Antimicrobial and Antibiofilm Activity of Curcumin-Loaded Electrospun Nanofibers for the Prevention of the Biofilm-Associated Infections.

Curcumin extracted from the rhizome of Curcuma Longa has been used in therapeutic preparations for centuries in different parts of the world. However, its bioactivity is limited by chemical instability, water insolubility, low bioavailability, and extensive metabolism. In this study, the coaxial electrospinning technique was used to produce both poly (ε-caprolactone) (PCL)-curcumin and core-shell nanofibers composed of PCL and curcumin in the core and poly (lactic acid) (PLA) in the shell. Morphology and physical properties, as well as the release of curcumin were studied and compared with neat PCL, showing the formation of randomly oriented, defect-free cylindrical fibers with a narrow distribution of the dimensions. The antibacterial and antibiofilm potential, including the capacity to interfere with the quorum-sensing mechanism, was evaluated on Pseudomonas aeruginosa PAO1, and Streptococcus mutans, two opportunistic pathogenic bacteria frequently associated with infections. The reported results demonstrated the ability of the Curcumin-loading membranes to inhibit both PAO1 and S. mutans biofilm growth and activity, thus representing a promising solution for the prevention of biofilm-associated infections. Moreover, the high biocompatibility and the ability to control the oxidative stress of damaged tissue, make the synthesized membranes useful as scaffolds in tissue engineering regeneration, helping to accelerate the healing process.

Ionic Liquid as Dispersing Agent of LDH-Carbon Nanotubes into a Biodegradable Vinyl Alcohol Polymer.

A Zn/Al layered double hydroxides (LDHs) hosting carbon nanotubes (80% of CNTs) was synthesized and dispersed into a commercial biodegradable highly amorphous vinyl alcohol polymer at different loading (i.e., 1; 3; 5; 10 wt%). In order to improve the degree of dispersion of the filler into the polymer matrix, an ionic liquid (IL) based on 1-hexadecyl-3-methylimidazolium dimethyl-5-sodiosulfoisophthalate was added to the composites' mixtures. Structural characterization of filler and polymeric composites was carried out. The analysis of thermal, mechanical and electrical properties of the composites, resulted improved compared to the unfilled material, allowed to hypothesize a good dispersion of the LDH-CNTs lamellar filler into the polymer matrix-assisted by the ionic liquid. This was demonstrated comparing electrical conductivity of composite at 5% of LDH-CNTs in the presence and in the absence of IL. The experimental results showed that the electrical conductivity of the sample with IL is four orders of magnitude higher than the one without IL. Furthermore, the percolation threshold of the whole system resulted very low-0.26% of LDH-CNTs loading, which is 0.21% of CNTs.

Multifunctional Bioactive Resin for Dental Restorative Materials.

Resin-based composites are widely used as dental restorative materials due to their excellent properties. They must have high modulus, high hardness, and be chemically inert while minimizing moisture uptake. To fulfill these higher standard prerequisites and properties, continuous improvements in each of their components are required. This study develops novel composites with multiple biofunctions. Light-cured Bis-GMA/TEGDMA dental resin (RK)/layered double hydroxide intercalated with fluoride ions (LDH-F)/calcium bentonite (Bt) hybrid composites were prepared. The loading ratio of LDH-F to Bt was varied, ranging from 2.5/2.5 to 10/10 parts per hundred RK and structural, mechanical, and biological properties were studied. The incorporation of even small mass fractions (e.g., 2.5 wt % of LDH-F and 2.5 wt % of Bt) in RK dental resin significantly improved the mechanical properties of the pristine resin. The synthetized materials showed antibacterial and antibiofilm effects against three bacterial strains isolated from healthy volunteers' saliva (Streptococcus spp., Bacteroides fragilis, and Staphylococcus epidermidis) without affecting its ability to induce dental pulp stem cells differentiation into odontoblast-like cells. The capability to balance between the antibiofilm activity and dental pulp stem cells differentiation in addition with improved mechanical properties make these materials a promising strategy in preventive and restorative dentistry.

PET and Active Coating Based on a LDH Nanofiller Hosting p-Hydroxybenzoate and Food-Grade Zeolites: Evaluation of Antimicrobial Activity of Packaging and Shelf Life of Red Meat.

Layered double hydroxide (LDH) nanofillers were considered as hosts of p-hydroxybenzoate as an antimicrobial molecule for active coating. A food grade resin with LDH-p-hydroxybenzoate and two different types of food grade zeolites was used to prepare active coatings for Polyethylene terephthalate (PET) trays. The release kinetics of the active molecule were followed using UV spectrophotometry and the experimental results were analyzed with the Gallagher-Corrigan model. The thermal properties of the coating mixtures and the PET coating were analyzed and found to be dependent on the coating's composition. On the basis of CO2 transmission rate and off-odors tests, the best coating composition was selected. Global migration in ethanol (10% v/v), acetic acid (3% w/v), and vegetable oil, and specific migration of p-hydroxybenzoic acid revealed the suitability of the material for food contact. Antimicrobial tests on the packaging demonstrated a good inhibition against Salmonella spp. and Campylobacter jejuni. Red meat was packed into the selected active materials and results were compared to uncoated PET packaging. Color tests (browning of the meat) and analysis of Enterobacteriaceae spp. and total viable count evolution up to 10 days of storage demonstrated the capability of the considered active packaging in prolonging the shelf life of red meat.

Fabrication and Characterization of Poly(lactic acid)/Poly(ε-caprolactone) Blend Electrospun Fibers Loaded with Amoxicillin for Tunable Delivering.

Blends of poly(lactic acid) (PLA) and poly(ε-caprolactone) (PCL), loaded with different amounts of Amoxicillin antibiotic (AMOX) were electrospun to investigate their release properties and obtain a controlled and tuneable release. The processing parameters for electrospinning were set up and reliable membranes were obtained. Morphology and thermal behaviour were found dependent on the component ratio as well as on the incorporated drug amount. A very different release kinetics of the two pristine polymers, very rapid for PCL and very slow for PLA, reflected in intermediate release time. However comparing the release amount with that predicted by the mixture rule a preferential incorporation of AMOX into PLA can be inferred.

Polymorphic solidification of Linezolid confined in electrospun PCL fibers for controlled release in topical applications.

Poly(ϵ-caprolactone) (PCL) membranes loaded with Linezolid, chemically N-[[(5S)-3-[3-fluoro-4-(4-morpholinyl)phenyl]-2-oxo-5-oxazolidinyl]methyl]acetamide (empirical formula C16H20FN3O4) have been prepared by electrospinning technique, at different Linezolid concentrations (0.5, 1, 2.5 and 5%, w/w). Structural characterization, morphological analysis and the study of the mechanical properties have been performed on loaded membranes and compared with neat PCL membranes. Linezolid embedded in the membranes is prevalently amorphous, with a low crystallinity showing a different polymorphic form respect to the usual Form I and Form II. The release kinetics of the drug were studied by spectrophotometric analysis (UV-vis). It allowed to discriminate between Linezolid molecules on the surface and encapsulated into the fibers. The antibacterial activity of the electrospun membranes was effective to inhibit Staphylococcus aureus. The properties of the loaded membranes and their capability for local delivery of the antibiotic make them good candidates as drug release devices for topical use.

Pectin functionalized with natural fatty acids as antimicrobial agent.

Several pectin derivatives were prepared by chemical modifications of the polysaccharide with natural fatty acids. The obtained biodegradable pectin-based materials, pectin-linoleate, pectin-oleate and pectin-palmitate, were investigated for their antimicrobial activity against several bacterial strains, Staphylococcus aureus and Escherichia coli. Good results were obtained for pectin-oleate and pectin-linoleate, which inhibit the growth of the selected microorganisms by 50-70%. They exert the better antimicrobial activity against S. aureus. Subsequently, the pectin-oleate and the pectin-linoleate samples were coated on polyethylene films and were assessed for their capacity to capture the oxygen molecules, reducing its penetration into the polymeric support. These results confirmed a possible application of the new materials in the field of active food packaging.

Effect of layered double hydroxide intercalated with fluoride ions on the physical, biological and release properties of a dental composite resin.

OBJECTIVES: The aim of this work was the preparation of a new fluoride-releasing dental material characterized by a release of fluoride relatively constant over time without any initial toxic burst effect. This type of delivery is obtained by a matrix controlled elution and elicits the beneficial effect of a low amount of fluoride on human dental pulp stem cells (hDPSCs) towards mature phenotype. METHODS: The modified hydrotalcite intercalated with fluoride ions (LDH-F), used as filler, was prepared via ion exchange procedure and characterized by X-ray diffraction and FT-IR spectroscopy. The LDH-F inorganic particles (0.7, 5, 10, 20wt.%) were mixed with a photo-activated Bis-GMA/TEGDMA (45/55wt/wt) matrix and novel visible-light cured composites were prepared. The dynamic thermo-mechanical properties were determined by dynamic mechanical analyzer. The release of fluoride ions in physiological solution was determined using a ionometer. Total DNA content was measured by a PicoGreen dsDNA quantification kit to assess the proliferation rate of hDPSCs. Alkaline phosphatase activity (ALP) was measured in presence of fluoride resins. RESULTS: Incorporation of even small mass fractions (e.g. 0.7 and 5wt.%) of the fluoride LDH in Bis-GMA/TEGDMA dental resin significantly improved the mechanical properties of the pristine resin, in particular at 37°C. The observed reinforcement increases on increasing the filler concentration. The release of fluoride ions resulted very slow, lasting months. ALP activity gradually increased for 28 days in hDPSCs cell grown, demonstrating that low concentrations of fluoride contributed to the cell differentiation. CONCLUSIONS: The prepared composites containing different amount of hydrotalcite filler showed improved mechanical properties, slow fluoride release and promoted hDPSCs cell proliferation and cell differentiation.

The role of carbon nanofiber defects on the electrical and mechanical properties of CNF-based resins.

Heat treatment of carbon nanofibers has proven to be an effective method in removing defects from carbon nanofibers, causing a strong increase in their structural perfection and thermal stability. It affects the bonding states of carbon atoms in the nanofiber structure and causes a significant transformation in the hybridization state of the bonded carbon atoms.Nanofilled resins made of heat-treated CNF show significant increases in their electrical conductivity even at low concentrations. This confirms that enhancement in the perfection of the fiber structure with consequent change in the morphological features plays a prominent role in affecting the electrical properties. Indeed heat-treated CNFs display a stiff structure and a smooth surface which tends to lower the thickness of the unavoidable insulating epoxy layer formed around the CNF which, in turn, plays a fundamental role in the electrical transport properties along the conducting clusters. This might be very beneficial in terms of electrical conductivity but might have negligible effect on the mechanical properties.

Enhanced in vitro antitumor activity of a titanocene complex encapsulated into polycaprolactone (PCL) electrospun fibers.

PURPOSE: The purpose of this work was to achieve detailed biomaterials characterization of a drug delivery system for local cancer treatment based on electrospun titanocene trichloride-loaded resorbable polycaprolactone (PCL) fibers. METHODS: The PCL fibers were characterized for their structural, morphologic and physical properties. The drug release kinetics of the titanocene complex was investigated at different concentrations, to obtain a set of correlations between structure and tuneable release. After exposing cancer cells directly onto the surface of PCL fibers, the anti-proliferative effects of titanocene-loaded PCL were assessed by: (i) counting viable cells via live/dead staining methods, and (ii) analyzing cell apoptosis. RESULTS AND CONCLUSIONS: Titanocene concentration influenced fiber diameters reduced for PCL filled with titanocene. X-ray analysis suggested that the titanocene, encapsulated into the PCL fibers, is not allowed to crystallize and exists as amorphous aggregates into the fibers. The titanocene release curves presented two stages unrelated to PCL degradation: an initial burst release followed by a release linear with time, extending for a very long time. All of the titanocene-loaded fibers revealed sustained drug release properties suggesting their potential clinical applicability for the treatment of local cancer diseases.

Preparation, characterization and antibacterial activity of poly(epsilon-caprolactone) electrospun fibers loaded with amoxicillin for controlled release in biomedical applications.

Amoxicillin (AMOX) was successfully encapsulated at different concentrations into poly(epsilon-caprolactone) (PCL) by the electrospinning technique, and mats of non-woven fibers were obtained and characterized in terms of morphology, in vitro release and antibacterial properties. The scanning electron microscopy evidenced the nanofibrous structure of the pristine PCL, composed of individual, uniform, and randomly oriented fibres with an average diameter ranging around 0.8 micron. The addition of amoxicillin at different concentrations (3, 5 and 7 wt%) did not evidence change in the fiber morphology. The release curves, for all samples, present mainly two stages: a first stage, quick as a "burst," is followed by a second slow stage. The burst was analyzed as a function of amoxicillin concentration and thickness of the membranes, and values corresponding to absence of burst were individuated. In the second stage the release was found very slow extending up to months for the most concentrated sample. The antibacterial activity of the electrospun fibers was effective to inhibit in different proportions Staphylococcus aureus, Enterococcus faecalis and Escherichia coli. The properties of the filled membranes and their capability for local delivery of antibiotics make them suitable for biomedical applications.

Deposition of LDH on plasma treated polylactic acid to reduce water permeability.

A simple and scalable deposition process was developed to prepare polylactic acid (PLA) coatings with enhanced water barrier properties for food packaging applications. This method based on electrostatic interactions between the positively charged layers of layered double hydroxides (LDHs) modified with ionic liquids (ILs) and the negatively charged plasma treated polylactic acid leads to homogeneous, stable, and highly durable coatings. Deposition of the LDH coatings increases the surface hydrophobicity of the neat PLA, which results to a decrease in water permeability by about 35%.

Solvent-free synthesis of modified pectin compounds promoted by microwave irradiation.

Microwave-assisted solvent-free modification of pectin was successfully accomplished, consisting in the esterification of several fatty acids by pectin alcoholic functions. The reaction was performed by simply mixing the reagents with a catalytic amount of the inorganic base (potassium carbonate) and irradiating the obtained mixture with microwaves for a short time (3-6 min). The replacement of the traditional heating with a microwave source allowed the development of a new synthetic protocol which provided increased yield of the final products, since it eliminates the small amount of degraded polysaccharide produced during traditional oil bath heating. The desired esters were fully characterized by FT-IR spectroscopy and thermogravimetric analysis.

Modified hydrotalcite-like compounds as active fillers of biodegradable polymers for drug release and food packaging applications.

This review treats the recent patents and related literature, mainly from the Authors laboratories, on biomedical and food packaging applications of nano-composites constituted of biodegradable polymers filled with micro or nano crystals of organically modified Layered Double Hydroxides of Hydrotalcite type. After a brief outline of the chemical and structural aspects of Hydrotalcite-like compounds (HTlc) and of their manipulation via intercalation of functional molecular anions to obtain materials for numerous, sometime unexpected applications, the review approaches the theme in three separated parts. Part 1 deals with the synthetic method used to prepare the pristine Mg-Al and Zn-Al HTlc and with the procedures of their functionalization with anti-inflammatory (diclofenac), antibacterial (chloramphenicol hemisuccinate), antifibrinolytic (tranexamic acid) drugs and with benzoates with antimicrobial activity. Procedures used to form (nano) composites of polycaprolactone, used as an example of biodegradable polymer, and functionalized HTlc are also reported. Part 2 discusses a patent and related papers on the preparation and biomedical use of a controlled delivery system of the above mentioned pharmacologically active substances. After an introduction dealing with the recent progress in the field of local drug delivery systems, the chemical and structural aspects of the patented system constituted of a biodegradable polymer and HTlc loaded with the active substances will be presented together with an extensive discussion of the drug release in physiological medium. Part 3 deals with a recent patent and related papers on chemical, structural and release property of antimicrobial species of polymeric films containing antimicrobial loaded HTlc able to act as active packaging for food products prolonging their shelf life.

Pectins filled with LDH-antimicrobial molecules: preparation, characterization and physical properties.

Nanohybrids of layered double hydroxide (LDH) with intercalated active molecules: benzoate, 2,4-dichlorobenzoate, para-hydroxybenzoate and ortho-hydroxybenzoate, were incorporated into pectins from apples through high energy ball milling in the presence of water. Cast films were obtained and analysed. X-ray diffraction analysis showed a complete destructuration of all nanohybrids in the pectin matrix. Thermogravimetric analysis showed a better thermal resistance of pectin in the presence of fillers, especially para-hydroxybenzoate and ortho-hydroxybenzoate. Mechanical properties showed an improvement of elastic modulus in particular for LDH-para-hydroxybenzoate nanohybrid, due probably to a better interaction between pectin matrix and nanohybrid layers. Barrier properties (sorption and diffusion) to water vapour showed improvement in the dependence on the intercalated active molecule, the best improvement was achieved for composites containing para-hydroxybenzoate molecules, suggesting that the interaction between the filler phase and the polymer plays an important role in sorption and diffusion phenomena. Incorporation of these active molecules gave antimicrobial properties to the composite films giving opportunities in the field of active packaging.

Physical and water sorption properties of chemically modified pectin with an environmentally friendly process.

A synthetic process was developed to modify pectin samples under solvent free conditions, obtaining pectin at increasing concentration of palmitic, oleic, and linoleic acids. The weight loss of the modified powders showed a degradation path very similar to the pure pectin, indicating that the pristine structure was preserved after the chemical modification. A decreasing mass of evaporating water on increasing the fatty acid concentration, in particular for the palmitic acid modification, indicated a reduced water sorption by the modified powders. Differential scanning calorimetry confirmed the thermogravimetric results and in addition indicated the crystallization of the lateral chains in the case of palmitic-acid-modified pectins. This result was confirmed by X-ray diffractograms of the palmitic acid samples, indicating the main crystallization of the form C, although possible orientation phenomena can be inferred. The sorption curves of either the pristine pectin or the modified samples showed a dual sorption behavior. The sorption curves were interpreted by the BET and GAB equations, both giving very similar results. Palmitic acid modification was very effective in reducing all sorption parameters, whereas in the case of oleic and linoleic acids, only at high concentrations was the hydrophobic influence detected.

Cure behavior and physical properties of epoxy resin-filled with multiwalled carbon nanotubes.

Different experimental procedures were investigated for incorporating Multi walled Carbon nanotubes (MWCNT) into epoxy diglycidil-ether bisphenol-A (DGEBA), cured with 4,4' diamine-dibenzyl-sulfone (DDS): (i) mechanical mixing for either 60 or 120 minutes; (ii) high energy ball milling for 30 minutes; (iii) ultrasonication for 20 minutes. The mechanical properties of the obtained samples were monitored and used in order to guide the selection of the most promising composite system. The best results were obtained by using, as method of incorporation of carbon nanotubes in the resin, sonication for 20 minutes. Moreover it was found that the presence of an accelerating agent, BF3, commonly used for the cure reaction, is ineffective in the presence of carbon nanotubes, besides leading to composites with a lower glass transition. Afterwards resins with increasing MWCNT concentration were prepared and the thermal properties analyzed, showing a slight improvement in either the glass transition or the degradation temperature. The electrical conductivity of the selected composite system is characterised by a percolation threshold (lower than 0.1%) comparable to that found for similar systems, but beyond threshold it assumes greater values. The conductivity is characterised by only a slight increase with temperature in the range 30-90 degrees C. The positive temperature coefficient can be related to a decrease of the small gaps separating the CNTs clusters. A simple power law describes the relation between the composite conductivity and CNT concentration near the percolation threshold. A dependence of all the parameters of the power law on the temperature has also been evidenced, and it should be properly considered and indicated when using such a model in describing the behaviour of composites near the percolation threshold.

New polymeric composites based on poly(-caprolactone) and layered double hydroxides containing antimicrobial species.

Benzoate (Bz), 2,4-dichlorobenzoate (BzDC), and p- and o-hydroxybenzoate (p- and o-BzOH) anions with antimicrobial activity have been intercalated into [Zn(0.65)Al(0.35)(OH)(2)](NO(3))(0.35).0.6H(2)O, layered double hydroxide (LDH), via anion-exchange reactions. The composition of the obtained intercalation compounds, determined by chemical, thermogravimetric, and ion chromatographic analyses, indicates that benzoate and benzoate derivative anions replace the nitrate counteranions, almost completely. Information on the interactions of the intercalated anions with the inorganic layer have been obtained from Fourier transform IR absorption spectroscopy and powder X-ray diffraction of the samples. It has been found that both the nature and the position of the aromatic ring substituents affect the value of the basal distance and the host-guest hydrogen bond network. Knowledge of the chemical composition, basal distance, and van der Waals dimensions of the guests has finally allowed the proposal of structural models of the intercalation compounds that have been used as fillers of poly(caprolactone), a biodegradable polymer. Films of polymeric composites were obtained by hot-pressing the powders of polymer and filler previously milled by a high-energy ball milling procedure. X-ray diffraction analysis and optical and scanning electron microscopy of the composites indicate that the LDH samples containing BzDC anions are delaminated into the polymeric matrix, whereas those containing p-BzOH anions maintain for the most part the crystal packing and give rise to microcomposites. Intermediate behavior was found for LDH modified with Bz and o-BzOH anions because exfoliated and partly intercalated composites were obtained. Preliminary antimicrobial tests indicate that the composites are able to inhibit the Saccharomyces cerevisiae growth of 40% in comparison with the growth in a pure culture medium. The composites can be studied as the model for "active packaging" systems because of the antimicrobial properties of the anions anchored to the LDH layer.

Development of nanostructured thermoregulating textile materials.

The present studies provide the first example of a nanostructured thermoregulating textile materials obtained by electrospinning process. Microencapsulated phase change material was dispersed into a Poly(epsilon-caprolactone) (PCL)-acetone solution and electrospun at ambient temperature. The morphological analysis showed that the addition of microcapsules to the PCL lead to the formation of nanofibers with a significantly lower average diameter with respect to that obtained by pure PCL in the same conditions. The resulting functional material has demonstrated to have the higher efficiency in the thermoregulating effect in comparison with more of the dispersion methods used up to now.

Effect of filler content and size on transport properties of water vapor in PLA/calcium sulfate composites.

Starting from calcium sulfate (gypsum) as fermentation byproduct of lactic acid production process, high-performance composites have been produced by melt-blending polylactide (PLA) and beta-anhydrite II (AII) filler, i.e., calcium sulfate hemihydrate previously dried at 500 degrees C. Characterized by attractive properties due to good filler dispersion throughout the polyester matrix and favorable interactions between components, these composites are interesting for potential use as biodegradable rigid packaging. The effect of filler content and mean particle diameter on the barrier properties such as sorption and diffusion to water vapor has been examined and compared to unfilled PLA. Even without additional treatments, the presence of the filler introduced constraints on molecular mobility in the permeable phase of amorphous PLA and the amount of solvent absorbed appears lower in the highly filled composites. Surprisingly, for PLA-30% AII compositions, by addition of filler characterized by high mean particle diameter (e.g., 43 microm) the thermodynamic diffusion parameter, D(0), decreased up to 2 orders of magnitude. The dimension of filler particles and their percentage in the continuous polymeric phase seem to be the most important parameters that determine the barrier properties of the PLA-AII composites to water vapor.