Cytotoxicity Evaluation of Ammonia-Modified Graphene Oxide Particles in Lung Cancer Cells and Embryonic Stem Cells

Potential toxicity of graphene oxide (GO) is a subject of increasing research interest in the recent years. Here, we have evaluated the cytotoxicity of ammonia-modified GO (GO-NH2) and pristine GO particles in human lung cancer cells, A549 and embryonic stem cells, Lep3 exposed to different particles concentrations (0.1, 1, 10, 20, and 50 μg/ml) for different times (24 and 48h). Compared with GO, GO-NH2 particles possessed smaller size, positive surface charge and higher thickness. An increased propensity to aggregation in cell cultures was also found for GO-NH2 particles. Cytotoxicity evaluation revealed that GO-NH2 particles are more toxic than pristine GO. Applied at concentrations of 10, 20 and 50 μg/ml for 24h they affect significantly cell morphology of viable embryonic stem cells whereas human lung cancer A549 cells seem to be relatively more resistant to short-time exposure. After 48h exposure however cell proliferation of A549 cells was strongly suppressed in a dose-dependent manner while the proliferation ability of embryonic stem cells was not affected. These results suggested that both GO particles exert different degree of cytotoxicity which is time, dose and cell dependent. In general, ammonia-modified GO particles are more toxic than the pristine GO which should be taken into account for future biomedical applications

Morphometric and Nanomechanical Features of Platelets from Women with Early Pregnancy Loss Provide New Evidence of the Impact of Inherited Thrombophilia

Pregnancy is associated with hypercoagulation states and increased thrombotic risk, especially in women with thrombophilia. We combine atomic force microscopy (AFM) and flow cytometry to examine the morphology and nanomechanics of platelets derived from women with early pregnancy loss (EPL) and control pregnant (CP) and non-pregnant (CNP) women. Both control groups exhibit similar morphometric parameters (height and surface roughness) and membrane stiffness of platelets. EPL patients’ platelets, on the other hand, are more activated than the control groups, with prominent cytoskeletal rearrangement. In particular, reduced membrane roughness (22.9 ± 6 nm vs. 39.1 ± 8 nm) (p < 0.05) and height (692 ± 128 nm vs. 1090 ± 131 nm) (p < 0.05), strong alteration in the membrane Young modulus, increased production of platelets’ microparticles, and higher expression of procoagulant surface markers, as well as increased occurrence of thrombophilia (FVL, FII20210A, PLA1/A2, MTHFR C677T or 4G/5G PAI-1) polymorphisms were found. We suggest that the carriage of thrombophilic mutations triggers structural and nanomechanical abnormalities in platelets, resulting in their increased activation. The activation state of platelets can be well characterized by AFM, and the morphometric and nanomechanical characteristics might serve as a new criterion for evaluation of the cause of miscarriage and offer the prospect of an innovative approach serving for diagnostic purposes

Hofmeister anions effect on the thickness and morphology of polyelectrolyte multilayers for biofunctionalization of cardiovascular stents

The effect of Hofmeister anions on the surface properties of polyelectrolyte multilayers built from hyaluronan and chitosan by layer-by-layer deposition is studied by ellipsometry and atomic force microscopy. The thickness, roughness and morphology of the resulting coatings were found to depend on the type of the anion. Relationship between the surface properties and the biological response of the polyelectrolyte multilayers is established by assessing the degree of protein (albumin) adsorption

Morphometric and Nanomechanical Screening of Peripheral Blood Cells with Atomic Force Microscopy for Label-Free Assessment of Alzheimer’s Disease, Parkinson’s Disease, and Amyotrophic Lateral Sclerosis

Neurodegenerative disorders (NDDs) are complex, multifactorial disorders with significant social and economic impact in today’s society. NDDs are predicted to become the second-most common cause of death in the next few decades due to an increase in life expectancy but also to a lack of early diagnosis and mainly symptomatic treatment. Despite recent advances in diagnostic and therapeutic methods, there are yet no reliable biomarkers identifying the complex pathways contributing to these pathologies. The development of new approaches for early diagnosis and new therapies, together with the identification of non-invasive and more cost-effective diagnostic biomarkers, is one of the main trends in NDD biomedical research. Here we summarize data on peripheral biomarkers, biofluids (cerebrospinal fluid and blood plasma), and peripheral blood cells (platelets (PLTs) and red blood cells (RBCs)), reported so far for the three most common NDDs—Alzheimer’s disease (AD), Parkinson’s disease (PD), and amyotrophic lateral sclerosis (ALS). PLTs and RBCs, beyond their primary physiological functions, are increasingly recognized as valuable sources of biomarkers for NDDs. Special attention is given to the morphological and nanomechanical signatures of PLTs and RBCs as biophysical markers for the three pathologies. Modifications of the surface nanostructure and morphometric and nanomechanical signatures of PLTs and RBCs from patients with AD, PD, and ALS have been revealed by atomic force microscopy (AFM). AFM is currently experiencing rapid and widespread adoption in biomedicine and clinical medicine, in particular for early diagnostics of various medical conditions. AFM is a unique instrument without an analog, allowing the generation of three-dimensional cell images with extremely high spatial resolution at near-atomic scale, which are complemented by insights into the mechanical properties of cells and subcellular structures. Data demonstrate that AFM can distinguish between the three pathologies and the normal, healthy state. The specific PLT and RBC signatures can serve as biomarkers in combination with the currently used diagnostic tools. We highlight the strong correlation of the morphological and nanomechanical signatures between RBCs and PLTs in PD, ALS, and AD

Composite polyelectrolyte multilayers for biofunctionalization of medical devices

Polyelectrolyte multilayer coatings (PEM) are prepared by alternative layer-by-layer deposition of cationic and anionic polyelectrolyte monolayers on charged surfaces. The thickness of the coatings ranges from nm to few μm. Their properties such as roughness, stiffness, surface charge and surface energy can be precisely tuned to fulfil different technical or biological requirements. The coating process is based on self-assembly of polyelectrolytes. Advantages of these coatings are their easy handling, no harsh chemistry and the possibility for coatings on complex geometries. The PEM coatings can be prepared from a variety of suitable polyelectrolytes. Their stability varies from very durable PEM coatings that are only soluble in strong solvents to quickly degradable, which may be applied as drug release system. One example of such a degradable PEM system is the one based on the polyelectrolyte pair Hyaluronan (HA) and Chitosan (CHI). These biopolymers originate from natural sources and show low toxicity towards human cells. However, HA/CHI multilayers show only weak adhesiveness for human umbilical vein endothelial cells (HUVEC). In this article, we summarize our approaches to enhance the HA/CHI multilayer by incorporation of a non-polymer substance –graphene oxide– to improve the cell adhesion and keep such properties as low cytotoxicity and biodegradability. Different approaches for incorporation of graphene oxide were performed and the cellular adhesion was tested by metabolic assay

Regulation of the growth, morphology, mechanical properties and biocompatibility of natural polysaccharide-based multilayers by Hofmeister anions

Herein the optimization of the physicochemical properties and surface biocompatibility of polyelectrolyte multilayers of the natural, biocompatible and biodegradable, linear polysaccharides hyaluronan and chitosan by Hofmeister anions was systematically investigated. We demonstrated that there is an interconnection between the bulk and surface properties of HA/Chi multilayers both varying in accordance with the arrangement of the anions in the Hofmeister series. Kosmotropic anions increased the hydration, thickness, micro- and macro-roughness, and hydrophilicity and improved the biocompatibility of the films by reduction (2 orders of magnitude) of the films stiffness and complete anti-thrombogenicity

Unprecedented formation of sterically stabilized phospholipid liposomes of cuboidal morphology

Sterically stabilized phospholipid liposomes of unprecedented cuboid morphology are formed upon introduction in the bilayer membrane of original polymers, based on polyglycidol bearing a lipid-mimetic residue. Strong hydrogen bonding in the polyglycidol sublayers creates attractive forces, which, facilitated by fluidization of the membrane, bring about the flattening of the bilayers and the formation of cuboid vesicles

Evaluation of antibacterial properties of polyelectrolyte multilayer coatings by norm tests

Medical implants play a central role in modern medicine and both, naturally derived and synthetic materials have been explored as biomaterials for such devices. However, when implanted into living tissue, most materials initiate a host response. In addition, implants often cause bacterial infections leading to complications. Polyelectrolyte multilayer (PEM) coatings can be used for functionalization of medical implants improving the implant integration and reducing foreign body reactions. Some PEMs are also known to show antibacterial properties. We developed a PEM coating suggesting that it can decrease the risk of bacterial infections occurring after implantation while being highly biocompatible. We applied two different standard tests for evaluating the PEM’s antibacterial properties, the ISO norm (ISO 22196) and one ASTM norm (ASTM E2180) test. We found a reduction of bacterial growth on the PEM but to a different degree depending on the testing method. This result demonstrates the need for defining proper method to evaluate antibacterial properties of surface coatings

Caractérisation basse fréquence de ferrofluides et de couches minces magnétiques entourées par une nappe de courant, selon un design original

Au cours de cette thèse j'ai d'abord étudié les propriétés magnéto-diélectriques de ferrofluides (essentiellement les variations de leur permittivité en présence d'un champ magnétique continu) puis la perméabilité de ces liquides magnétiques en utilisant la méthode du tore fendu. Parmi les objectifs du DIOM figure la réalisation de composants passifs hyperfréquences en couches minces. Celles-ci peuvent être obtenues au Laboratoire par pulvérisation cathodique radio-fréquence. Il est nécessaire de connaître leur perméabilité. Les méthodes décrites jusqu'ici dans la littérature sont bien adaptées à la mesure de couches à perméabilité élevée. La boucle de mesure, entourant la couche magnétique avec son substrat, est sensible à la contribution des deux milieux. Je propose une technique de mesure inductive ne prenant en compte que les caractéristiques de la couche déposée où la boucle de mesure n'entoure que la couche. Cette technique a été validée à plus grande échelle et des mesures préliminaires ont été effectuées sur des couches minces de BaM et de YIG déposées sur un substrat d'alumine. Ces mesures m'ont permis de détecter les différentes difficultés et sources d'erreurs et de les résoudre. Les dimensions du prototype sont relativement grandes (2cm X 5cm) ce qui, étant données les dimensions de la cible dont le diamètre est de 75mm, entraîne une certaine variation du profil de l'épaisseur des couches ; mais des essais n'utilisant que quelques rubans proches d'un bord de l'échantillon ont montré que cela n'entraînant pas d'erreur prohibitive. Ainsi une perméabilité relative voisine de 25 (YIG) est mesurable avec une précision de l'ordre de plus ou moins 10%During this thesis I initially studied the magneto-dielectric properties of ferrofluids (primary variations of their permittivity in the presence of a d-c magnetic field) then the permeability of these magnetic liquids by using the method of the split torus. Among the objectives of DIOM laboratory is the realization of ultra high frequencies passive components with thin layers. Those can be obtained at the Laboratory by radio frequency cathodic sputtering. It is necessary to know their permeability. The methods described up to now in the literature are well adapted to measurement of layers with high permeability. The measurement loop, surrounding the magnetic layer with its substrate, is sensitive to the contribution of both media. I propose an inductive measurement technique taking into account only the characteristics of the layer desposited where the loop surrounds exclusively the layer. This technique was validated on a large scale and preliminary measurements were carried out on thin layers of BaM and YIG deposited on alumina substrate. These measurements enabled to detect the various difficulties and sources of error and to solve them. The prototypes dimensions are relatively large (2cm X 5cm) involving, being given the dimensions of the target (75 mm diameter), a certain variation of the layers' thickness profile. However some tests involving close the edge strips showed that it didn't lead to prohibitary errors. Finally, I could measure a relative permeability of 25 for thin film of YIG, with a precision of about 10%ST ETIENNE-BU Sciences (422182103) / SudocSudocFranceF

Evaluation of antibacterial properties of polyelectrolyte multilayer coatings by norm tests

Medical implants play a central role in modern medicine and both, naturally derived and synthetic materials have been explored as biomaterials for such devices. However, when implanted into living tissue, most materials initiate a host response. In addition, implants often cause bacterial infections leading to complications. Polyelectrolyte multilayer (PEM) coatings can be used for functionalization of medical implants improving the implant integration and reducing foreign body reactions. Some PEMs are also known to show antibacterial properties. We developed a PEM coating suggesting that it can decrease the risk of bacterial infections occurring after implantation while being highly biocompatible. We applied two different standard tests for evaluating the PEM’s antibacterial properties, the ISO norm (ISO 22196) and one ASTM norm (ASTM E2180) test. We found a reduction of bacterial growth on the PEM but to a different degree depending on the testing method. This result demonstrates the need for defining proper method to evaluate antibacterial properties of surface coatings