Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication

Bone homeostasis seems to be controlled by delicate and subtle “cross talk” between the nervous system and “osteo-neuromediators” that control bone remodeling. The purpose of this study was to evaluate the effect of interactions between neuropeptides and human bone morphogenetic protein 2 (hBMP2) on human osteoblasts. We also investigated the effects of neuropeptides and hBMP2 on gap junction intercellular communication (GJIC). Osteoblasts were treated with neuropeptide Y (NPY), substance P (SP), or hBMP2 at three concentrations. At various intervals after treatment, cell viability was measured by the MTT assay. In addition, cellular alkaline phosphatase (ALP) activity and osteocalcin were determined by colorimetric assay and radioimmunoassay, respectively. The effects of NPY, SP and hBMP on GJIC were determined by laser scanning confocal microscopy. The viability of cells treated with neuropeptides and hBMP2 increased significantly in a time-dependent manner, but was inversely associated with the concentration of the treatments. ALP activity and osteocalcin were both reduced in osteoblasts exposed to the combination of neuropeptides and hBMP2. The GJIC of osteoblasts was significantly increased by the neuropeptides and hBMP2. These results suggest that osteoblast activity is increased by neuropeptides and hBMP2 through increased GJIC. Identification of the GJIC-mediated signal transduction capable of modulating the cellular activities of bone cells represents a novel approach to studying the biology of skeletal innervation.

Quantitative evaluation of experimental choroidal neovascularization by confocal scanning laser ophthalmoscopy: fluorescein angiogram parallels heparan sulfate proteoglycan expression

The objective of the present study was to develop a quantitative method to evaluate laser-induced choroidal neovascularization (CNV) in a rat model using Heidelberg Retina Angiograph 2 (HRA2) imaging. The expression of two heparan sulfate proteoglycans (HSPG) related to inflammation and angiogenesis was also investigated. CNV lesions were induced with argon laser in 21 heterozygous Zucker rats and after three weeks a fluorescein angiogram and autofluorescence exams were performed using HRA2. The area and greatest linear dimension were measured by two observers not aware of the protocol. Bland-Altman plots showed agreement between the observers, suggesting that the technique was reproducible. After fluorescein angiogram, HSPG (perlecan and syndecan-4) were analyzed by real-time RT-PCR and immunohistochemistry. There was a significant increase in the expression of perlecan and syndecan-4 (P < 0.0001) in retinas bearing CNV lesions compared to control retinas. The expression of these two HSPG increased with increasing CNV area. Immunohistochemistry demonstrated that the rat retina damaged with laser shots presented increased expression of perlecan and syndecan-4. Moreover, we observed that the overexpression occurred in the outer layer of the retina, which is related to choroidal damage. It was possible to develop a standardized quantitative method to evaluate CNV in a rat model using HRA2. In addition, we presented data indicating that the expression of HSPG parallels the area of CNV lesion. The understanding of these events offers opportunities for studies of new therapeutic interventions targeting these HSPG.

Avaliação da voz em pacientes submetidos à cordectomia com laser de CO2

OBJETIVO: Avaliar a voz de pacientes portadores de carcinomas iniciais glóticos submetidos à cordectomia com laser de CO2. MÉTODO: Foram avaliados 15 pacientes com diagnóstico de carcinoma espinocelular Tis e T1 glóticos. A avaliação foi feita por meio da análise perceptivo auditiva da voz, pela análise acústica computadorizada e videolaringoestroboscópica. Os pacientes responderam a um questionário de avaliação geral da voz e um protocolo de qualidade de vida relacionado à voz (QVV). Os resultados foram comparados aos de um grupo controle de indivíduos com laringes normais. RESULTADOS: A análise perceptivo-auditiva da voz revelou que a maioria dos pacientes submetidos à cordectomia apresentou algum grau de disfonia, à custa de rouquidão e soprosidade. Considerando os parâmetros acústicos analisados e os valores do grupo controle, observou-se a tendência de um pequeno incremento da freqüência fundamental, mas sem diferença estatisticamente significante; os valores de jitter, shimmer e da proporção harmônico-ruído apresentaram-se significantemente alterados. Os aspectos analisados na videolaringoestroboscopia mostraram-se melhores nas cordectomias menos extensas. Os escores do QVV sugerem que os pacientes tiveram um discreto impacto na qualidade de vida relacionada à voz. CONCLUSÕES: Apesar da presença de alterações na qualidade vocal dos pacientes submetidos à cordectomia com laser de CO2, os resultados funcionais tendem a ser bem aceitos pelos pacientes, com discreta repercussão na qualidade de vida.

Cicatrização da membrana timpânica na timpanocentese com laser de argônio comparado à técnica com microlanceta: estudo experimental em ratos

Otite média secretora (OMS) e otite média aguda recorrente (OMAR) podem necessitar tratamento cirúrgico para adequada ventilação da orelha média. A abertura clássica do tímpano (timpanocentese) requer incisão por microlanceta sob controle de microscópio cirúrgico e mantém-se patente por alguns dias. Estudos recentes sugerem que a timpanocentese feita por diferentes lasers pode permanecer permeável por maior tempo, o que possibilitaria a normalização da otite média. MATERIAL E MÉTODOS: Neste estudo experimental 34 ratos linhagem Wistar, albinos, machos adultos pesando cerca de 300g, foram anestesiados com cetamina 27 mg/kg e xilazina 2,7 mg/kg. A seguir, foram submetidos à timpanocentese incisional com microlanceta no ouvido direito (ML-OD), e à timpanocentese mediada por laser de argônio no ouvido esquerdo (LA-OE). RESULTADOS: Não houve diferença significativa no tempo de cicatrização das timpanocenteses feitas com laser de argônio ou microlanceta. Todas as timpanocenteses cicatrizaram em 10 dias. CONCLUSÃO: A timpanocentese com laser de argônio apresentou patência e cicatrização semelhantes à técnica clássica com microlanceta realizada em ratos Wistar sem enfermidades de orelha média.