Bio-Based Ceramic Membranes for Bacteria Removal from Water.

Bio-based ceramic membranes were elaborated from kaolinite clays, coconut husks and eggshells to retain E. coli bacteria present in water intended for human consumption. Their characterization and removal performances are investigated in this work. These bio-ceramic membranes were obtained by heating the formulation containing 75% clay, 15% coconut husk and 10% eggshell at 900 °C or 1000 °C, at different temperature rates, to give S1, S2 and S3 materials. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), mercury porosimetry and scanning electron microscopy (SEM) were used to characterize these membranes. Water flux density, bacterial removal and biofouling were also assessed. Water flux density was shown to depend on material porosity. Bacteria retention was 90% (with 1 log-removal) for S1, 80% (with 0.7 log-removal) for S2 and 100% (with 3.3 log-removal) for S3. Membranes S1 and S2 presented reversible biofouling, while no fouling was evidenced for S3 in the tested conditions. This work shows that the best bio-ceramic membrane in terms of bacterial removal and flux density was S3. Its water flux density was 2123 ± 72 L/h/m2 at an initial pressure of 0.2 bar. This material is particularly interesting because its production protocol is quite simple, fast and without the addition of chemical additives. Moreover, it can be used to efficiently remove bacteria from drinking water.

Mechanistic Study of the Kinetic Phenomena Influencing the Bacteriostatic Action of Silver Ions in Agar Bioassays.

Bacteriostatic action of a biocidal agent results from the cumulative impact of different kinetics, including those of bacterial growth, mass transfer of the agent and its antibacterial action against the targeted bacteria. Current studies on bacteriostatic effects always directly consider the combination of these kinetics at given times, without discrimination between each other. This work introduces a novel approach, consisting of first studying independently, by the experiment and the model, the different kinetics involved, and then in coupling these kinetics to obtain a model that will be confronted with experimental data. An agar diffusion test with silver ions against Escherichia coli bacteria was implemented herein to assess the relevance of this approach. This work achieved to characterize the different kinetics and to propose a dynamic model combining them, which fits the experimental data with a silver diffusivity in the biofilm fixed to 7.0 ± 0.1 × 10-12 m2 s-1. This study also proves that the diffusive phenomenon was limiting the bacteriostatic action of silver ions over the test duration.

Fabrication of 3D printed antimicrobial polycaprolactone scaffolds for tissue engineering applications.

Synthetic polymers are widely employed for bone tissue engineering due to their tunable physical properties and biocompatibility. Inherently, most of these polymers display poor antimicrobial properties. Infection at the site of implantation is a major cause for failure or delay in bone healing process and the development of antimicrobial polymers is highly desired. In this study, silver nanoparticles (AgNps) were synthesized in polycaprolactone (PCL) solution by in-situ reduction and further extruded into PCL/AgNps filaments. Customized 3D structures were fabricated using the PCL/AgNps filaments through 3D printing technique. As demonstrated by scanning electron microscopy, the 3D printed scaffolds exhibited interconnected porous structures. Furthermore, X-ray photoelectron spectroscopy analysis revealed the reduction of silver ions. Transmission electron microscopy along with energy-dispersive X-ray spectroscopy analysis confirmed the formation of silver nanoparticles throughout the PCL matrix. In vitro enzymatic degradation studies showed that the PCL/AgNps scaffolds displayed 80% degradation in 20 days. The scaffolds were cytocompatible, as assessed using hFOB cells and their antibacterial activity was demonstrated on Escherichia coli. Due to their interconnected porous structure, mechanical and antibacterial properties, these cytocompatible multifunctional 3D printed PCL/AgNps scaffolds appear highly suitable for bone tissue engineering.

Anti-bacterial dynamic hydrogels prepared from O-carboxymethyl chitosan by dual imine bond crosslinking for biomedical applications.

Imine dynamic hydrogels are synthesized via dual-imine bond crosslinking from O-carboxymethyl chitosan (CMCS) and a water soluble dynamer using a 'green' approach. Three dynamers are prepared through reaction of benzene-1,3,5-tricarbaldehyde and di-amino Jeffamine with molar mass of 500, 800 and 1900, respectively. Hydrogels, namely H500, H800 and H1900 are then obtained by mixing CMCS and dynamer aqueous solutions. FT-IR confirms the formation of hydrogels via imine bonding. H1900 presents larger pore size and higher storage modulus as compared to H500 and H800 due to the higher molar mass of Jeffamine linker. The hydrogels exhibit pH sensitive swelling behavior due to electrostatic attraction or repulsion in the pH range from 3 to 10. The highest swelling ratio is obtained at pH 8, reaching 7500% for H800. Self-healing of hydrogels is evidenced by rheological measurements with alternatively applied low and high strains, and by using a macroscopic approach with re-integration of injected filaments. Furthermore, the H1900 membrane exhibits outstanding antibacterial activity against an E. coli suspension at 108 CFU mL-1. Therefore, dynamic hydrogels synthesized from CMCS and Jeffamine present outstanding rheological, swelling, self-healing and antibacterial properties, and are most promising as healthcare material in wound dressing, drug delivery and tissue engineering.

Composites Based on Nanoparticle and Pan Electrospun Nanofiber Membranes for Air Filtration and Bacterial Removal.

Often, solid matter is separated from particle-laden flow streams using electrospun filters due to their high specific surface area, good ability to capture aerial particulate matter, and low material costs. Moreover, electrospinning allows incorporating nanoparticles to improve the filter's air filtration efficiency and bacterial removal. Therefore, a new, improved polyacrylonitrile (PAN) nanofibers membrane that could be used to remove air pollutants and also with antibacterial activity was developed. We engineered three different filters that are characterized by the different particles embedded in the PAN nanofibers: titanium dioxide (TiO2), zinc oxide (ZnO), and silver (Ag). Then, their filtration performance was assessed by quantifying the filtration of sodium chloride (NaCl) aerosol particles of 9 to 300 nm in diameter using a scanning mobility particle sizer. The TiO2_F filter displayed the smallest fiber diameter and the highest filtration efficiency (≈100%). Conversely, the Ag_F filter showed the highest quality factor (≈0.06 Pa-1) because of the lower air pressure drop. The resulting Ag_F nanofibers displayed a very good antibacterial activity using an Escherichia coli suspension (108 CFU/mL). Moreover, the quality factor of these membranes was higher than that of the commercially available nanofiber membrane for air filtration.

Impact of polyelectrolytes on lysozyme properties in colloidal dispersions.

In this work, we investigated the impact of different polyelectrolytes (polyacrylic acid, polystyrene sulfonate, dextran sulfate, and chondroitin sulfate) on lysozyme properties when they form colloidal complexes. To this aim, we characterized (i) the size and stability of the different polyelectrolyte-lysozyme complexes upon addition of NaCl (different concentrations) by diffusion light scattering, and (ii) the structure and accessibility of lysozyme active site in such complexes by fluorescence quenching and time resolved fluorescence analysis. We then used these results to explain the antibacterial activity variations among colloidal complexes and compared with free lysozyme. Our findings show that colloidal complexes that are more prone to swelling (i.e., lysozyme complexed with polystyrene sulfonate) are less stable upon salt addition. In these colloids, the enzymatic site is also more accessible. However, the antibacterial activity does not depend on the swelling properties because no large structural modification of the active site occurs.

Efficient nanoparticles removal and bactericidal action of electrospun nanofibers membranes for air filtration.

Human exposure to air pollution and especially to nanoparticles is increasing due to the combustion of carbon-based energy vectors. Fibrous filters are among the various types of equipment potentially able to remove particles from the air. Nanofibers are highly effective in this area; however, their utilization is still a challenge due to the lack of studies taking into account both nanoparticle collection efficiency and antibacterial effect. The aim of this work is to produce and evaluate novel silver/polyacrylonitrile (Ag/PAN) electrospun fibers deposited on a nonwoven substrate to be used as air filters to remove nanoparticles from the air and also showing antibacterial activity. In order to determine the optimum manufacturing conditions, the effects of several electrospinning process parameters were analyzed such as solution concentration, collector to needle distance, flow rate, voltage, and duration. Ag/PAN nanofibers were characterized by X-ray diffraction (XRD), Transmission Electron Microscopy (TEM), Fourier Transform Infra-Red spectroscopy (FTIR), Energy-dispersive X-ray spectroscopy (EDX), X-ray photoelectron spectroscopy (XPS), and Scanning Electron Microscopy (SEM). In addition, filtration performances were determined by measuring the pressure drop and collection efficiency of sodium chloride (NaCl) aerosol particles (9 to 300 nm diameters) using Scanning Mobility Particle Sizers (SMPS). Filters with high filtration efficiency (≈100%) and high-quality factor (≈0.05 Pa-1) were obtained even adding different concentrations of Ag nanoparticles (AgNPs) to PAN nanofibers. The resultant Ag/PAN nanofibers showed excellent antibacterial activity against 104 CFU/mL E. coli bacteria.

Dual role of layered double hydroxide nanocomposites on antibacterial activity and degradation of tetracycline and oxytetracyline.

The antibiotic intercalation inside the layered double hydroxide (LDH) layers was usually considered for water decontamination but rarely for drug delivery. Here, tetracycline (TCH) and oxytetracycline (OXY) were immobilized in Zn2Al-Cl LDH following two methods: co-precipitation and anionic exchange. The interfacial concentration of antibiotic varies from 0.04 to 0.5 depending the method of immobilization. The antibiotics are not intercalated in the interlayer space allowing their release in 10 Hours. The antibacterial activity against both E. coli and S. epidermidis revealed that the loaded antibiotics are still active but less efficient compared to the free ones. After exposition to UV light or to high temperature storage (30, 60 and 120 °C), their antibacterial activity significantly decreases due to their degradation especially when antibiotic is loaded on material by co-precipitation. These results are promise to reduce antibiotic contamination in waters.

Structure and antibacterial activity relationships of native and amyloid fibril lysozyme loaded on layered double hydroxide.

Lysozyme from hen egg white is composed by a unique linear chain of 129 amino acids. It is known to inhibit Gram positive bacteria and to form amyloid fibrils at low pH, under 75°C. This work investigates the effect of the fibrillation and/or adsorption onto a layered double hydroxide material on the antibacterial properties of lysozyme. The kinetics of adsorption follows a behavior of pseudo second order model. The X-ray diffraction and the Fourier transform infrared spectroscopy highlight that adsorption occurs only on the external surface of the material. Interestingly, the amyloid fibrils of lysozyme retain their antibacterial properties when they are adsorbed on the layered double hydroxide; even if their activity is lowered, the active site of the enzyme is not fully denatured and is still accessible. This is confirmed by the study of the tryptophan using time-resolved fluorescence spectroscopy.

Adsorption of nisin into layered double hydroxide nanohybrids and in-vitro controlled release.

Layered double hydroxide (LDH) nanohybrid intercalated biomolecules, including oligonecluotides, genes and peptides/proteins, have attracted particular attention since they exhibit improved safety and effectiveness as successful delivery biosystems. The current study specifically investigated the adsorption of nisin peptide and precisely the control of the release of the payload. Adsorption occurred from peptide solution in contact with zinc-aluminum LDH at room temperature, looking out over the influence of the Zn2+/Al3+ ratio, the anion exchange capacity, the nature of the intercalated anion, the host matrix, and the host morphology. Higher adsorption was obtained, around 80% of the loaded nisin and successful intercalation was verified by X-ray diffraction. The in-vitro release tests of the nisin from the biohybrid formulation was held over 25days in PBS medium (0.01M, pH7,4) and showed that no burst release phenomenon occurred at the beginning step, in addition, a sustained-time release of nisin was obtained compared with the free nisin. Therefore, these preliminary results are encouraging for the development of bioprotectors based on nisin intercalated LDH and being implemented in the food and medical industries.

Alkane biohydroxylation: Interests, constraints and future developments.

Alkanes constitute one of the vastest reserves of raw materials for the production of fine chemicals. This paper focuses on recent advances in alkane biohydroxylation, i.e. the bioactivation of alkanes into their corresponding alcohols. Enzyme and whole-cell biocatalysts have been reviewed. Process considerations to implement such biocatalysts in bioreactors at large scale by coupling the bioconversion with cofactor regeneration and product removal are also discussed.

Novel biocompatible electrospun gelatin fiber mats with antibiotic drug delivery properties.

The aim of this study was to synthesize stable gelatin electrospun mats (ESMs) (cross-linked by glutaraldehyde (GTA) vapors) with tunable drug release properties using pH as a stimulus. Gelatin ESMs loaded with rhodamine as a model drug were first synthesized. The in vitro release of rhodamine was characterized to understand the mechanisms of drug release and the effects of both cross-linker concentration and pH on drug release. An optimal cross-linker concentration of 5% was evidenced to provide ESMs allowing both sustainable release of drugs at pH 7 and burst release at pH 2. The release profiles were then fitted with a power law model to describe the release kinetics. The chlorhexidine antibiotic drug was finally loaded into the optimal electrospun mat and its bactericidal activity was demonstrated against Gram-negative (E. coli) and Gram-positive (S. epidermidis) bacteria by agar diffusion tests. This biocompatible material was shown to efficiently destroy bacterial biofilms and prevent bacterial growth within a short time (3 h), while maintaining its antibacterial activity up to at least 72 h. This study provides a promising material, which could treat infected sites and prevent infections, with tunable drug releasing properties using pH as a stimulus.

Dynamic Mechanisms of the Bactericidal Action of an Al2O3-TiO2-Ag Granular Material on an Escherichia coli Strain.

The bactericidal activity of an Al2O3-TiO2-Ag granular material against an Escherichia coli strain was confirmed by a culture-based method. In particular, 100% of microorganisms were permanently inactivated in 30 to 45 min. The present work aimed to investigate the mechanisms of the bactericidal action of this material and their dynamics on Escherichia coli using different techniques. Observations by transmission electron microscopy (TEM) at different times of disinfection revealed morphological changes in the bacteria as soon as they were put in contact with the material. Notably highlighted were cell membrane damage; cytoplasm detachment; formation of vacuoles, possibly due to DNA condensation, in association with regions exhibiting different levels of electron density; and membrane lysis. PCR and flow cytometry analyses were used to confirm and quantify the observations of cell integrity. The direct exposure of cells to silver, combined with the oxidative stress induced by the reactive oxygen species (ROS) generated, was identified to be responsible for these morphological alterations. From the first 5 min of treatment with the Al2O3-TiO2-Ag material, 98% of E. coli isolates were lysed. From 30 min, cell viability decreased to reach total inactivation, although approximately 1% of permeable E. coli cells and 1% of intact cells (10(5) genomic units·ml(-1)) were evidenced. This study demonstrates that the bactericidal effect of the material results from a synergic action of desorbed and supported silver. Supported silver was shown to generate the ROS evidenced.