A novel bifunctional multilayered nanofibrous membrane combining polycaprolactone and poly (vinyl alcohol) enriched with platelet lysate for skin wound healing.

Skin wound healing is a complex physiological process that involves various cell types, growth factors, cytokines, and other bioactive compounds. In this study, a novel dual-function multilayered nanofibrous membrane is developed for chronic wound application. The membrane is composed of five alternating layers of polycaprolactone (PCL) and poly (vinyl alcohol) (PVA) nanofibers (PCL-PVA) with a dual function: the PCL nanofibrous layers allow cell adhesion and growth, and the PVA layers enriched with incorporated platelet lysate (PCL-PVA + PL) serve as a drug delivery system for continuous release of bioactive compounds from PL into an aqueous environment. The material is produced using a needleless multi-jet electrospinning approach which can lead to homogeneous large-scale production. The bioactive PCL-PVA + PL membranes are cytocompatible and hemocompatible. A spatially compartmented co-culture of three cell types involved in wound healing - keratinocytes, fibroblasts and endothelial cells - is used for cytocompatibility studies. PCL-PVA + PL membranes enhance the proliferation of all cell types and increase the migration of both fibroblasts and endothelial cells. The membranes are also hemocompatible without any deleterious effect for thrombogenicity, hemolysis and coagulation. Thus, the beneficial effect of the PCL-PVA + PL membrane is demonstrated in vitro, making it a promising scaffold for the treatment of chronic wounds.

Novel lipophosphonoxin-loaded polycaprolactone electrospun nanofiber dressing reduces Staphylococcus aureus induced wound infection in mice.

Active wound dressings are attracting extensive attention in soft tissue repair and regeneration, including bacteria-infected skin wound healing. As the wide use of antibiotics leads to drug resistance we present here a new concept of wound dressings based on the polycaprolactone nanofiber scaffold (NANO) releasing second generation lipophosphonoxin (LPPO) as antibacterial agent. Firstly, we demonstrated in vitro that LPPO released from NANO exerted antibacterial activity while not impairing proliferation/differentiation of fibroblasts and keratinocytes. Secondly, using a mouse model we showed that NANO loaded with LPPO significantly reduced the Staphylococcus aureus counts in infected wounds as evaluated 7 days post-surgery. Furthermore, the rate of degradation and subsequent LPPO release in infected wounds was also facilitated by lytic enzymes secreted by inoculated bacteria. Finally, LPPO displayed negligible to no systemic absorption. In conclusion, the composite antibacterial NANO-LPPO-based dressing reduces the bacterial load and promotes skin repair, with the potential to treat wounds in clinical settings.

Fiber Optic Based Distributed Mechanical Vibration Sensing.

The distributed long-range sensing system, using the standard telecommunication single-mode optical fiber for the distributed sensing of mechanical vibrations, is described. Various events generating vibrations, such as a walking or running person, moving car, train, and many other vibration sources, can be detected, localized, and classified. The sensor is based on phase-sensitive optical time-domain reflectometry (ϕ-OTDR). Related sensing system components were designed and constructed, and the system was tested both in the laboratory and in the real deployment, with an 88 km telecom optical link, and the results are presented in this paper. A two-fiber sensor unit, with a double-sensing range was also designed, and its scheme is described. The unit was constructed and the initial measurement results are presented.

Laccase and horseradish peroxidase for green treatment of phenolic micropollutants in real drinking water and wastewater.

Biologically active micropollutants that contain diverse phenolic/aromatic structures are regularly present in wastewater effluents and are even found in drinking water. Advanced green technologies utilizing immobilized laccase and/or peroxidase, which target these micropollutants directly, may provide a reasonable alternative to standard treatments. Nevertheless, the use of these enzymes is associated with several issues that may prevent their application, such as the low activity of laccase at neutral and basic pH or the necessity of hydrogen peroxide addition as a co-substrate for peroxidases. In this study, the activity of laccase from Trametes versicolor and horseradish peroxidase was evaluated across a range of commonly used substrates (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), syringaldazine, and guaiacol). Moreover, conditions for their optimal performance were explored along with an assessment of whether these conditions accurately reflect the effectivity of both enzymes in the degradation of a mixture of bisphenol A, 17α-ethinylestradiol, triclosan, and diclofenac in tap drinking water and secondary wastewater effluent. Laccase and horseradish peroxidase showed optimal activity at strongly acidic pH if ABTS was used as a substrate. Correspondingly, the activities of both enzymes detected using ABTS in real waters were significantly enhanced by adding approximately 2.5% (v/v) of McIlvaine's buffer. Degradation of a mixture of micropollutants in wastewater with 2.5% McIlvaine's buffer (pH 7) resulted in a substantial decrease in estrogenic activity. Low degradation efficiency of micropollutants by laccase was observed in pure McIlvaine's buffer of pH 3 and 7, compared with efficient degradation in tap water of pH 7.5 without buffer. This study clearly shows that enzyme activity needs to be evaluated on micropollutants in real waters as the assessment of optimal conditions based on commonly used substrates in pure buffer or deionized water can be misleading.

Impact of Various Sterilization and Disinfection Techniques on Electrospun Poly-ε-caprolactone.

Electrospun materials made from biodegradable polycaprolactone are used widely in various tissue engineering and regenerative medicine applications because of their morphological similarity to the extracellular matrix. However, the main prerequisite for the use of such materials in clinical practice consists of the selection of the appropriate sterilization technique. This study is devoted to the study of the impact of traditional sterilization and disinfection methods on a nanofibrous polycaprolactone layer constructed by means of the needleless electrospinning technique. It was determined that hydrogen peroxide plasma treatment led to the loss of fibrous morphology and the creation of a foil. However, certain sterilization (ethylene oxide, gamma irradiation, and peracetic acid) and disinfection techniques (ethanol and UV irradiation) were found not to lead to a change in morphology; thus, the study investigates their impact on thermal properties, molecular weight, and interactions with a fibroblast cell line. It was determined that the surface properties that guide cell adhesion and proliferation were affected more than the bulk properties. The highest proliferation rate of fibroblasts seeded on nanofibrous scaffolds was observed with respect to gamma-irradiated polycaprolactone, while the lowest proliferation rate was observed following ethylene oxide sterilization.

The assessment of electrospun scaffolds fabricated from polycaprolactone with the addition of L-arginine.

Polycaprolactone (PCL) was electrospun with the addition of arginine (Arg), an α-amino acid that accelerates the healing process. The efficient needleless electrospinning technique was used for the fabrication of the nanofibrous layers. The materials produced consisted mainly of fibers with diameters of between 200 and 400 nm. Moreover, both microfibers and beads were present within the layers. Higher bead sizes were observed with the increased addition of arginine. The arginine content within the layers as well as the weight of the resultant electrospun materials were enhanced with the increased addition of arginine to the electrospinning solution (1, 5 and 10 wt%). The PCL + 1% Arg nanofibrous layer contained 5.67 ± 0.04% of arginine, the PCL + 5% Arg layer 22.66 ± 0.24% of arginine and the PCL + 10% Arg layer 37.33 ± 0.39% of arginine according to the results of the elemental analysis. A high burst release within 5 h of soaking was recorded for the PCL + 5% and PCL + 10% nanofibrous layers. However, the release rate of arginine from the PCL + 1% Arg was significantly slower, reaching a maximum level after 72 h of soaking. The resulting materials were hydrophobic. Hemocompatibility testing under static conditions revealed no effect on hemolysis following the addition of arginine and the prolongation of the prothrombin time with the increased addition of arginine, thus exerting an influence on the extrinsic and common pathway of coagulation activation. The results of the dynamic hemocompatibility assessment revealed that the numbers of blood cells and platelets were not affected significantly by the various electrospun samples during incubation. The TAT, ß-thromboglobulin and SC5-b9 concentrations were characterized by a moderate increase in the PCL group compared to those of the control group. The presence of arginine resulted in a decrease in the investigated hemocompatibility markers. The PMN elastase levels were comparable with respect to all the groups.

Glyphosate does not show higher phytotoxicity than cadmium: Cross talk and metabolic changes in common herb.

Toxicity of glyphosate (G) alone or in combination with cadmium (Cd) was studied in Matricaria chamomilla. Cadmium accumulated in shoots and roots in relation to prolonged exposure while glyphosate and aminomethylphosphonic acid (AMPA) were detected only in roots. After 7 days of exposure, root Cd and G accumulation was similar (56 µg G or 47 µg Cd/g DW in 1 µM treatments and 330 µg G or 321 µg Cd/g DW in 10 µM treatments). Despite this fact, Cd stimulated higher ROS formation and G rather suppressed nitric oxide while H2O2 content was elevated by Cd. Subsequent assay of antioxidative enzymes (SOD, CAT, and APX) showed only the impact of Cd. Non-enzymatic antioxidants revealed more pronounced impact of Cd on ascorbic acid and soluble phenols while non-protein thiols showed synergistic effect of G and Cd in roots. Surprisingly, G alone or in combination with Cd depleted shoot citrate and tartrate accumulation despite no detectable G in shoots. In the roots, Cd evoked expected increase in malate and citrate content while G rather suppressed Cd-induced elevation. These data indicate that glyphosate is less toxic than cadmium but even low G doses are able to induce metabolic changes.

Polyamide-Laccase Nanofiber Membrane for Degradation of Endocrine-Disrupting Bisphenol A, 17α-ethinylestradiol, and Triclosan.

Contamination of potable water by endocrine disrupting chemicals (EDCs) is a growing problem worldwide. One of the possible treatments is the utilization of laccase enzyme catalyzing oxidation of phenolic structures of EDC when anchored in a polymeric nanofiber membrane. Previous studies failed to develop a membrane with a sufficiently active enzyme, or the immobilization process was too complicated and time-consuming. Here, we established an elegant method for immobilizing Trametes versicolor laccase onto polyamide 6 nanofibers (PA6-laccase) via adsorption and glutaraldehyde crosslinking, promoting high enzyme activity and easier applicability in water treatment technology. This simple and inexpensive immobilization ensures both repeated use, with over 88% of initial activity retained after five ABTS catalytic cycles, and enhanced storage stability. PA6-laccase was highly effective in degrading a 50-µM EDC mixture, with only 7% of bisphenol A, 2% of 17α-ethinylestradiol, and 30% of triclosan remaining after a 24-h catalytic process. The PA6-laccase membrane can lead to the improvement of novel technologies for controlling of EDC contamination in potable water.

Electrospun vascular grafts fabricated from poly(L-lactide-co-ε-caprolactone) used as a bypass for the rabbit carotid artery.

The study involved the electrospinning of the copolymer poly(L-lactide-co-ε-caprolactone) (PLCL) into tubular grafts. The subsequent material characterization, including micro-computed tomography analysis, revealed a level of porosity of around 70%, with pore sizes of 9.34 ± 0.19 µm and fiber diameters of 5.58 ± 0.10 µm. Unlike fibrous polycaprolactone, the electrospun PLCL copolymer promoted fibroblast and endothelial cell adhesion and proliferation in vitro. Moreover, the regeneration of the vessel wall was detected following implantation and, after six months, the endothelialization of the lumen and the infiltration of arranged smooth muscle cells producing collagen was observed. However, the degradation rate was found to be accelerated in the rabbit animal model. The study was conducted under conditions that reflected the clinical requirements-the prostheses were sutured in the end-to-side fashion and the long-term end point of prosthesis healing was assessed. The regeneration of the vessel wall in terms of endothelialization, smooth cell infiltration and the presence of collagen fibers was observed after six months in vivo. A part of the grafts failed due to the rapid degradation rate of the PLCL copolymer.