Fabrication and characterization of Antheraea pernyi silk fibroin-blended P(LLA-CL) nanofibrous scaffolds for peripheral nerve tissue engineering

Abstract

Electrospun nanofibers have gained widespreading interest for tissue engineering application. In the present study, ApF/P(LLA-CL) nanofibrous scaffolds were fabricated via electrospinning. The feasibility of the material as tissue engineering nerve scaffold was investigated in vitro. The average diameter increased with decreasing the blend ratio of ApF to P(LLA-CL). Characterization of ¹³C NMR and FTIR clarified that there is no obvious chemical bond reaction between ApF and P(LLA-CL). The tensile strength and elongation at break increased with the content increase of P(LLA-CL). The surface hydrophilic property of nanofibrous scaffolds enhanced with the increased content of ApF. Cell viability studies with Schwann cells demonstrated that ApF/P(LLA-CL) blended nanofibrous scaffolds significantly promoted cell growth as compare to P(LLA-CL), especially when the weight ratio of ApF to P(LLA-CL) was 25:75. The present work provides a basis for further studies of this novel nanofibrous material (ApF/P(LLA-CL)) in peripheral nerve tissue repair or regeneration.

Keywords

ApF/P(LLA-CL) / electrospinning / nanofibers / scaffolds / Schwann cells / peripheral nerve tissue engineering

References

[1] Yuksel E, Choo J, Wettergreen M, . Challenges in soft tissue engineering. Seminars in Plastic Surgery, 2005, 19(3): 261–270

[2] Wang C Y, Liu J J, Fan C Y, . The effect of aligned core–shell nanofibres delivering NGF on the promotion of sciatic nerve regeneration. Journal of Biomaterials Science: Polymer Edition, 2012, 23(1–4): 167–184

[3] Beachley V, Wen X. Polymer nanofibrous structures: Fabrication, biofunctionalization, and cell interactions. Progress in Polymer Science, 2010, 35(7): 868–892

[4] Sabbatier G, Larrañaga A, Guay-Bégin A A, . Design, degradation mechanism and long-term cytotoxicity of poly(l-lactide) and poly(lactide-co-ε-caprolactone) terpolymer film and air-spun nanofiber scaffold. Macromolecular Bioscience, 2015, 15(10): 1392–1410

[5] Kim M S, Lee M H, Kwon B J, . Enhancement of human mesenchymal stem cell infiltration into the electrospun poly(lactic-co-glycolic acid) scaffold by fluid shear stress. Biochemical and Biophysical Research Communications, 2015, 463(1–2): 137–142

[6] Li D, Xia Y. Electrospinning of nanofibers: Reinventing the wheel? Advanced Materials, 2004, 16(14): 1151–1170

[7] Reneker D H, Chun I. Nanometre diameter fibres of polymer, produced by electrospinning. Nanotechnology, 1996, 7(3): 216–223

[8] Zhou H, Lawrence J G, Bhaduri S B. Fabrication aspects of PLA–CaP/PLGA–CaP composites for orthopedic applications: a review. Acta Biomaterialia, 2012, 8(6): 1999–2016

[9] Elsabee M Z, Naguib H F, Morsi R E. Chitosan based nanofibers. Materials Science and Engineering C, 2012, 32(7): 1711–1726

[10] Jayakumar R, Prabaharan M, Nair S V, . Novel chitin and chitosan nanofibers in biomedical applications. Biotechnology Advances, 2010, 28(1): 142–150

[11] Rnjak-Kovacina J, Wise S G, Li Z, . Tailoring the porosity and pore size of electrospun synthetic human elastin scaffolds for dermal tissue engineering. Biomaterials, 2011, 32(28): 6729–6736

[12] Lee K Y, Jeong L, Kang Y O, . Electrospinning of polysaccharides for regenerative medicine. Advanced Drug Delivery Reviews, 2009, 61(12): 1020–1032

[13] Sell S A, McClure M J, Garg K, . Electrospinning of collagen/biopolymers for regenerative medicine and cardiovascular tissue engineering. Advanced Drug Delivery Reviews, 2009, 61(12): 1007–1019

[14] Zhang X, Reagan M R, Kaplan D L. Electrospun silk biomaterial scaffolds for regenerative medicine. Advanced Drug Delivery Reviews, 2009, 61(12): 988–1006

[15] Tao W, Li M, Zhao C. Structure and properties of regenerated Antheraea pernyi silk fibroin in aqueous solution. International Journal of Biological Macromolecules, 2007, 40(5): 472–478

[16] Liu Y, Li Y, Li X, . The origin and dispersal of the domesticated Chinese oak silkworm, Antheraea pernyi, in China: a reconstruction based on ancient texts. Journal of Insect Science, 2010, 10(1): 180

[17] Kundu S C, Kundu B, Talukdar S, . Nonmulberry silk biopolymers. Biopolymers, 2012, 97(6): 455–467

[18] Patra C, Talukdar S, Novoyatleva T, . Silk protein fibroin from Antheraea mylitta for cardiac tissue engineering. Biomaterials, 2012, 33(9): 2673–2680

[19] Yukuhiro K, Kanda T, Tamura T. Preferential codon usage and two types of repetitive motifs in the fibroin gene of the Chinese oak silkworm, Antheraea pernyi. Insect Molecular Biology, 1997, 6(1): 89–95

[20] Tian H, Lin L, Chen J, . RGD targeting hyaluronic acid coating system for PEI–PBLG polycation gene carriers. Journal of Controlled Release, 2011, 155(1): 47–53

[21] Minoura N, Aiba S, Higuchi M, . Attachment and growth of fibroblast cells on silk fibroin. Biochemical and Biophysical Research Communications, 1995, 208(2): 511–516

[22] Kim B S, Mooney D J. Development of biocompatible synthetic extracellular matrices for tissue engineering. Trends in Biotechnology, 1998, 16(5): 224–230

[23] Sun B, Li J, Liu W, . Fabrication and characterization of mineralized P(LLA-CL)/SF three-dimensional nanoyarn scaffolds. Iranian Polymer Journal, 2015, 24(1): 29–40

[24] Huang C, Chen S, Lai C, . Electrospun polymer nanofibres with small diameters. Nanotechnology, 2006, 17(6): 1558–1563

[25] Zhu Y, Zhang J, Zheng Y, . Stable, superhydrophobic, and conductive polyaniline/polystyrene films for corrosive environments. Advanced Functional Materials, 2006, 16(4): 568–574

[26] Jun I, Jeong S, Shin H. The stimulation of myoblast differentiation by electrically conductive sub-micron fibers. Biomaterials, 2009, 30(11): 2038–2047

[27] Christopherson G T, Song H, Mao H Q. The influence of fiber diameter of electrospun substrates on neural stem cell differentiation and proliferation. Biomaterials, 2009, 30(4): 556–564

[28] McKee M G, Wilkes G L, Colby R H, . Correlations of solution rheology with electrospun fiber formation of linear and branched polyesters. Macromolecules, 2004, 37(5): 1760–1767

[29] Zhang K, Wang H, Huang C, . Fabrication of silk fibroin blended P(LLA-CL) nanofibrous scaffolds for tissue engineering. Journal of Biomedical Materials Research Part A, 2010, 93(3): 984–993

[30] Sezutsu H, Yukuhiro K. Dynamic rearrangement within the Antheraea pernyi silk fibroin gene is associated with four types of repetitive units. Journal of Molecular Evolution, 2000, 51(4): 329–338

[31] Freddi G, Gotoh Y, Mori T, . Chemical structure and physical properties of Antheraea assama silk. Journal of Applied Polymer Science, 1994, 52(6): 775–781

[32] Zong X, Kim K, Fang D, . Structure and process relationship of electrospun bioabsorbable nanofiber membranes. Polymer, 2002, 43(16): 4403–4412

[33] Xu Y, Wu J, Wang H, . Fabrication of electrospun poly(L-lactide-co-ε-caprolactone)/collagen nanoyarn network as a novel, three-dimensional, macroporous, aligned scaffold for tendon tissue engineering. Tissue Engineering Part C: Methods, 2013, 19(12): 925–936

[34] Lins L, Brasseur R. The hydrophobic effect in protein folding. FASEB Journal, 1995, 9(7): 535–540

[35] Zhong Z, Guo Q, Mi Y. Solid-state n. m. r. investigation of crosslinkable blends of novolac and poly(ε-caprolactone). Polymer, 1999, 40(1): 27–33

[36] Howe C, Sankar S, Tonelli A E. 13C n. m. r. observation of poly(l-lactide) in the narrow channels of its inclusion compound with urea. Polymer, 1993, 34(12): 2674–2676

[37] Nakazawa Y, Asakura T. High-resolution ¹³C CP/MAS NMR study on structure and structural transition of Antheraea pernyi silk fibroin containing poly(L-alanine) and Gly-rich regions. Macromolecules, 2002, 35(6): 2393–2400

[38] Altankov G, Grinnell F, Groth T. Studies on the biocompatibility of materials: fibroblast reorganization of substratum-bound fibronectin on surfaces varying in wettability. Journal of Biomedical Materials Research, 1996, 30(3): 385–391

[39] De Bartolo L, Morelli S, Bader A, . The influence of polymeric membrane surface free energy on cell metabolic functions. Journal of Materials Science: Materials in Medicine, 2001, 12(10): 959–963

[40] Lampin M, Warocquier-Clérout R, Legris C, . Correlation between substratum roughness and wettability, cell adhesion, and cell migration. Journal of Biomedical Materials Research, 1997, 36(1): 99–108

[41] Chen Z G, Wang P W, Wei B, . Electrospun collagen–chitosan nanofiber: a biomimetic extracellular matrix for endothelial cell and smooth muscle cell. Acta Biomaterialia, 2010, 6(2): 372–382

[42] Lutolf M P, Hubbell J A. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nature Biotechnology, 2005, 23(1): 47–55

Acknowledgements

This research was supported by the National Key Research Program of China (2016YFA0201702 of 2016YFA0201700), the National Natural Science Foundation of China (Grant Nos. 31470941 and 31271035), the Science and Technology Commission of Shanghai Municipality (Grant Nos. 15JC1490100 and 15441905100), the Ph. D. Programs Foundation of Ministry of Education of China (Grant No. 20130075110005), and the Yantai Double Hundred Talent Plan. The authors extend their appreciation to the International Scientific Partnership Program ISPP at King Saud University for funding this research work through ISPP# 0049.

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