RESUMEN
The superficial spinal dorsal horn is the first relay station of pain processing. It is also widely accepted that spinal synaptic processing to control the modality and intensity of pain signals transmitted to higher brain centers is primarily defined by inhibitory neurons in the superficial spinal dorsal horn. Earlier studies suggest that the construction of pain processing spinal neural circuits including the GABAergic components should be completed by birth, although major chemical refinements may occur postnatally. Because of their utmost importance in pain processing, we intended to provide a detailed knowledge concerning the development of GABAergic neurons in the superficial spinal dorsal horn, which is now missing from the literature. Thus, we studied the developmental changes in the distribution of neurons expressing GABAergic markers like Pax2, GAD65 and GAD67 in the superficial spinal dorsal horn of wild type as well as GAD65-GFP and GAD67-GFP transgenic mice from embryonic day 11.5 (E11.5) till postnatal day 14 (P14). We found that GABAergic neurons populate the superficial spinal dorsal horn from the beginning of its delineation at E14.5. We also showed that the numbers of GABAergic neurons in the superficial spinal dorsal horn continuously increase till E17.5, but there is a prominent decline in their numbers during the first two postnatal weeks. Our results indicate that the developmental process leading to the delineation of the inhibitory and excitatory cellular assemblies of pain processing neural circuits in the superficial spinal dorsal horn of mice is not completed by birth, but it continues postnatally.
Asunto(s)
Interneuronas/fisiología , Dolor/fisiopatología , Células del Asta Posterior/fisiología , Asta Dorsal de la Médula Espinal/fisiología , Animales , Neuronas GABAérgicas/fisiología , Ratones Transgénicos , Inhibición Neural/fisiología , Asta Dorsal de la Médula Espinal/embriología , Asta Dorsal de la Médula Espinal/crecimiento & desarrollo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
UNLABELLED: Mosaicplasty has become a well-accepted treatment modality for articular cartilage lesions in the knee. Postoperative bleeding remains potentially concerning. This study evaluates the porous poly(ethylene oxide)terephthalate/poly(butylene terephthalate) (PEOT/PBT) implants used for donor site filling. Empty donor sites were the controls. After 9 months, MRI, macroscopical and histological analysis were carried out. Treated defects did not cause postoperative bleeding. No adverse events or inflammatory response was observed. PEOT/PBT implants were well integrated. Empty controls occasionally showed protrusion of repair tissue at the defect margins. Surface stiffness was minimally improved compared to controls. Existing polymer fragments indicated considerable biodegradation. Histological evaluation of the filled donor sites revealed congruent fibrocartilaginous surface repair with proteoglycan-rich domains and subchondral cancellous bone formation with interspersed fibrous tissue in all implanted sites. The PEOT/PBT implants successfully reduce donor site morbidity and postoperative bleeding after mosaicplasty. LEVEL OF EVIDENCE: II.
Asunto(s)
Materiales Biocompatibles/uso terapéutico , Cartílago Articular/cirugía , Articulación de la Rodilla/cirugía , Poliésteres/uso terapéutico , Polietilenglicoles/uso terapéutico , Implantes Absorbibles , Adulto , Artroplastia/efectos adversos , Artroplastia/métodos , Materiales Biocompatibles/efectos adversos , Cartílago Articular/diagnóstico por imagen , Femenino , Estudios de Seguimiento , Humanos , Implantes Experimentales , Imagen por Resonancia Magnética , Masculino , Poliésteres/efectos adversos , Polietilenglicoles/efectos adversos , Tereftalatos Polietilenos , Hemorragia Posoperatoria/prevención & control , Radiografía , Andamios del Tejido , Trasplante Autólogo/efectos adversos , Trasplante Autólogo/métodos , Adulto JovenRESUMEN
One of the most promising applications for the restoration of small or moderately sized focal articular lesions is mosaicplasty (MP). Although recurrent hemarthrosis is a rare complication after MP, recently, various strategies have been designed to find an effective filling material to prevent postoperative bleeding from the donor site. The porous biodegradable polymer Polyactive (PA; a polyethylene glycol terephthalate - polybutylene terephthalate copolymer) represents a promising solution in this respect. A histological evaluation of the longterm PA-filled donor sites obtained from 10 experimental horses was performed. In this study, attention was primarily focused on the bone tissue developed in the plug. A computer-assisted image analysis and quantitative polarized light microscopic measurements of decalcified, longitudinally sectioned, dimethylmethylene blue (DMMB)- and picrosirius red (PS) stained sections revealed that the coverage area of the bone trabecules in the PA-filled donor tunnels was substantially (25%) enlarged compared to the neighboring cancellous bone. For this quantification, identical ROIs (regions of interest) were used and compared. The birefringence retardation values were also measured with a polarized light microscope using monochromatic light. Identical retardation values could be recorded from the bone trabeculae developed in the PA and in the neighboring bone, which indicates that the collagen orientation pattern does not differ significantly among these bone trabecules. Based on our new data, we speculate that PA promotes bone formation, and some of the currently identified degradation products of PA may enhance osteo-conduction and osteoinduction inside the donor canal.
Asunto(s)
Artroplastia/métodos , Trasplante Óseo/métodos , Cartílago/trasplante , Osteogénesis , Poliésteres/uso terapéutico , Polietilenglicoles/uso terapéutico , Animales , Materiales Biocompatibles , Caballos , Procesamiento de Imagen Asistido por Computador , Articulación de la Rodilla/cirugía , Oseointegración/efectos de los fármacosRESUMEN
The extent of tumor removal determines the effectiveness of postoperative oncotherapy. This is especially true for primary brain tumors, where peritumoral invasion usually makes radical resection impossible. The aim of the study was to determinate the specific expression pattern of invasion related molecules of different intracranial tumors and to identify molecules that are principally responsible for the peritumoral invasiveness of grade II astrocytoma mRNA expression of 26 extracellular matrix (ECM) molecules was determined in tissue samples from grade II astrocytoma, schwannoma, intracerebral metastases of non-small cell lung cancer and normal brain. Immunohistochemical staining for brevican, neurocan, tenascin-C and versican was also performed for each tumor group. Comparing astrocytoma to metastasis, schwannoma and normal brain; and metastasis and schwannoma to normal brain, 22, 17, 20, 21, and 19 molecules, respectively, were found to be significantly overexpressed at the mRNA level. Cluster analysis of mRNA expression showed a specific gene expression pattern for each histological group. Four molecules of 26 were found to be associated to astrocytoma. Immunohistochemical staining confirmed the results of the mRNA analysis at the protein level. Tumors of different origin have a specific invasive phenotype that can evidently determinate on gene expression level. This characteristic expression pattern of the invasion-related molecules might help to screen exact targets for anti-invasion drugs. In case of low-grade astrocytoma. brevican, neurocan, tenascin-C and versican were found to correlate principally with the invasive phenotype of low-grade astrocytoma, thus these molecules can potentially serve as targets for anti-invasion therapy in the future.
Asunto(s)
Astrocitoma/patología , Biomarcadores de Tumor/genética , Neoplasias Encefálicas/secundario , Encéfalo/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/secundario , Neoplasias Pulmonares/patología , Neurilemoma/patología , Astrocitoma/genética , Astrocitoma/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Humanos , Técnicas para Inmunoenzimas , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Invasividad Neoplásica , Neurilemoma/genética , Neurilemoma/metabolismo , Pronóstico , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVES: Ineffective surgical and radiotherapy of glioblastoma is mainly due to its intensive infiltrating behavior. Contrarily, brain metastases of anaplastic carcinomas are well-circumscribed intracerebral lesions that can be easily exstirpated in most cases. The molecules of the extracellular matrix (ECM) play a pivotal role in the peritumoral infiltration. In this study the mRNA expression of the ECM components was investigated in two types of intracerebral malignoma with different invasion activity. Our aim was to identify the ECM molecules that are responsible for the different intensity of peritumoral infiltration of tumors from different origin. METHODS: The mRNA expression of twenty-three ECM molecules was determined by quantitative reverse transcriptase polymerase chain reaction. Four pieces of glioblastoma and four pieces of intracerebral lung adenocarcinoma metastasis from neurosurgical operation were investigated. Immunohistochemical investigations were performed in case of five molecules. RESULTS: The mRNA expression of nine molecules (brevican, neurocan, neuroglycan-C, syndecan-1,2,4, tenascin-C, versican and matrix-metalloproteinase-[MMP]2) differed significantly by comparison of the two tumor types. By immunohistochemistry, neurocan, syndecan, versican and MMP-2 showed alteration in staining intensity according to the mRNA expression, while MMP-9 showed higher staining intensity in the metastatic tumor. CONCLUSIONS: The identified molecules can play an important role in the different infiltration activity of tumors from different origin. Thus these ECM-components could serve as targets for anti-invasion therapy in the future.
Asunto(s)
Adenocarcinoma/química , Biomarcadores de Tumor/análisis , Neoplasias Encefálicas/química , Neoplasias Encefálicas/patología , Matriz Extracelular/química , Matriz Extracelular/patología , Glioblastoma/química , Adenocarcinoma/secundario , Anciano , Biomarcadores de Tumor/genética , Neoplasias Encefálicas/secundario , Brevicano , Proteoglicanos Tipo Condroitín Sulfato/análisis , Femenino , Glioblastoma/secundario , Humanos , Inmunohistoquímica , Lectinas Tipo C/análisis , Neoplasias Pulmonares/patología , Masculino , Metaloproteinasa 2 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/análisis , Persona de Mediana Edad , Invasividad Neoplásica , Proteínas del Tejido Nervioso/análisis , Neurregulinas/análisis , Neurocano , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , Sindecanos/análisis , Tenascina/análisis , Versicanos/análisisRESUMEN
Tumor cell invasion into the surrounding brain tissue is mainly responsible for the failure of radical surgical resection and successful treatment, with tumor recurrence as microdisseminated disease. Epidermal growth factor receptors (EGFRs), integrins and their ligands in the extracellular matrix (ECM) predominantly participate in the invasion process, including the cell adhesion to the surrounding microenvironment and cell migration. The extent of infiltration of the surrounding brain tissue by malignant tumors strongly depends on the tumor cell type. Malignant gliomas show much more intensive peritumoral invasion than do metastatic tumors. In this study, the mRNA expression of 29 invasion-related molecules (18 cell membrane receptors or receptor subunits (EGFRs and integrins) and 11 ECM components: collagens, laminins and fibronectin) was investigated by quantitative reverse transcriptase-polymerase chain reaction. Fresh frozen human tissue samples from glioblastoma (GBM) and intracerebral bronchial adenocarcinoma metastases (five pieces from each) were evaluated. Significant differences were established in six of the 29 molecules (ErbB1, 2, 3, integrins alpha3, 7 and beta1). To confirm our results at the protein level, immunohistochemical analysis of nine molecules was performed. The staining intensity differed definitely in the case of ErbB1, 2 and integrins alpha3 and beta1. Determining the differences in invasion-related molecules in tumors of different origin can help identify the exact molecular mechanisms that facilitate peritumoral infiltration by glioblastoma cells. These results should allow the selection of target molecules for potential chemotherapeutic agents directed against highly invasive malignant gliomas.
Asunto(s)
Biomarcadores de Tumor/análisis , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patología , Regulación Neoplásica de la Expresión Génica , Actinas/análisis , Aminoácidos Diaminos/análisis , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/inmunología , Neoplasias Encefálicas/cirugía , Colágeno/análisis , Receptores ErbB/análisis , Fibronectinas/análisis , Glioblastoma/metabolismo , Glioblastoma/patología , Gliceraldehído-3-Fosfato Deshidrogenasas/análisis , Humanos , Inmunohistoquímica , Cadenas alfa de Integrinas/análisis , Cadenas beta de Integrinas/análisis , Antígeno Ki-67/análisis , Invasividad Neoplásica , ARN Mensajero/análisis , Receptor ErbB-2/análisis , Receptor ErbB-3/análisis , Receptor ErbB-4RESUMEN
The aim of the present study was to investigate the expression pattern of different cell adhesion molecules in corneal stromal dystrophies. Fifteen corneal buttons from patients diagnosed with three different types of stromal corneal dystrophies and healthy corneas were investigated. Paraffin embedded sections were stained immunohistochemically with monoclonal antibodies against human intercellular adhesion molecule-1 (ICAM-1), endothelial selectin (E-selectin) and endothelial cadherin (E-cadherin) using the avidin-biotin-peroxidase-complex technique. The sections were compared to normal eye bank controls. In corneas from granular dystrophy patients ICAM-1 was expressed focally in epithelial cells and in keratocytes, and expressed diffusely in endothelial cells. In corneas from macular dystrophy patients diffuse epithelial staining was observed and the stromal and endothelial expression was found to be similar to that of granular dystrophy. In lattice dystrophy, only the epithelial cells and endothelium were intensively positive for ICAM-1. E-selectin was not present on any layer of the corneal specimens. E-cadherin was observed only in the epithelium of all three types of corneal dystrophies. Normal corneas did not express any of the investigated adhesion molecules. We found different expression patterns of adhesion molecules in corneas from stromal dystrophies. Our results suggest that adhesion molecules may be involved in the pathogenesis of corneal stromal dystrophies.