Radu Hristu

Inhibitory Activity of

Undesired biofilm development is a major concern in many areas, especially in the medical field. The purpose of the present study was to comparatively investigate the antibiofilm efficacy of usnic acid, in soluble versus nanofluid formulation, in order to highlight the potential use of Fe3O4/oleic acid (FeOA) nanofluid as potential controlled release vehicle of this antibiofilm agent. The (+) -UA loaded into nanofluid exhibited an improved antibiofilm effect on S. aureus biofilm formation, revealed by the drastic decrease of the viable cell counts as well as by confocal laser scanning microscopy images. Our results demonstrate that FeOA nanoparticles could be used as successful coating agents for obtaining antibiofilm pellicles on different medical devices, opening a new perspective for obtaining new antimicrobial and antibiofilm surfaces, based on hybrid functionalized nanostructured biomaterials.

{\rm Fe}_{3} {\rm O}_{4}

The aim of the present study was to demonstrate that Fe3O4/oleic acid core/shell nanostructures could be used as systems for stabilizing the Eugenia carryophyllata essential oil (EO) on catheter surface pellicles, in order to improve their resistance to fungal colonization. EO microwave assisted extraction was performed in a Neo-Clevenger (related) device and its chemical composition was settled by GC-MS analysis. Fe3O4/oleic acid-core/shell nanoparticles (NP) were obtained by a precipitation method under microwave condition. High resolution transmission electron microscopy (HR-TEM) was used as a primary characterization method. The NPs were processed to achieve a core/shell/EO coated-shell nanosystem further used for coating the inner surface of central venous catheter samples. The tested fungal strains have been recently isolated from different clinical specimens. The biofilm architecture was assessed by confocal laser scanning microscopy (CLSM). Our results claim the usage of hybrid nanomaterial (core/shell/coated-shell) for the stabilization of E. carryophyllata EO, which prevented or inhibited the fungal biofilm development on the functionalized catheter, highlighting the opportunity of using these nanosystems to obtain improved, anti-biofilm coatings for biomedical applications.

/Oleic Acid/Usnic Acid—Core/Shell/Extra-Shell Nanofluid on S. aureus Biofilm Development

Biofilms formed by bacterial cells are associated with drastically enhanced resistance against most antimicrobial agents, contributing to the persistence and chronicization of the microbial infections and to therapy failure. The purpose of this study was to combine the unique properties of magnetic nanoparticles with the antimicrobial activity of three essential oils to obtain novel nanobiosystems that could be used as coatings for catheter pieces with an improved resistance to Staphylococcus aureus and Klebsiella pneumoniae clinical strains adherence and biofilm development. The essential oils of ylang ylang, patchouli and vanilla were stabilized by the interaction with iron oxide@C14 nanoparticles to be further used as coating agents for medical surfaces. Iron oxide@C14 was prepared by co-precipitation of Fe+2 and Fe+3 and myristic acid (C14) in basic medium. Vanilla essential oil loaded nanoparticles pelliculised on the catheter samples surface strongly inhibited both the initial adherence of S. aureus cells (quantified at 24 h) and the development of the mature biofilm quantified at 48 h. Patchouli and ylang-ylang essential oils inhibited mostly the initial adherence phase of S. aureus biofilm development. In the case of K. pneumoniae, all tested nanosystems exhibited similar efficiency, being active mostly against the adherence K. pneumoniae cells to the tested catheter specimens. The new nanobiosystems based on vanilla, patchouli and ylang-ylang essential oils could be of a great interest for the biomedical field, opening new directions for the design of film-coated surfaces with anti-adherence and anti-biofilm properties.

Hybrid Nanomaterial for Stabilizing the Antibiofilm Activity of Eugenia carryophyllata Essential Oil

A new method for high-resolution quantitative measurement of the dielectric function by using scattering scanning near-field optical microscopy (s-SNOM) is presented. The method is based on a calibration procedure that uses the s-SNOM oscillating dipole model of the probe-sample interaction and quantitative s-SNOM measurements. The nanoscale capabilities of the method have the potential to enable novel applications in various fields such as nano-electronics, nano-photonics, biology or medicine.

Efficiency of Vanilla, Patchouli and Ylang Ylang Essential Oils Stabilized by Iron Oxide@C14 Nanostructures against Bacterial Adherence and Biofilms Formed by Staphylococcus aureus and Klebsiella pneumoniae Clinical Strains

A reliable procedure for measuring parameters connected to surface roughness is needed to compare the gas sensing properties of various thin films or the effect of different fabrication procedures on the surface roughness and the sensing properties. In this article, we propose to investigate how the acquisition parameters specific to atomic force microscopy investigations such as pixel size, scan area and scan speed influence the roughness parameters, namely root mean square and surface area ratio, commonly used for characterizing the gas sensing properties of porphyrins and other materials. Microsc. Res. Tech. 2012.

High-resolution quantitative determination of dielectric function by using scattering scanning near-field optical microscopy

The robustness and distinctiveness of local features to various object or scene deformations and to modifications of the acquisition parameters play key roles in the design of many computer vision applications. In this paper we present the results of our experiments on the behavior of a recently developed technique for local feature detection and description, Speeded-Up Robust Features (SURF), regarding image modifications specific to Confocal Scanning Laser Microscopy (CSLM). We analyze the repeatability of detected SURF keypoints and the precision-recall of their matching under modifications of three important CSLM parameters: pinhole aperture, photomultiplier (PMT) gain and laser beam power. During any investigation by CSLM these three parameters have to be modified, individually or together, in order to optimize the contrast and the Signal Noise Ratio (SNR), being also inherently modified when changing the microscope objective. Our experiments show that an important amount of SURF features can be detected at the same physical locations in images collected at different values of the pinhole aperture, PMT gain and laser beam power, and further on can be successfully matched based on their descriptors. In the final part, we exemplify the potential of SURF in CSLM imaging by presenting a SURF-based computer vision application that deals with the mosaicing of images collected by this technique.

Influence of atomic force microscopy acquisition parameters on thin film roughness analysis

The dependence of the near-field signal on the dielectric function of a specific material proposes scattering-type near-field optical microscopy (s-SNOM) as a viable tool for material characterization studies. Our experiment shows that specific material identification by s-SNOM is not a straightforward task as parameters involved in the detection scheme can also influence material contrast measurements. More precisely, we demonstrate that s-SNOM contrast in a pseudo-heterodyne detection configuration depends on the oscillation amplitude of the reference mirror and that for reliable measurements of the contrast between different materials this aspect needs to be taken into consideration.

Influence of Confocal Scanning Laser Microscopy specific acquisition parameters on the detection and matching of Speeded-Up Robust Features

The surface properties of hydroxyapatite, including electric charge, can influence the biological response, tissue compatibility, and adhesion of biological cells and biomolecules. Results reported here help in understanding this influence by creating charged domains on hydroxyapatite thin films deposited on silicon using electron beam irradiation and investigating their shape, properties, and carbon contamination for different doses of incident injected charge by two methods. Photoluminescence laser scanning microscopy was used to image electrostatic charge trapped at pre-existing and irradiation-induced defects within these domains, while phase imaging in atomic force microscopy was used to image the carbon contamination. Scanning Auger electron spectroscopy and Kelvin probe force microscopy were used as a reference for the atomic force microscopy phase contrast and photoluminescence laser scanning microscopy measurements. Our experiment shows that by combining the two imaging techniques the effects of trapped charge and carbon contamination can be separated. Such separation yields new possibilities for advancing the current understanding of how surface charge influences mediation of cellular and protein interactions in biomaterials.

A study on the image contrast of pseudo-heterodyned scattering scanning near-field optical microscopy

Computer vision tasks such as recognition and classification of objects and structures or image registration and retrieval can provide significant information when applied to microscopy images. Recently developed techniques for the detection and description of local features make the extraction and description of local image features that are invariant to various changes possible. The invariance and robustness of feature detection and description techniques play a key role in the design and implementation of object recognition, image registration, or image mosaicing applications. The scale-invariant feature transform (SIFT) technique is a widely used method for the detection, description, and matching of image features. In this article we present the results of our experiments regarding the repeatability of SIFT features, and to the precision of the SIFT feature matching, under image modifications specific to confocal scanning laser microscopy (CSLM). We have analyzed the behavior of SIFT while changing the pinhole aperture, photomultiplier gain, laser beam power, and electronic zoom. Our experiments, conducted on CSLM images, show that the SIFT technique is able to match detected key points between images acquired at different values of the acquisition parameters with good precision and represents a consistent tool for computer vision applications in CSLM.

Surface Charge and Carbon Contamination on an Electron-Beam-Irradiated Hydroxyapatite Thin Film Investigated by Photoluminescence and Phase Imaging in Atomic Force Microscopy

Imaging tissue samples by polarization-resolved second harmonic generation microscopy provides both qualitative and quantitative insights into collagen organization in a label-free manner. Polarization-resolved second harmonic generation microscopy goes beyond simple intensity-based imaging by adding the laser beam polarization component and applying different quantitative metrics such as the anisotropy factor. It thus provides valuable information on collagen arrangement not available with intensity measurements alone. Current established approaches are limited to calculating the anisotropy factor for only a particular laser beam polarization and no general guidelines on how to select the best laser beam polarization have yet been defined. Here, we introduce a novel methodology for selecting the optimal laser beam polarization for characterizing tissues using the anisotropy in the purpose of identifying cancer signatures. We show that the anisotropy factor exhibits a similar laser beam polarization dependence to the second harmonic intensity and we combine it with the collagen orientation index computed by Fast Fourier Transform analysis of the recorded images to establish a framework for choosing the laser beam polarization that is optimal for an accurate interpretation of polarization-resolved second harmonic generation microscopy images and anisotropy maps, and hence a better differentiation between healthy and dysplastic areas. SHG image of skin tissue (a) and a selected area of interest for which we compute the SHG intensity (b) and anisotropy factor (c) dependence on the laser beam polarization and also the FFT spectrum (d) to evaluate the collagen orientation index.