Main-chain flexibility and hydrophobicity of ionenes strongly impact their antimicrobial activity: an extended study on drug resistance strains and Mycobacterium.

The spread of antibiotic-resistant pathogens and the resurgence of tuberculosis disease are major motivations to search for novel antimicrobial agents. Some promising candidates in this respect are cationic polymers, also known as synthetic mimics of antimicrobial peptides (SMAMPs), which act through the membrane-lytic mechanism. Development of resistance toward SMAMPs is less likely than toward currently employed antibiotics; however, further studies are needed to better understand their structure-activity relationship. The main objective of this work is to understand the cross-influence of hydrophobicity, main-chain flexibility, and the topology of ionenes (polycations containing a cationic moiety within the main-chain) on activity. To fulfill this goal, a library of ionenes was developed and compared with previously investigated molecules. The obtained compounds display promising activity against the model microorganisms and drug-resistance clinical isolates, including Mycobacterium tuberculosis. The killing efficiency was also investigated, and results confirm a strong effect of hydrophobicity, revealing higher activity for molecules possessing the flexible linker within the polymer main-chain.

The effect of surface morphology on endothelial and smooth muscle cells growth on blow-spun fibrous scaffolds.

This study aimed to analyze the growth of two types of blood vessel building cells: endothelial cells (ECs) and smooth muscle cells (SMCs) on surfaces with different morphology. Two types of materials, differing in morphology, were produced by the solution blow spinning technique. One-layer materials consisted of one fibrous layer with two fibrous surfaces. Bi-layer materials consisted of one fibrous-solid layer and one fibrous layer, resulting in two different surfaces. Additionally, materials with different average fiber diameters (about 200, 500, and 900 nm) were produced for each group. It has been shown that it is possible to obtain structures with a given morphology by changing the selected process parameters (working distance and polymer solution concentration). Both morphology (solid versus fibrous) and average fiber diameter (submicron fibers versus microfibers) of scaffolds influenced the growth of ECs. However, this effect was only visible after an extended period of culture (6 days). In the case of SMCs, it was proved that the best growth of SMCs is obtained for micron fibers (with an average diameter close to 900 nm) compared to the submicron fibers (with an average diameter below 900 nm).

Vascular Polyurethane Prostheses Modified with a Bioactive Coating-Physicochemical, Mechanical and Biological Properties.

This study describes a method for the modification of polyurethane small-diameter (5 mm) vascular prostheses obtained with the phase inversion method. The modification process involves two steps: the introduction of a linker (acrylic acid) and a peptide (REDV and YIGSR). FTIR and XPS analysis confirmed the process of chemical modification. The obtained prostheses had a porosity of approx. 60%, Young's Modulus in the range of 9-11 MPa, and a water contact angle around 40°. Endothelial (EC) and smooth muscle (SMC) cell co-culture showed that the surfaces modified with peptides increase the adhesion of ECs. At the same time, SMCs adhesion was low both on unmodified and peptide-modified surfaces. Analysis of blood-materials interaction showed high hemocompatibility of obtained materials. The whole blood clotting time assay showed differences in the amount of free hemoglobin present in blood contacted with different materials. It can be concluded that the peptide coating increased the hemocompatibility of the surface by increasing ECs adhesion and, at the same time, decreasing platelet adhesion. When comparing both types of peptide coatings, more promising results were obtained for the surfaces coated with the YISGR than REDV-coated prostheses.

Influence of lipid bilayer composition on the activity of antimicrobial quaternary ammonium ionenes, the interplay of intrinsic lipid curvature and polymer hydrophobicity, the role of cardiolipin.

Incorporation of hydrophobic component into amphiphilic polycations structure is frequently accompanied by an increase of antimicrobial activity. There is, however, a group of relatively hydrophilic polycations containing quaternary ammonium moieties along mainchain, ionenes, which also display strong antimicrobial and limited hemolytic properties. In this work, an influence of a hydrophobic side group length on antimicrobial mechanism of action is investigated in a series of novel amphiphilic ionenes. High antimicrobial activity was found by determination of minimum inhibitory concentration (MIC) and minimum bactericidal, and fungicidal concentration (MBC and MFC) in both growth media and a buffer. Biocompatibility was estimated by hemolytic and mammalian cells viability assays. Mechanistic studies were performed using large unilamellar vesicles (LUVs) with different lipid composition, as simplified models of cell membranes. The investigated ionenes are potent and selective antimicrobial molecules displaying a decrease of antimicrobial activity correlated with increase of hydrophobicity. Studies using LUVs revealed that the cardiolipin is an essential component responsible for the lipid bilayer permeabilization by investigated ionens. In contrast to relatively hydrophilic ionenes, more hydrophobic polymers showed an ability to stabilize membranes composed of lipids with negative spontaneous curvature in a certain range of polymer to lipid ratio. The results substantially contribute to the understanding of antimicrobial activity of the investigated class of polymers.

Cylindrical Polyurethane Scaffold Fabricated Using the Phase Inversion Method: Influence of Process Parameters on Scaffolds' Morphology and Mechanical Properties.

This work presents a method of obtaining cylindrical polymer structures with a given diameter (approx. 5 mm) using the phase inversion technique. As part of the work, the influence of process parameters (polymer hardness, polymer solution concentration, the composition of the non-solvent solution, process time) on the scaffolds' morphology was investigated. Additionally, the influence of the addition of porogen on the scaffold's mechanical properties was analyzed. It has been shown that the use of a 20% polymer solution of medium hardness (ChronoFlex C45D) and carrying out the process for 24 h in 0:100 water/ethanol leads to the achievement of repeatable structures with adequate flexibility. Among the three types of porogens tested (NaCl, hexane, polyvinyl alcohol), the most favorable results were obtained for 10% polyvinyl alcohol (PVA). The addition of PVA increases the range of pore diameters and the value of the mean pore diameter (9.6 ± 3.2 vs. 15.2 ± 6.4) while reducing the elasticity of the structure (Young modulus = 3.6 ± 1.5 MPa vs. 9.7 ± 4.3 MPa).

In vitro haemocompatibility assessment of acrylic acid deposited on solid, polyurethane substrate.

The main purpose of the work was to assess the haemocompatible properties of polyurethane discs with a modified surface dedicated to cardiovascular system regeneration. They were coated with acrylic acid-based material to inhibit the activation of the blood coagulation cascade. This coating improved the wettability of the material, leading to the prevention of protein adsorption on the surface. The blood-material interaction was analyzed in dynamic conditions with a specially designed tester, which helps to control blood-material interaction under high shear stress conditions. The corresponding numerical model of the tester was also developed by finite volume method (FVM). The 3D FVM model allows the determination of shear stresses applying different flow and boundary conditions representing blood-material interactions. The haemocompatibility analyses were performed through in vitro tests using a blood flow simulator. They revealed a low probability of activation of blood coagulation and low leukocyte activation. The original mechanical set-up to test the blood-material interaction helped to prove that acrylic acid-based coatings expressed good haemocompatible properties.

Fenton-type reaction grafting of polyvinylpyrrolidone onto polypropylene membrane for improving hemo- and biocompatibility.

Known techniques for modification of polypropylene membranes (PPm) often require modification of the membrane in its entire volume (i.e. at the manufacturing stage), which may affect its properties. In the present work, the authors proposed a simple method for PPm hydrophilization. The process involves a two-step Fenton-type reaction, with ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent and cumene hydroperoxide (CHP) as a source of free radicals. This hydrogel coating aims to enhance membrane hemocompatible and biocompatible properties. The biggest advantage of the proposed technique is the change of materials' surface properties, without interfering with its internal structure. Microscopic (SEM) and spectroscopic (FTIR-ATR) analyses confirmed the presence of hydrogel coating on PPm surfaces. Additionally, the evaluation of the surface density of the coating showed that the thickness of the coating increases with the reaction time and CHP concentration. The applied coatings significantly increase surface hydrophilicity (contact angle for PPm: 128.58°â€¯±â€¯0.52°, for all modified surfaces <53.31°â€¯±â€¯2.03°). The cytotoxicity test (XTT assay) proved biocompatibility of the PVP coating - cell viability remained above 90% for all variants tested. The modification resulted in a decrease in fibrinogen adsorption (of at least about 16%) and in a number of surface-adhered platelets. The assay evaluating the amount of secreted cell adhesion molecules (ICAM-1) showed a significant reduction (of at least about 50%) in the expression of ICAM-1 for all hydrogel-modified surfaces.

Hydrophilic Quaternary Ammonium Ionenes-Is There an Influence of Backbone Flexibility and Topology on Antibacterial Properties?

The antimicrobial properties of polycations are strongly affected by the structural features such as the backbone flexibility and topology (isomerism) through the polymer ability to attain proper conformation in interaction with the cell membrane. In this paper, a synthesis and biocidal properties evaluation of ionenes characterized by different backbone topology (isomerism) and flexibility are presented. The findings reveal influence of variation in topology on activity against different microorganisms, and general positive effect of improved flexibility. Furthermore, one of the obtained ionenes displays degradable properties in near physiological environment (phosphate-buffered saline pH 7.4, 37 °C). The degradation proceeds via Hofmann elimination reaction and the products are not of acidic character. For the first time a new class of degradable ionenes with a high antimicrobial potential is presented.

Polyvinylpyrrolidone (PVP) hydrogel coating for cylindrical polyurethane scaffolds.

The presented study describes a method for the preparation and modification of cylindrical polyurethane structures with polyvinylpyrrolidone (PVP) hydrogel coating. The modified polyurethane scaffolds were fabricated using the phase-inversion technique and intended to be used as a vascular prosthesis. The proposed modification method involves a two-step Fenton-type reaction. Physicochemical analysis (Fourier transform infrared spectroscopy, scanning electron microscopy) confirmed the presence of the hydrogel coating. The influence of PVP and polymerization initiator (cumene hydroperoxide) concentrations on hydrogel's properties were examined. The higher concentrations of reagent were used, the thicker coating was obtained. After modification, the material's surface becomes more hydrophilic in comparison to pristine polyurethane. Cytotoxicity assay (MTT test) confirmed that PVP-coating is not toxic. The introduction of hydrogel coating resulted in a significant decrease in the fibrinogen adsorbed to the material's surface as compared to a non-modified polymer. Platelet adhesion assay demonstrated almost no platelet adhesion to the modified surfaces.

Amphiphilic Polymethyloxazoline-Polyethyleneimine Copolymers: Interaction with Lipid Bilayer and Antibacterial Properties.

Polycations, mimicking activity of antibacterial peptides, belong to an important class of molecules investigated as a support or as an alternative to antibiotics. In this work, studies of modified linear amphiphilic statistical polymethyloxazoline (PMOX) and polyethyleneimine copolymers (PMOX_PEI) series are presented. Variation of PEI content in the structure results in controllable changes of polymeric aggregates zeta potential. The structure with the highest positive charge shows the best antimicrobial activity, well visible in tests against model Gram-positive and Gram-negative bacteria, fungi, and mycobacterium strains. The polymer toxicity is evaluated with MTT and hemolysis assay as a reference. Quartz crystal microbalance (QCM-D) is used to investigate interaction between polycations and a model lipid membrane. Polymer activity correlates well with molecular structure, showing that amphiphilic component is altering polymer behavior in contact with the lipid bilayer.

Determination of polyurethane-grafted peptide (GSGREDVGSG) using bicinchoninic acid assay.

The goal in the presented study was to develop a simple, fast and accurate method for measuring the surface density of a short peptide sequence bound to a polymeric substrate. We analyzed polyurethane samples chemically modified with acrylic acid and polyurethane-grafted peptide (GSGREDVGSG) and investigated the possibility of using the bicinchoninic acid (BCA) assay to determine surface density of the solid-supported peptide. We set the conditions (temperature, time) under which the test should be conducted. We also studied the interaction of the BCA reagent with polyurethane substrate and the effect of drying conditions as well as material type and form on the test response. We have proposed potential factors that might interfere with the BCA assay and chosen the proper control materials.

Electropolymerized hydrophilic coating on stainless steel for biomedical applications.

Current metal implants (e.g. stents) covered with drug-eluting coatings are not robust for long-term usage. Other types and methods of coatings are needed, especially ones that are not prone to activity loss in vivo. In this paper, the method of stainless steel (SS) coating with poly(ethylene glycol) dimethacrylate (PEGDMA) with the use of electropolymerization (EP) is presented. The application of a specific and simple reaction mixture enabled the production of SS-PEGDMA materials that possessed a homogenous surface. The polymer coating was durable for 28 days of constant washing. The resulting materials were non-toxic and haemolysis did not occur after incubation with blood. Moreover, because the coating filled up scratches present on bare SS and hydrophilized the SS surface, it reduced fibrinogen adsorption five times in comparison to SS and, unlike on SS, no platelet activation was detected. The presented method is a very promising candidate for scale up due to its simplicity and low cost.

Endothelialization of polyurethanes: Surface silanization and immobilization of REDV peptide.

The paper presents method for chemical immobilization of arginine-glutamic acid-aspartic acid-valine (REDV) peptide on polyurethane surface. The peptide has been covalently bonded using silanes as a spacer molecules. The aim of this work was to investigate the proposed modification process and assess its biological effectiveness, especially in contact with blood and endothelial cells. Physicochemical properties were examined in terms of wettability, atomic composition and density of introduced functional groups and peptide molecules. Experiments with blood showed that material coating reduced number of surface-adhered platelets and fibrinogen molecules. In contrast to polyurethane (PU), there were no blood components deposited on REDV-modified materials (PU-REDV); fibrinogen adsorption on PU-REDV surface has been strongly reduced compared to PU. Analysis of cell adhesion after 1, 2, 3, 4, and 5 days of culture showed a significant increase of the cell-coated area on PU-REDV compared to PU. However, an intense cell growth appeared also on the control surface modified without the addition of REDV. Thus, the positive effect of REDV peptide on the adhesion of HUVEC could not be unequivocally confirmed.