RESUMO
BACKGROUND: Nerve damage is consistently demonstrated after subepineural injection in animal studies, but not after purposeful injection in patients participating in clinical studies. There is a need to better visualise nerves in order to understand the structural changes that occur during subepineural injection. METHODS: We scanned the brachial plexuses of three anaesthetised pigs using micro-ultrasound imaging (55-22 MHz probe), inserted 21 gauge block needles into the radial, median, and axillary nerves, and injected two 0.5 ml boluses of saline into nerves at a rate of 12 ml min-1. Our objectives were to measure the area and diameter of nerves and fascicles, and to describe changes in nerve anatomy, comparing our findings with histology. RESULTS: Images were acquired at 42 sites across 18 nerves in three pigs and compared dimensions (geometric ratio; 95% confidence interval; P value). As expected, the nerve cross-sectional area was greater in the proximal brachial plexus compared with the mid-plexus (2.10; 1.07-4.11; P<0.001) and the distal plexus (2.64; 1.42-4.87; P<0.001). Nerve area expanded after 0.5 ml injection (2.13; 1.48-3.08; P<0.001). Using microultrasound, subepineural injection was characterised by nerve and fascicle rotation, uniform, or localised swelling and epineural rupture. Micro-ultrasound revealed a unique pattern suggestive of subperineural injection after a median nerve injection, and good face validity with histology. Histology showed epineural trauma and inflammation to the perineurium. CONCLUSION: We accurately identified fascicles and real-time structural changes to peripheral nerves using micro-ultrasound. This is the first study to visualise in vivo and in real-time the motion of nerves and fascicles in response to anaesthetic needle insertion and fluid injection.
Assuntos
Bloqueio do Plexo Braquial/métodos , Plexo Braquial/diagnóstico por imagem , Sistemas Computacionais , Transdutores , Ultrassonografia de Intervenção/métodos , Adjuvantes Anestésicos/administração & dosagem , Anestésicos Dissociativos/administração & dosagem , Animais , Plexo Braquial/efeitos dos fármacos , Masculino , Nervos Periféricos/diagnóstico por imagem , Nervos Periféricos/efeitos dos fármacos , SuínosRESUMO
The ventricular layer of the spinal cord is remodelled during embryonic development and ultimately forms the ependymal cell lining of the adult central canal, which retains neural stem cell potential. This anatomical transformation involves the process of dorsal collapse; however, accompanying changes in tissue organisation and cell behaviour as well as the precise origin of cells contributing to the central canal are not well understood. Here, we describe sequential localised cell rearrangements which accompany the gradual attrition of the spinal cord ventricular layer during development. This includes local breakdown of the pseudostratified organisation of the dorsal ventricular layer prefiguring dorsal collapse and evidence for a new phenomenon, ventral dissociation, during which the ventral-most floor plate cells separate from a subset that are retained around the central canal. Using cell proliferation markers and cell-cycle reporter mice, we further show that following dorsal collapse, ventricular layer attrition involves an overall reduction in cell proliferation, characterised by an intriguing increase in the percentage of cells in G1/S. In contrast, programmed cell death does not contribute to ventricular layer remodelling. By analysing transcript and protein expression patterns associated with key signalling pathways, we provide evidence for a gradual decline in ventral sonic hedgehog activity and an accompanying ventral expansion of initial dorsal bone morphogenetic protein signalling, which comes to dominate the forming the central canal lining. This study identifies multiple steps that may contribute to spinal cord ventricular layer attrition and adds to increasing evidence for the heterogeneous origin of the spinal cord ependymal cell population, which includes cells from the floor plate and the roof plate as well as ventral progenitor domains.
Assuntos
Proliferação de Células/fisiologia , Ventrículos Cerebrais/citologia , Medula Espinal/citologia , Animais , Apoptose/fisiologia , Proteínas Morfogenéticas Ósseas/metabolismo , Ciclo Celular/fisiologia , Ventrículos Cerebrais/metabolismo , Epêndima/citologia , Epêndima/metabolismo , Proteínas Hedgehog/metabolismo , Camundongos , Fatores de Transcrição SOXB1/metabolismo , Transdução de Sinais/fisiologia , Medula Espinal/metabolismoRESUMO
Cadaver preservation methods impact their utilization in anatomical research and teaching. Thiel-embalmed cadavers show flexibility, however, the cause remains poorly understood. This study aimed to (1) describe qualitative and quantitative histological differences between Thiel-embalmed and formalin-fixed skeletal muscle and tendon tissue; (2) investigate whether boric acid in Thiel solution is solely responsible for modification of tissues; and (3) explore whether the modifications observed could potentially explain the mechanisms underpinning flexibility of Thiel cadavers. Skeletal muscle and tendon samples were harvested from mice preserved using formalin, Thiel solution, or modified-Thiel solution (without boric acid). Using standard H&E and Gomori's trichrome histological methods, tissues were examined to determine whether differences were apparent between the preservative treatments. Differences were present between the Thiel and formalin-fixed tissues; formalin-fixed samples remained substantially more intact while Thiel-embalmed samples showed fiber fragmentation and lack of nuclei. The mean cell diameter of Thiel-embalmed muscle (24.4 µm) was significantly smaller (P < 0.005) than formalin-fixed muscle (40.7 µm). There was significantly greater (P < 0.005) fragmentation in Thiel-embalmed muscle (631.5 per 1 mm2 ) compared to formalin-fixed muscle (75.4 per 1 mm2 ). Samples embalmed using modified-Thiel showed a severe lack of integrity within internal tissue structure. This suggests that Thiel solution significantly alters tissue structure at cellular level, with quantitative data demonstrating measurable differences between Thiel and formalin-fixed specimens. While the precise mechanism for these alterations remains unknown, it is shown that boric acid is not the only component of Thiel responsible for degradation of internal tissue structure. Clin. Anat., 33:696-704, 2020. © 2019 Wiley Periodicals, Inc.
Assuntos
Ácidos Bóricos/química , Embalsamamento/métodos , Formaldeído/química , Músculo Esquelético/anatomia & histologia , Tendões/anatomia & histologia , Animais , Cadáver , CamundongosRESUMO
Primary loss and dysfunction of astrocytes may trigger demyelination, as seen in neuromyelitis optica, an inflammatory disease of the central nervous system. In most patients affected by this disease, injury to astrocytes is initiated by the action of autoantibodies targeting aquaporin 4 (AQP-4), a water channel on astrocytes. We show here that damage of astrocytes and subsequent demyelination can also occur in the absence of autoantibody-mediated mechanisms. Following injection of lipopolysaccharide into the white matter initial microglia activation is followed by a functional disturbance of astrocytes, mainly reflected by retraction of astrocytic foot processes at the glia limitans and loss of AQP-4 and connexins, which are involved in the formation of gap junctions between astrocytes and oligodendrocytes. Demyelination and oligodendrocyte degeneration in this model follows astrocyte pathology. Similar structural abnormalities were also seen in a subset of active lesions in multiple sclerosis. Our studies suggest that astrocyte injury may be an important early step in the cascade of lesion formation in brain inflammation.
Assuntos
Astrócitos/imunologia , Sistema Nervoso Central/patologia , Encefalomielite Autoimune Experimental , Imunidade Inata , Esclerose Múltipla/patologia , Animais , Aquaporina 4/metabolismo , Astrócitos/patologia , Células Cultivadas , Técnicas de Cocultura/métodos , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/etiologia , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Humanos , Proteínas de Membrana/metabolismo , Microscopia Confocal/métodos , Esclerose Múltipla/imunologia , Proteínas do Tecido Nervoso/metabolismo , Neuroglia/efeitos dos fármacos , Polissacarídeos/farmacologia , Ratos , Fatores de TempoRESUMO
Lesions obtained early in the course of multiple sclerosis (MS) have been studied immunocytochemically, and compared with the early stages of the experimental lesion induced in rats by the intraspinal injection of lipopolysaccharide. Large hemispheric or double hemispheric sections were examined from patients who had died in the course of acute or early relapsing multiple sclerosis. In MS patients exhibiting hypoxia-like lesions [Pattern III; Lucchinetti et al. Ann Neurol (2000) 47: 707-17], focal areas in the white matter showed mild oedema, microglial activation and mild axonal injury in the absence of overt demyelination. In such lesions T-cell infiltration was mild and restricted to the perivascular space. Myeloperoxidase and the inducible form of nitric oxide synthase were expressed primarily by microglia, and the activated form of these cells was associated with extracellular deposition of precipitated fibrin. In addition, these lesions showed up-regulation of proteins involved in tissue preconditioning. When active demyelination started, lesions were associated with massive T-cell infiltration and microglia and macrophages expressed all activation markers studied. Similar tissue alterations were found in rats in the pre-demyelinating stage of lesions induced by the focal injection of bacterial lipopolysaccharide into the spinal white matter. We suggest that the areas of microglial activation represent an early stage of tissue injury, which precedes the formation of hypoxia-like demyelinated plaques. The findings indicate that mechanisms associated with innate immunity may play a role in the formation of hypoxia-like demyelinating lesions in MS.
Assuntos
Encéfalo/imunologia , Esclerose Múltipla/imunologia , Linfócitos T/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Axônios/patologia , Encéfalo/patologia , Estudos de Casos e Controles , Citotoxicidade Imunológica , Encefalomielite Autoimune Experimental , Feminino , Humanos , Hipóxia/imunologia , Hipóxia/metabolismo , Imunidade Inata , Imuno-Histoquímica , Lipopolissacarídeos , Macrófagos/patologia , Masculino , Microglia/patologia , Microscopia de Fluorescência , Pessoa de Meia-Idade , Esclerose Múltipla/metabolismo , Esclerose Múltipla/patologia , Óxido Nítrico Sintase/metabolismo , Oligodendroglia/patologia , Ratos , Ratos Sprague-Dawley , Estatísticas não ParamétricasRESUMO
BACKGROUND: Although the glenoid labrum has an important role in shoulder stability, little is known about its composition, vascularity and innervation. The aims of this study were therefore to evaluate the histology, vascularity and innervation of the glenoid labrum. MATERIALS AND METHODS: Ten glenoid labrum specimens (three male, two female: mean age 81.2 years, range 76-90 years) were detached at the glenoid neck. Following decalcification, sections were cut through the whole thickness of each specimen perpendicular to the glenoid labrum at 12 radii corresponding to a clock face superimposed on the glenoid fossa. Then they were stained using haematoxylin and eosin, a silver nitrate protocol or subjected to immunohistochemistry using anti-protein gene protein 9.5 to demonstrate neuronal processes. RESULTS: The labrum was fibrocartilaginous, being more fibrous in its free margin. There was a variable distribution of blood vessels, being more vascular in its periphery, with many originating from the fibrous capsule and piercing the glenoid labrum. Immunohistochemistry revealed positive staining of nerve fibres within the glenoid labrum. CONCLUSION: The glenoid labrum is fibrocartilaginous, being more fibrous in its periphery, and is vascularized, with the anterosuperior aspect having a rich blood supply. Free sensory nerve fibres were also present; no encapsulated mechanoreceptors were observed. The presence of sensory nerve fibres in the glenoid labrum could explain why tears induce pain. It is postulated that these sensory fibres could play a role in glenohumeral joint proprioception.
Assuntos
Articulação do Ombro/irrigação sanguínea , Articulação do Ombro/inervação , Idoso , Idoso de 80 Anos ou mais , Cadáver , Feminino , Fibrocartilagem , Cavidade Glenoide , Humanos , Masculino , Escápula , Articulação do Ombro/patologiaRESUMO
BACKGROUND: Tears of the glenoid labrum are common after dislocation of the glenohumeral joint. The outcome for healing or surgical reconstruction of the glenoid labrum relies on the extent of its vascularization. This study aims to evaluate the glenoid labrum blood supply and to determine its regional vascularity. MATERIALS AND METHODS: A total of 140 shoulders (30 male and 40 female cadavers) were examined: mean age 81.5 years, range 53-101 years. All blood vessels around the glenohumeral joint were dissected and recorded. Ten specimens with the glenoid labrum and fibrous capsule attached were randomly selected and detached at the glenoid neck and subjected to decalcification. Sections (10-20 µm) were cut through the whole thickness of each specimen from the centre of the glenoid fossa perpendicular to the glenoid labrum at 12 radii corresponding to a clock face superimposed on the glenoid. Sections were stained using haematoxylin and eosin and then examined. RESULTS: The blood supply to the glenoid labrum is by direct branches from the second part of the axillary artery, subscapular, circumflex scapular and anterior circumflex humeral and posterior circumflex humeral arteries, as well as branches of muscular arteries supplying the surrounding muscles. CONCLUSION: This study shows that the glenoid labrum has a rich blood supply suggesting that, regardless of the types of the glenoid labrum lesions or their management, an excellent outcome for glenoid labrum healing and joint stability is possible. The observations also suggest that the blood supply to the glenoid labrum is sufficient, enabling its reattachment.
Assuntos
Cavidade Glenoide/irrigação sanguínea , Úmero/irrigação sanguínea , Escápula/irrigação sanguínea , Articulação do Ombro/irrigação sanguínea , Idoso , Idoso de 80 Anos ou mais , Cadáver , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Inflammation is a prominent feature of several disorders characterized by primary demyelination, but it is not clear whether a relationship exists between inflammation and myelin damage. We have found that substantial demyelination results from the focal inflammatory lesion caused by the injection of lipopolysaccharide (LPS; 200 ng) directly into the rat dorsal funiculus. Within 24 h, such injections caused a focal inflammatory response consisting of a substantial number of polymorphonuclear cells and ED1-positive and inducible nitric oxide synthase (iNOS)-positive macrophages/microglia. The number of inflammatory cells was substantially reduced by day 7. OX-52-positive T-cells were less frequently observed but were present in the meninges at 8 h, reached a maximum in the dorsal funiculus at 7 days, and were rare at 14 days. The inflammation was followed by the appearance of a large lesion of primary demyelination that encompassed up to approximately 75% of the cross-sectional area of the dorsal funiculus. Treatment with dexamethasone significantly reduced the number of cells expressing iNOS, but did not prevent the demyelination. By 28 days the lesions were largely remyelinated, usually by Schwann cells. These changes were not observed in control, saline-injected animals. We conclude that the intraspinal injection of LPS results in inflammation and subsequently in prominent demyelination. The mechanisms underlying the demyelination are not clear, but it is notable that it typically begins with disruption of the adaxonal myelin. Indeed, there is an early loss of myelin-associated glycoprotein within the lesion, despite the persistence of proteolipid protein. This combination is a feature of the pattern III lesion recently described in multiple sclerosis (Lucchinetti et al., 2000), and we therefore suggest that LPS-induced demyelination may serve as the first experimental model available for the study of this type of multiple sclerosis lesion.
Assuntos
Doenças Desmielinizantes/imunologia , Lipopolissacarídeos/farmacologia , Modelos Animais , Esclerose Múltipla , Animais , Doenças Desmielinizantes/microbiologia , Doenças Desmielinizantes/patologia , Dexametasona/uso terapêutico , Escherichia coli , Gânglios Espinais/imunologia , Gânglios Espinais/microbiologia , Gânglios Espinais/patologia , Glucocorticoides/uso terapêutico , Imuno-Histoquímica/métodos , Inflamação , Injeções Espinhais , Molécula 1 de Adesão Intercelular/análise , Interleucina-1/análise , Ativação de Macrófagos , Masculino , Microglia/imunologia , Infiltração de Neutrófilos , Óxido Nítrico Sintase/análise , Óxido Nítrico Sintase Tipo II , Ratos , Ratos Sprague-Dawley , Salmonella , Células de Schwann/patologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Fatores de TempoRESUMO
Many histological methods in forensic anthropology utilize combinations of traditional histomorphometric parameters which may not accurately describe the morphology of microstructural features. Here, we report the novel application of a geometric morphometric method suitable when considering structures without anatomically homologous landmarks for the quantification of complete secondary osteon size and morphology. The method is tested for its suitability in the measurement of intact secondary osteons using osteons digitized from transverse femoral diaphyseal sections prepared from two human individuals. The results of methodological testing demonstrate the efficacy of the technique when applied to intact secondary osteons. In providing accurate characterization of micromorphology within the robust mathematical framework of geometric morphometrics, this method may surpass traditional histomorphometric variables currently employed in forensic research and practice. A preliminary study of the intersectional histomorphometric variation within the femoral diaphysis is made using this geometric histomorphometric method to demonstrate its potential.
Assuntos
Diáfises/ultraestrutura , Fêmur/ultraestrutura , Ósteon/anatomia & histologia , Conceitos Matemáticos , Idoso de 80 Anos ou mais , Pontos de Referência Anatômicos , Remodelação Óssea , Feminino , Antropologia Forense , Análise de Fourier , Humanos , Processamento de Imagem Assistida por Computador , MicroscopiaRESUMO
Long-term disability in Guillain-Barré syndrome (GBS) is associated with axonal, and some neuronal, degeneration. Brain-derived neurotrophic factor (BDNF) can prevent neuronal death following damage to motor axons and we have therefore examined the ability of BDNF to ameliorate the effects of experimental autoimmune neuritis (EAN), a model of GBS. Treatment of Lewis rats with BDNF (10 mg/kg/day) did not significantly affect the neurological deficit, nor significantly improve survival, motor function or motor innervation. The weight of the urinary bladder was significantly increased in control animals with EAN, but remained similar to normal in animals treated with BDNF. With the exception of a possibly protective effect indicated by bladder weight, this study suggests that BDNF may not provide an effective therapy for GBS, at least in the acute phase of the disease.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/uso terapêutico , Neurite Autoimune Experimental/tratamento farmacológico , Animais , Anticorpos/análise , Fator Neurotrófico Derivado do Encéfalo/imunologia , Colina O-Acetiltransferase/metabolismo , Força da Mão , Histocitoquímica , Masculino , Atividade Motora/efeitos dos fármacos , Placa Motora/enzimologia , Placa Motora/ultraestrutura , Músculo Esquelético/enzimologia , Músculo Esquelético/ultraestrutura , Neurite Autoimune Experimental/fisiopatologia , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Endogâmicos Lew , Falha de Tratamento , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/patologiaRESUMO
We demonstrate that there are significantly more p75 neurotrophin receptor- (NTR)-expressing cells in olfactory ensheathing cell (OEC) primary cultures from olfactory nerve rootlets (ONR), but a greater proportion of O4 antigen- and PSA-NCAM-expressing cells in parallel cultures from the nerve fibre layer of the olfactory bulb (OB). By co-culturing adult rat retinal ganglion cells (RGCs) with OECs derived from either ONR or OB tissue, we compared their neurite regrowth-promoting properties. In phenotypically unsorted cultures, there is greater RGC neurite regrowth on ONR OECs compared to OB OECs. Following immunoselection of ONR cells for p75 NTR, there is increased RGC neurite regrowth on the enriched population compared to the unselected cell population or the p75 NTR depleted population. When p75 NTR-enriched cells from ONR and OB cultures are compared directly, tissue source-related differences are no longer observed. Our previous work implicated a pertussis toxin (PTx)-sensitive G protein-linked signalling pathway in OEC regulation of neurite regrowth. We show that this pathway probably operates in interactions between the p75 NTR-positive and -negative cells; separated populations lose the PTx-mediated enhancement of neurite regrowth-promoting properties seen in mixed cultures. Optimum neurite regrowth is observed when both phenotypes are present in cultures from either ONR or OB, and where glial G-protein signalling is disabled by PTx before co-culture with neurons. We thus propose that p75 NTR-positive cells, whilst being the more effective neurite regrowth promoting subpopulation in isolation, cooperate with negative cells to provide optimum support for axonal regrowth.