Stretchable hydrogels of chitosan/hyaluronic acid induced by polyelectrolyte complexation around neutral pH.

In this work, we propose the formation of stretchable hydrogels at neutral pH from the physical crosslinking of chitosan (CS) and hyaluronic acid (HA) by polyelectrolyte complexation. A mixture of CS (Mw ≈ 600 kg/mol, degree of acetylation ≈ 50 %) solution and HA (Mw ≈ 77 kg/mol) solution was prepared with an excess of salts screening the electrostatic interactions CS/HA. In a controlled manner, the polyelectrolyte complexation was induced through the progressive dialysis of the salted polymer mixture against a sodium acetate solution (AcONa, 0.01 M) for 7 days. Depending on [HA], various materials were obtained: viscous solutions at [HA] = 0.75 % (w/v); hydrogels at [HA] = 1.50-2.24 % (w/v) with Young modulus of 14 kPa and stretchable to 200 %. The small angle X-ray scattering characterization of the hydrogels revealed a multiscale organization related to the conformation of the polymers induced by the physical interactions. The dialysis process with AcONa was optimized by adding a dialysis step against a zinc acetate solution containing Zn2+. The combination of polyelectrolyte complexation between CS/HA and metal complexation between Zn2+ and the polymers led to an enhancement of the hydrogel stretchability up to 400 %.

Wet-Spun Chitosan-Sodium Caseinate Fibers for Biomedicine: From Spinning Process to Physical Properties.

We designed and characterized chitosan-caseinate fibers processed through wet spinning for biomedical applications such as drug delivery from knitted medical devices. Sodium caseinate was either incorporated directly into the chitosan dope or allowed to diffuse into the chitosan hydrogel from a coagulation bath containing sodium caseinate and sodium hydroxide (NaOH). The latter route, where caseinate was incorporated in the neutralization bath, produced fibers with better mechanical properties for textile applications than those formed by the chitosan-caseinate mixed collodion route. The latter processing method consists of enriching a pre-formed chitosan hydrogel with caseinate, preserving the structure of the semicrystalline hydrogel without drastically affecting interactions involved in the chitosan self-assembly. Thus, dried fibers, after coagulation in a NaOH/sodium caseinate aqueous bath, exhibited preserved ultimate mechanical properties. The crystallinity ratio of chitosan was not significantly impacted by the presence of caseinate. However, when caseinate was incorporated into the chitosan dope, chitosan-caseinate fibers exhibited lower ultimate mechanical properties, possibly due to a lower entanglement density in the amorphous phase of the chitosan matrix. A standpoint is to optimize the chitosan-caseinate composition ratio and processing route to find a good compromise between the preservation of fiber mechanical properties and appropriate fiber composition for potential application in drug release.

α-Substituted ketones as reagent for Passerini modification of carboxymethyl cellulose: Toward dually functionalized derivatives and thermo-sensitive chemical hydrogels.

The present works describes the Passerini modification of carboxymethyl cellulose (CMC) by using a library of nine α-substituted ketones derivatives, differing in their hydrophobicity and reactivity, conjointly with cyclohexyl isocyanide. The Passerini ligation, achieved in aqueous and mild conditions, was shown to be successful, leading to a large panel of dually functionalized CMC derivatives, in an eco-friendly manner. A particular attention was dedicated to the influence of the experimental parameters such as the stoichiometry, the nature of a co-solvent or the temperature, which allowed to tune the extent of modification. The reactivity of the ketone was proven to be governed by its i) compatibility with water, ii) sterical accessibility, and by iii) the presence of neighboring electron-withdrawing group. The resulting Passerini CMC products modified by methacrylate moieties (CMC-MA) were used as reactive macromonomer under a "grafting through" approach. The copolymerization of CMC-MA with oligoethylene glycol methacrylate (OEGMA) and diethylene glycol methacrylate (DEGMA) upon thermal radical reaction conditions enabled to generate tightly cross-linked chemical hydrogels, with a thermo-sensitive and thermo-reversible behavior, reflected by a macroscopical shrinkage/swelling response, and confirmed by SAXS analysis. Such chemical strategy paves the way toward multifunctional polysaccharide-based networks with potential utilizations as drug delivery devices, dye removals or actuators.

Grafted chitosan thin films of various degrees of acetylation as a reusable platform for the investigation of biological interactions.

Surface treatment by adhesive polymers is a promising solution to immobilize and study bacteria cells through microscopic assays and, for example, control their growth or determine their susceptibility to antibiotic treatment. The stability of such functional films in wet conditions is crucial, as the film degradation would compromise a persistent use of the coated devices. In this work, low roughness chitosan thin films of degrees of acetylation (DA) ranging from 0.5 % to 49 % were chemically grafted onto silicon and glass substrates and we have demonstrated how the physicochemical properties of the surfaces and the bacterial response were DA-dependent. A fully deacetylated chitosan film presented an anhydrous crystalline structure while the hydrated crystalline allomorph was the preferred structure at higher DA. Moreover, their hydrophilicity increased at higher DA, leading to higher film swelling. Low DA chitosan-grafted substrate favored bacterial growth away from the surface and could be envisioned as bacteriostatic surfaces. Contrarily, an optimum of Escherichia coli adhesion was found for substrates modified with chitosan of DA = 35 %: these surfaces are adapted for the study of bacterial growth and antibiotic testing, with the possibility of reusing the substrates without affecting the grafted film - ideal for limiting single-use devices.

Preparation of highly stable and ultrasmooth chemically grafted thin films of chitosan.

Chitosan-coated surfaces are of great interest for biomedical applications (antibacterial coatings, implants, would healing, single-cell microfluidics…). However, one major limitation of chitosan-based systems is the high solubility of the polymer under acidic aqueous conditions. Herein, we describe a simple procedure to prepare extremely smooth and stable chitosan coatings. In detail, chitosan films with a low degree of N-acetylation and of thicknesses varying from 40 nm to 10 µm were grafted onto epoxy-functionalized silicon wafers via an optimized water-temperature treatment (WTT). The formation of a grafted chitosan network insoluble in acidic aqueous media (pH 3.5) was evidenced and the films were stable for at least 2 days at pH 3.5. The film morphology and the swelling behavior were characterized by atomic force microscopy (AFM) and neutron reflectivity, which showed that the film roughness was extremely low. The physical cross-linking of the films was demonstrated using infrared spectroscopy, dynamic mechanical analysis (DMA) and wide-angle X-ray scattering (WAXS). Finally, we show that the swelling behavior of such films was largely influenced by the environmental conditions, such as the pH or ionic strength of the solution.

Regiospecific Grafting of Chitosan Oligomers Brushes onto Silicon Wafers.

The functionalization of surfaces using chitosan oligomers is of great interest for a wide range of applications in biomaterial and biomedical fields, as chitosan oligomers can provide various functional properties including biocompatibility, wetting, adhesion, and antibacterial activity. In this study, an innovative process for the regiospecific chemical grafting of reducing-end-modified chitosan oligomers brushes onto silicon wafers is described. Chitosan oligomers (COS) with well-defined structural parameters (average DP ~19 and DA ~0%) and bearing a 2,5-anhydro-d-mannofuranose (amf) unit at the reducing end were obtained via nitrous acid depolymerization of chitosan. After a silanization step where silicon wafers were modified with aromatic amine derivatives, grafting conditions were studied to optimize the reductive amination between aldehydes of amf-terminated COS and aromatic amines of silicon wafers. Functionalized surfaces were fully characterized by AFM, ATR-FTIR, ellipsometry, contact angle measurement, and ToF-SIMS techniques. Smooth surfaces were obtained with a COS layer about 3 nm thick and contact angle values between 72° and 76°. Furthermore, it was shown that the addition of the reducing agent NaBH3CN could positively improve the COS grafting density and/or led to a better stability of the covalent grafting to hydrolysis. Finally, this study also showed that this grafting process is also efficient for chitosan oligomers of higher DA (i.e., ~21%).

Cytocompatibility / Antibacterial Activity Trade-off for Knittable Wet-Spun Chitosan Monofilaments Functionalized by the In Situ Incorporation of Cu2+ and Zn2.

The wet spinning of cytocompatible, bioresorbable, and knittable chitosan (CTS) monofilaments would be advantageous for a variety of surgical applications. The complexation capacity of chitosan with Cu2+ or Zn2+ can be leveraged to enhance its antibacterial activity, but not at the expense of cytocompatibility. In this work, a wet-spinning process was adapted for the in situ incorporation of Cu2+ or Zn2+ with chitosan dopes to produce monofilaments at different drawing ratios (τtot) with various cation/glucosamine molar ratios, evaluated in the fibers (rCu,f and rZn,f). Cytocompatibility and antibacterial activity of wet-spun monofilaments were, respectively, quantified by in vitro live-dead assays on balb 3T3 and by different evaluations of the proliferation inhibition of Staphylococcus epidermidis (Gram+) and Escherichia coli (Gram-). Knittability was tested by a specific tensile test using a knitting needle and evaluated with an industrial knitting machine. It was found that rCu,f = 0.01 and rZn,f = 0.03 significantly increase the antibacterial activity without compromising cytocompatibility. Wet spinning with τtot = 1.6 allowed the production of knittable CTS-Cu monofilaments, as confirmed by knitting assays under industrial conditions.

Development of Breathable Pebax®/PEG Films for Optimization of the Shelf-Life of Fresh Agri-Food Products.

In this work, thin transparent breathable films were prepared for food packaging applications. The films were obtained by the solvent casting method from both the binary blends Pebax® MH1657 copolymer/ hydroxyl-terminated polyethylene glycol (PEGOH) and Pebax® MH1657/polyethylene glycol dimethyl ether (PEGDME) as well as the ternary blend Pebax® MH1657/PEGOH/PEGDME with a 50/50 and 37.5/62.5 PEGOH/PEGDME weight ratio for additive amounts comprised between 0 and 50 wt.%. The microstructure of these materials was investigated by differential scanning calorimetry (DSC) and wide-angle X-ray scattering (WAXS) analyses. Regardless of the PEG's nature, for a PEG amount inferior to 30 wt.%, the Pebax® and PEG phases were totally miscible. For higher amounts, a phase separation was obtained. In the presence of PEG, a decrease in crystallinity was obtained. The effects of the nature and amount of PEG on the thermo-mechanical, hydration, and gas (CO2, O2) transport properties were investigated. A study of the film's stability in terms of composition over time was also performed. From this work, a wide range of films could be proposed with a stable composition over time and adjustable mechanical and gas transport properties for the prolongation of the shelf-life of highly breathable fresh products.

Development of Bioinspired Functional Chitosan/Cellulose Nanofiber 3D Hydrogel Constructs by 3D Printing for Application in the Engineering of Mechanically Demanding Tissues.

Soft tissues are commonly fiber-reinforced hydrogel composite structures, distinguishable from hard tissues by their low mineral and high water content. In this work, we proposed the development of 3D printed hydrogel constructs of the biopolymers chitosan (CHI) and cellulose nanofibers (CNFs), both without any chemical modification, which processing did not incorporate any chemical crosslinking. The unique mechanical properties of native cellulose nanofibers offer new strategies for the design of environmentally friendly high mechanical performance composites. In the here proposed 3D printed bioinspired CNF-filled CHI hydrogel biomaterials, the chitosan serves as a biocompatible matrix promoting cell growth with balanced hydrophilic properties, while the CNFs provide mechanical reinforcement to the CHI-based hydrogel. By means of extrusion-based printing (EBB), the design and development of 3D functional hydrogel scaffolds was achieved by using low concentrations of chitosan (2.0-3.0% (w/v)) and cellulose nanofibers (0.2-0.4% (w/v)). CHI/CNF printed hydrogels with good mechanical performance (Young's modulus 3.0 MPa, stress at break 1.5 MPa, and strain at break 75%), anisotropic microstructure and suitable biological response, were achieved. The CHI/CNF composition and processing parameters were optimized in terms of 3D printability, resolution, and quality of the constructs (microstructure and mechanical properties), resulting in good cell viability. This work allows expanding the library of the so far used biopolymer compositions for 3D printing of mechanically performant hydrogel constructs, purely based in the natural polymers chitosan and cellulose, offering new perspectives in the engineering of mechanically demanding hydrogel tissues like intervertebral disc (IVD), cartilage, meniscus, among others.

Thermo-mechanical properties and blend behaviour of cellulose acetate/lactates and acid systems: Natural-based plasticizers.

This work brings together thermo-mechanical and structural information for plasticized cellulose acetate (CA) by lactates and octanoic acid. CA are processed with plasticizer due to their high Tg and their strong H-bonding network. We prepared CA / plasticizer blends by corotative twin screw extruder and by solvent casting methods. The study of the different relaxations and of the glassy zone modulus was performed by dynamic mechanical analysis (DMA). The miscibility range of cellulose acetate blends were identified by the analysis of the tan δ. Depending on the composition of the system, one or two transitions are noted, this last result indicates the presence of a phase rich in CA and another in plasticizer. To connect this information to crystallinity and molecular organization, X-ray diffraction analyses were carried out. The disappearance of crystallinity allows the plasticization of previously inaccessible zones, causing a glassy modulus drop of more than 1000 MPa.

Responsive Adsorption of N-Isopropylacrylamide Based Copolymers on Polymer Brushes.

We investigate the adsorption of pH- or temperature-responsive polymer systems by ellipsometry and neutron reflectivity. To this end, temperature-responsive poly (N-isopropylacrylamide) (PNIPAM) brushes and pH-responsive poly (acrylic acid) (PAA) brushes have been prepared using the "grafting onto" method to investigate the adsorption process of polymers and its reversibility under controlled environment. To that purpose, macromolecular brushes were designed with various chain lengths and a wide range of grafting density. Below the transition temperature (LCST), the characterization of PNIPAM brushes by neutron reflectivity shows that the swelling behavior of brushes is in good agreement with the scaling models before they collapse above the LCST. The reversible adsorption on PNIPAM brushes was carried out with linear copolymers of N-isopropylacrylamide and acrylic acid, P(NIPAM-co-AA). While these copolymers remain fully soluble in water over the whole range of temperature investigated, a quantitative adsorption driven by solvophobic interactions was shown to proceed only above the LCST of the brush and to be totally reversible upon cooling. Similarly, the pH-responsive adsorption driven by electrostatic interactions on PAA brushes was studied with copolymers of NIPAM and N,N-dimethylaminopropylmethacrylamide, P(NIPAM-co-MADAP). In this case, the adsorption of weak polycations was shown to increase with the ionization of the PAA brush with interactions mainly located in the upper part of the brush at pH 7 and more deeply adsorbed within the brush at pH 9.

In situ synthesis of Fe3O4 nanoparticles coated by chito-oligosaccharides: physico-chemical characterizations and cytotoxicity evaluation for biomedical applications.

Fe3O4 nanoparticles coated with chito-oligosaccharides (COS) were prepared in situ by a simple co-precipitation method through a mixing of iron ions (Fe3+ and Fe2+) and COS aqueous solutions followed by precipitation with ammonia. The impact of COS with different degree of polymerization (DP 10, 24 and 45) and degree of N-acetylation (DA) âˆ¼ 24% and 50% (exhibiting high solubility) on the synthesis and physical properties of the coated magnetic nanoparticles was evaluated. Several advantages were found when the magnetic nanoparticles were prepared in the presence of the studied COS, such as: preparation of functionalized magnetic nanoparticles with narrower size distributions and, consequently, higher saturation magnetization (an increase of up to 22%); and an expressive increasing in the concentration of COS-coated magnetic nanoparticles (up to twice) in the cell viability test in comparison with pure Fe3O4 nanoparticles. Furthermore, among the analyzed samples, the magnetic nanoparticles coated by COS with DA âˆ¼ 50% present a higher cytocompatibility. Our results allow envisioning various biomedical applications, valorizing the use of coated-magnetic nanoparticles for magnetic-field assisted drug delivery, enzyme or cell immobilization, or as a marker for specific cell tracking, among others.

Chitosan Films for Microfluidic Studies of Single Bacteria and Perspectives for Antibiotic Susceptibility Testing.

Single-cell microfluidics is a powerful method to study bacteria and determine their susceptibility to antibiotic treatment. Glass treatment by adhesive molecules is a potential solution to immobilize bacterial cells and perform microscopy, but traditional cationic polymers such as polylysine deeply affect bacterial physiology. In this work, we chemically characterized a class of chitosan polymers for their biocompatibility when adsorbed to glass. Chitosan chains of known length and composition allowed growth of Escherichia coli cells without any deleterious effects on cell physiology. Combined with a machine learning approach, this method could measure the antibiotic susceptibility of a diversity of clinical strains in less than 1 h and with higher accuracy than current methods. Finally, chitosan polymers also supported growth of Klebsiella pneumoniae, another bacterial pathogen of clinical significance.IMPORTANCE Current microfluidic techniques are powerful to study bacteria and determine their response to antibiotic treatment, but they are currently limited by their complex manipulation. Chitosan films are fully biocompatible and could thus be a viable replacement for existing commercial devices that currently use polylysine. Thus, the low cost of chitosan slides and their simple implementation make them highly versatile for research as well as clinical use.

Cellulose Nanofiber-Reinforced Chitosan Hydrogel Composites for Intervertebral Disc Tissue Repair.

The development of non-cellularized composites of chitosan (CHI) hydrogels, filled with cellulose nanofibers (CNFs) of the type nanofibrillated cellulose, was proposed for the repair and regeneration of the intervertebral disc (IVD) annulus fibrosus (AF) tissue. With the achievement of CNF-filled CHI hydrogels, biomaterial-based implants were designed to restore damaged/degenerated discs. The structural, mechanical and biological properties of the developed hydrogel composites were investigated. The neutralization of weakly acidic aqueous CNF/CHI viscous suspensions in NaOH yielded composites of physical hydrogels in which the cellulose nanofibers reinforced the CHI matrix, as investigated by means of microtensile testing under controlled humidity. We assessed the suitability of the achieved biomaterials for intervertebral disc tissue engineering in ex vivo experiments using spine pig models. Cellulose nanofiber-filled chitosan hydrogels can be used as implants in AF tissue defects to restore IVD biomechanics and constitute contention patches against disc nucleus protrusion while serving as support for IVD regeneration.

Lubrication and Wear Protection of Micro-Structured Hydrogels Using Bioinspired Fluids.

We report the fabrication and the use of a bioinspired synovial fluid acting as a lubricant fluid and antiwear agent at soft and porous chitosan hydrogel tribopairs. This synthetic synovial fluid is composed of sodium hyaluronate (HA) and a bottle-brush polymer (BB) having a polycationic attachment group and polyzwitterionic pendant chains. The 2.5%w/w chitosan hydrogel plugs are organized in a bilayered structure exposing a thin and dense superficial zone (SZ), covering a porous deep zone (DZ), and exhibiting microchannels perpendicularly aligned to the SZ. Using a low-load tribometer, the addition of HA lubricating solution at the hydrogel-hydrogel rubbing contact drastically decreased the coefficient of friction (CoF) from µ = 0.20 ± 0.01 to 0.04 ± 0.01 on the DZ configuration and from µ = 0.31 ± 0.01 to 0.08 ± 0.01 on the SZ surface when increasing the HA concentration from 0 to 1000 µg/mL and its molecular mass from 10 to 1500 kDa, similar to what was found when using the BB polymer alone. When combining the BB polymer and the 1500 kDa HA, the CoF remained stable at µ = 0.04 ± 0.01 for both studied contact configurations, highlighting the synergistic interaction of the two macromolecules. Hydrogel wear was characterized by assessing the final gel surface roughness by the means of an interferometer. Increasing HA concentration and molecular weight plus the addition of the BB polymer led to a dramatic surface wear protection with a final gel surface roughness of the hydrogels similar to the untested gels. In brief, the BB polymer in combination with high molecular weight HA is a potential lubricating fluid as well as a wear resistant agent for soft materials lubrication and wear protection.

Critical Role of Interfacial Diffusion and Diffuse Interphases Formed in Multi-Micro-/Nanolayered Polymer Films Based on Poly(vinylidene fluoride) and Poly(methyl methacrylate).

It is known that the macroscopic properties of multilayer polymer films are largely dominated by the diffuse interphase formed via interfacial diffusion between neighboring layers. However, not much is known about the origin of this effect. In this work, we reveal the role of interfacial diffusion and the diffuse interphase development in multilayer polymer films, based on a compatible poly(vinylidene fluoride)/poly(methyl methacrylate) system fabricated by forced-assembly micro-/nanolayer coextrusion. Interestingly, the layer morphology is found to prevail in all investigated multilayer films, even for the nanolayered system where the interdiffusion is substantial. It is also demonstrated that, in the presence of macromolecular and geometrical confinements, interfacial diffusion significantly alters the crystalline morphology and microstructure of the resulting micro-/nanolayered films, which leads to quantitatively different dielectric and rheological properties. More importantly, the combination of dielectric relaxation spectroscopy and energy-dispersive X-ray analysis further reveals that multiple diffuse interphases with various length scales exist in the multilayer structures. The presence of these multiple interphases is explained in terms of a proposed physical picture for the interdiffusion of fast-mode mechanism occurring in the coextrusion process, and their length scales (i.e., interphase thicknesses) are further mapped quantitatively. These findings provide new insights into the effects of interfacial diffusion and diffuse interphases toward tailoring interfaces/interphases in micro-/nanolayered polymer structures and for their advanced applications.

Bioinspired microstructures of chitosan hydrogel provide enhanced wear protection.

We describe the fabrication of physical chitosan hydrogels exhibiting a layered structure. This bilayered structure, as shown by SEM and confocal microscopy, is composed of a thin dense superficial zone (SZ), covering a deeper zone (DZ) containing microchannels orientated perpendicularly to the SZ. We show that such structure favors diffusion of macromolecules within the hydrogel matrix up to a critical pressure, σc, above which channels were constricted. Moreover, we found that the SZ provided a higher wear resistance than the DZ which was severely damaged at a pressure equal to the elastic modulus of the gel. The coefficient of friction (CoF) of the SZ remained independent of the applied load with µSZ = 0.38 ± 0.02, while CoF measured at DZ exhibited two regimes: an initial CoF close to the value found on the SZ, and a CoF that decreased to µDZ = 0.18 ± 0.01 at pressures higher than the critical pressure σc. Overall, our results show that internal structuring is a promising avenue in controlling and improving the wear resistance of soft materials such as hydrogels.

Dynamic Structuration of Physical Chitosan Hydrogels.

We studied the microstructure of physical chitosan hydrogels formed by the neutralization of chitosan aqueous solutions highlighting the structural gradients within thick gels (up to a thickness of 16 mm). We explored a high polymer concentrations range (Cp ≥ 1.0% w/w) with different molar masses of chitosan and different concentrations of the coagulation agent. The effect of these processing parameters on the morphology was evaluated mainly through small-angle light scattering (SALS) measurements and confocal laser scanning microscopy (CLSM) observations. As a result, we reported that the microstructure is continuously evolving from the surface to the bulk, with mainly two structural transitions zones separating three types of hydrogels. The first zone (zone I) is located close to the surface of the hydrogel and constitutes a hard (entangled) layer formed under fast neutralization conditions. It is followed by a second zone (zone II) with a larger thickness (∼3-4 mm), where in some cases large pores or capillaries (diameter ∼10 µm) oriented parallel to the direction of the gel front are present. Deeper in the hydrogel (zone III), a finer oriented microstructure, with characteristic sizes lower than 2-3 µm, gradually replace the capillary morphology. However, this last bulk morphology cannot be regarded as structurally uniform because the size of small micrometer-range-oriented pores continuously increases as the distance to the surface of the hydrogel increases. These results could be rationalized through the effect of coagulation kinetics impacting the morphology obtained during neutralization.

Highly stretchable hydrogels from complex coacervation of natural polyelectrolytes.

The controlled complex coacervation of oppositely charged hyaluronic acid (Mw ≈ 800-1000 kg mol-1) and chitosan (Mw ≈ 160 kg mol-1, degree of acetylation = 15%) led to hydrogels with controllable properties in terms of elasticity and strength. In this work, we performed desalting by dialysis of high ionic strength solutions of mixed polyelectrolytes and showed that the control of the pH during the polyelectrolyte assembly greatly impacts the mechanical properties of the hydrogel. First, for pHs from 5.5 to 7.5, a slight coacervation was observed due to low chitosan protonation and poor polyelectrolyte associations. Then, for pHs from 3.0 to 5.5, coacervation and syneresis led to free-standing and easy to handle hydrogels. Finally, for pHs from 2.0 to 3.0 (close to the pKa of the hyaluronic acid), we observed the unusual stretchability of these hydrogels that could arise from the pre-folding of hyaluronic acid chains while physical crosslinking was achieved by hyaluronic acid/chitosan polyelectrolyte complexation.

Modular construction of single-component polymer nanocapsules through a one-step surfactant-free microemulsion templated synthesis.

Formation of O/W surfactant-free microemulsions from water/oil/acetone ternary systems is exploited to construct precisely-defined shell-functionalized core-loaded nanocapsules with tunable diameters (ranging from 50 to 190 nm) in one step.