Nanoparticle uptake and their co-localization with cell compartments: a confocal Raman microscopy study at single cell level

Confocal Raman Microscopy, a non-invasive, non-destructive and label-free technique, was employed to study the uptake and localization of nanoparticles (NPs) in the Hepatocarcinoma human cell line HepG2 at the level of single cells. Cells were exposed to carbon nanotubes (CNTs) the surface of which was engineered with polyelectrolytes and lipid layers, aluminium oxide and cerium dioxide nanoparticles. Raman spectra deconvolution was applied to obtain the spatial distributions of NPs together with lipids/proteins in cells. The colocalization of the NPs with different intracellular environments, lipid bodies, protein and DNA, was inferred. Lipid coated CNTs associated preferentially with lipid rich regions, whereas polyelectrolyte coated CNTs were excluded from lipid rich regions. Al2O3 NPs were found in the cytoplasm. CeO2 NPs were readily taken up and have been observed all over the cell. Raman z-scans proved the intracellular distribution of the respective NPs

Tailored polyelectrolyte thin film multilayers to modulate cell adhesion

The layer-by-layer assembly of polyelectrolyte multilayers (PEMs) from natural or synthetic polyelectrolytes constitutes a very versatile and simple strategy to modify surfaces and modulate cell behavior. PEMs assembled from natural polyelectrolytes are very appealing for biological and medical applications due to their high biocompatibility. However, PEMs from natural polyelectrolytes display poor cell adhesion as they are soft materials with an elasticity modulus of a few kilopascal. In this report, the authors present results on the modulation of cell adhesion of different immortalized cell lines by PEMs. Two strategies are employed to vary cell adhesion: (1) a heterogeneous polyelectrolyte multilayer is assembled employing a rigid bottom block including a synthetic polyelectrolyte with a soft upper block of natural polyelectrolytes and (2) polyelectrolyte multilayers from natural polyelectrolytes are thermally annealed after assembly. The physicochemical characteristics of the PEMs change upon thermal treatment. Depending on the composition of the polyelectrolyte multilayer, cell adhesion may be enhanced or reduced. Based on the impact on PEM properties and cell adhesion caused by thermal annealing, a temperature gradient is applied to a PEM of poly-L-lysine/alginate to induce a spatial variation of PEM properties, resulting in a gradient in cell adhesion. The strategies shown here can be employed as simple alternatives to tailor PEM properties by means of fully biocompatible procedures.Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Concepts for Designing Tailored Thin Film Surfaces with Potential Biological Applications

The tailoring of surfaces with nanostructured thin films, where interfacial properties can be controlled at the nanoscale, offers multiple possibilities for biological applications. The design of thin films with appropriate properties to induce desired biological responses at cell level requires the convergence of research from physics, chemistry, material science, biology, and medicine. Here, we will discuss the main surface properties that determine the behavior of isolated cells, cell colonies, and tissues interacting with a material. Surface roughness, morphological features, stiffness, wettability, chemical nature, and protein-surface interaction characteristics, as well as spatiotemporal heterogeneities, are expected to contribute to the desired biological performance of a material. A brief review in relation to thin films for biological applications will be presented. We will focus on examples in which basic rather simple processes play a key role in determining the triggering of a particular biological cell phenotype

Kinetics of the thermal response of poly(N-isopropylacrylamide co methacrylic acid) hydrogel microparticles under different environmental stimuli: a time-lapse NMR study

Hypothesis: Hydrogels of N-isopropylacrylamide and methacrylic acid (P(NIPAm-co-MAA)) display pH sensitivity and complex positively charged molecules through carboxylate groups, while having a critical solution temperature at which they reduce in volume and dehydrate. We aimed to elucidate how the responsiveness of MAA to environmental changes alters PNIPAm hydrogels at the molecular level using nuclear magnetic resonance (NMR). Time-lapse NMR allows us to follow the evolution of NMR signal under a temperature stimulus, providing unique information on conformational freedom of the hydrogel polymers. Experiments: We used time-lapse NMR to follow the evolution of the NMR signal with time over a temperature change from 25 to 40 °C and to study the swelling/deswelling kinetics of P(NIPAm-co-MAA) microgels at different pH values and ionic strengths, and in the presence of positively charged molecules complexing carboxylate groups. Findings: At acid pH, hydrogel collapse is favored over neutral pH, and at basic pH the carboxylates remain steadily hydrated during temperature increase. Increasing ionic strength results in a faster, more effective collapse than decreasing pH. Complexation of medium-sized molecules with several charges (spermine, spermidine) causes a faster collapse than complexation with large molecular weight poly(allylamine) hydrochloride, but similar to the collapse effected by large poly(diallyldimethylammonium) chloride. This work opens new perspectives to using time-lapse NMR to study thermoresponsive systems that respond to multiple stimuli, with particular relevance in designing hydrogels for drug delivery.Fil: Martinez Moro, Marta. Basque Research and Technology Alliance; EspañaFil: Jenczyk, Jacek. Adam Mickiewicz University; PoloniaFil: Giussi, Juan Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas; ArgentinaFil: Jurga, Stefan. Adam Mickiewicz University; PoloniaFil: Moya, Sergio E.. Basque Research and Technology Alliance; Españ

Enhanced antiadhesive properties of chitosan/hyaluronic acid polyelectrolyte multilayers driven by thermal annealing : Low adherence for mammalian cells and selective decrease in adhesion for Gram-positive bacteria

The development of antifouling coatings with restricted cell and bacteria adherence is fundamental for many biomedical applications. A strategy for the fabrication of antifouling coatings based on the layer-by-layer assembly and thermal annealing is presented. Polyelectrolyte multilayers (PEMs) assembled from chitosan and hyaluronic acid were thermally annealed in an oven at 37 °C for 72 h. The effect of annealing on the PEM properties and topography was studied by atomic force microscopy, ζ-potential, circular dichroism and contact angle measurements. Cell adherence on PEMs before and after annealing was evaluated by measuring the cell spreading area and aspect ratio for the A549 epithelial, BHK kidney fibroblast, C2C12 myoblast and MC-3T3-E1 osteoblast cell lines. Chitosan/hyaluronic acid PEMs show a low cell adherence that decreases with the thermal annealing, as observed from the reduction in the average cell spreading area and more rounded cell morphology. The adhesion of S. aureus (Gram-positive) and E. coli (Gram-negative) bacteria strains was quantified by optical microscopy, counting the number of colony-forming units and measuring the light scattering of bacteria suspension after detachment from the PEM surface. A 20% decrease in bacteria adhesion was selectively observed in the S. aureus strain after annealing. The changes in mammalian cell and bacteria adhesion correlate with the changes in topography of the chitosan/hyaluronic PEMs from a rough fibrillar 3D structure to a smoother and planar surface after thermal annealing.Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Solvent Effects on the Structure−Property Relationship of Redox-Active Self-Assembled Nanoparticle−Polyelectrolyte−Surfactant Composite Thin Films: Implications for the Generation of Bioelectrocatalytic Signals in Enzyme-Containing Assemblies

The search for strategies to improve the performance of bioelectrochemical platforms based on supramolecular materials has received increasing attention within the materials science community, where the main objective is to develop low-cost and flexible routes using self-assembly as a key enabling process. Important contributions to the performance of such bioelectrochemical devices have been made based on the integration and supramolecular organization of redox-active polyelectrolyte−surfactant complexes on electrode supports. Here, we examine the influence of the processing solvent on the interplay between the supramolecular mesoorganization and the bioelectrochemical properties of redox-active self-assembled nanoparticle−polyelectrolyte−surfactant nanocomposite thin films. Our studies reveal that the solvent used in processing the supramolecular films and the presence of metal nanoparticles not only have a substantial influence in determining the mesoscale organization and morphological characteristics of the film but also have a strong influence on the efficiency and performance of the bioelectrochemical system. In particular, a higher bioelectrochemical response is observed when nanocomposite supramolecular films were cast from aqueous solutions. These observations seem to be associated with the fact that the use of aqueous solvents increases the hydrophilicity of the film, thus favoring the access of glucose, particularly at low concentrations. We believe that these results improve our current understanding of supramolecular nanocomposite materials generated via polyelectrolyte−surfactant complexes, in order to use the processing conditions as a variable to improve the performance of bioelectrochemical devices.Facultad de Ciencias ExactasInstituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Tailored polyelectrolyte thin film multilayers to modulate cell adhesion

The layer-by-layer assembly of polyelectrolyte multilayers (PEMs) from natural or synthetic polyelectrolytes constitutes a very versatile and simple strategy to modify surfaces and modulate cell behavior. PEMs assembled from natural polyelectrolytes are very appealing for biological and medical applications due to their high biocompatibility. However, PEMs from natural polyelectrolytes display poor cell adhesion as they are soft materials with an elasticity modulus of a few kilopascal. In this report, the authors present results on the modulation of cell adhesion of different immortalized cell lines by PEMs. Two strategies are employed to vary cell adhesion: (1) a heterogeneous polyelectrolyte multilayer is assembled employing a rigid bottom block including a synthetic polyelectrolyte with a soft upper block of natural polyelectrolytes and (2) polyelectrolyte multilayers from natural polyelectrolytes are thermally annealed after assembly. The physicochemical characteristics of the PEMs change upon thermal treatment. Depending on the composition of the polyelectrolyte multilayer, cell adhesion may be enhanced or reduced. Based on the impact on PEM properties and cell adhesion caused by thermal annealing, a temperature gradient is applied to a PEM of poly-L-lysine/alginate to induce a spatial variation of PEM properties, resulting in a gradient in cell adhesion. The strategies shown here can be employed as simple alternatives to tailor PEM properties by means of fully biocompatible procedures.Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Primer reporte de Ornithonyssus bursa (Berlese, 1888) (Acari: Mesostigmata: Macronyssidae) en República Dominicana

Se reporta por primera vez para República Dominicana el ácaro Ornithonyssus bursa (Berlese) (Acari: Mesostigmata: Macronyssidae) y un caso de dermatitis provocado por este ácaro a humanos. El caso estuvo relacionado a una infestación de ácaros originada en nidos de Passer domesticus L. Se sospecha que casos como este podrían estar ocurriendo con más frecuencia y ser confundidos con alergias

Fabrication of hybrid graphene oxide/polyelectrolyte capsules by means of layer-by-layer assembly on erythrocyte cell templates

A novel and facile method was developed to produce hybrid graphene oxide (GO)–polyelectrolyte (PE) capsules using erythrocyte cells as templates. The capsules are easily produced through the layer-by-layer technique using alternating polyelectrolyte layers and GO sheets. The amount of GO and therefore its coverage in the resulting capsules can be tuned by adjusting the concentration of the GO dispersion during the assembly. The capsules retain the approximate shape and size of the erythrocyte template after the latter is totally removed by oxidation with NaOCl in water. The PE/GO capsules maintain their integrity and can be placed or located on other surfaces such as in a device. When the capsules are dried in air, they collapse to form a film that is approximately twice the thickness of the capsule membrane. AFM images in the present study suggest a film thickness of approx. 30 nm for the capsules in the collapsed state implying a thickness of approx. 15 nm for the layers in the collapsed capsule membrane. The polyelectrolytes used in the present study were polyallylamine hydrochloride (PAH) and polystyrenesulfonate sodium salt (PSS). Capsules where characterized by transmission electron microscopy (TEM), atomic force microscopy (AFM), dynamic light scattering (DLS) and Raman microscopy, the constituent layers by zeta potential and GO by TEM, XRD, and Raman and FTIR spectroscopies

Strontium titanate (SrTiO3) mesoporous coatings for enhanced strontium delivery and osseointegration on bone implants

The incorporation of strontium (Sr) in titania enhances surface bioactivity and has a positive effect on pre-osteoblastic cell attachment, proliferation, and differentiation. Strontium titanate mesoporous films (SrTiMFs) with 30% pore volume and a 20% Sr molar content have been prepared by the evaporation induced self-assembly method. SrTiMFs display a large internal surface area available for exchange of Sr, which is released in cell media up to 44% within the first 8 h. SrTiMFs improve attachment of MC3T3-E1 pre-osteoblastic cells, which show larger filopodia and more elongated features than cells attached to plain mesoporous titania films (MTFs). SrTiMFs also display improved cell proliferation and differentiation rates indicating that overall Sr incorporation into mesoporous titania coatings can lead to enhanced osseointegration during the early stages of bone tissue formation.Fil: Escobar, Ane. Centro de Investigación Cooperativa en Biomateriales; EspañaFil: Muzzio, Nicolás Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas; Argentina. Centro de Investigación Cooperativa en Biomateriales; EspañaFil: Angelome, Paula Cecilia. Comisión Nacional de Energía Atómica. Gerencia del Área de Seguridad Nuclear y Ambiente. Gerencia de Química (CAC); Argentina. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bordoni, Andrea Veronica. Comisión Nacional de Energía Atómica. Gerencia del Área de Seguridad Nuclear y Ambiente. Gerencia de Química (CAC); Argentina. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Martínez, Angel. Centro de Investigación Cooperativa en Biomateriales; EspañaFil: Bindini, Elisa. Centro de Investigación Cooperativa en Biomateriales; EspañaFil: Coy, Emerson. Adam Mickiewicz University. Nanobiomedical Centre; PoloniaFil: Andreozzi, Patrizia. Centro de Investigación Cooperativa en Biomateriales; EspañaFil: Grzelczak, Marek. Donostia International Physic Center; . Basque Foundation for Science; EspañaFil: Moya, Sergio E.. Centro de Investigación Cooperativa en Biomateriales; Españ