Roxana Cristina Popescu

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

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.

Silver Nanocoatings for Reducing the Exogenous Microbial Colonization of Wound Dressings

The aim of this work was to obtain an antimicrobial coating (NanoAg) for polyester-nylon wound dressings (WDs) for reducing the risk of exogenous wound related infections. The as-prepared NanoAg-WDs were characterized by XRD (X-ray Diffraction), SEM (Scanning Electron Microscopy), TEM (Transmission Electron Microscopy), SAED (Selected Area Electron Diffraction) and IRM (InfraRed Microscopy). Biological characterization consisted of in vitro evaluation of the interaction with fibroblast cell cultures and in vivo biodistribution studies of AgNPs on mice models. Then, specimens of commercial WDs were immersed in a glucose and NaOH solution of silver nanoparticles, followed by the subsequent dropwise addition of AgNO3 solution. The antimicrobial efficiency of the NanoAg-WDs was assessed by in vitro qualitative and quantitative analyses on Staphylococcus aureus and Pseudomonas aeruginosa strains. The in vitro and in vivo studies demonstrated that the tested nanoparticles utilized to coat WDs have a good biocompatibility, allowing the normal development of cultured human cells and revealing a normal biodistribution within a mouse model, without toxic effects. The modified and viable cells count analyses proved that the modified WDs exhibit an improved inhibitory activity of microbial colonization, attachment and biofilm growth. The reported data recommend this type of coatings to obtain modified WDs with antibacterial properties, able to prevent the exogenous microbial contamination of the wound tissue, colonization and further biofilm development.

Metal based frameworks for drug delivery systems.

The attention for gold nanoparticles (AuNPs) has recently grown, especially regarding its applications in biomedicine, like cancer treatment and immunization by nano-vaccines. The interest for this type of nanoparticles is given by their ability to penetrate blood vessels and tissue barriers and to be targeted to a specific cell by means of specifically functionalized molecules. Moreover, AuNPs posses special properties which make them useful in concomitant diagnostic by medical imaging and tumor ablation by means of photothermal activation.

Fabrication, Characterization, and Evaluation of Bionanocomposites Based on Natural Polymers and Antibiotics for Wound Healing Applications

The aim of our research activity was to obtain a biocompatible nanostructured composite based on naturally derived biopolymers (chitin and sodium alginate) loaded with commercial antibiotics (either Cefuroxime or Cefepime) with dual functions, namely promoting wound healing and assuring the local delivery of the loaded antibiotic. Compositional, structural, and morphological evaluations were performed by using the thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and fourier transform infrared spectroscopy (FTIR) analytical techniques. In order to quantitatively and qualitatively evaluate the biocompatibility of the obtained composites, we performed the tetrazolium-salt (MTT) and agar diffusion in vitro assays on the L929 cell line. The evaluation of antimicrobial potential was evaluated by the viable cell count assay on strains belonging to two clinically relevant bacterial species (i.e., Escherichia coli and Staphylococcus aureus).

3D Biomimetic Magnetic Structures for Static Magnetic Field Stimulation of Osteogenesis

We designed, fabricated and optimized 3D biomimetic magnetic structures that stimulate the osteogenesis in static magnetic fields. The structures were fabricated by direct laser writing via two-photon polymerization of IP-L780 photopolymer and were based on ellipsoidal, hexagonal units organized in a multilayered architecture. The magnetic activity of the structures was assured by coating with a thin layer of collagen-chitosan-hydroxyapatite-magnetic nanoparticles composite. In vitro experiments using MG-63 osteoblast-like cells for 3D structures with gradients of pore size helped us to find an optimum pore size between 20–40 µm. Starting from optimized 3D structures, we evaluated both qualitatively and quantitatively the effects of static magnetic fields of up to 250 mT on cell proliferation and differentiation, by ALP (alkaline phosphatase) production, Alizarin Red and osteocalcin secretion measurements. We demonstrated that the synergic effect of 3D structure optimization and static magnetic stimulation enhances the bone regeneration by a factor greater than 2 as compared with the same structure in the absence of a magnetic field.

Fabrication and Cytotoxicity of Gemcitabine-Functionalized Magnetite Nanoparticles

Nanotechnology has been successfully used for the fabrication of targeted anti-cancer drug carriers. This study aimed to obtain Fe3O4 nanoparticles functionalized with Gemcitabine to improve the cytotoxic effects of the chemotherapeutic substance on cancer cells. The (un) functionalized magnetite nanoparticles were synthesized using a modified co-precipitation method. The nanoconjugate characterization was performed by XRD, SEM, SAED and HRTEM; the functionalizing of magnetite with anti-tumor substances has been highlighted through TGA. The interaction with biologic media has been studied by means of stability and agglomeration tendency (using DLS and Zeta Potential); also, the release kinetics of the drug in culture media was evaluated. Cytotoxicity of free-Gemcitabine and the obtained nanoconjugate were evaluated on human BT 474 breast ductal carcinoma, HepG2 hepatocellular carcinoma and MG 63 osteosarcoma cells by MTS. In parallel, cellular morphology of these cells were examined through fluorescence microscopy and SEM. The localization of the nanoparticles related to the cells was studied using SEM, EDX and TEM. Hemolysis assay showed no damage of erythrocytes. Additionally, an in vivo biodistribution study was made for tracking where Fe3O4@Gemcitabine traveled in the body of mice. Our results showed that the transport of the drug improves the cytotoxic effects in comparison with the one produced by free Gemcitabine for the BT474 and HepG2 cells. The in vivo biodistribution test proved nanoparticle accumulation in the vital organs, with the exception of spleen, where black-brown deposits have been found. These results indicate that our Gemcitabine-functionalized nanoparticles are a promising targeted system for applications in cancer therapy.

7 – Microorganisms: new trends in environment-friendly and energy-saving water purification

The current trend regarding the development and implementation of novel strategies for decontamination of polluted water became an important aspect of modern community. Given the current drawbacks of conventional technologies involved in the removal of water contaminants and the impressive potential of nanotechnology-based processes, promising unconventional approaches have been successfully developed. Moreover, the attention has been straightened toward ecofriendly novel technologies; either we are considering the beneficial role of microscopic organisms or the ecological aspects of such strategies. The aim of this chapter is to emphasize the impressive potential of both nonpathogenic and pathogenic microorganisms in water self-purification and water purification, respectively.

Chapter 2 – Toxicity of inorganic nanoparticles against prokaryotic cells

Nowadays, drug-resistant bacteria are a serious problem concerning the international health organizations, as high-dose treatments are necessary to overcome these infections, inducing higher toxicity, longer hospital stays, and increased mortality. Due to the progress that has been made in nanotechnology, some inorganic materials, that have already been known to possess antimicrobial properties, have been transformed into nanoparticles. Thus, silver, copper, or titanium dioxide nanoparticles were developed and administered as drinkable colloids, or transformed into nanostructured coatings in medical devices. Some of these are clinically approved and the mechanisms of toxicity are almost clearly elucidated, however, newly improved systems are continuously being developed, in order to overcome the shortcomings of the existing ones and to improve living standards. This chapter makes a summary of the existing studies regarding the mechanisms of toxicity for some inorganic nanoparticles against prokaryote cells. Also, some newly developed model systems are described, focusing on the biosynthesized nanoparticles, which are clearly a trend in this field.

Antibiofilm Coatings Based on PLGA and Nanostructured Cefepime-Functionalized Magnetite

The aim of our study was to obtain and evaluate the properties of polymeric coatings based on poly(lactic-co-glycolic) acid (PLGA) embedded with magnetite nanoparticles functionalized with commercial antimicrobial drugs. In this respect, we firstly synthesized the iron oxide particles functionalized (@) with the antibiotic Cefepime (Fe3O4@CEF). In terms of composition and microstructure, the as-obtained powdery sample was investigated by means of grazing incidence X-ray diffraction (GIXRD), thermogravimetric analysis (TGA), scanning and transmission electron microscopy (SEM and TEM, respectively). Crystalline and nanosized particles (~5 nm mean particle size) with spherical morphology, consisting in magnetite core and coated with a uniform and reduced amount of antibiotic shell, were thus obtained. In vivo biodistribution studies revealed the obtained nanoparticles have a very low affinity for innate immune-related vital organs. Composite uniform and thin coatings based on poly(lactide-co-glycolide) (PLGA) and antibiotic-functionalized magnetite nanoparticles (PLGA/Fe3O4@CEF) were subsequently obtained by using the matrix assisted pulsed laser evaporation (MAPLE) technique. Relevant compositional and structural features regarding the composite coatings were obtained by performing infrared microscopy (IRM) and SEM investigations. The efficiency of the biocompatible composite coatings against biofilm development was assessed for both Gram-negative and Gram-positive pathogens. The PLGA/Fe3O4@CEF materials proved significant and sustained anti-biofilm activity against staphylococcal and Escherichia coli colonisation.

Nanostructurated membranes for the microbiological purification of drinking water

Abstract The access for safe drinking water is a problem concerning both developed and developing countries, as pure water sources are getting limited. Worldwide, people are getting exposed to a variety of waterborne diseases or heavy metal poisoning. For this reason, new and improved methods for water purification are needed. This chapter is mainly focused on the removal of microorganisms from contaminated water by using different filtering membranes and discusses some examples of nanofiltering membranes from the scientific literature: membranes embedded with carbon nanotubes, respectively, membranes embedded with silver nanoparticles. Their properties and evaluation is discussed.