“…This neuron-like morphology is similar to that described by others [Clagett-Dame et al, 2006;Cheung et al, 2009]. Moreover, the in vitro reconstruction of the neuronal network obtained in this work on PCL membranes clearly indicates the neuronal differentiation, which is in ac- Bartolo et al, 2008;Morelli et al, 2010Morelli et al, , 2012b. To confirm that the observed morphological features were actually resulting from the neuronal differentiation of SH-SY5Y cells, the expression of specific neuronal markers, βIII-tubulin and synaptophysin, were investigated by immunocytochemical analysis.…”
Section: Discussionsupporting
confidence: 88%
“…The present study investigates the protective effect of didymin against H 2 O 2 -induced damage to the neuronal cells in a biohybrid membrane system model. Previous studies have demonstrated that semipermeable polymeric membranes in flat and hollow fiber configurations, thanks to their highly selective structural, physicochemical and transport properties, allow the successful in vitro reconstruction of neuronal tissue, reproducing a tissue model for studying metabolic diseases and drug effects [Woerly et al, 1996;Schmidt and Leach, 2003;Zhang et al, 2005;De Bartolo et al, 2008;Giusi et al, 2009;He et al, 2009;Morelli et al, 2010;Di Vito et al, 2011;Morelli et al, 2012b, c]. It was recently reported that polycaprolactone (PCL)-based membranes successfully supported outgrowth and differentiation of human neuronal cells [Morelli et al, 2012a].…”
In this study, the flavonoid didymin was administered in vitro in neuronal cells after hydrogen peroxide (H2O2)-induced injury (neurorescue) in order to investigate the effects of this natural molecule on cell damage in a neuronal membrane system. The results showed the effects of didymin in neuronal cells by using a polycaprolactone biodegradable membrane system as an in vitro model. Two major findings are presented in this study: first is the antioxidant property of didymin and, second, for the first time we provide evidence concerning its ability to rescue neuronal cells from oxidative damage. Didymin showed radical scavenging activities and it protected the neuronal cells against H2O2-induced neurotoxicity. Didymin increased cell viability, decreased intracellular reactive oxygen species generation, stimulated superoxide dismutase, catalase and glutathione peroxidase activity in neuronal cells which were previously insulted with H2O2. In addition, didymin strikingly inhibited H2O2-induced mitochondrial dysfunctions in terms of reduction of mitochondria membrane potential and the activation of cleaved caspase-3, and also decreased dramatically the H2O2-induced phosphorylation of c-Jun N-terminal kinase. Therefore, this molecule is capable of inducing recovery from oxidative damage, and promoting and/or restoring normal cellular conditions. Moreover, the mechanism underlying the protective effects of didymin in H2O2-injured neuronal cells might be related to the activation of antioxidant defense enzymes as well as to the inhibition of apoptotic features, such as p-JNK and caspase-3 activation. These data suggest that didymin may be a potential therapeutic molecule for the treatment of neurodegenerative disorders associated with oxidative stress.
“…This neuron-like morphology is similar to that described by others [Clagett-Dame et al, 2006;Cheung et al, 2009]. Moreover, the in vitro reconstruction of the neuronal network obtained in this work on PCL membranes clearly indicates the neuronal differentiation, which is in ac- Bartolo et al, 2008;Morelli et al, 2010Morelli et al, , 2012b. To confirm that the observed morphological features were actually resulting from the neuronal differentiation of SH-SY5Y cells, the expression of specific neuronal markers, βIII-tubulin and synaptophysin, were investigated by immunocytochemical analysis.…”
Section: Discussionsupporting
confidence: 88%
“…The present study investigates the protective effect of didymin against H 2 O 2 -induced damage to the neuronal cells in a biohybrid membrane system model. Previous studies have demonstrated that semipermeable polymeric membranes in flat and hollow fiber configurations, thanks to their highly selective structural, physicochemical and transport properties, allow the successful in vitro reconstruction of neuronal tissue, reproducing a tissue model for studying metabolic diseases and drug effects [Woerly et al, 1996;Schmidt and Leach, 2003;Zhang et al, 2005;De Bartolo et al, 2008;Giusi et al, 2009;He et al, 2009;Morelli et al, 2010;Di Vito et al, 2011;Morelli et al, 2012b, c]. It was recently reported that polycaprolactone (PCL)-based membranes successfully supported outgrowth and differentiation of human neuronal cells [Morelli et al, 2012a].…”
In this study, the flavonoid didymin was administered in vitro in neuronal cells after hydrogen peroxide (H2O2)-induced injury (neurorescue) in order to investigate the effects of this natural molecule on cell damage in a neuronal membrane system. The results showed the effects of didymin in neuronal cells by using a polycaprolactone biodegradable membrane system as an in vitro model. Two major findings are presented in this study: first is the antioxidant property of didymin and, second, for the first time we provide evidence concerning its ability to rescue neuronal cells from oxidative damage. Didymin showed radical scavenging activities and it protected the neuronal cells against H2O2-induced neurotoxicity. Didymin increased cell viability, decreased intracellular reactive oxygen species generation, stimulated superoxide dismutase, catalase and glutathione peroxidase activity in neuronal cells which were previously insulted with H2O2. In addition, didymin strikingly inhibited H2O2-induced mitochondrial dysfunctions in terms of reduction of mitochondria membrane potential and the activation of cleaved caspase-3, and also decreased dramatically the H2O2-induced phosphorylation of c-Jun N-terminal kinase. Therefore, this molecule is capable of inducing recovery from oxidative damage, and promoting and/or restoring normal cellular conditions. Moreover, the mechanism underlying the protective effects of didymin in H2O2-injured neuronal cells might be related to the activation of antioxidant defense enzymes as well as to the inhibition of apoptotic features, such as p-JNK and caspase-3 activation. These data suggest that didymin may be a potential therapeutic molecule for the treatment of neurodegenerative disorders associated with oxidative stress.
“…Micro-roughness is more controversial, because different cell types (e.g. epithelia versus fibroblasts) respond differently depending on the scale of surface roughness (or topography) [28–31]. Nano-roughness has been found to have significant effects on cell response, such as cell adhesion and proliferation [8, 18, 19].…”
This paper describes a high-throughput method for developing physically modified chitosan membranes to probe the cellular behavior of MDCK epithelial cells and HIG-82 fibroblasts adhered onto these modified membranes. To prepare chitosan membranes with micro/nanoscaled features, we have demonstrated an easy-to-handle, facile approach that could be easily integrated with IC-based manufacturing processes with mass production potential. These physically modified chitosan membranes were observed by scanning electron microscopy to gain a better understanding of chitosan membrane surface morphology. After MDCK cells and HIG-82 fibroblasts were cultured on these modified chitosan membranes for various culture durations (i.e. 1, 2, 4, 12 and 24 h), they were investigated to decipher cellular behavior. We found that both cells preferred to adhere onto a flat surface rather than on a nanopatterned surface. However, most (> 80%) of the MDCK cells showed rounded morphology and would suspend in the cultured medium instead of adhering onto the planar surface of negatively nanopatterned chitosan membranes. This means different cell types (e.g. fibroblasts versus epithelia) showed distinct capabilities/preferences of adherence for materials of varying surface roughness. We also showed that chitosan membranes could be re-used at least nine times without significant contamination and would provide us consistency for probing cell–material interactions by permitting reuse of the same substrate. We believe these results would provide us better insight into cellular behavior, specifically, microscopic properties and characteristics of cells grown under unique, nanopatterned cell-interface conditions.
“…In particular, at the leading edge of these processes are growth cones that recognize and translate a combination of chemical and physical cues into a specific trajectory towards a population of target cells. Therefore, in line with this concept and as reported previously (De Bartolo et al ., ; Morelli et al ., , ), the in vitro reconstruction of neuronal networks obtained in this study on the novel biodegradable membranes is a very important result, which clearly indicates cellular differentiation.…”
Semipermeable polymeric membranes with appropriate morphological, physicochemical and transport properties are relevant to inducing neural regeneration. We developed novel biodegradable membranes to support neuronal differentiation. In particular, we developed chitosan, polycaprolactone and polyurethane flat membranes and a biosynthetic blend between polycaprolactone and polyurethane by phase-inversion techniques. The biodegradable membranes were characterized in order to evaluate their morphological, physicochemical, mechanical and degradation properties. We investigated the efficacy of these different membranes to promote the adhesion and differentiation of neuronal cells. We employed as model cell system the human neuroblastoma cell line SHSY5Y, which is a well-established system for studying neuronal differentiation. The investigation of viability and specific neuronal marker expression allowed assessment that the correct neuronal differentiation and the formation of neuronal network had taken place in vitro in the cells seeded on different biodegradable membranes. Overall, this study provides evidence that neural cell responses depend on the nature of the biodegradable polymer used to form the membranes, as well as on the dissolution, hydrophilic and, above all, mechanical membrane properties. PCL-PU membranes exhibit mechanical properties that improve neurite outgrowth and the expression of specific neuronal markers.
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