RESUMEN
Nerve repair in several mm-long nerve gaps often requires an interventional technology. Microchannel scaffolds have proven effective for bridging nerve gaps and guiding axons in the peripheral nervous system (PNS). Nonetheless, fabricating microchannel scaffolds at this length scale remains a challenge and/or is time consuming and cumbersome. In this work, a simple computer-aided microdrilling technique was used to fabricate 10 mm-long agarose scaffolds consisting of 300 µm-microchannels and 85 µm-thick walls in less than an hour. The agarose scaffolds alone, however, did not exhibit adequate stiffness and integrity to withstand the mechanical stresses during implantation and suturing. To provide mechanical support and enable suturing, poly caprolactone (PCL) conduits were fabricated and agarose scaffolds were placed inside. A modified salt-leaching technique was developed to introduce interconnected porosity in PCL conduits to allow for tuning of the mechanical properties such as elastic modulus and strain to failure. It was shown that the PCL conduits were effective in stabilizing the agarose scaffolds in 10 mm-long sciatic nerve gaps of rats for at least 8 weeks. Robust axon ingress and Schwann cell penetration were observed within the microchannel scaffolds without using growth factors. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3392-3399, 2017.
Asunto(s)
Regeneración Tisular Dirigida/métodos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Regeneración Nerviosa , Poliésteres/química , Nervio Ciático/fisiología , Sefarosa/química , Andamios del Tejido/química , Animales , Módulo de Elasticidad , Masculino , Porosidad , Ratas Sprague-Dawley , Células de Schwann/citología , Nervio Ciático/citología , Nervio Ciático/lesionesRESUMEN
Schwann cells (SCs) detect injury to peripheral nerves and transform phenotypically to respond to injury and facilitate repair. Cell-signaling pathways and changes in gene expression that drive SC phenotypic transformation in injury have been described; however, the SC receptors that detect peripheral nervous system (PNS) injury have not been identified. LDL receptor-related protein 1 (LRP1) is a receptor for numerous ligands, including intracellular proteins released by injured cells and protein components of degenerated myelin. In certain cell types, including SCs, LRP1 is a cell-signaling receptor. Here, we show that binding of the LRP1 ligand, tissue-type plasminogen activator (tPA), to cultured rat SCs induces c-Jun phosphorylation, a central event in activation of the SC repair program. The response to tPA was blocked by the LRP1 antagonist, receptor-associated protein. c-Jun phosphorylation was also observed when cultured rat SCs were treated with a recombinant derivative of matrix metalloproteinase-9 that contains the LRP1 recognition motif (PEX). The ability of LRP1 to induce c-Jun phosphorylation and ERK1/2 activation was confirmed using cultures of human SCs. When tPA or PEX was injected directly into crush-injured rat sciatic nerves, c-Jun phosphorylation and ERK1/2 activation were observed in SCs in vivo. The ability of LRP1 to bind proteins released in the earliest stages of PNS injury and to induce c-Jun phosphorylation support a model in which SC LRP1 functions as an injury-detection receptor in the PNS.
Asunto(s)
Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Proteínas Proto-Oncogénicas c-jun/metabolismo , Células de Schwann/metabolismo , Neuropatía Ciática/metabolismo , Animales , Animales Recién Nacidos , Células Cultivadas , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Regulación de la Expresión Génica/efectos de los fármacos , Proteína Asociada a Proteínas Relacionadas con Receptor de LDL/farmacología , Metaloproteinasa 9 de la Matriz/farmacología , Endopeptidasa Neutra Reguladora de Fosfato PHEX/farmacología , Fosforilación/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Células de Schwann/efectos de los fármacos , Nervio Ciático/citología , Neuropatía Ciática/tratamiento farmacológico , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , Activador de Tejido PlasminógenoRESUMEN
NMDA receptors (NMDA-Rs) are ionotropic glutamate receptors, which associate with LDL-receptor-related protein-1 (LRP1) to trigger cell signaling in response to protein ligands in neurons. Here, we demonstrate for the first time that the NMDA-R is expressed by rat Schwann cells and functions independently and with LRP1 to regulate Schwann cell physiology. The NR1 (encoded by GRIN1) and NR2b (encoded by GRIN2B) NMDA-R subunits were expressed by cultured Schwann cells and upregulated in sciatic nerves following crush injury. The ability of LRP1 ligands to activate ERK1/2 (also known as MAPK3 and MAPK1, respectively) and promote Schwann cell migration required the NMDA-R. NR1 gene silencing compromised Schwann cell survival. Injection of the LRP1 ligands tissue-type plasminogen activator (tPA, also known as PLAT) or MMP9-PEX into crush-injured sciatic nerves activated ERK1/2 in Schwann cells in vivo, and the response was blocked by systemic treatment with the NMDA-R inhibitor MK801. tPA was unique among the LRP1 ligands examined because tPA activated cell signaling and promoted Schwann cell migration by interacting with the NMDA-R independently of LRP1, albeit with delayed kinetics. These results define the NMDA-R as a Schwann cell signaling receptor for protein ligands and a major regulator of Schwann cell physiology, which may be particularly important in peripheral nervous system (PNS) injury.
Asunto(s)
Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Células de Schwann/fisiología , Animales , Células Cultivadas , Maleato de Dizocilpina/farmacología , Sistema de Señalización de MAP Quinasas , Metaloproteinasa 9 de la Matriz/metabolismo , Traumatismos de los Nervios Periféricos/metabolismo , Interferencia de ARN , Ratas , Ratas Sprague-Dawley , Nervio Ciático , Transducción de Señal , Activador de Tejido Plasminógeno/metabolismoRESUMEN
PURPOSE: Patients with idiopathic carpal tunnel syndrome are commonly treated by steroid injections into the carpal tunnel. We administered triamcinolone (Tr) to chronic constriction injury model rats. We hypothesized that Tr administration would have both favorable behavioral effects and quantifiable immunohistological effects on compressed nerves. METHODS: Thirty-six male Wister rats were used. For rats to be treated with Tr, we loosely ligated their right sciatic nerves at 4 sites. Sham rats had their nerves exposed without ligation. On postoperative day 7, we reexposed their ligated nerves, after which we delivered either 0.1 mg of Tr (0.1-mg group), 0.5 mg of Tr (0.5-mg group), or normal saline (saline group) around the nerve fibers at the injured sites. Gait was analyzed, and allodynia was assessed with von Frey hairs, before surgery and on postoperative days 3, 7, 10, 14, and 21. The right sciatic nerve was resected and stained using hematoxylin-eosin, and the fourth and fifth lumbar dorsal root ganglia (DRG) were removed and assessed by immunohistochemistry for calcitonin gene-related peptide (CGRP) and activating transcription factor 3 (ATF3) on postoperative day 21. In addition, interleukin-1ß (IL-1ß) in sciatic nerve was quantified using enzyme-linked immunosorbent assays. RESULTS: Mechanical allodynia was significantly decreased in the 0.5-mg group compared with the saline group. In hematoxylin-eosin sections, the extent of inflammation-induced edema between the nerve fibers and infiltration of inflammatory cells was significantly reduced in the 0.5-mg group compared with the saline group. IL-1ß levels at the sciatic nerve in the 0.5-mg group were significantly lower than those in the saline group. CONCLUSIONS: Tr-treated chronic constriction injury rats exhibited significant alleviation of sensory disturbance, edema, inflammation, and pain-related peptide upregulation. These phenomena suggest the validity of Tr administration as a treatment affecting the nerve itself. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic I.
Asunto(s)
Síndromes de Compresión Nerviosa/tratamiento farmacológico , Triamcinolona/administración & dosificación , Triamcinolona/farmacología , Animales , Enfermedad Crónica , Modelos Animales de Enfermedad , Edema/tratamiento farmacológico , Ensayo de Inmunoadsorción Enzimática , Inflamación/tratamiento farmacológico , Inyecciones , Interleucina-1beta/metabolismo , Ligadura , Masculino , Ratas , Ratas Wistar , Nervio Ciático/lesiones , Nervio Ciático/metabolismo , Regulación hacia ArribaRESUMEN
PURPOSE: Adjacent segment degeneration (ASD) is one of the major complications of lumbar fusion. Several previous retrospective studies reported ASD after PLIF. However, few reports evaluated whether decompression surgery combined with fusion surgery increases the rate of complications in adjacent segments. The purpose of the current study was to investigate the degeneration in decompressed adjacent segments after PLIF. METHODS: A total of 23 patients (12 men, 11 women; average age, 58.6) who underwent PLIF surgery [1 level (n = 9), 2 levels (n = 8), 3 levels (n = 4), 4 levels (n = 2)] were included. Additional adjacent decompression above or below the level of interbody fusion was performed at 25 levels and no adjacent decompression was performed at 15 levels. We retrospectively investigated ASD by X-ray films of all 40 adjacent segments (above and below fusion level) and clinical outcomes of all 23 cases. RESULTS: Of the 40 adjacent segments, 19 (47.5%) showed ASD and 9 (22.5%) showed symptomatic ASD. In the 19 segments with ASD, ASD occurred in 16 of 25 (64.0%) segments at decompressed sites compared with 3 of 15 (20.0%) non-decompressed sites. The ratio of ASD in adjacent segments was significantly higher at decompressed sites than at non-decompressed sites (p < 0.01). CONCLUSION: ASD occurs frequently in association with additional decompression above or below the level of PLIF. In cases in which the adjacent segments require decompression, a surgical strategy that preserves as much of the posterior complex as possible should be selected.
Asunto(s)
Descompresión Quirúrgica/efectos adversos , Degeneración del Disco Intervertebral/etiología , Vértebras Lumbares/cirugía , Fusión Vertebral/efectos adversos , Adulto , Anciano , Femenino , Humanos , Degeneración del Disco Intervertebral/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Estudios Retrospectivos , Factores de Riesgo , Fusión Vertebral/métodos , Estenosis Espinal/cirugía , Resultado del TratamientoRESUMEN
STUDY DESIGN: We measured the response of the behavior and spinal glial activation to anti-nerve growth factor receptor (p75 neurotrophin receptor [p75NTR]) antibodies in the rat brachial plexus avulsion (BPA) model. OBJECTIVE: The aim of this study was to investigate the effect of anti-p75NTR antibodies on nociceptive behavior and activation of spinal microglia in the rat BPA model. SUMMARY OF BACKGROUND DATA: Tanezumab (anti-nerve growth factor antibody) treatment is associated with pain reduction and improvement in function, but with several complications. METHODS: Thirty male Wistar rats were used. In the BPA group, the C8-T1 roots were avulsed from the spinal cord with forceps at the lower trunk level and 10 µL of saline was applied locally (n = 10). In the anti-p75NTR group, the C8-T1 roots were avulsed and 10 µL of anti-p75NTR antibody was applied locally (n = 10). In a sham-operated group, the lower trunk was simply exposed (n = 10). Mechanical hyperalgesia and pain-induced walking patterns were measured using von Frey filaments (Stoelting, Wood Dale, IL) and the CatWalk gait analysis (Noldus Information Technology, the Netherlands) system every third day for 3 weeks. Activation of astrocytes and microglia was immunohistochemically examined in the spinal cord using anti-glial fibrillary acidic protein (GFAP) and anti-Iba1 antibodies both 7 and 21 days after surgery. RESULTS: Animals in the BPA group displayed significant mechanical hyperalgesia that continued through day 21 compared with animals in the sham-operated group, and mechanical hyperalgesia in the anti-p75NTR group was significantly improved 6 days after the operation. Regarding pain-induced gait analysis via CatWalk, animals in the BPA group displayed a significantly greater pain-like gait pattern than the p75 group for up to 3 weeks. Levels of GFAP-immunoreactive astrocytes and Iba1-immunoreactive microglia in the anti-p75NTR group were significantly reduced compared with the BPA group. CONCLUSION: Our results suggest that p75NTR contributes to neuropathic pain associated with BPA, and that inhibition of p75NTR reduces neuropathic pain. LEVEL OF EVIDENCE: N/A.
Asunto(s)
Anticuerpos/farmacología , Neuropatías del Plexo Braquial/fisiopatología , Ganglios Espinales/efectos de los fármacos , Dolor/prevención & control , Receptores de Factor de Crecimiento Nervioso/antagonistas & inhibidores , Animales , Anticuerpos/inmunología , Astrocitos/efectos de los fármacos , Astrocitos/metabolismo , Neuropatías del Plexo Braquial/metabolismo , Proteínas de Unión al Calcio/metabolismo , Modelos Animales de Enfermedad , Marcha/efectos de los fármacos , Marcha/fisiología , Ganglios Espinales/metabolismo , Ganglios Espinales/fisiopatología , Proteína Ácida Fibrilar de la Glía/metabolismo , Hiperalgesia/fisiopatología , Hiperalgesia/prevención & control , Inmunohistoquímica , Masculino , Proteínas de Microfilamentos/metabolismo , Microglía/efectos de los fármacos , Microglía/metabolismo , Microglía/fisiología , Proteínas del Tejido Nervioso , Dolor/fisiopatología , Ratas , Ratas Wistar , Receptores de Factores de Crecimiento , Receptores de Factor de Crecimiento Nervioso/inmunología , Receptores de Factor de Crecimiento Nervioso/metabolismo , Caminata/fisiologíaRESUMEN
STUDY DESIGN: Animal model of intravertebral disc (IVD) degeneration. OBJECTIVE: To examine production of inflammatory mediators in IVDs and neuropeptides in dorsal root ganglia (DRGs) in rat models of IVD compression and injury. SUMMARY OF BACKGROUND DATA: Sensory nerve fibers in IVDs and inflammatory mediator responses have been verified in animal models of IVD injury. However, the IVD injury in animals incompletely models degenerated human IVDs causing discogenic low back pain, because human IVDs are also subject to compression. METHODS: Experimental groups (controls, IVD injury, IVD compression, and their combination) of Sprague Dawley rats were prepared. Fluoro-Gold (FG; Fluorochrome, Denver, CO) was applied into coccygeal IVDs. Inflammatory mediators in IVDs, including nerve growth factor, tumor necrosis factor α, interleukin 1ß, and interleukin 6, were quantified using enzyme-linked immunosorbent assays. DRGs were immunostained for calcitonin gene-related peptide, activating transcription factor 3, and growth-associated phosphoprotein 43. RESULTS: The upregulation of inflammatory mediators was transient in the IVD injury group but delayed and long-lasting in the IVD compression group. When the IVD injury and compression were combined, the upregulation of inflammatory mediators was long-lasting through 8 weeks. The proportion of calcitonin gene-related peptide-immunoreactive neurons among Fluoro-Gold-labeled neurons remained significantly higher in the IVD injury, compression, and combination groups than in the controls. In contrast, increases in the proportions of activating transcription factor 3-immunoreactive or growth-associated phosphoprotein 43-immunoreactive neurons in the IVD injury group animals were transient but long-lasting in the compression and combination groups compared with controls. CONCLUSION: Disc injury in rats produces persistent increases in neuropeptides in DRGs but only transient increases in inflammatory mediators in IVDs. On the contrary, disc compression in rats produces a long-lasting increase in inflammatory mediators in IVDs and neuropeptides in DRGs. Moreover, disc compression induces persistent nerve injury and regeneration of the afferent fibers innervating IVDs.
Asunto(s)
Ganglios Espinales/metabolismo , Mediadores de Inflamación/metabolismo , Disco Intervertebral/metabolismo , Regeneración Nerviosa , Neuronas/metabolismo , Factor de Transcripción Activador 3/metabolismo , Animales , Distinciones y Premios , Péptido Relacionado con Gen de Calcitonina/metabolismo , Dolor Crónico/etiología , Dolor Crónico/metabolismo , Dolor Crónico/fisiopatología , Ensayo de Inmunoadsorción Enzimática , Proteína GAP-43/metabolismo , Ganglios Espinales/patología , Ganglios Espinales/fisiopatología , Humanos , Inmunohistoquímica , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Disco Intervertebral/inervación , Disco Intervertebral/patología , Degeneración del Disco Intervertebral/complicaciones , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/fisiopatología , Dolor de la Región Lumbar/etiología , Dolor de la Región Lumbar/metabolismo , Dolor de la Región Lumbar/fisiopatología , Masculino , Fibras Nerviosas/metabolismo , Fibras Nerviosas/patología , Neuronas/patología , Neuronas Aferentes/metabolismo , Neuronas Aferentes/patología , Neuropéptidos/metabolismo , Ratas , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
We present an operative technique and the results of intramedullary fixation of proximal phalangeal shaft fractures through a volar extra-tendon sheath approach. A J-shaped nail, which is a curved Kirschner wire sharply bent at the proximal end, was inserted from the palmar aspect of the proximal phalangeal base. Six fingers of the six patients (all male) were treated with this method. The mean age of the patients was 51 years (range, 20-69 years). There were four open and two closed fractures. All fingers attained successful fracture union and one of them had correction loss. No patient complained of pain at the final follow-up, and the average of total active motion was 223° (190° - 255°). This method may be an alternative for treatments of the proximal phalangeal shaft fractures because of its less invasive nature, although it does not offer anatomical reduction.
Asunto(s)
Traumatismos de los Dedos/cirugía , Falanges de los Dedos de la Mano/lesiones , Fijación Intramedular de Fracturas/métodos , Fracturas Óseas/cirugía , Tendones/cirugía , Adulto , Anciano , Traumatismos de los Dedos/diagnóstico por imagen , Traumatismos de los Dedos/fisiopatología , Articulaciones de los Dedos/diagnóstico por imagen , Articulaciones de los Dedos/fisiopatología , Articulaciones de los Dedos/cirugía , Falanges de los Dedos de la Mano/diagnóstico por imagen , Falanges de los Dedos de la Mano/cirugía , Estudios de Seguimiento , Curación de Fractura , Fracturas Óseas/diagnóstico por imagen , Fracturas Óseas/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Radiografía , Rango del Movimiento Articular , Estudios Retrospectivos , Resultado del Tratamiento , Adulto JovenRESUMEN
Although displaced fractures of the lateral aspect of the base of the proximal phalanx can be treated surgically, previously described approaches to the fracture are not necessarily easily performed. We describe a trans-web approach to the metacarpophalangeal joint and report 2 clinical cases. This technique allows the fracture fragments to be reduced and fixed with minimal risk of damage to the adjacent structures.