Purification of Natural Pigments Violacein and Deoxyviolacein Produced by Fermentation Using Yarrowia lipolytica.

Violacein and deoxyviolacein are bis-indole pigments synthesized by a number of microorganisms. The present study describes the biosynthesis of a mixture of violacein and deoxyviolacein using a genetically modified Y. lipolytica strain as a production chassis, the subsequent extraction of the intracellular pigments, and ultimately their purification using column chromatography. The results show that the optimal separation between the pigments occurs using an ethyl acetate/cyclohexane mixture with different ratios, first 65:35 until both pigments were clearly visible and distinguishable, then 40:60 to create a noticeable separation between them and recover the deoxyviolacein, and finally 80:20, which allows the recovery of the violacein. The purified pigments were then analyzed by thin-layer chromatography and nuclear magnetic resonance.

Multiscale-Engineered Muscle Constructs: PEG Hydrogel Micro-Patterning on an Electrospun PCL Mat Functionalized with Gold Nanoparticles.

The development of new, viable, and functional engineered tissue is a complex and challenging task. Skeletal muscle constructs have specific requirements as cells are sensitive to the stiffness, geometry of the materials, and biological micro-environment. The aim of this study was thus to design and characterize a multi-scale scaffold and to evaluate it regarding the differentiation process of C2C12 skeletal myoblasts. The significance of the work lies in the microfabrication of lines of polyethylene glycol, on poly(ε-caprolactone) nanofiber sheets obtained using the electrospinning process, coated or not with gold nanoparticles to act as a potential substrate for electrical stimulation. The differentiation of C2C12 cells was studied over a period of seven days and quantified through both expression of specific genes, and analysis of the myotubes' alignment and length using confocal microscopy. We demonstrated that our multiscale bio-construct presented tunable mechanical properties and supported the different stages skeletal muscle, as well as improving the parallel orientation of the myotubes with a variation of less than 15°. These scaffolds showed the ability of sustained myogenic differentiation by enhancing the organization of reconstructed skeletal muscle. Moreover, they may be suitable for applications in mechanical and electrical stimulation to mimic the muscle's physiological functions.

Diazonium Salt-Based Surface-Enhanced Raman Spectroscopy Nanosensor: Detection and Quantitation of Aromatic Hydrocarbons in Water Samples.

Here, we present a surface-enhanced Raman spectroscopy (SERS) nanosensor for environmental pollutants detection. This study was conducted on three polycyclic aromatic hydrocarbons (PAHs): benzo[a]pyrene (BaP), fluoranthene (FL), and naphthalene (NAP). SERS substrates were chemically functionalized using 4-dodecyl benzenediazonium-tetrafluoroborate and SERS analyses were conducted to detect the pollutants alone and in mixtures. Compounds were first measured in water-methanol (9:1 volume ratio) samples. Investigation on solutions containing concentrations ranging from 10-6 g L-1 to 10-3 g L-1 provided data to plot calibration curves and to determine the performance of the sensor. The calculated limit of detection (LOD) was 0.026 mg L-1 (10-7 mol L-1) for BaP, 0.064 mg L-1 (3.2 × 10-7 mol L-1) for FL, and 3.94 mg L-1 (3.1 × 10-5 mol L-1) for NAP, respectively. The correlation between the calculated LOD values and the octanol-water partition coefficient (Kow) of the investigated PAHs suggests that the developed nanosensor is particularly suitable for detecting highly non-polar PAH compounds. Measurements conducted on a mixture of the three analytes (i) demonstrated the ability of the developed technology to detect and identify the three analytes in the mixture; (ii) provided the exact quantitation of pollutants in a mixture. Moreover, we optimized the surface regeneration step for the nanosensor.

Tetrazine Click Chemistry for the Modification of 1-Hydroxy-1,1-methylenebisphosphonic Acids: Towards Bio-orthogonal Functionalization of Gold Nanoparticles.

Inverse electron demand Diels-Alder (iEDDA) was evaluated for the functionalization of gold nanoparticles. The reaction was first modelled with the free coating molecule 1-hydroxy-1,1-methylenebisphosphonate bearing an alkene functionality (HMBPene). A model tetrazine 3,6-dipyridin-2-yl-1,2,4,5-tetrazine (pyTz) was used, kinetic of the reaction was calculated and coupling products were analysed by NMR and HRMS. The reaction was then transposed at the nanoparticle surface. Gold nanoparticles bearing an alkene functionality were obtained using a one-pot methodology with HMBPene and the tetrazine click chemistry was evaluated at their surface using pyTz. The successful coupling was assessed by XPS measurements. This click-methodology was extended to the conjugation of a NIR probe at the NP surface.

Pd nanoparticles embedded in nanolignin (Pd@LNP) as a water dispersible catalytic nanoreactor for Cr(VI), 4-nitrophenol reduction and CC coupling reactions.

The use of water-dispersible and sustainable Pd nanocatalysts to reduce toxic heavy metal ions and catalyze important organic reactions has profound significance for the environmental remediation and the catalytic industry. In this work, a novel water-dispersible and recyclable Pd@LNPs nanoreactor composed of Pd nanoparticle cluster core and LNPs shell was developed in microwave reactor in aqueous solution. It turned out that Pd nanoparticles grew uniformly and stably inside LNPs nanosphere due to the coordinated binding and interaction between Pd and the functional groups in LNPs, which was significantly different from surface loading. The green and biodegradable LNPs nanospheres are not only used as reducing agents for Pd (II) and nanocarriers, but also act as individual nanocontainers to provide favorable sites for reactions and effectively control the entry and release of reactants and products. Furthermore, the excellent and efficient catalytic properties of Pd@LNPs were exhibited by CC coupling reactions and the reduction of Cr(VI) and 4-nitrophenol. The Pd@LNPs prepared in this study have the advantages of excellent dispersion, great recyclability, high turnover frequency and better green sustainability metrics. It will have a great significance for the development of the potential high-value of lignin and the progress in the field of bio-nanocatalysts.

Nanoparticles under the light: click functionalization by photochemical thiol-yne reaction, towards double click functionalization.

A light click away: The first application of the thiol-yne reaction to nanoparticle functionalization is described (see figure). This metal-free click chemistry approach is compatible with the addition of various molecules at the surface and can be combined with CuAAC methodology to perform chemoselective double functionalization.

Carbodiimide versus click chemistry for nanoparticle surface functionalization: a comparative study for the elaboration of multimodal superparamagnetic nanoparticles targeting αvß3 integrins.

Superparamagnetic fluorescent nanoparticles targeting αvß3 integrins were elaborated using two methodologies: carbodiimide coupling and click chemistries (CuACC and thiol-yne). The nanoparticles are first functionalized with hydroxymethylenebisphonates (HMBP) bearing carboxylic acid or alkyne functions. Then, a large number of these reactives functions were used for the covalent coupling of dyes, poly(ethylene glycol) (PEG), and cyclic RGD. Several methods were used to characterize the nanoparticle surface functionalization, and the magnetic properties of these contrast agents were studied using a 1.5 T clinical MRI. The affinity toward integrins was evidenced by solid-phase receptor-binding assay. In addition to their chemoselective natures, click reactions were shown to be far more efficient than the carbodiimide coupling. The grafting increase was shown to enhance targeting affinity to integrin without imparing MRI and fluorescent properties.

Microwave-Assisted Synthesis of Pd Nanoparticles into Wood Block (Pd@wood) as Efficient Catalyst for 4-Nitrophenol and Cr(VI) Reduction.

Palladium (Pd) nanoparticle catalysis has attracted increasing attention due to its efficient catalytic activity and its wide application in environmental protection and chemical synthesis. In this work, Pd nanoparticles (about 71 nm) were synthesized in aqueous solution by microwave-assisted thermal synthesis and immobilized in beech wood blocks as Pd@wood catalysts. The wood blocks were first hydrothermally treated with 10% NaOH solution to improve the internal structure and increase their porosity, thereby providing favorable attachment sites for the formed Pd nanoparticles. The stable deposition of Pd nanoparticle clusters on the internal channels of the wood blocks can be clearly observed. In addition, the catalytic performance of the prepared Pd@wood was investigated through two model reactions: the reduction of 4-nitrophenol and Cr(VI). The Pd@wood catalyst showed 95.4 g-1 s-1 M-1 of normalized rate constant knorm and 2.03 min-1 of the TOF, respectively. Furthermore, Pd nanoparticles are integrated into the internal structure of wood blocks by microwave-assisted thermal synthesis, which is an effective method for wood functionalization. It benefits metal nanoparticle catalysis in the synthesis of fine chemicals as well as in industrial wastewater treatment.

Additive manufacturing of microstructured reactors for organometallic catalytic reactions.

The use of Additive Manufacturing for the fabrication of chemical reactors for flow chemistry is a promising field as it can lead to several improvements over more standard equipment. In this work, two different reactors were fabricated and compared: a Honeycomb monolith reactor with straight channels and a Periodic Open Cell Structure reactor. The Honeycomb monolith reactor was used as an example of a standard reactor (not necessarily additive manufactured) while the Periodic Open Cell Structure is a promising new type of reactor, which improves some key features, such as contact surface area and porosity. The two reactors were manufactured by Stereolithography technology with a high temperature resin and their internal surfaces were chemically activated by the grafting of palladium. For the surface activation, a two-step procedure was developed, firstly using NaOH and in a second step an aqueous solution of Na2PdCl4. After activation, a heterogeneous catalytic reaction was used to characterize the performance of the two fabricated reactors. The chosen reaction was the Suzuki-Miyaura reaction, which is commonly used in the pharmaceutical industry. The experimental results showed that, for equal contact surface area, the new designed reactor had better performance compared to the standard geometry.

Fluid Mechanics of Droplet Spreading of Chitosan/PVA-Based Spray Coating Solution on Banana Peels with Different Wettability.

The spreading behavior of a coating solution is an important factor in determining the effectiveness of spraying applications. It determines how evenly the droplets spread on the substrate surface and how quickly they form a uniform film. Fluid mechanics principles govern it, including surface tension, viscosity, and the interaction between the liquid and the solid surface. In our previous work, chitosan (CS) film properties were successfully modified by blending with polyvinyl alcohol (PVA). It was shown that the mechanical strength of the composite film was significantly improved compared to the virgin CS. Here we propose to study the spreading behavior of CS/PVA solution on fresh bananas. The events upon droplet impact were captured using a high-speed camera, allowing the identification of outcomes as a function of velocity at different surface wettabilities (wetting and non-wetting) on the banana peels. The mathematical model to predict the maximum spreading factor, ßmax, was governed by scaling law analysis using fitting experimental data to identify patterns, trends, and relationships between ßmax and the independent variables, Weber (We) numbers, and Reynolds (Re) numbers. The results indicate that liquid viscosity and surface properties affect the droplet's impact and spreading behavior. The Ohnesorge (Oh) numbers significantly influenced the spreading dynamics, while the banana's surface wettability minimally influenced spreading. The prediction model reasonably agrees with all the data in the literature since the R2 = 0.958 is a powerful goodness-of-fit indicator for predicting the spreading factor. It scaled with ßmax=a+0.04We.Re1/3, where the "a" constants depend on Oh numbers.

A continuous flow mode with a scalable tubular reactor for the green preparation of stable alkali lignin nanoparticles assisted by ultrasound.

Lignin nanoparticles (LNPs) have become a hot topic recently because of their improved physicochemical properties and the excellent integration into various industrial sectors compared to lignin. However, the green large-scale production of stable LNPs severely restricts the high-value applications of LNPs. In this work, a simple and potentially scalable continuous-flow mode setup with a tubular flow reactor was designed for the green preparation of stable alkali LNPs assisted by ultrasound. When the flow rates of lignin solution and nitric acid solution were 8.00 mL/min and 2.67 mL/min respectively, and the length of the tube was 5.5 m, the average residence time of mixed solution was 62.2 s in the tubular reactor. Spheroid nanoparticles with an average size of 97.2 nm were obtained under this optimized condition. Furthermore, the results showed a better control of the mixing compared to the batch process, resulting in a homogeneous distribution of smaller particle sizes thus improving stability and UV-blocking properties. This is attributed to the better mixing and excellent mass transfer characteristics in the tube, which provides favorable conditions for the full contact and uniform dispersion of the mixed solution. More importantly, continuous flow mode makes it possible to prepare LNPs with excellent physicochemical properties on a large scale, which will bring great opportunities for the industrial production and application of LNPs.

Fucoidan-coated coils improve healing in a rabbit elastase aneurysm model.

BACKGROUND: Recanalization of coiled aneurysms remains unresolved. To limit aneurysm recanalization after embolization with coils, we propose an innovative approach to optimize aneurysm healing using fucoidan-coated coils. OBJECTIVE: To evaluate the short-term efficacy and long-term safety of the new coil system with conventional angiography, histology, and multiphoton microscopy for follow-up of fibrosis and neointima formation. METHODS: We conducted a feasibility study on rabbit elastase-induced aneurysms. Embolization was carried out with bare platinum coils, fucoidan-coated coils, or dextran-coated coils. Aneurysms were controlled after 1 month by digital subtraction angiography (DSA). Aneurysm samples were collected and processed for histological analysis. Aneurysm healing and fibrosis were measured by quantifying collagen according to the histological healing score by combining standard light microscopy and multiphoton imaging. We divided 27 rabbits into three groups: bare platinum group, fucoidan group, and dextran group as controls. RESULTS: Angiographic grading showed a trend toward less recanalization in the fucoidan group, although there were no significant differences among the three groups (P=0.21). Histological healing was significantly different according to the presence of more collagen in the neck area of aneurysms in the fucoidan group versus the bare platinum group (P=0.011), but not in the dextran group. Histological index was significantly better at the aneurysm neck in the fucoidan group than in the bare platinum group (P=0.004). Collagen organization index was also significantly better in the fucoidan group than in the bare platinum group (P=0.007). CONCLUSION: This proof-of-concept study demonstrated the feasibility and efficacy of treatment with fucoidan-coated coils to improve aneurysm healing. The results in this rabbit in vivo model showed that fucoidan-coated coils have the potential to improve healing following endovascular treatment.

Metal vs. Metal-Free Catalysts for Oxidation of 5-Hydroxymethylfurfural and Levoglucosenone to Biosourced Chemicals.

Lignocellulosic feedstocks, such as forestry biomass and agricultural crop residues, can be utilized to generate biofuels and biochemicals. Converting these organic waste materials into biochemicals is widely regarded as a remedial approach to develop a sustainable, clean, and green energy source. Nevertheless, are these methods sustainable and clean? Prior studies have shown that most such conversions use metals - including heavy metals or noble metals - as catalysts. In addition to the fact that many metals (e. g., aluminum, cobalt, titanium, platinum) have been listed as critical minerals, these methods suffer from high cost, deactivation, and leakage problems and the release of toxic wastes. This Review summarizes catalytic methods using metal and metal-free catalysts for the oxidation of the platform molecules 5-hydroxymethylfurfural and levoglucosenone and demonstrates the potential and effectiveness of metal-free catalysts.

The preparation of stable spherical alkali lignin nanoparticles with great thermal stability and no cytotoxicity.

In order to further develop the potential applications of lignin biomass, the research on lignin nanoparticles (LNPs) and their nanocomposites has attracted increasing attention. In this study, a facile and no chemical modification approach to prepare stable alkali lignin nanospheres is presented. The nanospheres around 85-125 nm were prepared through the π-π interactions between molecules in the self-assembly process. Lignin alkali was dissolved in ethylene glycol at different initial concentrations and subsequently ultrasound and dialysis treatment were conducted to prepare LNPs. The prepared LNPs had zeta potentials between -20 mV and -40 mV, and they were electrostatically stable over the pH range of 3 to 12 in aqueous solution. The chemical structure of LNPs was not significantly modified compared to lignin. Meanwhile the increased content of carboxyl and aliphatic hydroxyl groups in the LNPs structure was observed. Furthermore, the thermal stability and solubility in organic solvents (ethanol, acetone and THF) of LNPs were enhanced compared to those of lignin. In vitro cell viability evaluation indicated that the prepared LNPs had no cytotoxicity and excellent biocompatibility with mouse fibroblast. Therefore, we proposed here the production of high-quality and renewable LNPs, which will provide a novel perspective for multifunctional applications of bio-based nanomaterials.

Solvent-Free Production by Extrusion of Bio-Based Poly(glycerol-co-diacids) Sheets for the Development of Biocompatible and Electroconductive Elastomer Composites.

Faced with growing global demand for new potent, bio-based, biocompatible elastomers, the present study reports the solvent-free production of 13 pure and derived poly(glycerol-co-diacid) composite sheets exclusively using itaconic acid, sebacic acid, and 2,5-furandicarboxylic acid (FDCA) with glycerol. Herein, modified melt polycondensation and Co(II)-catalyzed polytransesterification were employed to produce all exploitable prepolymers, enabling the easy and rapid manufacturing of elastomer sheets by extrusion. Most of our samples were loaded with 4 wt% of various additives such as natural polysaccharides, synthetic polymers, and/or 25 wt% sodium chloride as porogen agents. The removal of unreacted monomers and acidic short oligomers was carried out by means of washing with NaHCO3 aqueous solution, and pH monitoring was conducted until efficient sheet surface neutralization. For each sheet, their surface morphologies were observed by Field-emission microscopy, and DSC was used to confirm their amorphous nature and the impact of the introduction of every additive. The chemical constitution of the materials was monitored by FTIR. Then, cytotoxicity tests were performed for six of our most promising candidates. Finally, we achieved the production of two different types of extrusion-made PGS elastomers loaded with 10 wt% PANI particulates and 4 wt% microcrystalline cellulose for adding potential electroconductivity and stability to the material, respectively. In a preliminary experiment, we showed the effectiveness of these materials as performant, time-dependent electric pH sensors when immersed in a persistent HCl atmosphere.

Modification of Physio-Mechanical Properties of Chitosan-Based Films via Physical Treatment Approach.

The premise of this work is the modification of the properties of chitosan-based film for possible use in food packaging applications. The biofilm was prepared via thermal and mechanical treatment through blending polymers with chitosan using Polyvinyl Alcohol (PVA) and loading different types of chemical agents, i.e., citric acid (CA), succinic acid (SA), and tetraethoxysilane (TEOS). The modification was carried out under high-speed homogenization at elevated temperature to induce physical cross-linkage of chitosan polymer chains without a catalyst. The findings showed that PVA improved the chitosan films' Tensile strength (TS) and elongation at break (Eb). The presence of chemicals caused an increase in the film strength for all samples prepared, in which a 5% w/w of chemical in the optimum composition CS/PVA (75/25) provided the maximum strength, namely, 33.9 MPa, 44.0 MPa, and 41.9 MPa, for CA-5, SA-5, and TEOS-5, respectively. The chemical agents also increased the water contact angles for all tested films, indicating that they promoted hydrophobicity. The chemical structure analysis showed that, by incorporating three types of chemical agents into the CS/PVA blend films, no additional spectral bands were found, indicating that no covalent bonds were formed. The thermal properties showed enhancement in melting peak and degradation temperature of the blend films, compared to those without chemical agents at the optimum composition. The X-ray diffraction patterns exhibited that PVA led to an increasing crystallization tendency in the blend films. The morphological observation proved that no irregularities were detected in CS/PVA blend films, representing high compatibility with both polymers.

Lignin Nanoparticles and Their Nanocomposites.

Lignin nanomaterials have emerged as a promising alternative to fossil-based chemicals and products for some potential added-value applications, which benefits from their structural diversity and biodegradability. This review elucidates a perspective in recent research on nanolignins and their nanocomposites. It summarizes the different nanolignin preparation methods, emphasizing anti-solvent precipitation, self-assembly and interfacial crosslinking. Also described are the preparation of various nanocomposites by the chemical modification of nanolignin and compounds with inorganic materials or polymers. Additionally, advances in numerous potential high-value applications, such as use in food packaging, biomedical, chemical engineering and biorefineries, are described.

Studies of the potential of a native natural biosorbent for the elimination of an anionic textile dye Cibacron Blue in aqueous solution.

This work is devoted to the adsorption of Cibacron Blue (CB) an anionic textile dye, on bean peel (BP) an agricultural waste with neither activation nor carbonization. The adsorption was realized in batch configuration at ambient temperature in acidic medium. The adsorbent was characterized by FTIR, SEM and BET analyses; the equilibrium isotherms and kinetics were also studied. It has been found that this waste could be used as a low-cost biosorbent for CB elimination under optimal working conditions. The rate of CB elimination reaches 95% on bean bark (3.6 g/L) at pH 2.2 and a reject concentration of 25 mg/L. The pseudo-second-order describes suitably the experimental data and the external diffusion is the rate-determining step. The Freundlich isotherm fits better the CB adsorption with a correlation coefficient (R2) of 0.94 and an RMSE = 1.5115. The negative enthalpy (ΔH) and free enthalpy (ΔG°) indicate a physical and spontaneous nature of the CB biosorption onto the biomaterial.

Cellulosic/Polyvinyl Alcohol Composite Hydrogel: Synthesis, Characterization and Applications in Tissue Engineering.

The biomedical field still requires composite materials for medical devices and tissue engineering model design. As part of the pursuit of non-animal and non-proteic scaffolds, we propose here a cellulose-based material. In this study, 9%, 18% and 36% dialdehyde-functionalized microcrystalline celluloses (DAC) were synthesized by sodium periodate oxidation. The latter was subsequently coupled to PVA at ratios 1:2, 1:1 and 2:1 by dissolving in N-methyl pyrrolidone and lithium chloride. Moulding and successive rehydration in ethanol and water baths formed soft hydrogels. While oxidation effectiveness was confirmed by dialdehyde content determination for all DAC, we observed increasing hydrolysis associated with particle fragmentation. Imaging, FTIR and XDR analysis highlighted an intertwined DAC/PVA network mainly supported by electrostatic interactions, hemiacetal and acetal linkage. To meet tissue engineering requirements, an interconnected porosity was optimized using 0-50 µm salts. While the role of DAC in strengthening the hydrogel was identified, the oxidation ratio of DAC showed no distinct trend. DAC 9% material exhibited the highest indirect and direct cytocompatibility creating spheroid-like structures. DAC/PVA hydrogels showed physical stability and acceptability in vivo that led us to propose our DAC 9%/PVA based material for soft tissue graft application.

Gold, Silver, and Iron Oxide Nanoparticle Incorporation into Silk Hydrogels for Biomedical Applications: Elaboration, Structure, and Properties.

Silk fibroin (SF) is a versatile material with biodegradable and biocompatible properties, which make it fit for broad biomedical applications. In this context, the incorporation of nanosized objects into SF allows the development of a variety of bionanocomposites with tailored properties and functions. Herein, we report a thorough investigation on the design, characterization, and biological evaluation of SF hydrogels incorporating gold, silver, or iron oxide nanoparticles. The latter are synthesized in aqueous media using a biocompatible ligand allowing their utilization in various biomedical applications. This ligand seems to play a pivotal role in nanoparticle dispersion within the hydrogel. Results show that the incorporation of nanoparticles does not greatly influence the mechanism of SF gelation and has a minor impact on the mechanical properties of the so-obtained bionanocomposites. By contrast, significant changes are observed in the swelling behavior of these materials, depending on the nanoparticle used. Interestingly, the main characteristics of these bionanocomposites, related to their potential use for biomedical purposes, show the successful input of nanoparticles, including antibacterial properties for gold and silver nanoparticles and magnetic properties for iron oxide ones.