Generation of hydroxyl radicals and singlet oxygen by particulate matter and its inorganic components.

Particulate matter (PM) can strongly affect redox biochemistry and therefore induce the response of the immune system and aggravate the course of autoimmune diseases. Nanoparticles containing transition metal compounds possessing semiconductor properties (TiO2, ZnO) may act as photocatalysts and accelerate the generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS). In this study, the NIST standard reference material, SRM 1648a, has been analyzed in terms of this consideration. Organic compounds present in SRM 1648a were removed by cold oxygen plasma treatment. Samples of SRM 1648a with removed organic content (<2% of organic carbon, <1% of nitrogen) were obtained within 2 h of this treatment. The treatment did not affect the morphology of the powder. The reference material and PM2.5 collected in Kraków are composed of smaller particles and nanoparticles forming aggregates. The efficiency of (photo)generation of hydroxyl radicals and singlet oxygen was compared for original and organics-free samples. The analyzed samples showed the highest activity towards ROS generation when exposed to UV-vis-NIR light, moderate under UV irradiation, and the lowest in dark. Data collected in the present study suggest that the organic fraction is mostly responsible for singlet oxygen generation, as almost twice higher efficiency of 1O2 generation was observed for the original NIST sample compared to the material without the organic fraction. However, particulate matter collected in Kraków was found to have a five times higher activity in singlet oxygen generation (compared for original NIST and Kraków dust samples).

Chitosan-based coatings in the prevention of intravascular catheter-associated infections.

Central venous access devices play an important role in patients with prolonged intravenous administration requirements. In the last years, the coating of these devices with bactericidal compounds has emerged as a potential tool to prevent bacterial colonization. Our study describes the modification of 3D-printed reservoirs and silicone-based catheters, mimicking central venous access devices, through different approaches including their coating with the well known biocompatible and bactericidal polymer chitosan, with the anionic polysaccharide alginate; also, plasma treated surfaces were included in the study to promote polymer adhesion. The evaluation of the antimicrobial action of those surface modifications compared to that exerted by a model antibiotic (ciprofloxacin) adsorbed on the surface of the devices was carried out. Surface characterization was developed by different methodologies and the bactericidal effects of the different coatings were assayed in an in vitro model of Staphylococcus aureus infection. Our results showed a significant reduction in the reservoir roughness (≤73%) after coating though no changes were observed for coated catheters which was also confirmed by scanning electron microscopy, pointing to the importance of the surface device topography for the successful attachment of the coating and for the subsequent development of bactericidal effects. Furthermore, the single presence of chitosan on the reservoirs was enough to fully inhibit bacterial growth exerting the same efficiency as that showed by the model antibiotic. Importantly, chitosan coating showed low cytotoxicity against human keratinocytes, human lung adenocarcinoma epithelial cells, and murine colon carcinoma cells displaying viability percentages in the range of the control samples (>95%). Chitosan-based coatings are proposed as an effective and promising solution in the prevention of microbial infections associated to medical devices.

Chemical composition of submicron and fine particulate matter collected in Krakow, Poland. Consequences for the APARIC project.

Submicron particulate matter containing particles with an aerodynamic diameter ≤1 µm (PM1) are not monitored continuously by Environmental Protection Agencies around the World and are seldom studied. Numerous studies have indicated that people exposed to ultrafine (≤100 nm), submicron and fine particulate matter containing particles with an aerodynamic diameter ≤2.5 µm (PM2.5), can suffer from respiratory track diseases, cardiovascular, immunological or heart diseases and others. Inorganic pollutants containing redox active transition metals and small gaseous molecules, are involved in the generation of reactive oxygen and reactive nitrogen species. Inhalation of this kind of particles can affect immune-toxicity. Environmental pollution may aggravate the course of autoimmune diseases, in particular influence the mechanisms of the autoimmune system. Important factors that influence the toxicity of particulate matter, are particle size distribution, composition and concentration. This report deals with the composition of PM1 and PM2.5 fractions collected in Krakow, Poland. In spring 2015, the mean concentrations of PM1 and PM2.5 were 19 ± 14 and 27 ± 19 µg/m3, respectively. The PM2.5 fraction contained approximately 70 ± 17% of submicron particulate matter. In spring 2016, the mean concentrations of PM1 and PM2.5 were 12 ± 5 and 22 ± 12 µg/m3, respectively. The PM2.5 fraction contained approximately 60 ± 15% of submicron particulate matter. The concentrations of the elements Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Br, Rb, Sr and Pb in both fractions were determined by X-ray fluorescence spectrometry. Most of the analyzed metals had higher concentrations in the fine fraction than in the submicron one. Concentrations of V and As were below the detection limit in both fractions, whereas concentrations of Mn and Ca were below the detection limits in the PM1 fraction. The results are discussed in terms of the consequences they may have on the APARIC project presently underway in Krakow.

Preparation and characterization of alginate/chitosan formulations for ciprofloxacin-controlled delivery.

In this work, alginate beads loaded with ciprofloxacin (AL_CP) and alginate beads loaded with ciprofloxacin and covered by chitosan (AL_CP_CS) were prepared by emulsification technique in combination with internal gelation method. Physicochemical characterization of the resulting formulations revealed the hydrodynamic diameter and Zeta potential ca. 160 nm and ca. -32 mV in the case of AL_CP and ca. 240 nm and ca. +14 mV in the case of AL_CP_CS (pH = 6.0), respectively. Kinetics of ciprofloxacin release from alginate/chitosan formulations was studied in different media (pH = 1.2, 6.8, and 7.4). Covering alginate core with a polycation such as chitosan moderates the drug release, resulting in a pH-sensitive hybrid controlled-release system. Herein, alginate beads with encapsulated ciprofloxacin covered with chitosan are proposed as an effective oral delivery system since the drug release from alginate core is limited in low pH solution (gastric conditions).

Bactericidal Effect of Gold-Chitosan Nanocomposites in Coculture Models of Pathogenic Bacteria and Human Macrophages.

The ability of pathogenic bacteria to develop resistance mechanisms to avoid the antimicrobial potential of antibiotics has become an increasing problem for the healthcare system. The search for more effective and selective antimicrobial materials, though not harmful to mammalian cells, seems imperative. Herein we propose the use of gold-chitosan nanocomposites as effective bactericidal materials avoiding damage to human cells. Nanocomposites were obtained by taking advantage of the reductive and stabilizing action of chitosan solutions on two different gold precursor concentrations. The resulting nanocomposites were added at different final concentrations to a coculture model formed by Gram-positive (Staphylococcus aureus) or Gram-negative (Escherichia coli) bacteria and human macrophages. Gold-chitosan colloids exhibited superior bactericidal ability against both bacterial models without showing cytotoxicity on human cells at the concentrations tested. Morphological and in vitro viability studies supported the feasibility of the infection model here described to test novel bactericidal nanomaterials. Flow cytometry and scanning electron microscopy analyses pointed to the disruption of the bacterial wall as the lethal mechanism. Data obtained in the present study suggest that gold-chitosan nanocomposites are powerful and promising nanomaterials for reducing bacteria-associated infections, respecting the integrity of mammalian cells, and displaying high selectivity against the studied bacteria.

Development of noncytotoxic silver-chitosan nanocomposites for efficient control of biofilm forming microbes.

Severe bacterial and fungal infections have become a major clinical and public health concern. Nowadays, additional efforts are needed to develop effective antimicrobial materials that are not harmful to human cells. This work describes the synthesis and characterization of chitosan-ascorbic acid-silver nanocomposites as films exhibiting high antimicrobial activity and non-cytotoxicity towards human cells. The reductive and stabilizing activity of both the biocompatible polymer chitosan and ascorbic acid were used in the synthesis of silver nanoparticles (AgNPs). Herein, we propose an improved composite synthesis based on medium average molecular weight chitosan with a high deacetylation degree, that together with ascorbic acid gave films with a uniform distribution of small AgNPs (<10 nm) exhibiting high antimicrobial activity against biofilm forming bacterial and fungal strains of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans. At the same time, the resulting solid nanocomposites showed, at the same doses, reduced or totally excluded cytotoxicity on mammalian somatic and tumoral cells. Data obtained in the present study suggest that adequately designed chitosan-silver nanocomposites are powerful and promising materials for reducing pathogenic microorganism-associated infections without harmful effects towards mammalian cells.

Study on inhibitory activity of chitosan-based materials against biofilm producing Pseudomonas aeruginosa strains.

Six antibiotic-resistant Pseudomonas aeruginosa strains, isolated from chronic diabetic foot infections, were chosen for studying the influence of different chitosan-based materials: chitosan solution and chitosan submicroparticles in both planktonic and 24 h-old biofilm-forming models. Chitosan solution occurred to be more effective in the reduction of bacterial populations than chitosan submicroparticles for both planktonic and biofilm-related Pseudomonas cells. It seems that the antimicrobial activity of the tested chitosan preparations depends on the individual bacterial strain susceptibility probably related to differences in the phenotypes and natural antioxidant abilities of Pseudomonas aeruginosa strains.

Development of noncytotoxic chitosan-gold nanocomposites as efficient antibacterial materials.

This work describes the synthesis and characterization of noncytotoxic nanocomposites either colloidal or as films exhibiting high antibacterial activity. The biocompatible and biodegradable polymer chitosan was used as reducing and stabilizing agent for the synthesis of gold nanoparticles embedded in it. Herein, for the first time, three different chitosan grades varying in the average molecular weight and deacetylation degree (DD) were used with an optimized gold precursor concentration. Several factors were analyzed in order to obtain antimicrobial but not cytotoxic nanocomposite materials. Films based on chitosan with medium molecular weight and the highest DD exhibited the highest antibacterial activity against biofilm forming strains of Staphylococcus aureus and Pseudomonas aeruginosa. The resulting nanocomposites did not show any cytotoxicity against mammalian somatic and tumoral cells. They produced a disruptive effect on the bacteria wall while their internalization was hindered on the eukaryotic cells. This selectivity and safety make them potentially applicable as antimicrobial coatings in the biomedical field.