Analysis of Nerve Endings in the Superior Labrum-Biceps Complex by Fluorescence Immunohistochemistry and Confocal Laser Microscopy / Análise de terminações nervosas no complexo labrum-bíceps superior usando imunohistoquímica de fluorescência e microscopia a laser confocal

Abstract Objectives The capsuloligamentous structures of the shoulder work as static stabilizers, together with the biceps and rotator cuff muscles, increasing the contact surface of the glenoid cavity. Free nerve endings and mechanoreceptors have been identified in the shoulder; however, there are a few studies that describe the presence of these nerves in the biceps' insertion. The present study aimed to describe the morphology and distribution of nerve endings using immunofluorescence with protein gene product 9.5 (PGP 9.5) and confocal microscopy. Methods Six labrum-biceps complexes from six fresh-frozen cadavers were studied. The specimens were coronally cut and prepared using the immunofluorescence technique. In both hematoxylin and eosin (H&E) and immunofluorescence, the organization of the connective tissue with parallel collagen fibers was described. Results In the H&E study, vascular structures and some nerve structures were visualized, which were identified by the elongated presence of the nerve cell. All specimens analyzed with immunofluorescence and confocal microscopy demonstrated poor occurrence of morphotypes of sensory corpuscles and free nerve endings. We identified free nerve endings located in the labrum and in the bicipital insertion, and sparse nerve endings along the tendon. Corpuscular endings with fusiform, cuneiform, and oval aspect were identified in the tendon. Conclusion These findings support the hypothesis that the generation of pain in the superior labral tear from Anterior to posterior (SLAP) lesions derives from the more proximal part of the long biceps cord and even more from the upper labrum. Future quantitative studies with a larger number of specimens may provide more information on these sensory systems.

Resumo Objetivos As estruturas capsulo-ligamentares do ombro funcionam como estabilizadores estáticos, juntamente com os músculos do bíceps e do manguito rotador, aumentando a superfície de contato da cavidade glenoide. Terminações nervosas livres e mecanorreceptores foram identificados no ombro; no entanto, existem alguns estudos que descrevem a presença desses nervos na inserção do bíceps. Este estudo teve como objetivo descrever a morfologia e distribuição de terminações nervosas utilizando imunofluorescência com protein gene product 9.5 (PGP 9.5) e microscopia confocal. Métodos Foram estudados seis complexos labrum-bíceps de seis cadáveres congelados frescos. Os espécimes foram cortados coronalmente e preparados pelo método de imunofluorescência. Tanto em hematoxilina e eosina (H&E) quanto em imunofluorescência, foi descrita a organização do tecido conjuntivo com fibras paralelas de colágeno. Resultados No estudo de H&E, foram visualizadas estruturas vasculares e algumas estruturas nervosas, que foram identificadas pela presença alongada da célula nervosa. Todas as amostras analisadas com imunofluorescência e microscopia confocal demonstraram baixa ocorrência de morfotipos de corpúsculos sensoriais e terminações nervosas livres. Identificamos terminações nervosas livres localizadas no labrum, inserção bicipital e terminações nervosas esparsas ao longo do tendão. Terminais corpusculares com aspecto fusiforme, cuneiforme e oval foram identificados no tendão. Conclusão Esses achados corroboram a hipótese de que a geração de dor nas lesões labrais superiores de anterior a posterior (SLAP, na sigla em inglês) deriva da parte mais proximal do cabo longo do bíceps e ainda mais do labrum superior. Estudos quantitativos futuros com um número maior de espécimes podem fornecer mais informações sobre esses sistemas sensoriais.

Análise dos mecanorreceptores e terminações nervosas livres da banda anterior doligamento glenoumeral inferior com imunofluorescência e microscopia confocal

A banda anterior do ligamento glenoumeral inferior(LGUI), possui uma importante função na estabilidade mecânica do ombro, sendo onsiderado o principal estabilizador estático do ombro, quando o braço encontra-se em abdução e rotação externa. Seis bandas anteriores do LGUI foram cuidadosamente dissecadas para avaliaçãoda morfologiados mecanorreceptorese terminações nervosas livres, buscando compreender a interação do sistema proprioceptivo e mecânico do ombro. Para imunihistoquimica foi utilizado um marcador tipo PGP9.5 (protein gene product)como anticorpo primário e Alexa Fluor 488 como anticorpo secundário, seguido de análise daslâminas pormicroscopia confocal de varredura a laser. Observou-se em todos os ligamentos corpúsculos de Meissnercom diâmetros queentre 30 e 65 μme comprimentos entre 80 e 400 μm,sendo estes descritos pela primeira vez no ligamento glenoumeral inferior;o corpúsculo dePacini, com diâmetro entre 40 e 80 μm e comprimento entre 100 e 180 μme terminações nervosas livres, onde as fibras apresentavam espessura entre 3 e 7 μm e comprimento entre 300 e 700 μm, além de terminações nervosas não classificadas, com formas irregulares, fusiformes e retangulares. Observamos que na banda anterior do ligamento glenoumeral inferior opredomínio de mecanorreceptores de adaptação rápida, tipo Meissner e Pacini. A densidade dos mecanorreceptores foi de aproximadamente 1,1026% ,sendo a densidadede 1,6102% quando a analise foi com profundidade entre 400 e 500 μme de 0,6018%...

The anterior band of the inferior glenohumeral ligament (IGHL) has an important role in the mechanical stability of the shoulder and is considered the main static stabilizer of the shoulder when the arm is in abduction and external rotation. Six anterior bands of IGHL were carefully dissected from the bone attachments to describe the morphology of the mechanoreceptors and free nerve endings and to elucidate the interaction between the proprioceptive system and shoulder mechanics. For immunostaining, a protein gene product 9.5 marker was used as the primary antibody and Alexa Fluor 488 was used as the secondary antibody, followed by image examination using confocal laser scanning microscopy. All the ligament samples contained Meissner corpuscles with a diameter between 30 and 65 μm and length between 80 and 400 μm. This is the first report of these corpuscles in IGHL. Furthermore, these ligaments contained Pacinian corpuscles, with a diameter between 40 and 80 μm and length between 100 and 180 μm, free nerve endings with fiber thickness between 3 and 7 μm and length between 300 and 700 μm, and unclassified nerve endings with irregular, spindle, and rectangular shapes. The mechanoreceptors were approximately 1.2% of the area of the ligament. There was a predominance of Meissner-and Pacini-type rapidly adapting mechanoreceptors.The density ofmechanoreceptorswas approximately 1.1026%, and the densitywhen the analysis was with a depthbetween 400 and 500μmwas 1.6102% and greaterdepthbetween 1500and 1850μm, was 0.6018%...

Effect of Pi-Zhen on 5-HT Content in Nerve Endings Tension Pain Rabbit Model / 世界科学技术-中医药现代化

This article was aimed to explore theeffect of Pi-Zhen(PZ) on 5-hydroxytryptamine(5-HT) in nerve end-ings tension painrabbitmodel. Forty-two healthy male big ear whiterabbits of three months old were randomly divided into 5 groups, which were the normal group,model group, drug group, Hao-Zhen (HZ) group, and PZ group. Nerve endings tension pain modelswere established. No operation was made in the normal group. The amount of 1 ml saline was injected to the operative sitebetween the shallow and deep fascia in each rabbit and then draw out at 5, 10, 20, 30, 40, 50, 60 min after operation. The 5-HT level in extracting solution of different groups was determined with ELISA method. The results showed that PZ interventioncan reducethe 5-HT level ofextracting solution in operative site; and with the increasing of time, the level of 5-HT was gradually reduced. Compared with the model group, the contents of 5-HT in the treatment group were significantly decreased at different time points(P<0.01). The content of 5-HT in the PZ group was significantly higher than that of the normal group at 5 min (P<0.01). Compared with the drug group, the contents of 5-HT in the HZ group and PZ group were significantly increased at 5, 10, 20, 30, 40 min (P<0.01). Compared with the normal group, the contents of 5-HT in the PZ group was significantly decreased at 40, 50, 60 min (P<0.05). Compared with the HZ group, the contents of 5-HT were significantly decreased at 20, 30, 40, 50, 60 min (P<0.01). There was no difference at other time points. It was concluded that PZ can ease pain through reducing the tension of local soft tissues, decreasing 5-HT releasing, and promoting its degradation.

Correlations between radiographic, magnetic resonance and histological examinations on the degeneration of human lumbar intervertebral discs / Correlação entre exame radiológico, ressonância magnética e exame histológico na degeneração dos discos intervertebrais lombares humanos

CONTEXT AND OBJECTIVE: There is controversy regarding which imaging method is best for identifying early degenerative alterations in intervertebral discs. No correlations between such methods and histological finds are presented in the literature. The aim of this study was to correlate the thickness of intervertebral discs measured on simple radiographs with the degree of degeneration seen on magnetic resonance images and the histological findings relating to nerve ends inside the discs. DESIGN AND SETTING: Cross-sectional correlation study on the lumbar spines of human cadavers, at Universidade Federal de São Paulo (Unifesp), São Paulo, Brazil. METHODS: Ten lumbar spinal columns were extracted from human cadavers and subjected to magnetic resonance imaging and simple radiography. They were classified according to the degree of disc degeneration seen on magnetic resonance, and the thickness of the discs was measured on radiographs. The intervertebral discs were then extracted, embedded in paraffin and analyzed immunohistochemically with protein S100, and the nerve fibers were counted and classified. RESULTS: No correlation was observed between the thickness of the intervertebral discs and the degree of degeneration seen on magnetic resonance images. Only the uppermost lumbar discs (L1/L2 and L2/L3) presented a correlation between their thickness and type I and IV nerve endings. CONCLUSION: Reduced disc thickness is unrelated to increased presence of nerve ends in intervertebral discs, or to the degree of disc degeneration.

CONTEXTO E OBJETIVO: Há controvérsia sobre qual o melhor método de imagem para identificar alterações degenerativas precoces do disco intervertebral. Falta na literatura correlação desses métodos com os achados histológicos. O objetivo deste estudo foi relacionar a altura dos discos intervertebrais medidos em radiografias simples com o grau de degeneração nas imagens de ressonância magnética e os achados histológicos das terminações nervosas encontradas no interior do disco. TIPO DE ESTUDO E LOCAL: Estudo transversal de correlação em coluna lombar de cadáveres humanos, na Universidade Federal de São Paulo (Unifesp), São Paulo, Brasil. MÉTODOS: Dez colunas lombares foram retiradas de cadáveres humanos e submetidas a imagens de ressonância magnética e radiografias simples. Foram classificadas de acordo com o grau de degeneração dos discos pela ressonância e mensuradas as alturas dos discos nas radiografias. Os discos intervertebrais foram retirados, incluídos em parafina e foi realizado estudo imunoistoquímico com proteína S100; as fibras nervosas foram contadas e classificadas. RESULTADOS: Não foi observada correlação entre a altura dos discos intervertebrais com o grau de degeneração nas imagens de ressonância magnética. Apenas os discos lombares altos (L1/L2 e L2/L3) apresentaram correlação entre a altura e as terminações nervosas dos tipos I e IV. CONCLUSÃO: A diminuição da altura dos discos não está relacionada ao aumento de terminações nervosas nos discos intervertebrais e nem com o grau de degeneração dos discos.

Quantificação digital da imunoexpressão de receptores adrenérgicos e terminações nervosas no detrusor de portadores da síndrome de prune belly / Digital quantification of the imunoexpression of adrenergic receptors and nervous terminations in the detrusor of patients with prune belly syndrome

INTRODUÇÃO: A síndrome de prune belly (PBS) é caracterizada por uma tríade com flacidez da parede abdominal, criptorquidia bilateral e malformações do trato urinário que compreende bexiga de capacidade aumentada, com complacência elevada, hipossensibilidade, hipocontratilidade, com divertículo ou fístula uracal e resíduo pós-miccional elevado. Alguns autores recomendam tratamento clínico, porém outros propõe correção cirúrgica, com reconstrução da via urinária incluindo ureteroplastia e cistoplastia redutoras, orquidopexia e abdominoplastia. Mesmo após a cirurgia, alguns doentes necessitam de cateterismo limpo intermitente. A inervação vesical determina seu funcionamento, mediado por neuroreceptores na junção neuromuscular. Os adrenoreceptores a1 estão relacionados à contratilidade detrusora e o b3 ao seu relaxamento, e certas condições como obstrução infravesical levam à hiperexpressão de receptores a1. O objetivo da presente pesquisa é verificar se no detrusor de doentes com PBS há alteração na densidade de terminações nervosas, hiper ou hipoexpressão de receptores adrenérgicos a1a, a1b, a1d e b3 e proporção anormal dos tecidos muscular e conectivo. MÉTODO Trata-se de estudo retrospectivo de caso-controle que envolveu 14 espécimes de detrusor de doentes com PBS operados entre 1985 a 2005 no Hospital das Clínicas da FMUSP. Dois grupos foram constituídos como controle: 13 fragmentos de bexiga de doentes submetidos à prostatectomia radical no Departamento de Urologia da Universidade de Mainz, com urodinâmica pré-operatória normal (GC1), e cinco fragmentos de bexiga de crianças submetidas à necrópsia no SVOC-USP, sem anomalias neurológicas e de trato urinário. A coloração de van Gieson foi realizada para análise da proporção músculo/tecido conectivo, e a reação imunohistoquímica para os anticorpos policlonais anti-proteína S100 e antiadrenoreceptores a1a, a1b, a1d e b3. A coloração castanho foi considerada evidência da expressão do adrenoreceptor...

INTRODUCTION: Prune belly syndrome (PBS) is charactherized by a triad of abdominal wall flaccidity, bilateral criptorchidism and urinary tract malformation, that includes a large-capacity bladder, with high detrusor compliance, low sensibility and contractility, associated to urachal diverticulum or fistula and elevated post void residual volumes. Some autors recommend clinical treatment, but others propose surgery correction, with urinary tract reconstruction, including reductive ureteroplasty and cystoplasty, orchidopexy and abdominoplasty. Even after surgery, some patients need intermittent catheterism. The detrusor innervation determines its function, mediated by neuroceptors at the neuromuscular junction. The a1 adrenoceptors are related to detrusor contractility and b3 to relaxation, and some conditions, like infravesical obstruction, lead to a1 adrenoceptor up-regulation. The objective of this work is to verify whether, in the detrusor from patients with PBS, there is altered nerve density, up or down-regulation of a1a, a1b, a1d and b3 adrenergic receptors and if there is an abnormal proportion between muscle and connective tissue. MATERIALS AND METHODS A retrospective case-control study was performed involving 14 detrusor specimens from patients with PBS, who underwent surgical treatment between 1985 an 2005 at University of São Paulo, Medical School Hospital. Two groups were taken as control: 13 bladder fragments from patients who underwent radical prostatectomy at Department of Urology of Mainz University, with normal urodynamic study prior to the surgery (GC1) and 5 bladder fragments from children submitted to autopsy at SVOC-USP, with no neurological or urinary tract malformation (GC2). Staining was performed using the van Gieson dye to analyse the proportion between muscle and connective tissue, and immunohistochemical reaction was employed, with polyclonal antibodies against S100 protein, as well as a1a, a1b, a1d and b3 adrenoceptors. Brown...

Anatomic Motor Point Localization of the Biceps Brachii and Brachialis Muscles

Injection of the neurolytic agents into motor points of the biceps brachii or brachialis muscles is an effective treatment of spasticity of the elbow flexors in many stroke survivors. Accurate localization of the motor points of each muscle is necessary for enhancing the efficacy of motor point blocks. To identify the precise locations of the motor points (terminal nerve endings) of the biceps brachii and brachialis muscles in relation to anatomic surface landmarks for motor point blocks, we dissected 23 limbs from 12 cadavers. A reference line was defined as a line connecting the coracoid process with the lateral epicondyle of the humerus. The location of the motor points of the biceps brachii and brachialis muscles was identified in reference to the reference line. The motor point of the biceps brachii muscle was found to be approximately half of the reference line. In the brachialis muscle, the location of the motor point was 70% of the reference line from the coracoid process and 2 cm medial to the line. The results are expected to facilitate effective localization of the motor point block of these muscles in selective motor nerve block.

Influence of Hindlimb-unloading on the Morphology of Nerve Endings in Soleus Muscle Spindles / 航天医学与医学工程

Objective To investigate changes induced by hindlimb unloading in morphology of nerve endings in soleus muscle spindles.Method Hindlimb unloading model was simulated by tail suspension in female rats.Fast silver-staining was used to observe morphology of nerve endings of single muscle spindle in 7 d,14 d,21 d and 28 d tail-suspended and control rats.Result Nerve endings of muscle spindles in control rats were intact and clear,and there was no obvious abnormality in single soleus muscle spindle of 7 d tail-suspended rats.Slight fragmentation of nerve endings was observed in Equatorial Region of soleus muscle spindle in 14 d tail-suspended rats,while reduction in density and disruption of nerve endings were observed in soleus muscle spindle in 21 d tail-suspended rats.Serious degeneration and distortion of nerve endings were found in soleus muscle spindle in 28 d tail-suspended rats.Conclusion Hindlimb-unloading can induce time-related changes in morphology of nerve endings of muscle spindle in rats soleus muscle.

Ultrastructural Changes of Myotendinous Nerve Endings following Recession or Resection Procedures of Extraocular Muscle Surgeries in Cats

To verify the postoperative ultrastructural changes of the myotendinous nerve endings of feline extraocular muscles, which are known as proprioceptors. Sixteen recti of four cats were used and divided into three groups. In group A, eight lateral recti were recessed. In group B, four medial recti were resected by 10 mm from insertion to include the myotendinous junction. In group C, four medial recti were resected by 4 mm of muscle bellies only, without disturbing the myotendinous junction. Four weeks after surgery, specimens were examined with electron microscopy. In group A, overall neural structures were well maintained with slight axonal degeneration. In group B, only muscle fibers were observed without any regeneration of neural sprouts. In group C, axonal disintegration and shrinkage were evident. These results indicate that myotendinous nerve endings can be damaged in strabismus surgery, and that resection was more invasive than recession in disrupting myotendinous nerve endings.

Sensory Innervation in Human Hip Joint Capsule and Pseudocapsule After Total Hip Replacement: A Morphological Investigation / 대한정형외과학회잡지

PURPOSE: To identify sensory nerve endings in the human hip joint capsule and in the pseudocapsule after total hip replacement. MATERIALS AND METHODS: Ten hip joint capsules from patients undergoing bipolar hip replacement for acute femoral neck fracture, and six pseudocapsules from patients undergoing revision hip surgery for failed total hip replacement were harvested and stained in bulk using a modified gold-chloride method. Sensory nerve endings were identified using the criteria described by Freeman and Wyke. RESULTS: Three morphologically distinct types of nerve endings were identified in the normal human hip joint capsules; type I Ruffini corpuscles, type II Pacinian corpuscles and type IV free nerve endings. In contrast, no proprioceptive nerve endings (type I and II receptors) were observed in pseudocapsular tissues. A small number of type IV receptors were noted in the pseudocapsule, but these were significantly fewer in number than in normal hip capsular tissue. CONCLUSION: The pseudocapsule that forms after hip replacement surgery may protect joint stability through a mechanical check-rein effect rather than through a proprioceptive feedback mechanism.

The neurohistology of the rotator cuff and capsule of shoulder / 中华骨科杂志

Objective The articular neurohistology is a new approach to explore stabilized and chronic pain mechanisms for joint research. Furthermore, the distribution of mechanoreceptors and their functional condition play an important role in the movement and stability of joint. The animal experimental study suggest that there is a various characteristics of the type and amount of mechanoreceptors in different joints. Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs as joint mechanoreceptors are widely distributed on nearly all of human joint, and have impact on stabilization of joint. The present investigates the neurohistological features of Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs in the rotator cuff and the capsule, and explores their behaviors for maintaining stability of the shoulder joint. Methods Six rotator cuffs and capsules of shoulder joint were obtained from cadavers died of the accident. The tendinous tissue of subscapularis, supraspinatus、infraspinatus, and teres minis were resected from the interface of tendon and muscle to great and lesser tuberosity of humerus, the capsule tissues of glenohumeral were harvested as well. Then, the tissues were cut into a small fragments fixed with the formalin buffer solution, and embed in paraffin. The section was made along the longitudinal axis of the tendon at 150 ?m interval with Leitz 1516 machine. The modified Bielschowsky staining and the S-100 immuohistochemical staining were adopted to observe the mechanoreceptors of Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs. The distribution density and size of the sensitive nerve endings were analyzed with Leica imaging process system. Results The tendinous part of rotator cuff could be divided into three layers: synovial, intermediate and articular layer. The morphology of the these three types of mechanoreceptors, Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs, could be identified clearly in the every layer of the tendious tissues under microscopic examination. Pacini corpuscle was ellipse and had multi layer capsule with the thickness of 20-40 ?m; Ruffini corpuscle was columnar and 350-550 ?m in size; Most of Golgi tendon organs were found at the interface of tendon and muscle. Among of them, the Pacini corpuscle and Ruffini corpuscle were much more than the Golgi tendon organs in rotator cuff and capsule of shoulder. The results of immuohistochemical staining and the counting of nerve ending assisted by the computer showed the number of sensory nerve endings in the subscapularis, supraspinatus and anterior capsule of the shoulder were much more than in the infraspinatus, teres minis and posterior capsule. The diameter of nerve bundle in the subscapularis, supraspinatus and anterior capsule was thicker than in the other periarticular structures. In periarticular structures, the mean value of nerve fiber counting were 25 in infraspinatus, 25 in supraspinatus, 13 in subscapularis, 10 in teres minis, 22 in anterior capsule and 12 in posterior capsule. Conclusion Shoulder capsule and rotator cuff are rich in the three type of mechanoreceptors, Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs. However, there are a different distribution density and size of them in rotator cuff and capsule of shoulder, but the three of mechanoreceptors are mainly concentrated in the subscapularis, supraspinatus and the anterior capsule. S-100 immuohistochemical staining is much more sensitive than the modified Bielschowsky silver staining in displaying the innervations of the rotator cuff and capsule. The characters of the sensory nerve ending distribution in shoulder imply that the innervations of the shoulder play an important role in keeping the stability and mobility of the shoulder.

Morphometric study of the plasticity of nerve fibers of spared spinal dorsal root and its terminals in rats / 第三军医大学学报

The number of nerve fibers in the spared dorsal root of the L4 spinal nerve and the number of nerve terminals in the Lamina Ⅲ of the segment of the spinal cord between L2 and S1 were determined with electron microscopic morphometry,and the distribution and density of nerve fibers in the spinal cord were observed with silver staining.It was found that the distribution and density of nerve fibers in the spinal cord showed no difference between the operated and the control sides under optical microscopy;morphometric analysis revealed a significant increase of the number of unmyelinated fibers and a tendency to increase of their diameter;and the number of both simple and complex terminals in the Lamina Ⅲ of the spinal cord could return to the normal level on the 35th day after operation with a significant increase of their diameter.The importance of these changes was preliminarily discussed.