RESUMO
The cell-free extract, a crude enzyme (cytosolic and membrane fraction) obtained from an environmental isolate, Bacillus pumilus sp. BAB-3706 worked as excellent in reducing as well as stabilizing agent and facilitated the formation of stable colloidal selenium nanoparticles (SeNPs). Resulting nanoparticles were characterized using UV-vis spectrophotometer, TEM, EDAX, FT-IR and XRD, respectively. A working electrode was modified by coating the surface of indium tin oxide (ITO) with colloidal SeNPs. Successive additions of H2O2 (100 to 600 µM) in conventional three electrodes system, cyclic voltammeter with potential scan rate 25.0 mV/s, in 0.1 M phosphate buffer solution (PBS) yielded increase in current. A perpetual amperometric response at fixed potential (-1.0 V) and at selected time interval of 100 s showed different magnitude of current at every addition of H2O2. The linear range of detection of H2O2 was from 5 to 600 mM (R(2) = 0.9965), while the calculated limit of detection was found to be 3.00 µM. The current study suggested that microbial SeNPs can be used for fabrication of low cost, sensitive H2O2 biosensor.
Assuntos
Técnicas Biossensoriais , Peróxido de Hidrogênio/análise , Nanopartículas Metálicas/química , Selênio/química , Bacillus/químicaRESUMO
Inflammatory response in the dental pulp can alter the collagen matrix formation by dental pulp stem cells and lead to a delay or poor healing of the pulp. This inflammatory response is mediated by cytokines, including interleukin-1ß and tumor necrosis factor-α. In this study, it is hypothesized that suppressing the actions of these inflammatory cytokines by knocking down the activity of transcription factor Nuclear Factor-κB will lead to dental pulp stem cell differentiation into odontoblasts and the production of collagen. Here, the role of Nuclear Factor-κB signaling and its reduction was examined during odontogenic behavior in the presence of these cytokines. The results showed a significant increase in Nuclear Factor-κB gene expression and p65 protein expression by interleukin-1ß and tumor necrosis factor-α. Nuclear Factor-κB activation in the presence of these cytokines decreased significantly in a dose-dependent manner by a Nuclear Factor-κB inhibitor (MG132) and p65 siRNA. Down-regulation of Nuclear Factor-κB activity also enhanced the gene expression of the odontoblastic markers (dentin sialophosphoprotein, Nestin, and alkaline phosphatase) and displayed an odontoblastic cell morphology indicating the promotion of odontogenic differentiation of dental pulp stem cells. Finally, dental pulp stem cells exposed to reduced Nuclear Factor-κB activity resulted in a significant increase in collagen (I)-α1 expression in the presence of these cytokines. In conclusion, a decrease in Nuclear Factor-κB in dental pulp stem cells in the presence of inflammatory cytokines enhanced odontoblastic differentiation and collagen matrix formation.
Assuntos
Diferenciação Celular/genética , Colágeno/genética , Polpa Dentária/citologia , NF-kappa B/genética , Odontoblastos/citologia , Odontoblastos/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismo , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/metabolismo , Citocinas/farmacologia , Matriz Extracelular/metabolismo , Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Mediadores da Inflamação/farmacologia , Leupeptinas/farmacologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , NF-kappa B/metabolismo , Odontoblastos/efeitos dos fármacos , Interferência de RNA , RNA Interferente Pequeno/genética , Células-Tronco/efeitos dos fármacosRESUMO
Poly(butylene adipate-co-terephthalate) (PBAT) and Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) are biopolymers that have the potential to be used in applications of bone healing. In this study, it is hypothesized that the polymer blend has the combined strength and osteoconductivity to support osteoblast collagen formation. PBAT (PBAT 100), and a blend with 20% PHBV (PBAT 80) were extruded in the form of fibers and then knitted in the form of mesh. These were tested in the warp as well as weft direction for the tensile properties; these showed that the weft direction had higher performance than the warp. The individual fibers were kept in phosphate buffered saline (PBS) over the period of 8 weeks and were tested for the storage and loss modulus using a dynamic mechanical analyser (DMA). The results indicated that mechanical relaxation strength showed a decrease and then an increase. In vitro osteoconductivity studies were done by using differentiating osteoblasts (MC3T3-E1 subclone 4 cells). Environmental Scanning Electron Microscopy (ESEM) showed that pre-soaking the samples in α-MEM for two weeks resulted in cell attachment and growth. X-ray diffraction (XRD) was used to determine the change in structure of polymers due to in vitro degradation for two weeks. Raman spectroscopy showed that all scaffolds supported the formation of a collagenous network over the scaffold surfaces. For a combination of knittable manufacturing, mechanical performance and osteoconductivity, blends offer an effective route.
Assuntos
Materiais Biocompatíveis/farmacologia , Osseointegração/efeitos dos fármacos , Poliésteres/farmacologia , Animais , Varredura Diferencial de Calorimetria , Linhagem Celular , Módulo de Elasticidade/efeitos dos fármacos , Camundongos , Microscopia Eletrônica de Varredura , Análise Espectral Raman , Estresse Mecânico , Resistência à Tração/efeitos dos fármacos , Difração de Raios XRESUMO
Ischemic conditioning has long held promise for preventing ischemic-reperfusion (I-R) injury. Although a number of studies have evaluated the effects of brief repeated episodes of ischemia before a prolonged ischemic episode on the cardiovascular system using clinical endpoints, more sensitive techniques by which to measure its effects are lacking. Since endothelial function is sensitive to I-R injury, flow mediated dilation of the brachial artery has been proposed for this purpose, but has significant limitations. Hyperemia normally decreases carotid to radial pulse wave velocity (PWV). Accordingly, we sought to determine the effects of I-R injury and ischemic conditioning on the hyperemic change (Δ) in PWV. We induced hyperemia by release of arterial cuff occlusion before and after ipsilateral arm I-R injury (7.5min occlusion) in 25 healthy males, age 29±6 years. The protocol was repeated on 2 occasions in combination with either pre- or post- conditioning stimuli (3× 30s contralateral arm occlusions). Hyperemia resulted in a significant decrease (-13.7%, p<.001) before but not after prolonged ischemia (-0.88%, p=0.40). I-R along with either pre- or post-ischemic conditioning restored the PWV decline (pre: -11.0%, p<0.001; post: -9.9%, p<0.001). In conclusion, 7.5min ischemia blunts the normal PWV decline produced by hyperemia. Remote pre- and post-conditioning restores this response. This technique may be useful for the assessment of novel treatment strategies and mechanisms underlying remote pre- and post-ischemic conditioning in protecting the cardiovascular system.
Assuntos
Artérias Carótidas/fisiopatologia , Hiperemia/fisiopatologia , Pós-Condicionamento Isquêmico , Precondicionamento Isquêmico , Fluxo Pulsátil , Artéria Radial/fisiopatologia , Traumatismo por Reperfusão/prevenção & controle , Extremidade Superior/irrigação sanguínea , Adulto , Análise de Variância , Humanos , Masculino , Manometria , New York , Traumatismo por Reperfusão/fisiopatologia , Fatores de Tempo , Adulto JovemRESUMO
Aging is associated with increased central aortic systolic pressure (CSP) and pulse pressure which are predictive of cardiovascular events. Mechanisms implicated for higher central pressures include a higher forward incident pressure wave (P1), higher augmented pressure (AP), and shorter reflected wave round trip travel time (Tr). African-Americans (AA) have more frequent and deleterious blood pressure elevation. Using applanation tonometry, we studied the association of age and CSP with P1 and AP in 900 AA subjects. Data showed that in subjects ≤50 years old, CSP was mediated by AP but not P1 or Tr, whereas in those >50, CSP was mediated by both AP and P1 and to a lesser extent by Tr. Predictive models were significant (R(2) = 0.97) for both age groups. In conclusion, wave reflection is the primary determinant of CSP in younger AA, while in older subjects, CSP is mediated by both the magnitude and timing of wave reflection as well as aortic impedance.