Chitosan tubes enriched with fresh skeletal muscle fibers for delayed repair of peripheral nerve defects
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摘要
Nerve regeneration after delayed nerve repair is often unsuccessful. Indeed, the expression of genes associated with regeneration, including neurotrophic and gliotrophic factors, is drastically reduced in the distal stump of chronically transected nerves; moreover, Schwann cells undergo atrophy, losing their ability to sustain regeneration. In the present study, to provide a three-dimensional environment and trophic factors supporting Schwann cell activity and axon re-growth, we combined the use of an effective conduit(a chitosan tube) with a promising intraluminal structure(fresh longitudinal skeletal muscle fibers). This enriched conduit was used to repair a 10-mm rat median nerve gap after 3-month delay and functional and morphometrical analyses were performed 4 months after nerve reconstruction. Our data show that the enriched chitosan conduit is as effective as the hollow chitosan conduit in promoting nerve regeneration,and its efficacy is not statistically different from the autograft, considered the "gold standard" technique for nerve reconstruction. Since hollow tubes not always lead to good results after long defects(> 20 mm), we believe that the conduit enriched with fresh muscle fibers could be a promising strategy to repair longer gaps, as muscle fibers create a favorable three-dimensional environment and release trophic factors. All procedures were approved by the Bioethical Committee of the University of Torino and by the Italian Ministry of Health(approval number: 864/2016/PR) on September 14, 2016.
Nerve regeneration after delayed nerve repair is often unsuccessful. Indeed, the expression of genes associated with regeneration, including neurotrophic and gliotrophic factors, is drastically reduced in the distal stump of chronically transected nerves; moreover, Schwann cells undergo atrophy, losing their ability to sustain regeneration. In the present study, to provide a three-dimensional environment and trophic factors supporting Schwann cell activity and axon re-growth, we combined the use of an effective conduit(a chitosan tube) with a promising intraluminal structure(fresh longitudinal skeletal muscle fibers). This enriched conduit was used to repair a 10-mm rat median nerve gap after 3-month delay and functional and morphometrical analyses were performed 4 months after nerve reconstruction. Our data show that the enriched chitosan conduit is as effective as the hollow chitosan conduit in promoting nerve regeneration,and its efficacy is not statistically different from the autograft, considered the "gold standard" technique for nerve reconstruction. Since hollow tubes not always lead to good results after long defects(> 20 mm), we believe that the conduit enriched with fresh muscle fibers could be a promising strategy to repair longer gaps, as muscle fibers create a favorable three-dimensional environment and release trophic factors. All procedures were approved by the Bioethical Committee of the University of Torino and by the Italian Ministry of Health(approval number: 864/2016/PR) on September 14, 2016.
Nerve regeneration after delayed nerve repair is often unsuccessful. Indeed, the expression of genes associated with regeneration, including neurotrophic and gliotrophic factors, is drastically reduced in the distal stump of chronically transected nerves; moreover, Schwann cells undergo atrophy, losing their ability to sustain regeneration. In the present study, to provide a three-dimensional environment and trophic factors supporting Schwann cell activity and axon re-growth, we combined the use of an effective conduit(a chitosan tube) with a promising intraluminal structure(fresh longitudinal skeletal muscle fibers). This enriched conduit was used to repair a 10-mm rat median nerve gap after 3-month delay and functional and morphometrical analyses were performed 4 months after nerve reconstruction. Our data show that the enriched chitosan conduit is as effective as the hollow chitosan conduit in promoting nerve regeneration,and its efficacy is not statistically different from the autograft, considered the "gold standard" technique for nerve reconstruction. Since hollow tubes not always lead to good results after long defects(> 20 mm), we believe that the conduit enriched with fresh muscle fibers could be a promising strategy to repair longer gaps, as muscle fibers create a favorable three-dimensional environment and release trophic factors. All procedures were approved by the Bioethical Committee of the University of Torino and by the Italian Ministry of Health(approval number: 864/2016/PR) on September 14, 2016.
引文
Bak M,Gutkowska ON,Wagner E,Gosk J(2017)The role of chitin and chitosan in peripheral nerve reconstruction.Polim Med 47:43-47.
Battiston B,Tos P,Cushway TR,Geuna S(2000a)Nerve repair by means of vein filled with muscle grafts I.Clinical results.Microsurgery 20:32-36.
Battiston B,Tos P,Geuna S,Giacobini-Robecchi MG,Guglielmone R(2000b)Nerve repair by means of vein filled with muscle grafts.II.Morphological analysis of regeneration.Microsurgery 20:37-41.
Evriviades D,Jeffery S,Cubison T,Lawton G,Gill M,Mortiboy D(2011)Shaping the military wound:issues surrounding the reconstruction of injured servicemen at the Royal Centre for Defence Medicine.Philos Trans R Soc Lond B Biol Sci 366:219-230.
Faroni A,Mobasseri SA,Kingham PJ,Reid AJ(2015)Peripheral nerve regeneration:experimental strategies and future perspectives.Adv Drug Deliv Rev 82-83:160-167.
Geuna S(2000)Appreciating the difference between design-based and model-based sampling strategies in quantitative morphology of the nervous system.J Comp Neurol 427:333-339.
Geuna S,Tos P,Battiston B,Guglielmone R(2000)Verification of the two-dimensional disector,a method for the unbiased estimation of density and number of myelinated nerve fibers in peripheral nerves.Ann Anat 182:23-34.
Gonzalez-Perez F,Cobianchi S,Geuna S,Barwig C,Freier T,Udina E,Navarro X(2015)Tubulization with chitosan guides for the repair of long gap peripheral nerve injury in the rat.Microsurgery 35:300-308.
Gordon T,Tetzlaff W(2015)Regeneration-associated genes decline in chronically injured rat sciatic motoneurons.Eur J Neurosci 42:2783-2791.
Griffin MF,Malahias M,Hindocha S,Khan WS(2014)Peripheral nerve injury:principles for repair and regeneration.Open Orthop J8:199-203.
Grinsell D,Keating CP(2014)Peripheral nerve reconstruction after injury:a review of clinical and experimental therapies.Biomed Res Int 2014:698256.
Haastert-Talini K,Geuna S,Dahlin LB,Meyer C,Stenberg L,Freier T,Heimann C,Barwig C,Pinto LF,Raimondo S,Gambarotta G,Samy SR,Sousa N,Salgado AJ,Ratzka A,Wrobel S,Grothe C(2013)Chitosan tubes of varying degrees of acetylation for bridging peripheral nerve defects.Biomaterials 34:9886-9904.
Hallgren A,Bjorkman A,Chemnitz A,Dahlin LB(2013)Subjective outcome related to donor site morbidity after sural nerve graft harvesting:a survey in 41 patients.BMC Surg 13:39.
Hoke A(2006)Mechanisms of Disease:what factors limit the success of peripheral nerve regeneration in humans?Nat Clin Pract Neurol2:448-454.
Hoke A(2011)A(heat)shock to the system promotes peripheral nerve regeneration.J Clin Invest 121:4231-4234.
Jessen KR,Mirsky R(2016)The repair Schwann cell and its function in regenerating nerves.J Physiol 594:3521-3531.
Jones S,Eisenberg HM,Jia X(2016)Advances and future applications of augmented peripheral nerve regeneration.Int J Mol Sci 17.pii:E1494.
Manoli T,Schulz L,Stahl S,Jaminet P,Schaller HE(2014)Evaluation of sensory recovery after reconstruction of digital nerves of the hand using muscle-in-vein conduits in comparison to nerve suture or nerve autografting.Microsurgery 34:608-615.
Marcoccio I,Vigasio A(2010)Muscle-in-vein nerve guide for secondary reconstruction in digital nerve lesions.J Hand Surg Am 35:1418-1426.
Meyer C,Stenberg L,Gonzalez-Perez F,Wrobel S,Ronchi G,Udina E,Suganuma S,Geuna S,Navarro X,Dahlin LB,Grothe C,Haastert-Talini K(2016)Chitosan-film enhanced chitosan nerve guides for long-distance regeneration of peripheral nerves.Biomaterials76:33-51.
Michalski B,Bain JR,Fahnestock M(2008)Long-term changes in neurotrophic factor expression in distal nerve stump following denervation and reinnervation with motor or sensory nerve.J Neurochem105:1244-1252.
Montoya GJ,Sutachan JJ,Chan WS,Sideris A,Blanck TJ,Recio-Pinto E(2009)Muscle-conditioned media and cAMP promote survival and neurite outgrowth of adult spinal cord motor neurons.Exp Neurol 220:303-315.
Moore AM,Wagner IJ,Fox IK(2015)Principles of nerve repair in complex wounds of the upper extremity.Semin Plast Surg 29:40-47.
Papalia I,Geuna S,Tos PL,Boux E,Battiston B,Stagno D’Alcontres F(2003)Morphologic and functional study of rat median nerve repair by terminolateral neurorrhaphy of the ulnar nerve.J Reconstr Microsurg 19:257-264.
Pfister BJ,Gordon T,Loverde JR,Kochar AS,Mackinnon SE,Cullen DK(2011)Biomedical engineering strategies for peripheral nerve repair:surgical applications,state of the art,and future challenges.Crit Rev Biomed Eng 39:81-124.
Raimondo S,Nicolino S,Tos P,Battiston B,Giacobini-Robecchi MG,Perroteau I,Geuna S(2005)Schwann cell behavior after nerve repair by means of tissue-engineered muscle-vein combined guides.JComp Neurol 489:249-259.
Ronchi G,Nicolino S,Raimondo S,Tos P,Battiston B,Papalia I,Varejao AS,Giacobini-Robecchi MG,Perroteau I,Geuna S(2009)Functional and morphological assessment of a standardized crush injury of the rat median nerve.J Neurosci Methods 179:51-57.
Ronchi G,Cillino M,Gambarotta G,Fornasari BE,Raimondo S,Pugliese P,Tos P,Cordova A,Moschella F,Geuna S(2017)Irreversible changes occurring in long-term denervated Schwann cells affect delayed nerve repair.J Neurosurg 127:843-856.
Ronchi G,Fornasari BE,Crosio A,Budau CA,Tos P,Perroteau I,Battiston B,Geuna S,Raimondo S,Gambarotta G(2018)Chitosan Tubes Enriched with Fresh Skeletal Muscle Fibers for Primary Nerve Repair.Biomed Res Int 2018:9175248.
Sendtner M,Stockli KA,Thoenen H(1992)Synthesis and localization of ciliary neurotrophic factor in the sciatic nerve of the adult rat after lesion and during regeneration.J Cell Biol 118:139-148.
Shapira Y,Tolmasov M,Nissan M,Reider E,Koren A,Biron T,Bitan Y,Livnat M,Ronchi G,Geuna S,Rochkind S(2016)Comparison of results between chitosan hollow tube and autologous nerve graft in reconstruction of peripheral nerve defect:An experimental study.Microsurgery 36:664-671.
Stenberg L,Stossel M,Ronchi G,Geuna S,Yin Y,Mommert S,Martensson L,Metzen J,Grothe C,Dahlin LB,Haastert-Talini K(2017)Regeneration of long-distance peripheral nerve defects after delayed reconstruction in healthy and diabetic rats is supported by immunomodulatory chitosan nerve guides.BMC Neurosci 18:53.
St??el M,Wildhagen VM,Helmecke O,Metzen J,Pfund CB,Freier T,Haastert-Talini K(2018)Comparative Evaluation of Chitosan Nerve Guides with Regular or Increased Bendability for Acute and Delayed Peripheral Nerve Repair:A Comprehensive Comparison with Autologous Nerve Grafts and Muscle-in-Vein Grafts.Anat Rec(Hoboken)301:1697-1713.
Sulaiman W,Gordon T(2013)Neurobiology of peripheral nerve injury,regeneration,and functional recovery:from bench top research to bedside application.Ochsner J 13:100-108.
Sunderland S(1990)The anatomy and physiology of nerve injury.Muscle Nerve 13:771-784.
Battiston B,Tos P,Cushway TR,Geuna S(2000a)Nerve repair by means of vein filled with muscle grafts I.Clinical results.Microsurgery 20:32-36.
Battiston B,Tos P,Geuna S,Giacobini-Robecchi MG,Guglielmone R(2000b)Nerve repair by means of vein filled with muscle grafts.II.Morphological analysis of regeneration.Microsurgery 20:37-41.
Evriviades D,Jeffery S,Cubison T,Lawton G,Gill M,Mortiboy D(2011)Shaping the military wound:issues surrounding the reconstruction of injured servicemen at the Royal Centre for Defence Medicine.Philos Trans R Soc Lond B Biol Sci 366:219-230.
Faroni A,Mobasseri SA,Kingham PJ,Reid AJ(2015)Peripheral nerve regeneration:experimental strategies and future perspectives.Adv Drug Deliv Rev 82-83:160-167.
Geuna S(2000)Appreciating the difference between design-based and model-based sampling strategies in quantitative morphology of the nervous system.J Comp Neurol 427:333-339.
Geuna S,Tos P,Battiston B,Guglielmone R(2000)Verification of the two-dimensional disector,a method for the unbiased estimation of density and number of myelinated nerve fibers in peripheral nerves.Ann Anat 182:23-34.
Gonzalez-Perez F,Cobianchi S,Geuna S,Barwig C,Freier T,Udina E,Navarro X(2015)Tubulization with chitosan guides for the repair of long gap peripheral nerve injury in the rat.Microsurgery 35:300-308.
Gordon T,Tetzlaff W(2015)Regeneration-associated genes decline in chronically injured rat sciatic motoneurons.Eur J Neurosci 42:2783-2791.
Griffin MF,Malahias M,Hindocha S,Khan WS(2014)Peripheral nerve injury:principles for repair and regeneration.Open Orthop J8:199-203.
Grinsell D,Keating CP(2014)Peripheral nerve reconstruction after injury:a review of clinical and experimental therapies.Biomed Res Int 2014:698256.
Haastert-Talini K,Geuna S,Dahlin LB,Meyer C,Stenberg L,Freier T,Heimann C,Barwig C,Pinto LF,Raimondo S,Gambarotta G,Samy SR,Sousa N,Salgado AJ,Ratzka A,Wrobel S,Grothe C(2013)Chitosan tubes of varying degrees of acetylation for bridging peripheral nerve defects.Biomaterials 34:9886-9904.
Hallgren A,Bjorkman A,Chemnitz A,Dahlin LB(2013)Subjective outcome related to donor site morbidity after sural nerve graft harvesting:a survey in 41 patients.BMC Surg 13:39.
Hoke A(2006)Mechanisms of Disease:what factors limit the success of peripheral nerve regeneration in humans?Nat Clin Pract Neurol2:448-454.
Hoke A(2011)A(heat)shock to the system promotes peripheral nerve regeneration.J Clin Invest 121:4231-4234.
Jessen KR,Mirsky R(2016)The repair Schwann cell and its function in regenerating nerves.J Physiol 594:3521-3531.
Jones S,Eisenberg HM,Jia X(2016)Advances and future applications of augmented peripheral nerve regeneration.Int J Mol Sci 17.pii:E1494.
Manoli T,Schulz L,Stahl S,Jaminet P,Schaller HE(2014)Evaluation of sensory recovery after reconstruction of digital nerves of the hand using muscle-in-vein conduits in comparison to nerve suture or nerve autografting.Microsurgery 34:608-615.
Marcoccio I,Vigasio A(2010)Muscle-in-vein nerve guide for secondary reconstruction in digital nerve lesions.J Hand Surg Am 35:1418-1426.
Meyer C,Stenberg L,Gonzalez-Perez F,Wrobel S,Ronchi G,Udina E,Suganuma S,Geuna S,Navarro X,Dahlin LB,Grothe C,Haastert-Talini K(2016)Chitosan-film enhanced chitosan nerve guides for long-distance regeneration of peripheral nerves.Biomaterials76:33-51.
Michalski B,Bain JR,Fahnestock M(2008)Long-term changes in neurotrophic factor expression in distal nerve stump following denervation and reinnervation with motor or sensory nerve.J Neurochem105:1244-1252.
Montoya GJ,Sutachan JJ,Chan WS,Sideris A,Blanck TJ,Recio-Pinto E(2009)Muscle-conditioned media and cAMP promote survival and neurite outgrowth of adult spinal cord motor neurons.Exp Neurol 220:303-315.
Moore AM,Wagner IJ,Fox IK(2015)Principles of nerve repair in complex wounds of the upper extremity.Semin Plast Surg 29:40-47.
Papalia I,Geuna S,Tos PL,Boux E,Battiston B,Stagno D’Alcontres F(2003)Morphologic and functional study of rat median nerve repair by terminolateral neurorrhaphy of the ulnar nerve.J Reconstr Microsurg 19:257-264.
Pfister BJ,Gordon T,Loverde JR,Kochar AS,Mackinnon SE,Cullen DK(2011)Biomedical engineering strategies for peripheral nerve repair:surgical applications,state of the art,and future challenges.Crit Rev Biomed Eng 39:81-124.
Raimondo S,Nicolino S,Tos P,Battiston B,Giacobini-Robecchi MG,Perroteau I,Geuna S(2005)Schwann cell behavior after nerve repair by means of tissue-engineered muscle-vein combined guides.JComp Neurol 489:249-259.
Ronchi G,Nicolino S,Raimondo S,Tos P,Battiston B,Papalia I,Varejao AS,Giacobini-Robecchi MG,Perroteau I,Geuna S(2009)Functional and morphological assessment of a standardized crush injury of the rat median nerve.J Neurosci Methods 179:51-57.
Ronchi G,Cillino M,Gambarotta G,Fornasari BE,Raimondo S,Pugliese P,Tos P,Cordova A,Moschella F,Geuna S(2017)Irreversible changes occurring in long-term denervated Schwann cells affect delayed nerve repair.J Neurosurg 127:843-856.
Ronchi G,Fornasari BE,Crosio A,Budau CA,Tos P,Perroteau I,Battiston B,Geuna S,Raimondo S,Gambarotta G(2018)Chitosan Tubes Enriched with Fresh Skeletal Muscle Fibers for Primary Nerve Repair.Biomed Res Int 2018:9175248.
Sendtner M,Stockli KA,Thoenen H(1992)Synthesis and localization of ciliary neurotrophic factor in the sciatic nerve of the adult rat after lesion and during regeneration.J Cell Biol 118:139-148.
Shapira Y,Tolmasov M,Nissan M,Reider E,Koren A,Biron T,Bitan Y,Livnat M,Ronchi G,Geuna S,Rochkind S(2016)Comparison of results between chitosan hollow tube and autologous nerve graft in reconstruction of peripheral nerve defect:An experimental study.Microsurgery 36:664-671.
Stenberg L,Stossel M,Ronchi G,Geuna S,Yin Y,Mommert S,Martensson L,Metzen J,Grothe C,Dahlin LB,Haastert-Talini K(2017)Regeneration of long-distance peripheral nerve defects after delayed reconstruction in healthy and diabetic rats is supported by immunomodulatory chitosan nerve guides.BMC Neurosci 18:53.
St??el M,Wildhagen VM,Helmecke O,Metzen J,Pfund CB,Freier T,Haastert-Talini K(2018)Comparative Evaluation of Chitosan Nerve Guides with Regular or Increased Bendability for Acute and Delayed Peripheral Nerve Repair:A Comprehensive Comparison with Autologous Nerve Grafts and Muscle-in-Vein Grafts.Anat Rec(Hoboken)301:1697-1713.
Sulaiman W,Gordon T(2013)Neurobiology of peripheral nerve injury,regeneration,and functional recovery:from bench top research to bedside application.Ochsner J 13:100-108.
Sunderland S(1990)The anatomy and physiology of nerve injury.Muscle Nerve 13:771-784.