Electrospun biocomposite nanofibrous scaffolds for neural tissue engineering.

Bridging of nerve gaps after injury is a major problem in peripheral nerve regeneration. Considering the potential application of a bio-artificial nerve guide material, polycaprolactone (PCL)/chitosan nanofibrous scaffolds was designed and evaluated in vitro using rat Schwann cells (RT4-D6P2T) for nerve tissue engineering. PCL, chitosan, and PCL/chitosan nanofibers with average fiber diameters of 630, 450, and 190 nm, respectively, were fabricated using an electrospinning process. The surface chemistry of the fabricated nanofibers was determined using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Simple blending of PCL with chitosan proved an easy and efficient method for fabricating PCL/chitosan nanofibrous scaffolds, whose surface characteristics proved more hydrophilic than PCL nanofibers. Evaluation of mechanical properties showed that the Young's modulus and strain at break of the electrospun PCL/chitosan nanofibers were better than those of the chitosan nanofibers. Results of cell proliferation studies on nanofibrous scaffolds using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay showed 48% more cell proliferation on PCL/chitosan scaffolds than on PCL scaffolds after 8 days of culture. PCL/chitosan scaffolds showed better cell proliferation than PCL scaffolds and maintained their characteristic cell morphology, with spreading bipolar elongations to the nanofibrous substrates. This electrospun nanofibrous matrix thus proved of specific interest in tissue engineering for peripheral nerve regeneration.

[1]  Xiaosong Gu,et al.  The interaction of Schwann cells with chitosan membranes and fibers in vitro. , 2004, Biomaterials.

[2]  J. Feijen,et al.  Interaction of cultured human endothelial cells with polymeric surfaces of different wettabilities. , 1985, Biomaterials.

[3]  R. Bunge,et al.  Isolation and functional characterization of Schwann cells derived from adult peripheral nerve , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[4]  Wan-Ju Li,et al.  Biological response of chondrocytes cultured in three-dimensional nanofibrous poly(ϵ-caprolactone) scaffolds† , 2003 .

[5]  S. Frostick,et al.  Schwann cells, neurotrophic factors, and peripheral nerve regeneration , 1998, Microsurgery.

[6]  J. Bain Peripheral nerve and neuromuscular allotransplantation: Current status , 2000, Microsurgery.

[7]  K. Marra,et al.  Peptide-surface modification of poly(caprolactone) with laminin-derived sequences for adipose-derived stem cell applications. , 2006, Biomaterials.

[8]  G. Evans,et al.  Peripheral nerve injury: A review and approach to tissue engineered constructs , 2001, The Anatomical record.

[9]  Z. Xiufang,et al.  Surface Modification and Characterization of Chitosan Film Blended with Poly-L-Lysine , 2004, Journal of biomaterials applications.

[10]  M. Kotaki,et al.  Aligned biodegradable nanofibrous structure: a potential scaffold for blood vessel engineering. , 2004, Biomaterials.

[11]  H. B. Williams,et al.  Nerve Injuries and Their Repair: A Critical Appraisal , 1991 .

[12]  Y. Hsieh,et al.  Chitosan bicomponent nanofibers and nanoporous fibers. , 2006, Carbohydrate research.

[13]  K. Kataoka,et al.  Immobilization of laminin peptide in molecularly aligned chitosan by covalent bonding. , 2005, Biomaterials.

[14]  E J Wood,et al.  The effect of chitin and chitosan on the proliferation of human skin fibroblasts and keratinocytes in vitro. , 2001, Biomaterials.

[15]  A. Zalewski,et al.  Rejection of nerve allografts after cessation of immunosuppression with cyclosporin A. , 1981, Transplantation.

[16]  Xiaosong Gu,et al.  Biocompatibility evaluation of silk fibroin with peripheral nerve tissues and cells in vitro. , 2007, Biomaterials.

[17]  Gianluca Ciardelli,et al.  Materials for peripheral nerve regeneration. , 2006, Macromolecular bioscience.

[18]  Orawan Suwantong,et al.  In vitro biocompatibility of electrospun poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) fiber mats. , 2007, International journal of biological macromolecules.

[19]  D. Kaplan,et al.  The crosslinking of chitosan fibers , 1992 .

[20]  C. Domenici,et al.  Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/poly(epsilon-caprolactone) blends for tissue engineering applications in the form of hollow fibers. , 2008, Journal of biomedical materials research. Part A.

[21]  V. V. van Hinsbergh,et al.  Direct grafting of RGD-motif-containing peptide on the surface of polycaprolactone films , 2006, Journal of biomaterials science. Polymer edition.

[22]  J. Terzis,et al.  Historical and Basic Science Review: Past, Present, and Future of Nerve Repair , 1997, Journal of reconstructive microsurgery.

[23]  Roy M. Smeal,et al.  Substrate Curvature Influences the Direction of Nerve Outgrowth , 2005, Annals of Biomedical Engineering.

[24]  Shen‐guo Wang,et al.  Enhanced cell affinity of poly (D,L-lactide) by combining plasma treatment with collagen anchorage. , 2002, Biomaterials.

[25]  S. Ichinose,et al.  Hydroxyapatite-coated tendon chitosan tubes with adsorbed laminin peptides facilitate nerve regeneration in vivo , 2003, Brain Research.

[26]  K. Healy,et al.  Osteogenic Cell Attachment to Degradable Polymers , 1991 .

[27]  A. Mikos,et al.  Electrospinning of polymeric nanofibers for tissue engineering applications: a review. , 2006, Tissue engineering.

[28]  Y. Gong,et al.  Physical, mechanical and degradation properties, and Schwann cell affinity of cross-linked chitosan films , 2005, Journal of biomaterials science. Polymer edition.

[29]  P Connolly,et al.  Cell guidance by micropatterned adhesiveness in vitro. , 1992, Journal of cell science.

[30]  F. Grinnell Cellular adhesiveness and extracellular substrata. , 1978, International review of cytology.

[31]  R. Tuan,et al.  A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells. , 2005, Biomaterials.

[32]  T. Gordon,et al.  The cellular and molecular basis of peripheral nerve regeneration , 1997, Molecular Neurobiology.

[33]  S. Madihally,et al.  Blending chitosan with polycaprolactone: effects on physicochemical and antibacterial properties. , 2006, Biomacromolecules.

[34]  G. Stevens,et al.  Controllable surface modification of poly(lactic-co-glycolic acid) (PLGA) by hydrolysis or aminolysis I: physical, chemical, and theoretical aspects. , 2004, Biomacromolecules.

[35]  P. Richardson Neurotrophic factors in regeneration , 1991, Current Opinion in Neurobiology.

[36]  R. Martini Expression and functional roles of neural cell surface molecules and extracellular matrix components during development and regeneration of peripheral nerves , 1994, Journal of neurocytology.

[37]  Giovanni Vozzi,et al.  Blends of Poly-(ε-caprolactone) and Polysaccharides in Tissue Engineering Applications , 2005 .