RESUMEN
The current development of nanobiotechnologies requires a better understanding of cell-surface interactions on the nanometre scale. Recently, advances in nanoscale patterning and detection have allowed the fabrication of appropriate substrates and the study of cell-substrate interactions. In this review we discuss the methods currently available for nanoscale patterning and their merits, as well as techniques for controlling the surface chemistry of materials at the nanoscale without changing the nanotopography and the possibility of truly characterizing the surface chemistry at the nanoscale. We then discuss the current knowledge of how a cell can interact with a substrate at the nanoscale and the effect of size, morphology, organization and separation of nanofeatures on cell response. Moreover, cell-substrate interactions are mediated by the presence of proteins adsorbed from biological fluids on the substrate. Many questions remain on the effect of nanotopography on protein adsorption. We review papers related to this point. As all these parameters have an influence on cell response, it is important to develop specific studies to point out their relative influence, as well as the biological mechanisms underlying cell responses to nanotopography. This will be the basis for future research in this field. An important topic in tissue engineering is the effect of nanoscale topography on bacteria, since cells have to compete with bacteria in many environments. The limited current knowledge of this topic is also discussed in the light of using topography to encourage cell adhesion while limiting bacterial adhesion. We also discuss current and prospective applications of cell-surface interactions on the nanoscale. Finally, based on questions raised previously that remain to be solved in the field, we propose future directions of research in materials science to help elucidate the relative influence of the physical and chemical aspects of nanotopography on bacteria and cell response with the aim of contributing to the development of nanobiotechnologies.
Asunto(s)
Bacterias/metabolismo , Adhesión Celular/fisiología , Nanoestructuras/química , Nanotecnología/métodos , Animales , Materiales Biocompatibles , Línea Celular , Citoesqueleto/metabolismo , Humanos , Propiedades de SuperficieRESUMEN
Contact guidance is generally evaluated by measuring the orientation angle of cells. However, statistical analyses are rarely performed on these parameters. Here we propose a statistical analysis based on a new parameter sigma, the orientation parameter, defined as the dispersion of the distribution of orientation angles. This parameter can be used to obtain a truncated Gaussian distribution that models the distribution of the data between -90 degrees and +90 degrees. We established a threshold value of the orientation parameter below which the data can be considered to be aligned within a 95% confidence interval. Applying our orientation parameter to cells on grooves and using a modelling approach, we established the relationship sigma=alpha(meas)+(52 degrees -alpha(meas))/(1+C(GDE)R) where the parameter C(GDE) represents the sensitivity of cells to groove depth, and R the groove depth. The values of C(GDE) obtained allowed us to compare the contact guidance of human osteoprogenitor (HOP) cells across experiments involving different groove depths, times in culture and inoculation densities. We demonstrate that HOP cells are able to identify and respond to the presence of grooves 30, 100, 200 and 500 nm deep and that the deeper the grooves, the higher the cell orientation. The evolution of the sensitivity (C(GDE)) with culture time is roughly sigmoidal with an asymptote, which is a function of inoculation density. The sigma parameter defined here is a universal parameter that can be applied to all orientation measurements and does not require a mathematical background or knowledge of directional statistics.
Asunto(s)
Nanotecnología , Células Cultivadas , Colorantes Fluorescentes , HumanosRESUMEN
OBJECTIVE: Patients undergoing anti-tumour necrosis factor-alpha (TNF-alpha) treatment often develop autoantibodies. Apoptotic cell antigens are a potential initiating stimulus for autoantibodies. Our goal was to verify whether anti-cytokine therapy causes the release of nucleosomes, a major autoantigen generated during cell death. DESIGN: Laboratory research study with comparison group. SETTING: Clinical Immunology Unit and Lab, H San Raffaele University Hospital, Italy. SUBJECTS: Eleven healthy controls and 87 rheumatic patients were studied, including 51 with rheumatoid arthritis (RA) and 33 patients with systemic lupus erythematosus (SLE). INTERVENTIONS: Vein blood samples were taken via the antecubital vein. Blood was retrieved from 11 patients before and after injection of anti-TNF-alpha humanized antibodies. Nucleosomes were measured with an enzyme-linked immunosorbent assay. Cell death induced by anti-TNF-alpha antibodies and by the soluble cytokine was assessed in vitro. MAIN OUTCOME MEASURES: Nucleosome level by treatment. RESULTS: Enzyme-linked immunosorbent assay effectively detected nucleosomes either released by dying cells in vitro or circulating in the plasma. SLE but not RA patients had circulating nucleosomes at the steady state. Eight of 11 patients had significantly higher levels of plasma nucleosomes after infliximab. Minute amounts of TNF-alpha enabled infliximab to induce cell death in vitro. CONCLUSIONS: The accumulation of nucleosomes possibly fosters the development of autoantibodies in subjects with appropriate genetic backgrounds.