Photocrosslinkable chitosan based hydrogels for neural tissue engineering.

Hydrogel based scaffolds for neural tissue engineering can provide appropriate physico-chemical and mechanical properties to support neurite extension and facilitate transplantation of cells by acting as 'cell delivery vehicles'. Specifically, in situ gelling systems such as photocrosslinkable hydrogels can potentially conformally fill irregular neural tissue defects and serve as stem cell delivery systems. Here, we report the development of a novel chitosan based photocrosslinkable hydrogel system with tunable mechanical properties and degradation rates. A two-step synthesis of amino-ethyl methacrylate derivitized, degradable, photocrosslinkable chitosan hydrogels is described. When human mesenchymal stem cells were cultured in photocrosslinkable chitosan hydrogels, negligible cytotoxicity was observed. Photocrosslinkable chitosan hydrogels facilitated enhanced neurite differentiation from primary cortical neurons and enhanced neurite extension from dorsal root ganglia (DRG) as compared to agarose based hydrogels with similar storage moduli. Neural stem cells (NSCs) cultured within photocrosslinkable chitosan hydrogels facilitated differentiation into tubulin positive neurons and astrocytes. These data demonstrate the potential of photocrosslinked chitosan hydrogels, and contribute to an increasing repertoire of hydrogels designed for neural tissue engineering.

[1]  Xiaojun Yu,et al.  Tissue-engineered scaffolds are effective alternatives to autografts for bridging peripheral nerve gaps. , 2003, Tissue engineering.

[2]  Yingjun Wang,et al.  In Situ Fabrication of Nano-hydroxyapatite in a Macroporous Chitosan Scaffold for Tissue Engineering , 2009, Journal of biomaterials science. Polymer edition.

[3]  Y. Gong,et al.  Preparation of cross-linked carboxymethyl chitosan for repairing sciatic nerve injury in rats , 2009, Biotechnology Letters.

[4]  Erin B Lavik,et al.  A library of tunable poly(ethylene glycol)/poly(L-lysine) hydrogels to investigate the material cues that influence neural stem cell differentiation. , 2009, Journal of biomedical materials research. Part A.

[5]  Nic D. Leipzig,et al.  Differentiation of neural stem cells in three-dimensional growth factor-immobilized chitosan hydrogel scaffolds. , 2011, Biomaterials.

[6]  R V Bellamkonda,et al.  The influence of physical structure and charge on neurite extension in a 3D hydrogel scaffold. , 1998, Journal of biomaterials science. Polymer edition.

[7]  Ravi V Bellamkonda,et al.  The role of aligned polymer fiber-based constructs in the bridging of long peripheral nerve gaps. , 2008, Biomaterials.

[8]  S. Nicoll,et al.  Characterization of novel photocrosslinked carboxymethylcellulose hydrogels for encapsulation of nucleus pulposus cells. , 2010, Acta biomaterialia.

[9]  S. Weiss,et al.  BDNF enhances the differentiation but not the survival of CNS stem cell- derived neuronal precursors , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[10]  M. Dadsetan,et al.  Stimulation of neurite outgrowth using positively charged hydrogels. , 2009, Biomaterials.

[11]  M. Dadsetan,et al.  Effect of hydrogel porosity on marrow stromal cell phenotypic expression. , 2008, Biomaterials.

[12]  Efstathios Karathanasis,et al.  Tumor Vascular Permeability to a Nanoprobe Correlates to Tumor-Specific Expression Levels of Angiogenic Markers , 2009, PloS one.

[13]  R. Midha,et al.  Analysis of upper and lower extremity peripheral nerve injuries in a population of patients with multiple injuries. , 1998, The Journal of trauma.

[14]  Rajiv Midha,et al.  Peripheral nerve regeneration through a synthetic hydrogel nerve tube. , 2005, Restorative neurology and neuroscience.

[15]  Robert Langer,et al.  Peritoneal application of chitosan and UV-cross-linkable chitosan. , 2006, Journal of biomedical materials research. Part A.

[16]  D. K. Cullen,et al.  Collagen-Dependent Neurite Outgrowth and Response to Dynamic Deformation in Three-Dimensional Neuronal Cultures , 2007, Annals of Biomedical Engineering.

[17]  Diane Hoffman-Kim,et al.  Topography, cell response, and nerve regeneration. , 2010, Annual review of biomedical engineering.

[18]  David F Meaney,et al.  Matrices with compliance comparable to that of brain tissue select neuronal over glial growth in mixed cortical cultures. , 2006, Biophysical journal.

[19]  S. Nicoll,et al.  Development of photocrosslinked methylcellulose hydrogels for soft tissue reconstruction. , 2009, Acta biomaterialia.

[20]  Mark W. Grinstaff,et al.  SYNTHESIS OF A NOVEL POLYSACCHARIDE HYDROGEL , 1999 .

[21]  Young-tae Kim,et al.  In situ gelling hydrogels for conformal repair of spinal cord defects, and local delivery of BDNF after spinal cord injury. , 2006, Biomaterials.

[22]  Y. Gong,et al.  Degradation of covalently cross-linked carboxymethyl chitosan and its potential application for peripheral nerve regeneration , 2007 .

[23]  Christine E Schmidt,et al.  Neural tissue engineering: strategies for repair and regeneration. , 2003, Annual review of biomedical engineering.

[24]  R. Shi,et al.  Rapidly photo-cross-linkable chitosan hydrogel for peripheral neurosurgeries. , 2011, Biomacromolecules.

[25]  S. Pricl,et al.  Flow properties of N-(carboxymethyl) chitosan aqueous systems in the sol and gel domains. , 1990, International journal of biological macromolecules.

[26]  B. Amsden,et al.  Methacrylated glycol chitosan as a photopolymerizable biomaterial. , 2007, Biomacromolecules.

[27]  Zhangqi Feng,et al.  Cellular compatibility of RGD-modified chitosan nanofibers with aligned or random orientation , 2010, Biomedical materials.

[28]  Mark W. Tibbitt,et al.  Hydrogels as extracellular matrix mimics for 3D cell culture. , 2009, Biotechnology and bioengineering.

[29]  Jason B Shear,et al.  The effects of hyaluronic acid hydrogels with tunable mechanical properties on neural progenitor cell differentiation. , 2010, Biomaterials.

[30]  Robert Langer,et al.  Stimulation of neurite outgrowth by neurotrophins delivered from degradable hydrogels. , 2006, Biomaterials.

[31]  R. Bellamkonda,et al.  The polarity and magnitude of ambient charge influences three-dimensional neurite extension from DRGs. , 2000, Journal of biomedical materials research.

[32]  David A Stenger,et al.  Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices , 2001, Neuroscience Letters.

[33]  Ze Zhang,et al.  Synthesis and property studies of N‐carboxymethyl chitosan , 2011 .

[34]  R V Bellamkonda,et al.  Dorsal root ganglia neurite extension is inhibited by mechanical and chondroitin sulfate‐rich interfaces , 2001, Journal of neuroscience research.

[35]  R V Bellamkonda,et al.  Polylysine-functionalised thermoresponsive chitosan hydrogel for neural tissue engineering. , 2007, Biomaterials.

[36]  Wei He,et al.  Nanoscale neuro-integrative coatings for neural implants. , 2005, Biomaterials.

[37]  Glenn D Prestwich,et al.  In situ crosslinkable hyaluronan hydrogels for tissue engineering. , 2004, Biomaterials.

[38]  R. Gilbert,et al.  Fabrication and characterization of tunable polysaccharide hydrogel blends for neural repair. , 2011, Acta biomaterialia.

[39]  M. LaPlaca,et al.  Variations in rigidity and ligand density influence neuronal response in methylcellulose-laminin hydrogels. , 2011, Acta biomaterialia.

[40]  R V Bellamkonda,et al.  Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures. , 2001, Biomaterials.

[41]  M. Grinstaff,et al.  Photocrosslinkable polysaccharides for in situ hydrogel formation. , 2001, Journal of biomedical materials research.

[42]  T. Georgiou,et al.  Synthesis, characterization, and DNA adsorption studies of ampholytic model conetworks based on cross-linked star copolymers. , 2008, Biomacromolecules.

[43]  Charles Tator,et al.  Chitosan implants in the rat spinal cord: biocompatibility and biodegradation. , 2011, Journal of biomedical materials research. Part A.

[44]  Wei He,et al.  Simple agarose-chitosan gel composite system for enhanced neuronal growth in three dimensions. , 2009, Biomacromolecules.

[45]  Spinal cord injury facts and figures at a glance. , 2013, The journal of spinal cord medicine.

[46]  Christine E Schmidt,et al.  Photocrosslinked hyaluronic acid hydrogels: natural, biodegradable tissue engineering scaffolds. , 2003, Biotechnology and bioengineering.

[47]  Michelle C LaPlaca,et al.  Thermoreversible laminin-functionalized hydrogel for neural tissue engineering. , 2006, Journal of biomedical materials research. Part A.

[48]  Ravi V Bellamkonda,et al.  Peripheral nerve regeneration: an opinion on channels, scaffolds and anisotropy. , 2006, Biomaterials.

[49]  Ravi V Bellamkonda,et al.  Differences between the effect of anisotropic and isotropic laminin and nerve growth factor presenting scaffolds on nerve regeneration across long peripheral nerve gaps. , 2008, Biomaterials.

[50]  Jinghua Hao,et al.  RNA extraction from polysaccharide-based cell-laden hydrogel scaffolds. , 2008, Analytical biochemistry.

[51]  F. Cui,et al.  Viability and differentiation of neural precursors on hyaluronic acid hydrogel scaffold , 2009, Journal of neuroscience research.

[52]  Y. Thomann,et al.  Synthesis and characterization of anionic amphiphilic model conetworks of 2-butyl-1-octyl-methacrylate and methacrylic acid: effects of polymer composition and architecture. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[53]  S. Nair,et al.  Novel chitin and chitosan nanofibers in biomedical applications. , 2010, Biotechnology advances.

[54]  Nic D. Leipzig,et al.  The effect of substrate stiffness on adult neural stem cell behavior. , 2009, Biomaterials.

[55]  Kristi S Anseth,et al.  Photocrosslinking of gelatin macromers to synthesize porous hydrogels that promote valvular interstitial cell function. , 2009, Tissue engineering. Part A.

[56]  Ravi V Bellamkonda,et al.  Anisotropic scaffolds facilitate enhanced neurite extension in vitro. , 2006, Journal of biomedical materials research. Part A.

[57]  Vivian Charles McAlister,et al.  Prevention of experimental postoperative peritoneal adhesions by N,O-carboxymethyl chitosan. , 1996, Surgery.

[58]  P. Dario,et al.  Polymer electret guidance channels enhance peripheral nerve regeneration in mice , 1989, Brain Research.

[59]  C. Laurencin,et al.  Biologically active chitosan systems for tissue engineering and regenerative medicine. , 2008, Current topics in medicinal chemistry.

[60]  Nic D. Leipzig,et al.  Functional immobilization of interferon-gamma induces neuronal differentiation of neural stem cells. , 2009, Journal of biomedical materials research. Part A.

[61]  Charles Tator,et al.  Effects of Dibutyryl Cyclic-AMP on Survival and Neuronal Differentiation of Neural Stem/Progenitor Cells Transplanted into Spinal Cord Injured Rats , 2011, PloS one.

[62]  R. Bellamkonda,et al.  Targeted downregulation of N‐acetylgalactosamine 4‐sulfate 6‐O‐sulfotransferase significantly mitigates chondroitin sulfate proteoglycan‐mediated inhibition , 2011, Glia.

[63]  Eben Alsberg,et al.  Photocrosslinked alginate hydrogels with tunable biodegradation rates and mechanical properties. , 2009, Biomaterials.

[64]  Wim E Hennink,et al.  The effect of photopolymerization on stem cells embedded in hydrogels. , 2009, Biomaterials.

[65]  M. Shoichet,et al.  Peptide surface modification of methacrylamide chitosan for neural tissue engineering applications. , 2007, Journal of biomedical materials research. Part A.

[66]  Ravi S Kane,et al.  The influence of hydrogel modulus on the proliferation and differentiation of encapsulated neural stem cells. , 2009, Biomaterials.

[67]  Y. Ozeki,et al.  Controlled release of paclitaxel from photocrosslinked chitosan hydrogels and its subsequent effect on subcutaneous tumor growth in mice. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[68]  M. Rinaudo,et al.  Substituent distribution on O,N-carboxymethylchitosans by 1H and 13C n.m.r. , 1992, International journal of biological macromolecules.

[69]  Jochen Guck,et al.  Viscoelastic properties of individual glial cells and neurons in the CNS , 2006, Proceedings of the National Academy of Sciences.

[70]  I. Kwon,et al.  Fabrication of a pure porous chitosan bead matrix: influences of phase separation on the microstructure , 2002, Journal of biomaterials science. Polymer edition.

[71]  S. Madihally,et al.  Porous chitosan scaffolds for tissue engineering. , 1999, Biomaterials.