Lesion-Induced Changes in the Production of Newly Synthesized and Secreted Apo-E and Other Molecules Are Independent of the Concomitant Recruitment of Blood-Borne Macrophages into Injured Peripheral Nerves

Peripheral nerve injury produces Wallerian degeneration characterized by a change in the composition of resident nonneuronal cells: macrophages are recruited from the circulation to join Schwann, fibroblast, and endothelial cells. At the same time, the nonneuronal cell population exhibits, as a whole, alterations in synthesis and secretion of diffusible molecules, some of which are instrumental in nerve repair mechanisms. In this study, we determined whether changes in the production of secreted molecules depend on the concomitant modification in cell composition. Therefore, we studied the secretion of newly synthesized molecules by defined cell populations of intact nerves, intact nerve explants undergoing in vitro axonal degeneration, in vivo degenerating nerves, and recruited cells. Nerves were incubated in serum-free, [35S]methionine-containing media. Secreted, radioactively labeled proteins were precipitated from the medium and analyzed by gel electrophoresis. Reduced production of 43-, 46-, and 48-kDa proteins and increased production of 33-34-, 37-, 49-, 59-, and 67-kDa proteins were detected in in situ degenerating nerves. High-density ultracentrifugation and immunoblot analysis revealed that the 33-34-kDa protein is apolipoprotein-E (apo-E). Similar alterations in the production of these molecules were detected in intact nerve explants from which blood-borne cells were excluded. Apo-E, 37-, 49-, 59-, and 67-kDa proteins were also produced in frozen nerves that lacked the intact nerve nonneuronal cell population. Instead, these preparations contained blood-borne cells, primarily macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electron microscopy of severed motor fibers in the crayfish

Summary Cut and crushed crayfish claw nerves were examined with the electron microscope at intervals up to 6 months after lesion. In sections 1 centimeter distal to the lesion there were no signs of degeneration among the giant motor axons even after many months. Swelling of glial wrappings was observed within 48 hours of nerve severance and was particularly notable in the innermost glial layer, the adaxonal layer. Golgi elements, rough endoplasmic reticulum, and mitochondria accumulated in the glia. These changes were perhaps indicative of a greater supportive role required by the severed axons. Regeneration from the proximal stumps of the giant axons began within one week and had proceeded across the lesion gap by 4 weeks. Axon sprouts appeared to travel toward the terminals within the glial sheaths of the distal giant axon segments. Before regeneration was complete, as determined by a simple behaviour test, the regenerating axons occupied increasing proportions of the sheath space. After regeneration was complete occasional degenerations were seen among the sprouts. These degenerations may have occurred in regenerating axons which had grown to the incorrect muscles. The original distal giant axons probably degenerated, as well, after regeneration was complete. There was no evidence of rehealing of proximal and distal segments of the axons.This work was supported by NIH postdoctoral fellowship number 1F2 NB 32, 723 N RB awarded to RHN and grants in aid from the Multiple Sclerosis Society, The American Cancer Society and The National Institutes of Health.     

Whole-body vibration therapy does not improve the peripheral nerve regeneration in experimental model

Objectives:Whole-body vibration (WBV) is commonly used to improve motor function, balance and functional performance, but its effects on the body are not fully understood. The main objective was to evaluate the morphometric and functional effects of WBV in an experimental nerve regeneration model.Methods:Wistar rats were submitted to unilateral sciatic nerve crush and treated with WBV (4-5 weeks), started at 3 or 10 days after injury. Functional performances were weekly assessed by sciatic functional index, horizontal ladder rung walking and narrow beam tests. Nerve histomorphometry analysis was assessed at the end of the protocol.Results:Injured groups, sedentary and WBV started at 3 days, had similar functional deficits. WBV, regardless of the start time, did not alter the histomorphometry parameters in the regeneration process.Conclusions:The earlier therapy did not change the expected and natural recovery after the nerve lesion, but when the WBV starts later it seems to impair function parameter of recovery.     

Physical properties and biocompatibility of a porous chitosan-based fiber-reinforced conduit for nerve regeneration

Porous fiber-reinforced chitosan nerve conduits were fabricated from chitosan yarns and a chitosan solution by combining an industrial braiding method with a mold casting/lyophilization technique. The conduits were permeable to molecules ranging in molecular size from 180 Da (glucose) to 66,200 Da (BSA). The compressive load of the reinforced conduits was significantly higher than that of a non-reinforced control conduit at equal levels of strain. The tensile strength of the reinforced conduits was also increased from 0.41 ± 0.17 to 3.69 ± 0.64 MPa. An in vitro cytotoxicity test showed the conduits were not cytotoxic to Neuro-2a cells. Preliminary in vivo implantation testing indicated that the conduits were compatible with the surrounding tissue. Aijun Wang and Qiang Ao contributed equally to this work.     

植入血管束的血管化人工神经导管对受损神经功能恢复影响的实验研究

目的:研究植入血管束的血管化人工神经导管修复SD大鼠长段坐骨神经缺损对神经功能恢复的影响。方法:将18只成年雌性SD大鼠制成14mm的大鼠坐骨神经缺损模型后,随机分为3组(每组12条神经),分别采用不同的修复方法。A组:自体神经移植组(自体组);B组:普通PGLA神经导管移植组(导管组);C组:植入自体血管束的普通PGLA神经导管移植组(血管化导管组)。观察术后大鼠后肢皮肤溃疡面积;检测术后6周、12周时步态变化和肌电图。结果:术后各组SD大鼠均出现后肢溃疡,血管化导管组SD大鼠后肢溃疡愈合较导管组早2周。血管化导管组步态检测SFI明显优于导管组,与自体神经移植组无明显差异。肌电图检测表明血管化导管组无论是神经传导速度,还是动作电位振幅均明显大于导管组(P<0.05),与自体神经移植组无明显差异(P>0.05)。结论:植入血管束的血管化人工神经导管能有效地促进受损神经的功能恢复。     

Acceleration of recovery from sciatic nerve damage by the ACTH(4–9) analog Org 2766: Different routes of administration

Functional recovery following a sciatic nerve crush in rats was investigated by measuring the reflex withdrawal of the hindpaw to a hot air stream. The ACTH(4–9) analog Org 2766 accelerated recovery when administered subcutaneously (two-daily injections: 10 μg/animal; minipumps: 20–40 μg/animal per 24 hr; biodegradable microspheres: 40 μg/animal per 24 hr), but oral administration (1.5–20 mg/animal daily, in the drinking water; 1.5–15 mg/animal daily, by gavage) was not effective.     

大白鼠交感节前神经再生的研究Ⅰ.生化、组化与功能观察

用液氮骤冻造成大白鼠交感节前神经变性后,通过神经末梢乙酰胆碱含量、胆碱酯酶活性测定以及电刺激交感干时外周反应等研究其再生规律。结果表明冻伤后3周内再生过程进展迅速,神经结构与功能均有相当程度的恢复;3周后再生过程转慢,直至一年时各指标仍远未达到正常。这证明交感节前神经的再生过程不同于中枢及其它外周神经而独具特征。     

Effect of Neurodynamics Nerve Flossing on Femoral Neuropathy in Haemophilic Patients: A randomized controlled study

Objective:Heamophilia is considered a disorder in both children and adolescents which may affect their quality of life seriously than their normal peers; this study investigated the impact of the Neurodynamics Nerve Flossing Technique (NFT) on femoral neuropathy in patients with haemophilia.Method:Thirty haemophilic children with Femoral Neuropathy were randomly allocated into two equivalent groups; the study group which received Neurodynamics NFT of the femoral nerve and conventional therapy program, and the control group which received only the conventional therapy program, three sessions/week for 12 weeks. Femoral nerve motor conduction velocity (MCV) and level of pain sensation according to the Visual Analogue Scale (VAS), were assessed pre and post interventions.Results:post-treatment comparison between both groups revealed that there was a significant increase in femoral nerve MCV and reduction of pain sensation of the study group compared to the control group (p<0.05).Conclusion:Neurodynamics Nerve flossing had a clear effect in the treatment of femoral neuropathy in children with haemophilia.     

Composition of the medial and posterior articular nerves of the mouse knee joint

The number and the distribution of fiber size in the medial (MAN) and posterior (PAN) articular nerves of the mouse knee joint were studied by electron microscopy. The MAN contained 75 &#45 28 nerve fibers consisting of 63 &#45 24 unmyelinated and 12 &#45 6 myelinated fibers. The PAN was composed of 195 &#45 50 nerve fibers, namely 129 &#45 28 unmyelinated and 66 &#45 24 myelinated fibers. A skewed unimodal distribution of the unmyelinated nerve fiber diameters was seen in both nerves ranging from 0.1 to 1.2 &#119 m with a maximum between 0.3 and 0.6 &#119 m. The myelinated nerve fibers in the MAN ranged from 1 to 8 &#119 m with a peak between 2 and 5 &#119 m. In the PAN, their diameters ranged from 1 to 12 &#119 m with a clearly visible peak at 4-5 &#119 m and a plateau at 8-9 &#119 m that may represent a second maximum. These data show that the knee joint innervation of the mouse is comparable to those of the cat and rat concerning the types of nerve fibers and the composition of the two nerves. However, in relation to the much smaller area of tissue to be innervated the total number of primary afferents is considerable smaller in the mouse.     

NGF电纺纤维的体外缓释研究

目的:研究担载神经生长因子(NGF)的静电纺丝纤维的表征,考察NGF电纺纤维对于周围神经修复的效果。方法:将NGF水溶液分散于PLLA溶液,通过W/O乳液法制备静电纺丝纤维,对纤维的形态、力学性能等进行表征,Elisa方法测定NGF的体外释放动力学,Alamer Blue法检测试剂来考察纤维释放液对于PC12细胞增殖的影响。结果:NGF电纺纤维具备良好的形态和力学性质,直径为500-900 nm,纤维具备三维多孔结构。纤维的最大拉伸应力为2.50±0.41 MPa。电纺纤维中NGF在体外可有效释放9天,累积释放量接近3000 pg。细胞活性实验结果显示,第1、3、5、7天释放液的荧光强度与对照组相比有显著差异。结论:担载NGF的乳液法静电纺丝纤维有促进缺损周围神经修复的潜质。     

How is information transmitted in a nerve?

In the last 15 years, a debate has emerged about the validity of the famous Hodgkin-Huxley model for nerve impulse. Mechanical models have been proposed. This note reviews the experimental properties of the nerve impulse and discusses the proposed alternatives. The experimental data, which rule out some of the alternative suggestions, show that while the Hodgkin-Huxley model may not be complete, it nevertheless includes essential features that should not be overlooked in the attempts made to improve, or supersede, it.     

担载神经生长因子(NGF)的乳液法涂层纤维体外缓释研究

目的:目前周围神经修复中,神经导管是研究热点,本文研究乳液法涂层纤维制备的神经导管在神经修复中应用的可能性。方法:本文采用乳液法制备担载NGF的丝素-聚乳酸(PLLA)涂层电纺纤维,观察纤维的形态,测定NGF的体外释放动力学参数,并考察纤维释放液对于PC12细胞增殖的影响。结果:担载NGF的涂层纤维具备类似于细胞外基质(ECM)的三维结构和多孔形态;涂层纤维中NGF体外有效缓释10天;细胞实验中,在含有释放液的培养基中生长的PC12细胞,与空白对照组相比,荧光强度平均多了2000-4000个荧光强度,所以释放液可以更好地促进PC12细胞的增殖。结论:担载NGF的乳液法涂层纺丝纤维具备促进缺损周围神经修复的条件,可以进一步研究在动物体内修复缺损周围神经中的效果,为以后的临床应用打下基础。     

Branching patterns of rabbit oculomotor and trochlear nerves demonstrated by Sihler's stain technique

A modified Sihler's stain technique was used to visualize the branching patterns of oculomotor and trochlear nerves. The levator palpebrae, superior rectus, inferior rectus, medial rectus, inferior oblique, superior oblique and tensor trochlea muscles were isolated from the eyes of normal rabbits and processed using modified Sihler's technique. The distributions and terminal ramifications of the oculomotor and trochlear nerves were observed. Two distinct divisions and terminal branches of the oculomotor nerve were shown in detail together with the trochlear nerve distribution. The application of Sihler's technique enables researchers to trace nerve branching within relatively transparent muscles, whereas the nerve fibers are counterstained and clearly visible. This technique could be useful for detailed studies of the motor control of extraocular muscles.     

Investigating the mechanical shear-plane between core and sheath elements of peripheral nerves

The mechanical architecture of rat sciatic nerve has been described as a central core surrounded by a sheath, although the way in which these structures contribute to the overall mechanical properties of the nerve is unknown. We have studied the retraction responses of the core and sheath following transection, together with their tensile properties and the interface between them. Nerves were harvested and maintained at their in situ tension and then either transected entirely, through the sheath only, or through an exposed section of the core. The retraction of each component was measured within 5 min and again after 45 min. Post mortem loss of retraction was tested 0 min or 60 min after excision. For fresh nerves, immediate retraction was 12.68% (whole nerve), 5.35% (sheath) and 4% (core), with a total retraction of 15%, 7.21% and 5.26% respectively. For stored nerves, immediate retraction was 5.33% (whole nerve) and 5.87% (sheath), with an extension of 0.78% for core, and a total retraction of 6.71% and 7.87% and an extension of 1.74%, respectively. Tensile extension and pullout force profiles were obtained for the sheath, the core and the interface between them. These showed a consistent hierarchy of break strengths that would, under increasing load, result in failure of the interface, then the core and finally the sheath. These data reflect the contributions of material tension and fluid swelling pressure to total retraction, and the involvement of an energy-dependent process that runs down rapidly post mortem. This study increases our understanding of the composite nature of peripheral nerve tissue architecture and quantifies the material properties of the distinct elements that contribute to overall mechanical function.Preliminary forms of this study were presented at: (1) British Matrix Biology Society Meeting, Manchester, March 2001; (2) Tissue and Cell Engineering Society Meeting, Keele, United Kingdom, September 2001.Financial support was provided by the EU framework 5 programme in neural tissue engineering (QLK3-CT-1999-00625).     

神经生长因子治疗急性颅脑损伤的效果及对患者神经功能的影响

目的:研究神经生长因子在急性颅脑损伤中的治疗效果及对神经功能的影响。方法:选取2014年8月至2015年7月本院收治的82例急性颅脑损伤患者,随机分为观察组和对照组,每组41例。对照组采取常规对症治疗,观察组在对照组基础上采用神经生长因子治疗。观察并比较两组患者治疗前后血清S100β,白介素-6(IL-6),髓鞘碱性蛋白(MBP)及神经元特异性烯醇化酶(NSE)水平的变化情况以及临床疗效。结果:观察组总有效率高于对照组,差异具有统计学意义(P0.05)。与治疗前比较,两组患者治疗后血清S100β及IL-6水平均降低,差异具有统计学意义(P0.05);与对照组比较,观察组患者治疗后血清S100β及IL-6水平较低,差异具有统计学意义(P0.05);与治疗前比较,两组患者治疗后血清MBP及NSE水平均降低,差异具有统计学意义(P0.05);与对照组比较,观察组患者治疗后血清MBP及NSE水平较低,差异具有统计学意义(P0.05)。结论:神经生长因子治疗急性颅脑损伤的效果显著,能够改善患者免疫功能和神经功能,值得临床推广应用。     

蛇毒神经生长因子促进周围神经再生的诱发电位观察

目的：研究蛇毒神经生长因子（sNGF)对大鼠坐骨神经损伤后诱发电位的影响,评价蛇毒神经生长因子在促进周围神经再生中的作用。方法：建立大鼠坐骨神经钳夹模型,局部滴加药物和术后肌注sNGF,通过脊髓诱发电位（SEP）,运动诱发电位（MEP）评定,观察坐骨神经修复情况。结果：sNGF治疗后可使伤后SEP,MEP提早出现,结论：蛇毒提取的NGF对大鼠坐骨神经损伤修复具有促进作用。     

Lack of physiological stimulation induces decreased proteolytic activity in nerve terminals

The effect of optic nerve transsection on proteolytic degradation of axonally transported proteins in the superior colliculus of the rabbit was studied. Proteolysis of labeled proteins was determined in vitro in small pieces of the superior colliculus. Within 2 hours after sectioning the optic nerve there was a decreased degradation of slowly transported labeled proteins in the nerve terminals in the superior colliculus.Special Issue dedicated to Prof. Holger Hydén.     

Stability of periodic travelling wave solutions of a nerve conduction equation

Summary Nagumo's nerve conduction equation has travelling wave solutions of pulse type and periodic wave type. We consider the stability of the latter ones. We denote byL(c) the minimum spatial period of a periodic travelling wave solution whose propagation speed isc. It is shown that this travelling wave solution is unstable ifL′(c)<0.     

三维取向PLGA神经导管促进坐骨神经再生的实验研究

目的:探讨应用改进静电纺丝技术一次成型制备三维(3D)取向聚乳酸与聚羟基乙酸共聚物(PLGA)纳米神经导管的可行性,检测其对坐骨神经再生的促进作用。方法:应用改进的静电纺丝技术制备无缝取向PLGA纳米神经导管,通过扫描电镜和透射电镜检测支架的纳米结构;分别制备取向和非取向纳米纤维支架修复13mm坐骨神经缺损模型。36只成年SD大鼠随机分为3组(每组12只),A组:非取向PLGA神经导管组(阴性对照);B组:取向PLGA神经导管组,C组:自体神经移植组(阳性对照),于术后3月通过大体观察、行走足印分析、腓肠肌萎缩率、电生理检测、组织形态学检测、透射电镜检测及图像分析,评价无缝取向PLGA纳米神经导管修复坐骨神经缺损的效果。结果:神经导管修复神经缺损三月后,大体观察显示神经导管结构完整,无坍塌和断裂;各组再生神经均有通过神经导管长入远端。B组与C组的腓肠肌萎缩率和神经电传导速度无统计学差异(P0.05),均优于A组。B组与C组再生神经纤维数量及成熟程度均要明显优于A组。结论:无缝取向PLGA纳米神经导管能够诱导并促进神经再生,提高坐骨神经再生的质量,有望成为自体神经移植的替代物。     

三维取向PLGA神经导管促进坐骨神经再生的实验研究

目的：探讨应用改进静电纺丝技术一次成型制备三维（3D）取向聚乳酸与聚羟基乙酸共聚物（PLGA）纳米神经导管的可行性,检测其对坐骨神经再生的促进作用。方法：应用改进的静电纺丝技术制备无缝取向PLGA纳米神经导管,通过扫描电镜和透射电镜检测支架的纳米结构;分别制备取向和非取向纳米纤维支架修复13mm坐骨神经缺损模型。36只成年SD大鼠随机分为3组（每组12只）,A组：非取向PLGA神经导管组（阴性对照）;B组：取向PLGA神经导管组,C组：自体神经移植组（阳性对照）,于术后3月通过大体观察、行走足印分析、腓肠肌萎缩率、电生理检测、组织形态学检测、透射电镜检测及图像分析,评价无缝取向PLGA纳米神经导管修复坐骨神经缺损的效果。结果：神经导管修复神经缺损三月后,大体观察显示神经导管结构完整,无坍塌和断裂;各组再生神经均有通过神经导管长入远端。B组与C组的腓肠肌萎缩率和神经电传导速度无统计学差异（P〈0.05）,均优于A组。B组与C组再生神经纤维数量及成熟程度均要明显优于A组。结论：无缝取向PLGA纳米神经导管能够诱导并促进神经再生,提高坐骨神经再生的质量,有望成为自体神经移植的替代物。