Controlled release of neurotrophin-3 from fibrin gels for spinal cord injury.

The goal of this work was to assess the feasibility of using affinity-based delivery systems to release neurotrophin-3 (NT-3) in a controlled manner from fibrin gels as a therapy for spinal cord injury. A heparin-based delivery system (HBDS) was used to immobilize NT-3 within fibrin gels via non-covalent interactions to slow diffusion-based release of NT-3, thus allowing cell-activated degradation of fibrin to mediate release. The HBDS consists of three components: immobilized linker peptide, heparin and NT-3. The linker peptide contained a Factor XIIIa substrate and was covalently cross-linked to fibrin during polymerization. This immobilized linker peptide sequesters heparin within fibrin gels, and sequestered heparin binds NT-3, preventing its diffusion. Mathematical modeling was performed to examine the effect of heparin concentration on the fraction of NT-3 initially bound to fibrin. In vitro release studies confirmed that heparin concentration modulates diffusion-based release of NT-3. Fibrin gels containing the HBDS and NT-3 stimulated neural outgrowth from chick dorsal root ganglia by up to 54% versus unmodified fibrin, demonstrating that the NT-3 released is biologically active. In a preliminary in vivo study, fibrin gels containing the HBDS and NT-3 showed increased neural fiber density in spinal cord lesions versus unmodified fibrin at 9 days.

[1]  M. Tuszynski,et al.  Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury , 2003, Experimental Neurology.

[2]  S. Olson,et al.  Binding of high affinity heparin to antithrombin III. Stopped flow kinetic studies of the binding interaction. , 1981, The Journal of biological chemistry.

[3]  A. Tessler,et al.  Neurotrophic agents in fibrin glue mediate adult dorsal root regeneration into spinal cord. , 1999, Neurosurgery.

[4]  W. Tetzlaff,et al.  BDNF and NT‐3, but not NGF, Prevent Axotomy‐induced Death of Rat Corticospinal Neurons In Vivo , 1996, The European journal of neuroscience.

[5]  R. Fletterick,et al.  Crystal structure of neurotrophin-3 homodimer shows distinct regions are used to bind its receptors. , 1998, Biochemistry.

[6]  B. Olwin,et al.  Activating and inhibitory heparin sequences for FGF-2 (basic FGF). Distinct requirements for FGF-1, FGF-2, and FGF-4. , 1993, The Journal of biological chemistry.

[7]  J. Mcdonald,et al.  Spinal-cord injury , 2002, The Lancet.

[8]  S. McMahon,et al.  NT‐3 promotes growth of lesioned adult rat sensory axons ascending in the dorsal columns of the spinal cord , 1999, The European journal of neuroscience.

[9]  Yihai Cao,et al.  Spinal Cord Repair in Adult Paraplegic Rats: Partial Restoration of Hind Limb Function , 1996, Science.

[10]  W. Gispen,et al.  Collagen Containing Neurotrophin-3 (NT-3) Attracts Regrowing Injured Corticospinal Axons in the Adult Rat Spinal Cord and Promotes Partial Functional Recovery , 1998, Experimental Neurology.

[11]  J. Hubbell,et al.  Development of fibrin derivatives for controlled release of heparin-binding growth factors. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[12]  B. J. Baumgartner,et al.  Neurotrophin-3 Expressed In Situ Induces Axonal Plasticity in the Adult Injured Spinal Cord , 2003, The Journal of Neuroscience.

[13]  Robert B. Harris,et al.  Heparin binding domain peptides of antithrombin III: Analysis by isothermal titration calorimetry and circular dichroism spectroscopy , 1994, Protein science : a publication of the Protein Society.

[14]  J. Hubbell,et al.  Effects of fibrinolysis on neurite growth from dorsal root ganglia cultured in two‐ and three‐dimensional fibrin gels , 1996, The Journal of comparative neurology.

[15]  C. Henderson.  Role of neurotrophic factors in neuronal development , 1996, Current Opinion in Neurobiology.

[16]  E. Edelman,et al.  Kinetics of basic fibroblast growth factor binding to its receptor and heparan sulfate proteoglycan: a mechanism for cooperactivity. , 1992, Biochemistry.

[17]  M. Murray,et al.  Neurotrophin‐3 prevents death of axotomized Clarke's nucleus neurons in adult rat , 1998, The Journal of comparative neurology.

[18]  S. Mörk,et al.  Class III β-Tubulin Isotype: A Key Cytoskeletal Protein at the Crossroads of Developmental Neurobiology and Tumor Neuropathology , 2003, Journal of child neurology.

[19]  S. Zigmond.  Consequences of chemotactic peptide receptor modulation for leukocyte orientation , 1981, The Journal of cell biology.

[20]  F. Gage,et al.  Cellular Delivery of Neurotrophin-3 Promotes Corticospinal Axonal Growth and Partial Functional Recovery after Spinal Cord Injury , 1997, The Journal of Neuroscience.

[21]  H. Buettner,et al.  Neuronal plasminogen activators: cell surface binding sites and involvement in neurite outgrowth , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[22]  J. Kurtzke,et al.  Epidemiology of spinal cord injury , 1975, Experimental Neurology.

[23]  B. Stokes,et al.  Brain-Derived Neurotrophic Factor Stimulates Hindlimb Stepping and Sprouting of Cholinergic Fibers after Spinal Cord Injury , 1998, Experimental Neurology.

[24]  M. Saarma,et al.  Neurotrophins as in vitro growth cone guidance molecules for embryonic sensory neurons , 1997, Cell and Tissue Research.

[25]  A. Davies.  The role of neurotrophins in the developing nervous system. , 1994, Journal of neurobiology.

[26]  Yves-Alain Barde,et al.  Neurotrophin-3 enhances sprouting of corticospinal tract during development and after adult spinal cord lesion , 1994, Nature.

[27]  P. Gettins,et al.  Examination, by 1H-n.m.r. spectroscopy, of the binding of a synthetic, high-affinity heparin pentasaccharide to human antithrombin III. , 1989, Carbohydrate research.

[28]  T. Tamaki,et al.  Factor XIII‐mediated cross‐linking of NH2‐terminal peptide of α2‐plasmin inhibitor to fibrin , 1983 .

[29]  M Sobel,et al.  Structure-function relations of antithrombin III-heparin interactions as assessed by biophysical and biological assays and molecular modeling of peptide-pentasaccharide-docked complexes. , 1996, Archives of biochemistry and biophysics.

[30]  X. Cao,et al.  Investigating the synergistic effect of combined neurotrophic factor concentration gradients to guide axonal growth , 2003, Neuroscience.

[31]  Göran Lundborg,et al.  Spatial‐Temporal progress of peripheral nerve regeneration within a silicone chamber: Parameters for a bioassay , 1983, The Journal of comparative neurology.

[32]  M. Tuszynski,et al.  NT-3 gene delivery elicits growth of chronically injured corticospinal axons and modestly improves functional deficits after chronic scar resection , 2003, Experimental Neurology.

[33]  J. Hubbell,et al.  Cross-linking exogenous bifunctional peptides into fibrin gels with factor XIIIa. , 1999, Bioconjugate chemistry.

[34]  M. Brenner,et al.  Controlled release of nerve growth factor enhances sciatic nerve regeneration , 2003, Experimental Neurology.

[35]  S. Kridel,et al.  Requirement of Lysine Residues Outside of the Proposed Pentasaccharide Binding Region for High Affinity Heparin Binding and Activation of Human Antithrombin III* , 1996, The Journal of Biological Chemistry.

[36]  J. Hubbell,et al.  Controlled release of nerve growth factor from a heparin-containing fibrin-based cell ingrowth matrix. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[37]  R. Linhardt,et al.  Nature of the interaction of heparin with acidic fibroblast growth factor. , 1993, Biochemistry.