RESUMEN
Magnesium (Mg) ions are the most abundant intracellular divalent cations and play a pivotal role in numerous cellular processes. Biodegradable Mg-containing materials, including scaffolds, are promising candidates for orthopedic applications. Here, we investigated the effect of Mg ions on the cellular properties of osteoblasts. Cytotoxicity tests on osteoblasts confirmed that no cytotoxic effects were found up to a supplementing Mg ion concentration of 10 mM. Mg ions at a concentration of 5 mM increased the migration and invasiveness of osteoblasts. To investigate the stimulatory effect of Mg ions on cell motility in scaffolds, we fabricated 10 wt% Mg ion-containing polycaprolactone (PCL) scaffolds, using the wire-network molding (WNM) technique. Mg ion-containing scaffolds persistently released Mg ions at a concentration of 5 mM in the media after pre-incubation. Furthermore, increased cell motility was confirmed in Mg ion-containing scaffolds by quantification of genomic DNA and protein content. Our results provide an important basis for the function of Mg ions and their effect on cell motility, and propose a novel role for Mg ions in scaffold applications.
Asunto(s)
Movimiento Celular/efectos de los fármacos , Magnesio/química , Osteoblastos/metabolismo , Andamios del Tejido/química , Materiales Biocompatibles/farmacología , Sustitutos de Huesos/farmacología , Cationes , ADN/química , Humanos , Ensayo de Materiales , Microscopía Fluorescente , Osteogénesis/efectos de los fármacos , Poliésteres/química , Porosidad , Ingeniería de Tejidos/métodosRESUMEN
Oriental natural plants have been used as medical herbs for the treatment of various diseases for over 2,000 years. In this study, we evaluated the effect of several natural plants on the preservation of male fertility by assessing the ability of plant extracts to stimulate spermatogonial stem cell (SSC) proliferation by using a serum-free culture method. In vitro assays showed that Petasites japonicus extracts, especially the butanol fraction, have a significant effect on germ cells proliferation including SSCs. The activity of SSCs cultured in the presence of the Petasites japonicus butanol fraction was confirmed by normal colony formation and spermatogenesis following germ cell transplantation of the treated SSCs. Our findings could lead to the discovery of novel factors that activate SSCs and could be useful for the development of technologies for the prevention of male infertility.
Asunto(s)
Petasites/química , Extractos Vegetales/farmacología , Espermatogonias/efectos de los fármacos , Espermatogonias/metabolismo , Células Madre/efectos de los fármacos , Células Madre/metabolismo , Animales , Técnicas de Cultivo de Célula , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Medio de Cultivo Libre de Suero , Fertilidad/efectos de los fármacos , Masculino , RatonesRESUMEN
Basal and activity-dependent cerebral blood flow changes are coordinated by the action of critical processes, including cerebral autoregulation, endothelial-mediated signaling, and neurovascular coupling. The goal of our study was to determine whether astrocytes contribute to the regulation of parenchymal arteriole (PA) tone in response to hemodynamic stimuli (pressure/flow). Cortical PA vascular responses and astrocytic Ca(2+) dynamics were measured using an in vitro rat/mouse brain slice model of perfused/pressurized PAs; studies were supplemented with in vivo astrocytic Ca(2+) imaging. In vitro, astrocytes responded to PA flow/pressure increases with an increase in intracellular Ca(2+). Astrocytic Ca(2+) responses were corroborated in vivo, where acute systemic phenylephrine-induced increases in blood pressure evoked a significant increase in astrocytic Ca(2+). In vitro, flow/pressure-evoked vasoconstriction was blunted when the astrocytic syncytium was loaded with BAPTA (chelating intracellular Ca(2+)) and enhanced when high Ca(2+) or ATP were introduced to the astrocytic syncytium. Bath application of either the TRPV4 channel blocker HC067047 or purinergic receptor antagonist suramin blunted flow/pressure-evoked vasoconstriction, whereas K(+) and 20-HETE signaling blockade showed no effect. Importantly, we found TRPV4 channel expression to be restricted to astrocytes and not the endothelium of PA. We present evidence for a novel role of astrocytes in PA flow/pressure-evoked vasoconstriction. Our data suggest that astrocytic TRPV4 channels are key molecular sensors of hemodynamic stimuli and that a purinergic, glial-derived signal contributes to flow/pressure-induced adjustments in PA tone. Together our results support bidirectional signaling within the neurovascular unit and astrocytes as key modulators of PA tone.
Asunto(s)
Arteriolas/fisiología , Astrocitos/fisiología , Circulación Cerebrovascular/fisiología , Canales Catiónicos TRPV/biosíntesis , Vasoconstricción/fisiología , Animales , Encéfalo/irrigación sanguínea , Encéfalo/fisiología , Homeostasis/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Técnicas de Cultivo de Órganos , Ratas , Ratas WistarRESUMEN
An understanding of the signalling events underlying neurovascular coupling mechanisms in the brain is a crucial step in the development of novel therapeutic approaches for the treatment of cerebrovascular-associated disorders. In this study we present an enhanced in vitro brain slice preparation from male Wistar rat cortical slices that incorporates haemodynamic variables (flow and pressure) into parenchymal arterioles resulting in the development of myogenic tone (28% from maximum dilatation). Moreover, we characterized flow-induced vascular responses, resulting in various degrees of vasoconstrictions and the response to 10 mM K(+) or astrocytic activation with the mGluR agonist, t-ACPD (100 µM), resulting in vasodilatations of 33.6±4.7% and 38.6±4.6%, respectively. Using fluorescence recovery, we determined perfusate velocity to calculate diameter changes under different experimental pH conditions. Using this approach, we demonstrate no significant differences between diameter changes measured using videomicroscopy or predicted from the velocity values obtained using fluorescence recovery after photobleaching. The model is further validated by demonstrating our ability to cannulate arterioles in two brain regions (cortex and supraoptic nucleus of the hypothalamus). Altogether, we believe this is the first study demonstrating successful cannulation and perfusion of parenchymal arterioles while monitoring/estimating luminal diameter and pressure under conditions where flow rates are controlled.
Asunto(s)
Arteriolas/fisiología , Corteza Cerebral/irrigación sanguínea , Hipotálamo/irrigación sanguínea , Animales , Presión Sanguínea , Corteza Cerebral/fisiología , Hipotálamo/fisiología , Masculino , Microcirculación , Ratas , Ratas Wistar , Flujo Sanguíneo RegionalRESUMEN
The objective is to evaluate the changes in gait patterns with rhythmic auditory stimulation (RAS) in adults with cerebral palsy (CP). Fourteen CP with bilateral spasticity participated in this study. A repeated-measures analysis of gait was performed in the presence and absence of RAS. Thirty healthy controls were also recruited. Each subject walked 10 m at their comfortable walking speed. Temporospatial data and kinematic parameters of gait were analyzed without RAS and with RAS. RAS was provided using a combination of a metronome beat set to the individual's cadence and rhythmic cueing from a live keyboard playing. Kinematic parameters, gait deviation index (GDI) as a measure of overall gait pathology, and asymmetry of temporospatial data were assessed. Gait analysis revealed that anterior tilt of pelvis and hip flexion during a gait cycle was significantly changed with RAS (p < 0.05), whereas there were no statistical differences in knee, ankle, and foot kinematic parameters. Additionally, the GDI exhibited a modest, but a statistically significant, improvement with RAS (p < 0.05). Based on ambulatory status, household ambulators showed that side-to-side asymmetry of step length as well as the GDI was significantly attenuated with RAS (p < 0.05). Walking with RAS resulted in kinematic changes of the pelvic and hip movement in spastic CP. Especially, the application of RAS immediately ameliorated overall gait pathology as well as temporospatial asymmetry in household ambulators. Therefore, RAS may be one of the therapeutic tools for gait training in adults with CP.
Asunto(s)
Estimulación Acústica , Marcha , Espasticidad Muscular/rehabilitación , Adolescente , Adulto , Articulación del Tobillo/fisiopatología , Fenómenos Biomecánicos , Estudios de Casos y Controles , Parálisis Cerebral/rehabilitación , Femenino , Articulaciones del Pie/fisiopatología , Trastornos Neurológicos de la Marcha/rehabilitación , Articulación de la Cadera/fisiopatología , Humanos , Articulación de la Rodilla/fisiopatología , MasculinoRESUMEN
This paper describes recent research and development activities on the diagnostic instruments for personalized healthcare services in Seoul National University. Utilizing the state-of-the-art information technologies (IT), various diagnostic medical instruments have been integrated into a personal wearable device and a home telehealthcare system. We developed a wrist-worn integrated health monitoring device (WIHMD) which performs the measurements of non-invasive blood pressure (NIBP), pulse oximetry (SpO2), electrocardiogram (ECG), respiration rate, heart rate, and body surface temperature and the detection of falls to determine the onset of emergency situation. The WIHMD also analyzes the acquired bio-signals and transmits the resultant data to a healthcare service center through a commercial cellular phone. Two different kinds of IT-based blood glucometer have been developed using a cellular phone and PDA(personal digital assistant) as a main unit. A blood glucometer was also integrated within a wrist pressure measurement module which is interfaced with a cellular phone via Telecommunications Technology Association (TTA) standard in order to provide users with easiness in measuring and handling two important health parameters. Non-intrusive bio-signal measurement systems were developed for the ease of home use. One can measure his ECG on a bed while he is sleeping; measure his ECG, body temperature, bodyfat ratio and weight on a toilet seat; measure his ECG on a chair; and estimate the degree of activity by motion analysis using a camera. Another integrated diagnostic system for home telehealthcare services has been developed to include a 12 channels ECG, a pressure meter for NIBP, a blood glucometer, a bodyfat meter and a spirometer. It is an expert system to analyze the measured health data and based on the diagnostic result, the system provides an appropriate medical consultation. The measured data can be either stored on the system or transmitted to the central server through the internet. We have installed the developed systems on a model house for the performance evaluation and confirmed the possibility of the system as an effective tool for the personalized healthcare services.