm Bull 19:1518–1520

    Article  Google Scholar 

  23. Lo S-F, Nalawade SM, Mulabagal V, Matthew S, Chen C-L, Kuo C-L, Tsay H-S (2004) In vitro propagation by asymbiotic seed germination and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity studies of tissue culture raised plants of three medicinally important species of Dendrobium. Biol Pharm Bull 27:731–735

    Article  Google Scholar 

  24. Murphy AR, Kaplan DL (2009) Biomedical applications of chemically-modified silk fibroin. J Mater Chem 19:6443–6450

    Article  Google Scholar 

  25. Lowery JL, Datta N, Rutledge GC (2010) Effect of fiber diameter, pore size and seeding method on growth of human dermal fibroblasts in electrospun poly (ɛ-caprolactone) fibrous mats. Biomaterials 31:491–504

    Article  Google Scholar 

  26. Chen M, Patra PK, Warner SB, Bhowmick S (2007) Role of fiber diameter in adhesion and proliferation of NIH 3T3 fibroblast on electrospun polycaprolactone scaffolds. Tissue Eng 13:579–587

    Article  Google Scholar 

  27. Um IC, Kweon H, Park YH, Hudson S (2001) Structural characteristics and properties of the regenerated silk fibroin prepared from formic acid. Int J Biol Macromol 29:91–97

    Article  Google Scholar 

  28. Konishi T, Kondo M, Kurokawa M (1967) Study on the structure of silk fibroin under enzymatic hydrolysis. Sen’i Gakkaishi 23:64–69

    Article  Google Scholar 

  29. Konishi T, Kurokawa M (1968) The structure of silk fibroin-α. Sen’i Gakkaishi 24:550–554

    Article  Google Scholar 

  30. Tsukada M (1986) Effect of α-chymotrypsin on the structure of silk fibroin. J Sericult Sci Jap 55:126–130

    Google Scholar 

  31. Cheng Q, Lee BL-P, Komvopoulos K, Li S (2013) Engineering the microstructure of electrospun fibrous scaffolds by microtopography. Biomacromolecules 14:1349–1360

    Article  Google Scholar 

  32. Morikawa M, Kimura T, Murakami M, Katayama K, Terada S, Yamaguchi A (2009) Rat islet culture in serum-free medium containing silk protein sericin. J Hepatobiliary Pancreat 16:223–228

    Article  Google Scholar 

  33. Tsubouchi K, Igarashi Y, Takasu Y, Yamada H (2005) Sericin enhances attachment of cultured human skin fibroblasts. Biosci Biotech Biochem 69:403–405

    Article  Google Scholar 

  34. Kim S-h, Kang K-a, Zhang R, Piao M-j, Ko D-o, Wang Z-h, Chae S-w, Kang S-s, Lee K-h, Kang H-k (2008) Protective effect of esculetin against oxidative stress-induced cell damage via scavenging reactive oxygen species. Acta Pharm Sin 29:1319–1326

    Article  Google Scholar 

  35. Dehpour AA, Ebrahimzadeh MA, Seyed Fazel N, Seyed Mohammad N (2009) Antioxidant activity of the methanol extract of Ferula assafoetida and its essential oil composition. Grasas Aceites 60:405–412

    Article  Google Scholar 

  36. Zhaorigetu S, Yanaka N, Sasaki M, Watanabe H, Kato N (2003) Inhibitory effects of silk protein, sericin on UVB-induced acute damage and tumor promotion by reducing oxidative stress in the skin of hairless mouse. J Photochem Photobiol, B 71:11–17

    Article  Google Scholar 

  37. Lee J-C, Lim K-T, Jang Y-S (2002) Identification of Rhus verniciflua Stokes compounds that exhibit free radical scavenging and anti-apoptotic properties. Biochim Biophys Acta 1570:181–191

    Article  Google Scholar 

  38. Sill TJ, von Recum HA (2008) Electrospinning: applications in drug delivery and tissue engineering. Biomaterials 29:1989–2006

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by the First-step R&D Program (No. C0352351) funded by the Small and Medium Business Administration (SMBA, Korea) and the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (NRF-2015R1A2A2A03002680).

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Correspondence to Kyu Oh Kim.

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Kim, H.H., Park, Y.H., Yoon, K.J. et al. Fabrication of nanofibrous silkworm gland three-dimensional scaffold containing micro/nanoscale pores and study of its effects on adipose tissue-derived stem cell growth. J Mater Sci 51, 9267–9278 (2016). https://doi.org/10.1007/s10853-016-0173-4

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  • DOI: https://doi.org/10.1007/s10853-016-0173-4

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