Carboxymethylated and Sulfated Furcellaran from Furcellaria lumbricalis and Its Immobilization on PLA Scaffolds.

This study involved the creation of highly porous PLA scaffolds through the porogen/leaching method, utilizing polyethylene glycol as a porogen with a 75% mass ratio. The outcome achieved a highly interconnected porous structure with a thickness of 25 µm. To activate the scaffold's surface and improve its hydrophilicity, radiofrequency (RF) air plasma treatment was employed. Subsequently, furcellaran subjected to sulfation or carboxymethylation was deposited onto the RF plasma treated surfaces with the intention of improving bioactivity. Surface roughness and water wettability experienced enhancement following the surface modification. The incorporation of sulfate/carboxymethyl group (DS = 0.8; 0.3, respectively) is confirmed by elemental analysis and FT-IR. Successful functionalization of PLA scaffolds was validated by SEM and XPS analysis, showing changes in topography and increases in characteristic elements (N, S, Na) for sulfated (SF) and carboxymethylated (CMF). Cytocompatibility was evaluated by using mouse embryonic fibroblast cells (NIH/3T3).

Sulfation of furcellaran and its effect on hemocompatibility in vitro.

In this study, furcellaran (FUR) obtained from Furcellaria lumbricalis was firstly employed for sulfation via various methods, including SO3-pyridine (SO3∙Py) complex in different aprotic solvents, chlorosulfonic acid and sulfuric acid with a "coupling" reagent N,N'-Dicyclohexylcarbodiimide. Structural characterization through FT-IR, GPC, XPS and elemental analyses confirmed the successful synthesis of 6-O-sulfated FUR derivates characterized by varying degrees of sulfation (DS) ranging from 0.15 to 0.91 and molecular weight (Mw) spanning from12.5 kDa to 2.7 kDa. In vitro clotting assays, partial thromboplastin time (aPTT), thrombin time (TT), and prothrombin time (PT) underscored the essential role of sulfate esters in conferring anticoagulant activity whereas FUR prepared via chlorosulfonic acid with DS of 0.91 reached 311.4 s in aPPT showing almost 4-fold higher anticoagulant activity than native FUR at the concentration 2 mg/mL. MTT test showed all tested samples decreased cell viability in a dose dependent manner while all of them are non-cytotoxic up to the concentration of 0.1 mg/mL. Furthermore, sulfated derivates deposited onto polyethylene terephthalate surface presented substantial decrease in platelet adhesion, as well as absence of the most activated platelet stages. These findings support the pivotal role of O-6 FUR sulfates in enhancing hemocompatibility and provide valuable insights for a comparative assessment of effective sulfating approaches.

Fucoidan- and Ciprofloxacin-Doped Plasma-Activated Polymer Coatings on Biodegradable Zinc: Hemocompatibility and Drug Release.

Blood-contacting medical devices such as biodegradable metallic bone implant materials are expected to show excellent hemocompatibility both in vitro and in vivo. Different approaches are being studied and used to modify biomaterial surfaces for enhanced biocompatibility and hemocompatibility. However, the composition of degradable biomaterial must address several drawbacks at once. Iron-reinforced zinc material was used as a metallic substrate with improved mechanical properties when compared with those of pure zinc. Poly(lactic) acid (PLA) or polyethylenimine (PEI) was selected as a polymeric matrix for further doping with antibiotic ciprofloxacin (CPR) and marine-sourced polysaccharide fucoidan (FU), which are known for their antibacterial and potential anticoagulant properties, respectively. Radiofrequency air plasma was employed to induce metallic/polymer-coated surface activation before further modification with FU/CPR. Sample surface morphology and composition were studied and evaluated (contact angle measurements, AFM, SEM, and FT-IR) along with the hemolysis ratio and platelet adhesion test. Successful doping of the polymer layer by FU/CRP was confirmed. While PEI induced severe hemolysis over 12%, the PLA-coated samples exhibited even lower hemolysis (∼2%) than uncoated samples while the uncoated samples showed the lowest platelet adhesion. Moreover, gradual antibiotic release from PLA determined by the electrochemical methods using screen-printed carbon electrodes was observed after 24, 48, and 72 h, making the PLA-coated zinc-based material an attractive candidate for biodegradable material design.

Comparison of Plasma-Polymerized Thin Films Deposited from 2-Methyl-2-oxazoline and 2-Ethyl-2-oxazoline: I Film Properties.

Poly(2-oxazoline) is a promising new class of polymeric materials due to their antibiofouling properties and good biocompatibility. Poly(2-oxazoline) coatings can be deposited on different substrates via plasma polymerization, which can be more advantageous than other coating methods. The aim of this study is to deposit poly(2-oxazoline) coatings using a surface dielectric barrier discharge burning in nitrogen at atmospheric pressure using 2-methyl-2-oxazoline and 2-ethyl-2-oxazoline vapours as monomers and compare the film properties. For the comparison, the antibacterial and cytocompatibility tests were peformed according to ISO norms. The antibacterial tests showed that all the deposited films were highly active against Staphylococcus aureus and Escherichia coli bacteria. The chemical composition of the films was studied using FTIR and XPS, and the film surface's properties were studied using AFM and surface energy measurement. The cytocompatibility tests showed good cytocompatibility of all the deposited films. However, the films deposited from 2-methyl-2-oxazoline exhibit better cytocompatibility. This difference can be explained by the different chemical compositions and surface morphologies of the films deposited from different monomers.

Antibacterial Thin Films Deposited from Propane-Butane Mixture in Atmospheric Pressure Discharge.

Antibacterial coatings on biomedical instruments are of great interest because they can suppress bacterial colonization on these instruments. In this study, antibacterial polymeric thin coatings were deposited on teflon substrates using atmospheric pressure plasma polymerization from a propane-butane mixture. The plasma polymerization was performed by means of surface dielectric barrier discharge burning in nitrogen at atmospheric pressure. The chemical composition of plasma polymerized propane-butane films was studied by energy-dispersive X-ray spectroscopy (EDX) and FTIR. The film surface properties were studied by SEM and by surface energy measurement. The EDX analysis showed that the films consisted of carbon, nitrogen and oxygen from ambient air. The FTIR analysis confirmed, in particular, the presence of alkyl, nitrile, acetylene, imide and amine groups. The deposited films were hydrophilic with a water contact angle in the range of 13-23°. The thin film deposited samples were highly active against both S. aureus and E. coli strains in general. On the other hand, the films were cytocompatible, reaching more than 80% of the cell viability threshold compared to reference polystyrene tissue.

Effect of Saccharides Coating on Antibacterial Potential and Drug Loading and Releasing Capability of Plasma Treated Polylactic Acid Films.

More than half of the hospital-associated infections worldwide are related to the adhesion of bacteria cells to biomedical devices and implants. To prevent these infections, it is crucial to modify biomaterial surfaces to develop the antibacterial property. In this study, chitosan (CS) and chondroitin sulfate (ChS) were chosen as antibacterial coating materials on polylactic acid (PLA) surfaces. Plasma-treated PLA surfaces were coated with CS either direct coating method or the carbodiimide coupling method. As a next step for the combined saccharide coating, CS grafted samples were immersed in ChS solution, which resulted in the polyelectrolyte complex (PEC) formation. Also in this experiment, to test the drug loading and releasing efficiency of the thin film coatings, CS grafted samples were immersed into lomefloxacin-containing ChS solution. The successful modifications were confirmed by elemental composition analysis (XPS), surface topography images (SEM), and hydrophilicity change (contact angle measurements). The carbodiimide coupling resulted in higher CS grafting on the PLA surface. The coatings with the PEC formation between CS-ChS showed improved activity against the bacteria strains than the separate coatings. Moreover, these interactions increased the lomefloxacin amount adhered to the film coatings and extended the drug release profile. Finally, the zone of inhibition test confirmed that the CS-ChS coating showed a contact killing mechanism while drug-loaded films have a dual killing mechanism, which includes contact, and release killing.

Furcellaran Surface Deposition and Its Potential in Biomedical Applications.

Surface coatings of materials by polysaccharide polymers are an acknowledged strategy to modulate interfacial biocompatibility. Polysaccharides from various algal species represent an attractive source of structurally diverse compounds that have found application in the biomedical field. Furcellaran obtained from the red algae Furcellaria lumbricalis is a potential candidate for biomedical applications due to its gelation properties and mechanical strength. In the present study, immobilization of furcellaran onto polyethylene terephthalate surfaces by a multistep approach was studied. In this approach, N-allylmethylamine was grafted onto a functionalized polyethylene terephthalate (PET) surface via air plasma treatment. Furcellaran, as a bioactive agent, was anchored on such substrates. Surface characteristics were measured by means of contact angle measurements, X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). Subsequently, samples were subjected to selected cell interaction assays, such as antibacterial activity, anticoagulant activity, fibroblasts and stem cell cytocompatibility, to investigate the Furcellaran potential in biomedical applications. Based on these results, furcellaran-coated PET films showed significantly improved embryonic stem cell (ESC) proliferation compared to the initial untreated material.

False positive effect of sulfur sources used in growing and processing of vine (Vitis Vinifera L.) leaves on the results of dithiocarbamate analysis based on carbon disulfide measurement.

Vine leaves, which are produced fresh, brined or fermented from the leaves of Vitis Vinifera in Türkiye are an important food. Sulfur is used as a pesticide and sulfur compounds can be used as additives during the growing and processing of the vine leaves. These sulfur sources cause positive results on carbon disulfide (CS2) measurements by GC-MS. Therefore, the main objective of the present study was to investigate the effects of residues of sulfur or sulfur compounds on dithiocarbamate analysis methods based on CS2 measurement. For this, vine leaves were produced by controlled agricultural production and processed as brine under controlled conditions. The sulfur dioxide (SO2) and dithiocarbamate analysis were carried out on the vine leave obtained by applying sulfur spraying in agricultural treatments and brined vine leaves produced by adding sodium metabisulfite (SM), and control samples of each stage. SO2 was not detected in any of the samples in this study. SO2 residues did not occur in the vine leaves as a result of the sulfur spraying application and therefore did not have a false positive effect on dithiocarbamate analysis. However, approximately 0.15 mg kg-1 false positive dithiocarbamate was detected, which is thought to originate from natural sulfur in the vine leaves. The effect of SM, which was used in low concentration in the production of brined vine leaves, on dithiocarbamate results was limited. Even if SM was not used, the total false positive dithiocarbamate result in the brined vine leaves production process was approximately determined as 0.20 mg kg-1. This study showed that the dithiocarbamates analysis method based on CS2 measurement may lead to false positive results in brined vine leaves since sulfur compounds are found naturally in vine leaves.

Controlled release of enrofloxacin by vanillin-crosslinked chitosan-polyvinyl alcohol blends.

In transdermal drug delivery applications uniform drug distribution and sustained release are of great importance to decrease the side effects. In this direction in the present research, vanillin crosslinked chitosan (CS) and polyvinyl alcohol (PVA) blend based matrix-type transdermal system was prepared by casting and drying of aqueous solutions for local delivery of enrofloxacin (ENR) drug. Subsequently, the properties including the morphology, chemical structure, thermal behavior, tensile strength, crosslinking degree, weight uniformity, thickness, swelling and drug release of the CS-PVA blend films before and after crosslinking were characterized. In vitro drug release profiles showed the sustained release of ENR by the incorporation of vanillin as a crosslinker into the CS-PVA polymer matrix. Furthermore, the release kinetic profiles revealed that the followed mechanism for all samples was Higuchi and the increase of vanillin concentration in the blend films resulted in the change of diffusion mechanism from anomalous transport to Fickian diffusion. Overall, the obtained results suggest that the investigated vanillin crosslinked CS-PVA matrix-type films are potential candidates for transdermal drug delivery system.

Plasma Mediated Chlorhexidine Immobilization onto Polylactic Acid Surface via Carbodiimide Chemistry: Antibacterial and Cytocompatibility Assessment.

The development of antibacterial materials has great importance in avoiding bacterial contamination and the risk of infection for implantable biomaterials. An antibacterial thin film coating on the surface via chemical bonding is a promising technique to keep native bulk material properties unchanged. However, most of the polymeric materials are chemically inert and highly hydrophobic, which makes chemical agent coating challenging Herein, immobilization of chlorhexidine, a broad-spectrum bactericidal cationic compound, onto the polylactic acid surface was performed in a multistep physicochemical method. Direct current plasma was used for surface functionalization, followed by carbodiimide chemistry to link the coupling reagents of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) and N-Hydroxysuccinimide (NHs) to create a free bonding site to anchor the chlorhexidine. Surface characterizations were performed by water contact angle test, X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM). X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM). The antibacterial activity was tested using Staphylococcus aureus and Escherichia coli. Finally, in vitro cytocompatibility of the samples was studied using primary mouse embryonic fibroblast cells. It was found that all samples were cytocompatible and the best antibacterial performance observed was the Chlorhexidine immobilized sample after NHs activation.

Atmospheric Pressure Plasma Polymerized 2-Ethyl-2-oxazoline Based Thin Films for Biomedical Purposes.

Polyoxazoline thin coatings were deposited on glass substrates using atmospheric pressure plasma polymerization from 2-ethyl-2-oxazoline vapours. The plasma polymerization was performed in dielectric barrier discharge burning in nitrogen at atmospheric pressure. The thin films stable in aqueous environments were obtained at the deposition with increased substrate temperature, which was changed from 20 ∘C to 150 ∘C. The thin film deposited samples were highly active against both S. aureus and E. coli strains in general. The chemical composition of polyoxazoline films was studied by FTIR and XPS, the mechanical properties of films were studied by depth sensing indentation technique and by scratch tests. The film surface properties were studied by AFM and by surface energy measurement. After tuning the deposition parameters (i.e., monomer flow rate and substrate temperature), stable films, which resist bacterial biofilm formation and have cell-repellent properties, were achieved. Such antibiofouling polyoxazoline thin films can have many potential biomedical applications.

Atmospheric Pressure Plasma Polymerized Oxazoline-Based Thin Films-Antibacterial Properties and Cytocompatibility Performance.

Polyoxazolines are a new promising class of polymers for biomedical applications. Antibiofouling polyoxazoline coatings can suppress bacterial colonization of medical devices, which can cause infections to patients. However, the creation of oxazoline-based films using conventional methods is difficult. This study presents a new way to produce plasma polymerized oxazoline-based films with antibiofouling properties and good biocompatibility. The films were created via plasma deposition from 2-methyl-2-oxazoline vapors in nitrogen atmospheric pressure dielectric barrier discharge. Diverse film properties were achieved by increasing the substrate temperature at the deposition. The physical and chemical properties of plasma polymerized polyoxazoline films were studied by SEM, EDX, FTIR, AFM, depth-sensing indentation technique, and surface energy measurement. After tuning of the deposition parameters, films with a capacity to resist bacterial biofilm formation were achieved. Deposited films also promote cell viability.

Chitosan-collagen based film for controlled delivery of a combination of short life anesthetics.

The present research was undertaken to develop a chitosan-collagen film for controlled delivery of combinations of local anesthetics. The film has been prepared by casting which is a versatile, rapid and low-cost approach distinguished by high reproducibility. The mechanical, morphological, and physicochemical properties of the films and the impact of the drug loading were evaluated. We showed that the formulations have a good combination of strength and flexibility with high water permeability. Surface morphology investigation indicates a variation in roughness depending on the loaded compound. Release studies were performed in controlled environments and the data processed by the Higuchi model to assess the dynamics of the release. The local anesthetics, lidocaine, tetracaine, and benzocaine, were uniformly distributed within the matrix and released in a rate and magnitude specific for the drug concentration and combination tunable in a range time from 6 h to 24 h. The films dissolve completely in the physiological environment within 24 h without leaving any toxic metabolites as both of the components are recognized as safe. In vitro cytotoxicity and cell proliferation tests performed on human dermal fibroblast demonstrate the biocompatibility and lack of cytotoxicity of the prepared formulations.

(PVA/Chitosan/Fucoidan)-Ampicillin: A Bioartificial Polymeric Material with Combined Properties in Cell Regeneration and Potential Antibacterial Features.

Chitosan, fucoidan, and polyvinyl alcohol are categorized as polymers with biomedical applications. Ampicillin, on the other hand, is considered as an important antibiotic that has shown effectivity in both gram-positive and gram-negative micro-organisms. The aforementioned polymers possess unique properties that are considered desirable for cell regeneration although they exhibit drawbacks that can affect their final application. Therefore, films of these biomaterials were prepared and they were characterized using FTIR, SEM, XRD, degree of swelling and solubility, and MTT assay. The statistical significance of the experiments was determined using a two-way analysis of variance (ANOVA) with p < 0.05. The characterization techniques demonstrated that the obtained material exhibits properties suitable for cell regeneration, and that a higher concentration of natural polymers promotes cells proliferation to a greater extent. The presence of PVA, on the other hand, is responsible for matrix stability and dictates the degree of swelling and solubility. The SEM images demonstrated that neither aggregations nor clusters were formed, which is favorable for the biological properties without detrimental to the morphological and physical features. Cell viability was comparatively similar in samples with and without antibiotic, and the physical and biological properties were not negatively affected. Indeed, the inherent bactericidal effect of chitosan was reinforced by the presence of ampicillin. The new material is an outstanding candidate for cell regeneration as a consequence of the synergic effect that each component provides to the blend.

Antibacterial Activity and Cytotoxicity of Immobilized Glucosamine/Chondroitin Sulfate on Polylactic Acid Films.

Polylactic acid (PLA) is one of the most produced polymeric materials, due to its exceptional chemical and mechanical properties. Some of them, such as biodegradability and biocompatibility, make them attractive for biomedical applications. Conversely, the major drawback of PLA in the biomedical field is their vulnerability to bacterial contamination. This study focuses on the immobilization of saccharides onto the PLA surface by a multistep approach, with the aim of providing antibacterial features and evaluting the synergistic effect of these saccharides. In this approach, after poly (acrylic acid) (PAA) brushes attached non-covalently to the PLA surface via plasma post-irradiation grafting technique, immobilization of glucosamine (GlcN) and chondroitin sulfate (ChS) to the PAA brushes was carried out. To understand the changes in surface properties, such as chemical composition, surface topography and hydrophilicity, the untreated and treated PLA films were analyzed using various characterization techniques (contact angle, scanning electron microscopy, X-ray photoelectron spectroscopy). In vitro cytotoxicity assays were investigated by the methyl tetrazolium test. The antibacterial activity of the PLA samples was tested against Escherichia coli and Staphylococcus aureus bacteria strains. Plasma-treated films immobilized with ChS and GlcN, separately and in combination, demonstrated bactericidal effect against the both bacteria strains and also the results revealed that the combination has no synergistic effect on antibacterial action.

Anticoagulant Polyethylene Terephthalate Surface by Plasma-Mediated Fucoidan Immobilization.

Biomaterial-based blood clot formation is one of the biggest drawbacks of blood-contacting devices. To avoid blood clot formation, their surface must be tailored to increase hemocompatibility. Most synthetic polymeric biomaterials are inert and lack bonding sites for chemical agents to bond or tailor to the surface. In this study, polyethylene terephthalate was subjected to direct current air plasma treatment to enhance its surface energy and to bring oxidative functional binding sites. Marine-sourced anticoagulant sulphated polysaccharide fucoidan from Fucus vesiculosus was then immobilized onto the treated polyethylene terephthalate (PET) surface at different pH values to optimize chemical bonding behavior and therefore anticoagulant performance. Surface properties of samples were monitored using the water contact angle; chemical analyses were performed by FTIR and X-ray photoelectron spectroscopy (XPS) and their anticoagulant activity was tested by means of prothrombin time, activated partial thromboplastin time and thrombin time. On each of the fucoidan-immobilized surfaces, anticoagulation activity was performed by extending the thrombin time threshold and their pH 5 counterpart performed the best result compared to others.

Polysaccharides based microspheres for multiple encapsulations and simultaneous release of proteases.

The work is focused on the development of microspheres based on the combination of two polysaccharides; chitosan and alginic acid with the aim to allocate, hold, release and protect environmentally sensible molecules. The microspheres were prepared using a solvent-free, low cost and scalable approach and two enzymes; trypsin and protease from Aspergillus Oryzae have been used as a model to evaluate the microspheres peculiarities. The proteins were encapsulated during the microspheres preparation. The relationship between the polysaccharides weight ratio and the morphology, stability and ability of the carrier to allocate the enzymes has been evaluated. The enzymatic activity and the release kinetics were assessed in different conditions to assess the impact of the external environment. Obtained results demonstrate the efficacy of the prepared microspheres to preserve the activity of relevant bioactive compounds which are highly relevant in food, cosmetic and pharmaceutic, but the application is limited due to their high sensibility.

A novel multistep method for chondroitin sulphate immobilization and its interaction with fibroblast cells.

Polymeric biomaterials are widely used in medical applications owing to their low cost, processability and sufficient toughness. Surface modification by creating a thin film of bioactive agents is promising technique to enhance cellular interactions, regulate the protein adsorption and/or avoid bacterial infections. Polyethylene is one of the most used polymeric biomaterial but its hydrophobic nature impedes its further chemical modifications. Plasma treatment is unique method to increase its hydrophilicity by incorporating hydrophilic oxidative functional groups and tailoring the surface by physical etching. Furthermore, grafting of polymer brushes of amine group containing monomers onto the functionalized surface lead to strongly immobilized bioactive agents at the final step. Chondroitin sulphate is natural polysaccharide mainly found in connective cartilage tissue which used as a bioactive agent to immobilize onto polyethylene surface by multistep method in this study.

A New Route of Fucoidan Immobilization on Low Density Polyethylene and Its Blood Compatibility and Anticoagulation Activity.

Beside biomaterials' bulk properties, their surface properties are equally important to control interfacial biocompatibility. However, due to the inadequate interaction with tissue, they may cause foreign body reaction. Moreover, surface induced thrombosis can occur when biomaterials are used for blood containing applications. Surface modification of the biomaterials can bring enhanced surface properties in biomedical applications. Sulfated polysaccharide coatings can be used to avoid surface induced thrombosis which may cause vascular occlusion (blocking the blood flow by blood clot), which results in serious health problems. Naturally occurring heparin is one of the sulfated polysaccharides most commonly used as an anticoagulant, but its long term usage causes hemorrhage. Marine sourced sulfated polysaccharide fucoidan is an alternative anticoagulant without the hemorrhage drawback. Heparin and fucoidan immobilization onto a low density polyethylene surface after functionalization by plasma has been studied. Surface energy was demonstrated by water contact angle test and chemical characterizations were carried out by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Surface morphology was monitored by scanning electron microscope and atomic force microscope. Finally, their anticoagulation activity was examined for prothrombin time (PT), activated partial thromboplastin time (aPTT), and thrombin time (TT).

Mechanical properties, structural and texture evolution of biocompatible Ti-45Nb alloy processed by severe plastic deformation.

Biocompatible ß Ti-45Nb (wt%) alloys were subjected to different methods of severe plastic deformation (SPD) in order to increase the mechanical strength without increasing the low Young׳s modulus thus avoiding the stress shielding effect. The mechanical properties, microstructural changes and texture evolution were investigated, by means of tensile, microhardness and nanoindentation tests, as well as TEM and XRD. Significant increases of hardness and ultimate tensile strength up to a factor 1.6 and 2, respectively, could be achieved depending on the SPD method applied (hydrostatic extrusion - HE, high pressure torsion - HPT, and rolling and folding - R&F), while maintaining the considerable ductility. Due to the high content of ß-stabilizing Nb, the initial lattice structure turned out to be stable upon all of the SPD methods applied. This explains why with all SPD methods the apparent Young׳s modulus measured by nanoindentation did not exceed that of the non-processed material. For its variations below that level, they could be quantitatively related to changes in the SPD-induced texture, by means of calculations of the Young׳s modulus on basis of the texture data which were carefully measured for all different SPD techniques and strains. This is especially true for the significant decrease of Young׳s modulus for increasing R&F processing which is thus identified as a texture effect. Considering the mechanical biocompatibility (percentage of hardness over Young׳s modulus), a value of 3-4% is achieved with all the SPD routes applied which recommends them for enhancing ß Ti-alloys for biomedical applications.