A composite poly‐hydroxybutyrate–glial growth factor conduit for long nerve gap repairs

There is considerable evidence that peripheral nerves have the potential to regenerate in an appropriate microenvironment. We have developed a novel artificial nerve guide composed of poly 3‐hydroxybutyrate (PHB) filled with glial growth factor (GGF) suspended in alginate hydrogel. Gaps of 2–4 cm in rabbit common peroneal nerve were bridged using a PHB conduit containing either GGF in alginate hydrogel (GGF) or alginate alone (Alginate), or with an empty PHB conduit (Empty). Tissues were harvested 21, 42 and 63 days post‐operatively. Schwann cell and axonal regeneration were assessed using quantitative immunohistochemistry. At 21 days, addition of GGF increased significantly the distance of axonal and Schwann cells regeneration in comparison with that observed in Alginate and Empty conduits for both gap lengths. The axons bridged the 2‐cm GGF conduits gap by 63 days, with a comparable rate of regeneration seen in 4‐cm conduits. Schwann cells and axonal regeneration quantity was similar for both gap lengths in each group. However, at all time points the quantity of axonal and Schwann cells regeneration in GGF grafts was significantly greater than in both Alginate and Empty conduits, the latter showing better regeneration than Alginate conduits. The results indicate an inhibitory effect of alginate on regeneration, which is partially reversed by the addition of GGF to the conduits. In conclusion, GGF stimulates a progressive and sustainable regeneration increase in long nerve gap conduits.

[1]  T. Gilchrist,et al.  In vitro nerve repair--in vivo. The reconstruction of peripheral nerves by entubulation with biodegradeable glass tubes--a preliminary report. , 1998, British journal of plastic surgery.

[2]  G. Brunelli,et al.  Different conduits in peripheral nerve surgery. , 1994, Microsurgery.

[3]  S. Mackinnon,et al.  The Role of Conduits in Nerve Repair: A Review , 1996, Reviews in the neurosciences.

[4]  Dellon Al,et al.  Reeducation of sensation in the hand following nerve suture. , 1982 .

[5]  M. Wiberg,et al.  Addition of fibronectin to alginate matrix improves peripheral nerve regeneration in tissue-engineered conduits. , 2003, Tissue engineering.

[6]  J. Fawcett,et al.  Muscle basal lamina: a new graft material for peripheral nerve repair. , 1986, Journal of neurosurgery.

[7]  G. Brunelli,et al.  Invited review: Different conduits in peripheral nerve surgery , 1994 .

[8]  R. Mirsky,et al.  Origin and early development of Schwann cells , 1998, Microscopy research and technique.

[9]  S. Mackinnon,et al.  A Study of Nerve Regeneration Across Synthetic (Maxon) and Biologic (Collagen) Nerve Conduits For Nerve Gaps Up to 5 Cm in the Primate , 1990, Journal of reconstructive microsurgery.

[10]  D. Finkelstein,et al.  Leukemia inhibitory factor enhances the regeneration of transected rat sciatic nerve and the function of reinnervated muscle , 1997, Journal of neuroscience research.

[11]  Lingzhi Fan,et al.  The glucose oxidase-DAB-nickel method in peroxidase histochemistry of the nervous system , 1988, Neuroscience Letters.

[12]  Göran Lundborg,et al.  Nerve regeneration in silicone chambers: Influence of gap length and of distal stump components , 1982, Experimental Neurology.

[13]  Mark Ellisman,et al.  Nerve regeneration through artificial tubular implants , 1989, Progress in Neurobiology.

[14]  Y. Yarden,et al.  Neu differentiation factor is a neuron-glia signal and regulates survival, proliferation, and maturation of rat schwann cell precursors , 1995, Neuron.

[15]  W. Richardson,et al.  Glial cell development : basic principles and clinical relevance , 1997 .

[16]  A. Dellon,et al.  Reeducation of sensation in the hand following nerve suture. , 1982, Clinical orthopaedics and related research.

[17]  G. Terenghi,et al.  Differential effects of NT‐3 on reinnervation of the fast extensor digitorum longus (EDL) and the slow soleus muscle of rat , 2000, The European journal of neuroscience.

[18]  G Lundborg,et al.  A 25-year perspective of peripheral nerve surgery: evolving neuroscientific concepts and clinical significance. , 2000, Journal of Hand Surgery-American Volume.

[19]  R. Koller,et al.  Role of a muscle target organ on the regeneration of motor nerve fibres in long nerve grafts: a synopsis of experimental and clinical data. , 1996, Microsurgery.

[20]  C. Heath,et al.  The development of bioartificial nerve grafts for peripheral-nerve regeneration. , 1998, Trends in biotechnology.

[21]  R. Brown,et al.  Neurotrophin‐3 Delivered Locally via Fibronectin Mats Enhances Peripheral Nerve Regeneration , 1997, The European journal of neuroscience.

[22]  X. Navarro,et al.  Influence of Collagen and Laminin Gels Concentration on Nerve Regeneration after Resection and Tube Repair , 1998, Experimental Neurology.

[23]  D. Wise,et al.  Influence of glial growth factor and Schwann cells in a bioresorbable guidance channel on peripheral nerve regeneration. , 2000, Tissue engineering.

[24]  Yoshihisa Suzuki,et al.  Cat peripheral nerve regeneration across 50 mm gap repaired with a novel nerve guide composed of freeze-dried alginate gel , 1999, Neuroscience Letters.

[25]  D. Erdmann,et al.  The Use of Cultured Schwann Cells in Nerve Repair in a Rabbit Hind-Limb Model , 1996, Journal of reconstructive microsurgery.

[26]  R. Mirsky,et al.  Schwann cells and their precursors emerge as major regulators of nerve development , 1999, Trends in Neurosciences.

[27]  S. Hall THE EFFECT OF INHIBITING SCHWANN CELL MITOSIS ON THE RE‐INNERVATION OF ACELLULAR AUTOGRAFTS IN THE PERIPHERAL NERVOUS SYSTEM OF THE MOUSE , 1986, Neuropathology and applied neurobiology.

[28]  P. M. Galletti,et al.  Collagen- and laminin-containing gels impede peripheral nerve regeneration through semipermeable nerve guidance channels , 1987, Experimental Neurology.

[29]  E. Anton,et al.  Glial Growth Factor 2, a Soluble Neuregulin, Directly Increases Schwann Cell Motility and Indirectly Promotes Neurite Outgrowth , 1996, The Journal of Neuroscience.

[30]  C. Woolf,et al.  Reciprocal Schwann cell-axon interactions , 1993, Current Opinion in Neurobiology.

[31]  Tatsuo Nakamura,et al.  Nerve regeneration across a 25-mm gap bridged by a polyglycolic acid-collagen tube: a histological and electrophysiological evaluation of regenerated nerves , 1996, Brain Research.

[32]  M. Wiberg,et al.  A resorbable nerve conduit as an alternative to nerve autograft in nerve gap repair. , 1999, British journal of plastic surgery.

[33]  G. Terenghi,et al.  Peripheral nerve regeneration and neurotrophic factors , 1999, Journal of anatomy.

[34]  M. Milek,et al.  Results of Nerve Grafting in Severe Soft Tissue Injuries , 1984, Clinical orthopaedics and related research.

[35]  Robert A. Brown,et al.  Targeted Delivery of Nerve Growth Factor via Fibronectin Conduits Assists Nerve Regeneration in Control and Diabetic Rats , 1995, The European journal of neuroscience.

[36]  C. Doré,et al.  Increased axonal regeneration over long nerve gaps using autologous nerve‐muscle sandwich grafts , 1995, Microsurgery.

[37]  B. Strauch,et al.  Determining the maximal length of a vein conduit used as an interposition graft for nerve regeneration. , 1996, Journal of reconstructive microsurgery.

[38]  M. Glasby,et al.  The Limit of Graft Length in the Experimental use of Muscle Grafts for Nerve Repair , 1993, Journal of hand surgery.

[39]  R. C. Wray,et al.  The results of nerve grafting in the wrist and hand. , 1980 .

[40]  M. Schachner,et al.  Restricted localization of L1 and N‐CAM at sites of contact between Schwann cells and neurites in culture , 1994, Glia.

[41]  G. Terenghi,et al.  Effects of delayed re‐innervation on the expression of c‐erbB receptors by chronically denervated rat Schwann cells in vivo , 1997, Glia.

[42]  G. Lundborg,et al.  Temporal changes of neuronotrophic activities accumulating in vivo within nerve regeneration chambers , 1983, Experimental Neurology.

[43]  M. Wiberg,et al.  Poly-3-hydroxybutyrate (PHB): a resorbable conduit for long-gap repair in peripheral nerves. , 2002, British journal of plastic surgery.

[44]  L. Austin,et al.  Controlled release of leukaemia inhibitory factor (LIF) to tissues. , 1997, Growth factors.

[45]  W. Shaw,et al.  In vitro slow release profile of endothelial cell growth factor immobilized within calcium alginate microbeads. , 1995, Artificial cells, blood substitutes, and immobilization biotechnology.

[46]  J. Trachtenberg,et al.  Schwann cell apoptosis at developing neuromuscular junctions is regulated by glial growth factor , 1996, Nature.