Injectable and photocurable macromonomers synthesized using a heterometallic magnesium-titanium metal-organic catalyst for elastomeric polymer networks.

Injectable and in situ photocurable biomaterials are receiving a lot of attention due to their ease of application via syringe or dedicated applicator and ability to be used in laparoscopic and robotic minimally invasive procedures. The aim of this work was to synthesize photocurable ester-urethane macromonomers using a heterometallic magnesium-titanium catalyst, magnesium-titanium(iv) butoxide for elastomeric polymer networks. The progress of the two-step synthesis of macromonomers was monitored using infrared spectroscopy. The chemical structure and molecular weight of the obtained macromonomers were characterized using nuclear magnetic resonance spectroscopy and gel permeation chromatography. The dynamic viscosity of the obtained macromonomers was evaluated by a rheometer. Next, the photocuring process was studied under both air and argon atmospheres. Both the thermal and dynamic mechanical thermal properties of the photocured soft and elastomeric networks were investigated. Finally, in vitro cytotoxicity screening of polymer networks based on ISO10993-5 revealed high cell viability (over 77%) regardless of curing atmosphere. Overall, our results indicate that this heterometallic magnesium-titanium butoxide catalyst can be an attractive alternative to commonly used homometallic catalysts for the synthesis of injectable and photocurable materials for medical applications.

Enhancement of neutrophil chemotaxis by trans-anethole-treated Staphylococcus aureus strains.

This study aimed to analyze the chemotactic response of differentiated HL-60 neutrophil-like (dHL-60) cells to trans-anethole (TA)-treated Staphylococcus aureus strains. Special attention was paid to evaluate the influence of TA on the chp gene expression level, as well as molecular docking and molecular dynamics (MD) simulation studies on interactions of TA with chemotaxis inhibitory protein of S. aureus (CHIPS). The following parameters were studied: susceptibility to TA using the agar diffusion method, the chp gene detection and its expression under TA influence, and clonal diversity of S. aureus strains using molecular techniques. Furthermore, a chemotactic response of dHL-60 cells to TA-treated S. aureus using Boyden chamber assay was detected and molecular modeling using both the docking methodology and unbiased MD simulations was conducted. It was found that TA showed antibacterial activity against all strains. Three genotypes and one unique pattern were distinguished among the strains. 50% of the isolates were chp-positive. It was observed that TA reduced/inhibited chp gene expression in most S. aureus strains. Enhanced chemotactic response of dHL-60 cells to TA-treated S. aureus strains was also noted. This correlation was similar for both chp-positive and chp-negative strains. Both molecular docking and MD simulations studies confirmed that TA is preferentially bound in the complement component 5a/CHIPS interface interaction region and can interfere with any processes exploiting this binding cavity. It has been proven that dHL-60 cells exhibited a higher chemotactic response to TA-treated S. aureus strains in comparison to non-treated bacteria, regardless of the achieved expression of the chp gene or its lack. Nevertheless, further analyses are required to understand this mechanism better.

Enhancement of Inhibition of the Pseudomonas sp. Biofilm Formation on Bacterial Cellulose-Based Wound Dressing by the Combined Action of Alginate Lyase and Gentamicin.

Bacterial biofilms generally contribute to chronic infections, including wound infections. Due to the antibiotic resistance mechanisms protecting bacteria living in the biofilm, they are a serious problem in the wound healing process. To accelerate the wound healing process and avoid bacterial infection, it is necessary to select the appropriate dressing material. In this study, the promising therapeutic properties of alginate lyase (AlgL) immobilised on BC membranes for protecting wounds from Pseudomonas aeruginosa infection were investigated. The AlgL was immobilised on never dried BC pellicles via physical adsorption. The maximum adsorption capacity of AlgL was 6.0 mg/g of dry BC, and the equilibrium was reached after 2 h. The adsorption kinetics was studied, and it has been proven that the adsorption was consistent with Langmuir isotherm. In addition, the impact of enzyme immobilisation on bacterial biofilm stability and the effect of simultaneous immobilisation of AlgL and gentamicin on the viability of bacterial cells was investigated. The obtained results showed that the AlgL immobilisation significantly reduced the amount of polysaccharides component of the P. aeruginosa biofilm. Moreover, the biofilm disruption by AlgL immobilised on BC membranes exhibited synergism with the gentamicin, resulting in 86.5% more dead P. aeruginosa PAO-1 cells.

Argon plasma-modified bacterial cellulose filters for protection against respiratory pathogens.

In this work, we present novel, sustainable filters based on bacterial cellulose (BC) functionalized with low-pressure argon plasma (LPP-Ar). The "green" production process involved BC biosynthesis by Komagataeibacter xylinus, followed by simple purification, homogenization, lyophilization, and finally LPP-Ar treatment. The obtained LPP-Ar-functionalized BC-based material (LPP-Ar-BC-bM) showed excellent antimicrobial and antiviral properties against both Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria, and an enveloped bacteriophage phage Φ6, with no cytotoxicity versus murine fibroblasts in vitro. Further, filters consisting of three layers of LPP-Ar-BC-bM had >99 % bacterial and viral filtration efficiency, while maintaining sufficiently low airflow resistance (6 mbar at an airflow of 95 L/min). Finally, as a proof-of-concept, we were able to prepare 80 masks with LPP-Ar-BC-bM filter and ~85 % of volunteer medical staff assessed them as "good" or "very good" in terms of comfort. We conclude that our novel sustainable, biobased, biodegradable filters are suitable for respiratory personal protective equipment (PPE), such as surgical masks and respirators.

Alginate Biofunctional Films Modified with Melanin from Watermelon Seeds and Zinc Oxide/Silver Nanoparticles.

Bioactive films find more and more applications in various industries, including packaging and biomedicine. This work describes the preparation, characterization and physicochemical, antioxidant and antimicrobial properties of alginate films modified with melanin from watermelon (Citrullus lanatus) seeds at concentrations of 0.10%, 0.25% and 0.50% w/w and with silver and zinc oxide nanoparticles (10 mM film casting solutions for both metal nanoparticles). Melanin served as the active ingredient of the film and as a nanoparticle stabilizer. The additives affected the color, antioxidant (~90% ABTS and DPPH radicals scavenging for all melanin modified films) and antimicrobial activity (up to 4 mm grow inhibition zones of E. coli and S. aureus for both zinc oxide and silver nanoparticles), mechanical (silver nanoparticles addition effected two-fold higher tensile strength), thermal and barrier properties for water and UV-vis radiation. The addition of ZnONP resulted in improved UV barrier properties while maintaining good visible light transmittance, whereas AgNP resulted in almost complete UV barrier and reduced visible light transmittance of the obtained films. What is more, the obtained films did not have an adverse effect on cell viability in cytotoxicity screening. These films may have potential applications in food packaging or biomedical applications.

Silicone polyether surfactant enhances bacterial cellulose synthesis and water holding capacity.

The versatility and unique properties of bacterial cellulose (BC) motivate research into enhancing its synthesis. Here a silicone polyether surfactant (SPS) was synthesized and tested as a non-nutritional additive to the cultivation media of Komagataeibacter xylinus. The addition of SPS to the Hestrin-Schramm (HS) medium resulted in a concentration-dependent decrease in surface tension from 59.57 ± 0.37 mN/m to 30.05 ± 0.41 mN/m (for 0.1% addition) that was correlated with an increased yield of BC, up to 37% wet mass for surfactant concentration close to its critical micelle concentration (0.008%). Physicochemical characterization of bacterial cellulose obtained in presence of SPS, showed that surfactant is not incorporated into BC structure and has a moderate effect on its crystallinity, thermal stability. Moreover, the water holding capacity was enhanced by over 40%. Importantly, obtained BC did not affect L929 murine fibroblast cell viability. We conclude that SPS provides an eco-friendly approach to increasing BC yield in static culture, enabling more widespread industrial and biomedical applications.

Novel Naproxen Salts with Increased Skin Permeability.

The paper presents the synthesis, full identification, and characterization of new salts-L-proline alkyl ester naproxenates [ProOR][NAP], where R was a chain from ethyl to butyl (including isopropyl). All obtained compounds were characterized by Nuclear Magnetic Resonance (NMR), Fourier transform infrared spectroscopy (FTIR), X-ray powder diffractometry (XRD), and in vitro dissolution studies. The specific rotation, phase transition temperatures (melting point), and thermal stability were also determined. In addition, their lipophilicity, permeability, and accumulation in pigskin were determined. Finally, toxicity against mouse L929 fibroblast cells was tested. The obtained naproxen derivatives showed improved solubility and higher absorption of drug molecules by biological membranes. Their lipophilicity was lower and increased with the increase in the alkyl chain of the ester. The derivative with isopropyl ester had the best permeability through pigskin. The use of L-proline isopropyl ester naproxenate increased the permeation of naproxen through the skin almost four-fold. It was also shown that the increase in permeability is not associated with additional risk: all compounds had a similar effect on cell viability as the parent naproxen.

Potato Juice, a Starch Industry Waste, as a Cost-Effective Medium for the Biosynthesis of Bacterial Cellulose.

In this work, we verified the possibility of valorizing a major waste product of the potato starch industry, potato tuber juice (PJ). We obtained a cost-effective, ecological-friendly microbiological medium that yielded bacterial cellulose (BC) with properties equivalent to those from conventional commercial Hestrin-Schramm medium. The BC yield from the PJ medium (>4 g/L) was comparable, despite the lack of any pre-treatment. Likewise, the macro- and microstructure, physicochemical parameters, and chemical composition showed no significant differences between PJ and control BC. Importantly, the BC obtained from PJ was not cytotoxic against fibroblast cell line L929 in vitro and did not contain any hard-to-remove impurities. The PJ-BC soaked with antiseptic exerted a similar antimicrobial effect against Staphylococcus aureus and Pseudomonas aeruginosa as to BC obtained in the conventional medium and supplemented with antiseptic. These are very important aspects from an application standpoint, particularly in biomedicine. Therefore, we conclude that using PJ for BC biosynthesis is a path toward significant valorization of an environmentally problematic waste product of the starch industry, but also toward a significant drop in BC production costs, enabling wider application of this biopolymer in biomedicine.

Evaluation of Chemical Changes in Laboratory-Induced Colistin-Resistant Klebsiella pneumoniae.

This study evaluates the electrical potential and chemical alterations in laboratory-induced colistin-resistant Klebsiella pneumoniae, as compared to the susceptible strain using spectroscopic analyses. The minimal inhibitory concentration (MIC) of colistin, ζ-potential and chemical composition analysis of K. pneumoniae strains are determined. The results obtained for the K. pneumoniaeCol-R with induced high-level colistin resistance (MIC = 16.0 ± 0.0 mg/L) are compared with the K. pneumoniaeCol-S strain susceptible to colistin (MIC = 0.25 ± 0.0 mg/L). Fourier transform infrared (FTIR) and Raman spectroscopic studies revealed differences in bacterial cell wall structures and lipopolysaccharide (LPS) of K. pneumoniaeCol-R and K. pneumoniaeCol-S strains. In the beginning, we assumed that the obtained results could relate to a negative charge of the bacterial surface and different electrostatic interactions with cationic antibiotic molecules, reducing the affinity of colistin and leading to its lower penetration into K. pneumoniaeCol-R cell. However, no significant differences in the ζ-potential between the K. pneumoniaeCol-R and K. pneumoniaeCol-S strains are noticed. In conclusion, this mechanism is most probably associated with recognisable changes in the chemical composition of the K. pneumoniaeCol-R cell wall (especially in LPS) when compared to the susceptible strain.

Superabsorbent crosslinked bacterial cellulose biomaterials for chronic wound dressings.

In this work, we present a novel ex situ modification of bacterial cellulose (BC) polymer, that significantly improves its ability to absorb water after drying. The method involves a single inexpensive and easy-to-perform process of BC crosslinking, using citric acid along with catalysts, such as disodium phosphate, sodium bicarbonate, ammonium bicarbonate or their mixtures. In particular, the mixture of disodium phosphate and sodium bicarbonate was the most promising, yielding significantly greater water capacity (over 5 times higher as compared to the unmodified BC) and slower water release (over 6 times as compared to the unmodified BC). Further, our optimized crosslinked BC had over 1.5x higher water capacity than modern commercial dressings dedicated to highly exuding wounds, while exhibiting no cytotoxic effects against fibroblast cell line L929 in vitro. Therefore, our novel BC biomaterial may find application in super-absorbent dressings, designed for chronic wounds with imbalanced moisture level.

Whey Protein Concentrate/Isolate Biofunctional Films Modified with Melanin from Watermelon (Citrullus lanatus) Seeds.

Valorization of food industry waste and plant residues represents an attractive path towards obtaining biodegradable materials and achieving "zero waste" goals. Here, melanin was isolated from watermelon (Citrullus lanatus) seeds and used as a modifier for whey protein concentrate and isolate films (WPC and WPI) at two concentrations (0.1% and 0.5%). The modification with melanin enhanced the ultraviolet (UV) blocking, water vapor barrier, swelling, and mechanical properties of the WPC/WPI films, in addition to affecting the apparent color. The modified WPC/WPI films also exhibited high antioxidant activity, but no cytotoxicity. Overall, the effects were melanin concentration-dependent. Thus, melanin from watermelon seeds can be used as a functional modifier to develop bioactive biopolymer films with good potential to be exploited in food packaging and biomedical applications.

Glycoside hydrolase (PelAh) immobilization prevents Pseudomonas aeruginosa biofilm formation on cellulose-based wound dressing.

Bacterial cellulose (BC) is recognized as a wound dressing material well-suited for chronic wounds; however, it has no intrinsic antimicrobial activity. Further, the formation of biofilms can limit the effectiveness of the pre-saturation of BC with antimicrobial agents. Here, to hinder biofilm formation by P. aeruginosa, we immobilized the hydrolytic domain of PelA (a glycohydrolase involved in the synthesis of biofilm polysaccharide Pel) on the surface of BC. The immobilization of 32.35 ±â€¯1.05 mg PelAh per g BC membrane resulted in an eight-fold higher P. aeruginosa cell detachment from BC membrane, indicating reduced biofilm matrix stability. Further, 1D and 2D infrared spectroscopy analysis indicated systematic reduction of polysaccharide biofilm elements, confirming the specificity of immobilized PelAh. Importantly, BC-PelAh was not cytotoxic towards L929 fibroblast cells. Thus, we conclude that PelAh can be used in BC wound dressings for safe and specific protection against biofilm formation by P. aeruginosa.

Architectured helically coiled scaffolds from elastomeric poly(butylene succinate) (PBS) copolyester via wet electrospinning.

Electrospinning is one of the most investigated methods used to produce polymeric fiber scaffolds that mimic the morphology of native extracellular matrix. These structures have been extensively studied in the context of scaffolds for tissue regeneration. However, the compactness of materials obtained by traditional electrospinning, collected as two-dimensional non-woven scaffolds, can limit cell infiltration and tissue ingrowth. In addition, for applications in smooth muscle tissue engineering, highly elastic scaffolds capable of withstanding cyclic mechanical strains without suffering significant permanent deformations are preferred. In order to address these challenges, we report the fabrication of microscale 3D helically coiled scaffolds (referred as 3D-HCS) by wet-electrospinning method, a modification of the traditional electrospinning process in which a coagulation bath (non-solvent system for the electrospun material) is used as the collector. The present study, for the first time, successfully demonstrates the feasibility of using this method to produce various architectures of 3D helically coiled scaffolds (HCS) from segmented copolyester of poly (butylene succinate-co-dilinoleic succinate) (PBS-DLS), a thermoplastic elastomer. We examined the role of process parameters and propose a mechanism for the HCS formation. Fabricated 3D-HCS showed high specific surface area, high porosity, and good elasticity. Further, the marked increase in cell proliferation on 3D-HCS confirmed the suitability of these materials as scaffolds for soft tissue engineering.

Double-Emulsion Copolyester Microcapsules for Sustained Intraperitoneal Release of Carboplatin.

Despite on-going medical advances, ovarian cancer survival rates have stagnated. In order to improve IP delivery of platinum-based antineoplastics, we aimed to develop a sustained drug delivery system for carboplatin (CPt). Toward this aim, we pursued a double emulsion process for obtaining CPt-loaded microcapsules composed of poly(ethylene terephthalate-ethylene dilinoleate) (PET-DLA) copolymer. We were able to obtain PET-DLA microspheres in the targeted size range of 10-25 µm (median: 18.5 µm), to reduce intraperitoneal clearance by phagocytosis and lymphoid transit. Empty microspheres showed the lack of toxicity in vitro. The double emulsion process yielded 2.5% w/w CPt loading and obtained microcapsules exhibited sustained (>20 day) zero-order release. The encapsulated CPt was confirmed to be bioavailable, as the microcapsules demonstrated efficacy against human ovarian adenocarcinoma (SK-OV-3) cells in vitro. Following intraperitoneal injection in mice, we did not observe adhesions, only mild, clinically-insignificant, local inflammatory response. Tissue platinum levels, monitored over 14 days using atomic absorption spectroscopy, revealed low burst and reduced systemic uptake (plasma, kidney), as compared to neat carboplatin injection. Overall, the results demonstrate the potential of the developed microencapsulation system for long-term intraperitoneal sustained release of carboplatin for the treatment of ovarian cancer.

Biofunctional catheter coatings based on chitosan-fatty acids derivatives.

Multifunctional and biofunctional coatings for medical devices are an attractive strategy towards tailoring the interactions of the device with the body, thereby influencing the host response, and the susceptibility to microbial colonization. Here we describe the development of a coating process to yield amphiphilic, lubricious coatings, resistant to bacterial colonization, based on chitosan. Chitosan-fatty acid derivatives were obtained by simultaneous N,O-acylation of chitosan with either linoleic, α-linolenic, or dilinoleic acid. Chemical characterization of new materials was carried out using 1H NMR, FTIR, and XPS. Surface properties of coated polyester samples were studied using SEM and contact angle measurements, which indicated that the incorporation of hydrophobic constituents into chitosan macromolecules led to a decrease of both surface roughness and water contact angle. Importantly, tribological testing demonstrated that these new coatings decrease the coefficient of friction due to the self-organization of fatty acid (from 0.53 for the neat chitosan to 0.35 for chitosan-fatty acid derivative). Meanwhile, preliminary bacterial colonization tests indicated significant-over 80%-reduction in E. coli colonization following coating with chitosan-linoleic and chitosan-α-linolenic derivatives. Finally, cytotoxicity and hemocompatibility studies confirmed that all amphiphilic chitosan-fatty acid derivatives were non-toxic and non-hemolytic. Collectively, our results demonstrate the potential of the developed coating strategy, particularly the chitosan-linoleic and chitosan-α-linolenic acid derivatives, for applications as biofunctional catheter coatings.

Bio-Based PBT-DLA Copolyester as an Alternative Compatibilizer of PP/PBT Blends.

The aim of this work was to assess whether synthesized random copolyester, poly(butylene terephthalate-r-butylene dilinoleate) (PBT-DLA), containing bio-based components, can effectively compatibilize polypropylene/poly(butylene terephthalate) (PP/PBT) blends. For comparison, a commercial petrochemical triblock copolymer, poly(styrene-b-ethylene/butylene-b-styrene) (SEBS) was used. The chemical structure and block distribution of PBT-DLA was determined using nuclear magnetic resonance spectroscopy and gel permeation chromatography. PP/PBT blends with different mass ratios were prepared via twin-screw extrusion with 5 wt% of each compatibilizer. Thermogravimetric analysis, differential scanning calorimetry, and dynamic mechanical analysis were used to assess changes in phase structure of PP/PBT blends. Static tensile testing demonstrated marked improvement in elongation at break, to ~18% and ~21% for PBT-DLA and SEBS, respectively. Importantly, the morphology of PP/PBT blends compatibilized with PBT-DLA copolymer showed that it is able to act as interphase modifier, being preferentially located at the interface. Therefore, we conclude that by using polycondensation and monomers from renewable resources, it is possible to obtain copolymers that efficiently modify blend miscibility, offering an alternative to widely used, rubber-like petrochemical styrene compatibilizers.

Adsorption of Fibrinogen and Fibronectin on Elastomeric Poly(butylene succinate) Copolyesters.

Proteins adsorbed onto biomaterial surfaces facilitate cell-material interactions, including adhesion and migration. Of particular importance are provisional matrix components, fibrinogen (Fg) and fibronectin (Fn), which play an important role in the wound-healing process. Here, to assess the potential of a series of elastomeric poly(butylene succinate) (PBS) copolymers for soft tissue engineering and regenerative medicine applications, we examined the adsorption of Fg and Fn. We prepared spin-coated thin films of the poly(butylene succinate) homopolymer and a series of elastomeric poly(butylene succinate) copolymers with butylene succinate (PBS, hard segment) to succinate-dimer linoleic diol unit (dilinoleic succinate (DLS), soft segments) weight ratios of 70:30, 60:40, and 50:50. X-ray diffraction was used to assess crystallinity, whereas the obtained thin films were characterized using a quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy. Protein adsorption was assessed using QCM-D, followed by data analysis using viscoelastic modeling. On all three copolymers, we observed robust adsorption of both key provisional matrix proteins. Importantly, for both proteins, viscoelastic modeling determined that the adlayers were 30-40 nm thick and had low shear modulus values (<25 kPa), thus indicating soft orientations (end-on for Fg) or conformations (open for Fn) of the hydrated proteins. Overall, our results are very encouraging, as they predict excellent cell adhesion and migration, key features enabling tissue integration of potential PBS-DLS scaffolds.

Enzymatic Degradation of Poly(butylene succinate) Copolyesters Synthesized with the Use of Candida antarctica Lipase B.

Biodegradable polymers are an active area of investigation, particularly ones that can be produced from sustainable, biobased monomers, such as copolymers of poly(butylene succinate) (PBS). In this study, we examine the enzymatic degradation of poly(butylene succinate-dilinoleic succinate) (PBS-DLS) copolymers obtained by "green" enzymatic synthesis using lipase B from Candida antarctica (CALB). The copolymers differed in their hard to soft segments ratio, from 70:30 to 50:50 wt %. Enzymatic degradation was carried out on electrospun membranes (scaffolds) and compression-moulded films using lipase from Pseudomomas cepacia. Poly(ε-caprolactone) (PCL) was used as a reference aliphatic polyester. The degradation process was monitored gravimetrically via water uptake and mass loss. After 24 days, approx. 40% mass loss was observed for fibrous materials prepared from the PBS-DLS 70:30 copolymer, as compared to approx. 10% mass loss for PBS-DLS 50:50. Infrared spectroscopy (FTIR) and size exclusion chromatography (SEC) analysis were used to examine changes in chemical structure. Differential scanning calorimetry (DSC) and scanning light microscopy (LSM) revealed changes in degree of crystallinity, and changes in surface morphology, consistent with a surface erosion mechanism. We conclude that the obtained copolymers are suitable for tissue engineering applications thanks to tuneable degradation and lack of acidification during breakdown.

A biofunctionalizable ink platform composed of catechol-modified chitosan and reduced graphene oxide/platinum nanocomposite.

We present an ink platform for a printable polymer-graphene nanocomposite that is intended for the development of modular biosensors. The ink consists of catechol-modified chitosan and reduced graphene oxide decorated with platinum nanoparticles (rGO-Pt). We modified the chitosan with catechol groups, in order to obtain adhesive properties and improve solubility. Dispersions of rGO-Pt in ethylene glycol were admixed with an aqueous solution of modified chitosan to yield an ink that is suitable for non-contact piezoelectric printing using a commercial microplotter (Sonoplot GIX Microplotter Desktop). As a proof of concept, printed patterns were biofunctionalized with DNA oligonucleotide probes for Streptococcus agalactiae (Group B streptococcus) using glutaraldehyde as a linker. Confocal microscopy revealed the successful hybridization of complementary polymerase chain reaction (PCR) products and low non-specific binding. Our results demonstrate that catechol-modified chitosan/rGO-Pt nanocomposites can be used as inks for piezoelectric printing and facilitate the attachment of biorecognition elements for biosensor applications.

The effect of carbon nanoparticles on biological properties of polyester nanocomposites.

The aim of present study was to determine the hemocompatibility, cellular response of endothelial cells and bacterial adhesion to a new polyester nanocomposite. The carbon nanoparticle nanocomposite was prepared via in situ polymerization of monomers to obtain material of hardness 55 Sh D similar to polyurethanes used in medical applications, for example, in heart-assisting devices. The carbon nanoparticle-containing polyester exhibits markedly reduced bacterial colonization, as compared to commercially available polyurethanes. Further the nanocomposite possesses markedly improved hemocompatibility, as determined by flow cytometry, and robust endothelialization. Possible explanations for these beneficial properties include surface nanoroughness of carbon nanoparticle-containing nanocomposites and presence of fatty acid sequences within polymer structure.