Drug delivery based on a supramolecular chemistry approach by using chitosan hydrogels.

Microbial infections are a serious healthcare related problem, causing several complications and even death. That is why, the development of new drug delivery systems with prolonged effect represents an interesting research topic. This study presents the synthesis and characterization of new hydrogels based on chitosan and three halogenated monoaldehydes. Further, the hydrogels were used as excipients for the development of drug delivery systems (DDS) by the incorporation of fluconazole, an antifungal drug. The systems were structurally characterized by Fourier Transformed Infrared Spectroscopy and Nuclear Magnetic Resonance, both methods revealing the formation of the imine linkages between chitosan and the aldehydes. The samples presented a high degree of ordering at supramolecular level, as demonstrated by WXRD and POM and a good water-uptake, reaching a maximum of 1.6 g/g. The obtained systems were biodegradable, loosing between 38 and 49 % from their initial mass in the presence of lysozyme in 21 days. The ability to release the antifungal drug in a sustained manner for seven days, along with the high values of the inhibition zone diameter, reaching a maximum of 64 mm against Candida parapsilosis for the chlorine containing sample, recommend these systems as promising materials for bioapplications.

Mesoporous chitosan nanofibers loaded with norfloxacin and coated with phenylboronic acid perform as bioabsorbable active dressings to accelerate the healing of burn wounds.

The paper reports new chitosan-based nanofibers, designed to address the healing of burn wounds. To this aim, mesoporous chitosan fiber mats were prepared by electrospinning using poly(ethylene oxide) as sacrificial additive, followed by loading with norfloxacin and coating with an antifungal agent via dynamic imine bonds. Dynamic vapor sorption experiment proved intra-fiber mesopores around 2.7 nm, and UV-vis, FTIR, and NMR spectroscopy confirmed the norfloxacin embedding and the imination reaction. SEM, AFM and POM techniques displayed semicrystalline nanofibers with average diameter around 170 nm entangled into a non-woven mat. Their mesoporous nature favored a rapid adsorption of fluids up to 17 g/g, and a biodegradation rate fitting the wound healing rate, i.e. up to 30 % mass loss in media of pH characteristic to wound exudate and total degradation in that characteristic to normal dermis. The composite fibers released the NFX and 2FPBA in a controlled manner, and showed antimicrobial activity against gram positive, gram negative and fungal strains. They had no cytotoxic effect on normal human dermal fibroblasts, and showed biocompatibility on experimental rats. The investigation of wound healing ability on second/third-degree burn model in rats revealed wound closure and total restoration of the fully functional dermis and epidermis.

New Hydrogels Nanocomposites Based on Chitosan, 2-Formylphenylboronic Acid, and ZnO Nanoparticles as Promising Disinfectants for Duodenoscopes Reprocessing.

New hydrogels nanocomposites, based on iminoboronate hydrogels and ZnO nanoparticles (ZnO-NPs), were obtained and characterised in order to develop a new class of disinfectants able to fight the nosocomial infections produced by duodenoscopes investigation procedures. The formation of the imine linkages between chitosan and the aldehyde was demonstrated using NMR and FTIR spectroscopy, while the supramolecular architecture of the developed systems was evaluated via wide-angle X-ray diffraction and polarised optical microscopy. The morphological characterisation of the systems via scanning electron microscopy revealed the highly porous structure of the materials, in which no ZnO agglomeration could be observed, indicating the very fine and homogenous encapsulation of the nanoparticles into the hydrogels. The newly synthetised hydrogels nanocomposites was proven to have synergistic antimicrobial properties, being very efficient as disinfectants against reference strains as: Enterococcus faecalis, Klebsiella pneumoniae, and Candida albicans.

New Hydrogels and Formulations Based on Piperonyl-Imino-Chitosan Derivatives.

Candida infections have been always a serious healcare related problem. The present study reports the preparation of hydrogels and formulations based on piperonyl-imino-chitosan derivatives and Amphotericin B drug for the treatment of Candida infections. The hydrogels were obtained by the imination reaction of chitosan with piperonal monoaldehyde, followed by the self-assembling of the resulted imines, while the formulations were obtained by an in situ hydrogelation method of chitosan with piperonal in the presence of Amphotericin B antifungal drug. The structural characterization of both hydrogels and formulations by Fourier transform infrared spectroscopy and Nuclear magnetic resonance spectroscopy revealed the formation of imine units between the reagents, while their supramolecular characterization using polarized optical microscopy and wide angle X-ray diffraction demonstrated that hydrophilic/hydrophobic segregation is the process which governed the formation of gel like systems. The systems were further investigated from the point of view of their further applications revealing that they were biodegradable, presented high swelling ability and were able to release the antifungal drug in a sustained manner, presenting promising antifungal activity against five Candida strains.

Citryl-Imino-Chitosan Xerogels as Promising Materials for Mercury Recovery from Waste Waters.

The present study reported the obtention of xerogels based on chitosan and citral and their use as materials for mercury ion recovery from aqueous solutions, this being a serious problem related to the environment. The systems were prepared by the acid condensation of chitosan with citral, followed by the lyophilization of the resulting hydrogels, in order to obtain highly porous solid materials. The structural, morphological and supramolecular characterization of the systems was performed using 1H-NMR and FTIR spectroscopy, scanning electron microscopy and wide-angle X-ray diffraction. The ability of the obtained materials to be used for the recovery of mercury from aqueous solutions revealed the high potential of the xerogels to be used in this sense, the analysis of the materials post mercury absorption experiments revealing that this ability is predominantly conferred by the imine linkages which act as coordinating moieties for mercury ions.

TEGylated Phenothiazine-Imine-Chitosan Materials as a Promising Framework for Mercury Recovery.

This paper reports new solid materials based on TEGylated phenothiazine and chitosan, with a high capacity to recover mercury ions from aqueous solutions. They were prepared by hydrogelation of chitosan with a formyl derivative of TEGylated phenothiazine, followed by lyophilization. Their structural and supramolecular characterization was carried out by 1H-NMR and FTIR spectroscopy, as well as X-ray diffraction and polarized light microscopy. Their morphology was investigated by scanning electron microscopy and their photophysical behaviour was examined by UV/Vis and emission spectroscopy. Swelling evaluation in different aqueous media indicated the key role played by the supramolecular organization for their hydrolytic stability. Mercury recovery experiments and the analysis of the resulting materials by X-ray diffraction and FTIR spectroscopy showed a high ability of the studied materials to bind mercury ions by coordination with the sulfur atom of phenothiazine, imine linkage, and amine units of chitosan.

Biocompatible drug delivery systems able to co-deliver antifungal and antiviral agents.

The present study reports the synthesis and characterization of 12 drug delivery systems (DDS) for the co-delivery of antifungal and antiviral agents. The systems were obtained by an in situ hydrogelation method of 6 chitosan oligomers with values of the polymerization degree between 14 and 51, with 2-formylphenylboronic acid, in the presence of tenofovir. The structural characterization by NMR and FTIR spectroscopy demonstrated the formation of imine linkages, while WXRD revealed the 3D layered architecture of the systems. SEM and POM images demonstrated the uniform distribution of tenofovir into the matrix, while the Zeta potential measurements revealed the strong interactions which develop between system components. The obtained DDSs presented biodegradability, hemocompatibility and in vivo biocompatibility, which along with their ability to release both the drug and the antifungal aldehyde make them promising materials for the treatment of HIV infection and its associated co-infections' symptoms.

Biocompatible Chitosan-Based Hydrogels for Bioabsorbable Wound Dressings.

Supramolecular hydrogels based on chitosan and monoaldehydes are biomaterials with high potential for a multitude of bioapplications. This is due to the proper choice of the monoaldehyde that can tune the hydrogel properties for specific practices. In this conceptual framework, the present paper deals with the investigation of a hydrogel as bioabsorbable wound dressing. To this aim, chitosan was cross-linked with 2-formylphenylboronic acid to yield a hydrogel with antimicrobial activity. FTIR, NMR, and POM procedures have characterized the hydrogel from a structural and supramolecular point of view. At the same time, its biocompatibility and antimicrobial properties were also determined in vitro. Furthermore, in order to assess the bioabsorbable character, its biodegradation was investigated in vitro in the presence of lysosome in media of different pH, mimicking the wound exudate at different stages of healing. The biodegradation was monitored by gravimetrical measurements, SEM microscopy and fractal analyses of the images. The fractal dimension values and the lacunarity of SEM pictures were accurately calculated. All these successful investigations led to the conclusion that the tested materials are at the expected high standards.

Iminoboronate-chitooligosaccharides hydrogels with strong antimicrobial activity for biomedical applications.

The paper reports hydrogels prepared from chitooligosaccharides with different polymerization degrees (14 to 51), by crosslinking with 2-formylphenylboronicacid in three molar ratios of their functionalities. The structural, morphological and supramolecular characterization confirmed a hydrogelation mechanism based on self-assembling of newly formed imine units and porous morphology. Rheological measurements confirmed the formation of thixotropic hydrogels, and swelling tests indicated mass equilibrium swelling values up to 25 in water and 9 in phosphate buffer saline. The monitoring of enzymatic degradability demonstrated the enhancing of biodegradation rate as long as the polymerization degrees of the oligomers decreased, the mass loss increasing from 16% to 43%. In vivo and ex-vivo biocompatibility investigation on experimental mice showed no cytotoxic effect, and in vitro antimicrobial tests revealed remarkable antimicrobial properties on nine strains, with a maximum inhibition diameter of 49 mm on Aspergilius brasiliensis and very good results on Cladosporium cladosporioides, Penicillium crysogenum and different Candida species.

Amphiphilic chitosan-g-poly(trimethylene carbonate) - A new approach for biomaterials design.

The paper presents the synthesis and characterization of poly(trimethylene carbonate) grafted chitosan as a new water soluble biopolymer suitable for in vivo applications. The synthesis was performed via ring-opening polymerization of 1,3-dioxan-2-one (trimethylene carbonate) (TMC) monomer, initiated by the functional groups of chitosan in the presence of toluene as solvent/swelling agent. By varying the molar ratio between the glucosamine units of chitosan and TMC, a series of chitosan derivatives with different content of poly(trimethylene carbonate) chains was synthetized. The structural characterization of the polymers was realized by FTIR and 1H NMR spectroscopy and their solubility was assessed in water and in organic solvents as well. The biocompatibility was investigated by MTS assay on Normal Human Dermal Fibroblasts, and the biodegradability was evaluated in lysozyme buffer solution. Further, the surface properties of the polymer films were analyzed by polarized optical microscopy, atomic force microscopy and water-to-air contact angle measurements. It was established that, by 5% substitution of chitosan with poly(trimethylene carbonate) chains having an average polymerization degree of 7, a water soluble polymer can be attained. Compared to the pristine chitosan, it has improved biocompatibility in solution and moderate wettability and higher biodegradability rate in solid state, pointing its suitability for in vivo applications.

Theoretical model for the diclofenac release from PEGylated chitosan hydrogels.

Controlled drug delivery systems are of utmost importance for the improvement of drug bioavailability while limiting the side effects. For the improvement of their performances, drug release modeling is a significant tool for the further optimization of the drug delivery systems to cross the barrier to practical application. We report here on the modeling of the diclofenac sodium salt (DCF) release from a hydrogel matrix based on PEGylated chitosan in the context of Multifractal Theory of Motion, by means of a fundamental spinor set given by 2 × 2 matrices with real elements, which can describe the drug-release dynamics at global and local scales. The drug delivery systems were prepared by in situ hydrogenation of PEGylated chitosan with citral in the presence of the DCF, by varying the hydrophilic/hydrophobic ratio of the components. They demonstrated a good dispersion of the drug into the matrix by forming matrix-drug entities which enabled a prolonged drug delivery behavior correlated with the hydrophilicity degree of the matrix. The application of the Multifractal Theory of Motion fitted very well on these findings, the fractality degree accurately describing the changes in hydrophilicity of the polymer. The validation of the model on this series of formulations encourages its further use for other systems, as an easy tool for estimating the drug release toward the design improvement. The present paper is a continuation of the work 'A theoretical mathematical model for assessing diclofenac release from chitosan-based formulations,' published in Drug Delivery Journal, 27(1), 2020, that focused on the consequences induced by the invariance groups of Multifractal Diffusion Equations in correlation with the drug release dynamics.

Hydrogels Based on Imino-Chitosan Amphiphiles as a Matrix for Drug Delivery Systems.

This paper reports new formulations based on chitosan, citral, and diclofenac sodium salt (DCF). The central idea was to encapsulate an anionic drug into a polycationic hydrogel matrix in order to increase the intermolecular forces between them and thus to ensure slower drug release, while citral was used as a penetration enhancer to assure efficient delivery of the drug. Hydrogels without drug were also synthesized and used as a reference. The structure, morphology, and supramolecular architecture of the drug delivery systems were evaluated by FTIR spectroscopy, scanning electron microscopy, polarized optical microscopy, and wide-angle X-ray diffraction. The drug release kinetics was monitored in vitro by UV-VIS spectroscopy, in physiological conditions, while the enzymatic and hydrolytic degradability of the hydrogels were evaluated in the presence of lysozyme and phosphate buffer saline (PBS), at 37 °C. All of the data revealed that the anionic DCF was strongly anchored into the polycationic matrix and the drug was slowly released over 7 days. Moreover, the release rate can be controlled by simple variation of the molar ratio between the polycationic chitosan and lipophilic citral.

Citryl-imine-PEG-ylated chitosan hydrogels - Promising materials for drug delivery applications.

The present paper focuses on the synthesis and characterization of new hydrogels and drug delivery systems, designed for local therapy. The hydrogels were obtained by reacting PEG-ylated chitosan derivatives with citral in different molar ratios of their functionalities. The drug delivery systems were obtained by the in situ hydrogelation of PEG-ylated chitosan derivatives with citral, in the presence of a hydrophilic anti-inflammatory drug, diclofenac sodium salt. The hydrogels and the drug delivery systems were characterized from the structural, supramolecular and morphological points of view by FTIR spectroscopy, wide angle X-ray diffraction, polarized optical microscopy and scanning electron microscopy. The in vitro release kinetics of the drug has been monitored in physiological conditions, while the in vivo release was evaluated by the somatic pain model on rats. The in vitro enzymatic degradability of the hydrogels was evaluated in the presence of lysozyme, leading to a significant mass loss of 47% in 21 days. All the findings, recommend the investigated materials as promising candidates for local drug delivery applications.

Poly(vinyl alcohol boric acid)-Diclofenac Sodium Salt Drug Delivery Systems: Experimental and Theoretical Studies.

The main aim of the paper was to simulate the drug release by a multifractal theoretical model, as a valuable method to assess the drug release mechanism. To do this, drug delivery films were prepared by mixing poly(vinyl alcohol boric acid) (PVAB) and diclofenac (DCF) sodium salt drug in different mass ratios from 90/10 to 70/30, in order to obtain drug delivery systems with different releasing rates. The different drug content of the three systems was confirmed by energy-dispersive spectroscopy (EDAX) analysis, and the encapsulation particularities were investigated by scanning electron microscopy (SEM), atomic force microscopy (AFM), and polarized optical microscopy (POM) techniques. The ability of the PVAB matrix to anchor the DCF was assessed by Fourier transform infrared (FTIR) spectroscopy. The in vitro release of the diclofenac sodium salt from the formulations was investigated in biomimetic conditions (pH = 7.4 and 37°C) by UV-Vis spectroscopy, measuring the absorbance of the drug at 275 nm and fitting the results on a previously drawn calibration curve. An estimation of the drug release kinetics was performed by fitting three traditional mathematical models on experimental release data. Further, the drug delivery was simulated by the fractal theory of motion, in which the release dynamics of the polymer-drug complex system is described through various Riccati-type "regimes." To explain such dynamics involved multifractal self-modulation in the form of period doubling, quasiperiodicity, intermittency, etc., as well as multifractal self-modulation of network type. Standard release dynamics were explained by multifractal behaviors of temporary kink type. The good correlation between the traditional mathematical models and the new proposed theoretical model demonstrated the validity of the multifractal model for the investigation of the drug release.

Polyvinyl alcohol boric acid - A promising tool for the development of sustained release drug delivery systems.

The paper deals with the design and investigation of the morphology, in vitro drug release and biocompatibility of some new formulations based on polyvinyl alcohol boric acid (PVAB) and diclofenac sodium salt (DCF), with the aim to explore the ability of PVAB to act as a matrix for controlled drug delivery systems. A series of three formulations was obtained by mixing the drug and the polymeric matrix in different mass ratios, with high drug content from 10% w/w to 30% w/w. Their structural and supramolecular characterization, performed by FTIR spectroscopy and X-ray diffraction, revealed important physical interactions between the drug and the polymeric matrix. The morphological data, obtained by X-ray diffraction, polarized optical microscopy and scanning electron microscopy revealed the presence of the drug into the PVAB polymeric matrix, as micrometric polycrystals with a mean diameter in the range 10-15 µm, depending on the drug/polymer ratio. The investigation of their surface peculiarities indicated highly hydrophilic surfaces with a water to air contact angle between 29.9 and 41.4 deg and a surface free energy of 45.6-54.2 N/m2. The in vitro release kinetics was monitored by UV-VIS spectroscopy and the cytotoxic effect was investigated in vitro on fibroblasts and HeLa cells. The PVAB proved excellent cytocompatibility, a relative cell viability of the fibroblasts higher than 90% being recorded for concentrations of PVAB up to 7.5% w/v. The drug has been strongly anchored into the electron deficient PVAB matrix, fact which led to its prolonged release up to 5 days. These findings recommend PVAB as a versatile tool for the development of sustained release drug delivery systems with real chances to cross the gap from theory to applications.

Dynamic constitutional chemistry towards efficient nonviral vectors.

Dynamic constitutional chemistry has been used to design nonviral vectors for gene transfection. Their design has been thought in order to fulfill ab initio the main requirements for gene therapy. As building blocks were used hyperbranched PEI as hydrophilic part and benzentrialdehyde and a diamine linear siloxane as hydrophobic part, connected through reversible imine linkages. The obtaining of the envisaged structures has been confirmed by NMR and FTIR spectroscopy. The dynamic synthesized amphiphiles proved to be able to self-assemble in nano-sized spherical entities as was demonstrated by TEM and DLS, characterized by a narrow dimensional polydispersity. Agarose gel electrophoresis proved the ability of the synthesized compounds to bind DNA, while TEM revealed the spherical morphology of the formed polyplexes. As a proof of the concept, the nonviral vectors promoted an efficient transfection on HeLa cells, demonstrating that dynamic constitutional chemistry can be an important tool in the development of this domain.

Drug delivery systems based on biocompatible imino-chitosan hydrogels for local anticancer therapy.

A series of drug delivery systems were prepared by chitosan hydrogelation with citral in the presence of an antineoplastic drug: 5-fluorouracil. The dynamic covalent chemistry of the imine linkage allowed the obtaining of supramolecular tridimensional architectures in which the drug has been homogenously dispersed. Fourier-transform infrared spectroscopy (FTIR), wide-angle X-ray diffraction (WXRD) and polarized light microscopy (POM) measurements were used in order to follow the hydrogelation and drug encapsulation processes. The ability of the prepared systems to release the drug has been investigated by UV-Vis spectroscopy using a calibration curve and by fitting the results with different mathematic models. To mimic the behavior of the hydrogel matrix in bio-environmental conditions in view of applications, their enzymatic degradability was monitored in the presence of lysozyme. The in vivo side effects of the systems, in terms of their influence on the blood elements, biochemical and immune parameters were monitored on white Swiss mice by intraperitoneal administration of the injectable obtained hydrogels. All the characteristics of the obtained systems, such as micro-porous morphology, uniform drug encapsulation, enzymatic degradability, lack of side effects, other than the one of the drug itself, along with their ability to release the drug in a sustained manner proved that these material meet the requirements for the development of drug delivery systems, making them suitable for being applied in intraperitoneal chemotherapy.

Development of biocompatible glycodynameric hydrogels joining two natural motifs by dynamic constitutional chemistry.

The paper focusses on the synthesis of novel hydrogels by joining natural biodegradable compounds with the aim to achieve biocompatible materials for bio related applications. The hydrogels were prepared from chitosan and citral by constitutional dynamic chemistry, incorporating both molecular and supramolecular dynamic features. The hydrophobic flexible citral has been reversible immobilized onto the hydrophilic chitosan backbone via imine bonds to form amphiphilic glycodynamers, which further self-ordered through supramolecular interactions into a 3D-network of biodynameric hydrogel. The synthetic pathway has been demonstrated by NMR and FTIR spectroscopy, X-ray diffraction and polarized light microscopy. Studies of the hydrogel morphology revealed a 3D porous microstructure, whose pores size correlated with the crosslinking degree. Rheological investigations evidenced high elasticity, thermo-responsiveness and thixotropic behavior. As a proof of the concept, the hydrogels proved in vivo biocompatibility on laboratory mice. The paper successfully implements the constitutional dynamic chemistry in generation of chitosan high performance hydrogels.

Dual crosslinked iminoboronate-chitosan hydrogels with strong antifungal activity against Candida planktonic yeasts and biofilms.

Chitosan based hydrogels are a class of cross-linked materials intensely studied for their biomedical, industrial and environmental application, but their biomedical use is limited because of the toxicity of different organic crosslinkers. To overcome this disadvantage, a new strategy to produce supramolecular chitosan hydrogels using low molecular weight compounds able to form covalent linkages and H-bonds to give a dual crosslinking is proposed. For this purpose we used 2-formylphenylboronic acid, which brings the advantage of imine stabilization via iminoboronate formation and potential antifungal activity due to the presence of boric acid residue. FTIR and NMR spectroscopy indicated that the gelling process took place by chemo-physical crosslinking forming a dual iminoboronate-chitosan network. Further, X-ray diffraction demonstrated a three-dimensional nanostructuring of the iminoboronate network with consequences on the micrometer-scale morphology and on the improvement of mechanical properties, as demonstrated by SEM and rheological investigation. The hydrogels proved strong antifungal activity against Candida planktonic yeasts and biofilms, promising to be a friendly treatment of the recurrent vulvovaginitis infections.

Dynameric Frameworks for DNA Transfection.

The design of performant nonviral vectors for efficient cellular DNA uptake represents a grand challenge. The variability of DNA targets and of the transfected cells limits the discovery of highly active nonviral vectors. Dynamic constitutional strategy presented here, combining easy synthesis and rapid screening, enables the selection of dynameric frameworks, DFs, for DNA transfection. On the basis of reversible recombination of cationic heads and of hydrophilic/hydrophobic network-constitutive building blocks, the multivalent core-shell dendritic architectures with an optimal diameter of 100 nm may be adaptively generated in the presence of DNA targets. The fittest DFs simultaneously exhibit optimal DNA binding, superior transfection yield to standard transfection SuperFect agent and preserve high HEK 293T cell viability. The present results constitutes an important advancement toward novel biologically friendly, low-cost, and efficient nonviral vectors.