Validation of a Janus role of methotrexate-based PEGylated chitosan nanoparticles in vitro

Recently, methotrexate (MTX) has been used to target to folate (FA) receptor-overexpressing cancer cells for targeted drug delivery. However, the systematic evaluation of MTX as a Janus-like agent has not been reported before. Here, we explored the validity of using MTX playing an early-phase cancer-specific targeting ligand cooperated with a late-phase therapeutic anticancer agent based on the PEGylated chitosan (CS) nanoparticles (NPs) as drug carriers. Some advantages of these nanoscaled drug delivery systems are as follows: (1) the NPs can ensure minimal premature release of MTX at off-target site to reduce the side effects to normal tissue; (2) MTX can function as a targeting ligand at target site prior to cellular uptake; and (3) once internalized by the target cell, the NPs can function as a prodrug formulation, releasing biologically active MTX inside the cells. The (MTX + PEG)-CS-NPs presented a sustained/proteases-mediated drug release. More importantly, compared with the PEG-CS-NPs and (FA + PEG)-CS-NPs, the (MTX + PEG)-CS-NPs showed a greater cellular uptake. Furthermore, the (MTX + PEG)-CS-NPs demonstrated a superior cytotoxicity compare to the free MTX. Our findings therefore validated that the MTX-loaded PEGylated CS-NPs can simultaneously target and treat FA receptor-overexpressing cancer cells.     

Interests of chitosan nanoparticles ionically cross-linked with tripolyphosphate for biomedical applications

The process of ionic gelation is one of the easiest ways to develop chitosan nanoparticles reported so far in the literature. Its success is mainly due to its one-shot synthesis, and to the mild environment required to produce the nanoparticles. The nanoparticle formation all along this process has been therefore thoroughly studied to lead to particles with a nanometric size, a narrow size distribution, and a spherical shape that are ideal for biomedical uses. The purpose of this review is to compile the biomedical applications that have been considered in the literature for these chitosan nanoparticles prepared by ionic gelation using tripolyphosphate as ionic agent. Their intrinsic biological properties such as non-toxicity, antimicrobial activity, mucoadhesivity and haemocompatibility are firstly discussed and compared to those of chitosan solutions. Then, the different bioactive species (drugs and biomacromolecules) incorporated in these chitosan nanoparticles, their maximal incorporation efficiency, their loading capacity, and their principal associated biomedical applications are presented.     

Extraction of chitosan from Aspergillus niger mycelium and synthesis of hydrogels for controlled release of betahistine

Chitosan was extracted from the fungus Aspergillus niger, an alternative source of chitin that is widely available as a byproduct of the industrial production of citric acid. Chitosan with deacetylation degree (DD) of 73.6% was characterized by elemental analysis, capillary viscometry (molecular weight of 1.9 × 10⁵ g/mol), Fourier transform infrared (FTIR), nuclear magnetic resonance (¹H NMR, ¹³C NMR and ¹⁵N NMR) spectroscopies, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). Fungal chitosan was crosslinked with glutaraldehyde and glutaric acid to obtain hydrogels. Chitosan hydrogels were characterized by FTIR and by scanning electron microscopy (SEM), which showed that these materials have irregular, polydisperse, and interconnected pores. Kinetic studies of the release of betahistine from the swollen hydrogels showed a Fickian diffusion mechanism. Finally, hydrolytic degradation of chitosan hydrogels under simulated physiological conditions (pH 7.4 and 37 °C) was investigated as well as in vivo biocompatibility tests using New Zealand white rabbits as animal models.     

Preparation and characterisation of laminated hydroxyapatite/chitosan composite hydrogels

Abstract

Chitosan (CS) has been proposed for the use of electrically modulated drug delivery. However, the gel fatigue makes CS hydrogel difficult to achieve precise and prolonged drug release. In this study, laminated hydroxyapatite (HAp)/CS composite hydrogels were prepared via solution intercalation method. Cyclic electrostimulation test revealed that the fatigue of neat CS was significantly improved by incorporation of laminated HAp. It is detected by transmission electron microscopy that laminated HAp distributed disorderly in the CS matrix, and the dimension of the HAp lamella is ～150 nm. Furthermore, the in vitro cytotoxicity of 2HAp/CS was evaluated by optical microscopy, scanning electron microscopy and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The prepared composite hydrogels exhibit improved swelling fatigue property and good biocompatibility, which will further provide a wide range of potential application in the drug delivery.     

A comparison of the structure, thermal properties, and biodegradability of polycaprolactone/chitosan and acrylic acid grafted polycaprolactone/chitosan

The effects of replacing PCL with acrylic acid grafted PCL (PCL-g-AA) on the structure and properties of a PCL/chitosan composite were investigated. The properties of both PCL-g-AA/chitosan and PCL/chitosan were examined and compared using FTIR, ¹H and ¹³C nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), and a biodegradation test. With PCL-g-AA in the composite, compatibility with chitosan and, consequently, the properties of the blend were both much improved due to the formation of ester and imide groups that conferred better dispersion and homogeneity of chitosan in the matrix. Moreover, PCL-g-AA/chitosan had a lower melt temperature and was, therefore, more easily processed than PCL/chitosan. Resistance to water was higher in the PCL-g-AA/chitosan blend, and consequently so was its resistance to biodegradation in soil and in an enzymatic environment. Nevertheless, weight loss of blends buried in soil or exposed to an enzymatic environment indicates that both blends were biodegradable, especially at high levels of chitosan content. Both blends suffered deterioration in tensile strength and elongation at break after exposure to soil or enzymatic environments.     

Synthesis,characterization, swelling-deswelling properties of Novel nanoparticle-hydrogel containing core chitosan and their cyclohexanone-crosslinked counterparts

Novel Chitosan-cyclohexanone Mannich based hydrogel nanoparticles (CCMb1-4) were prepare via the reaction of chitosan with different concentration of 2, 6-bis (piperidin-1-ylmethyl) cyclohexanone. 2HCl (1%, 5%, 10% and 15% wt/wt) at 70–80°C for 7 h. These hydrogels were subjected to equilibrium swelling studies at room temperature in solutions of pH 2, 4, 6 and 8. The (CCMb1-4) showed maximum percent swellability at pH = 2.0. Furthermore, the swelling of the (CCMb1-4) followed Fickian diffusion. This preliminary investigation of chitosan-based interpolymeric hydrogels showed that they may be exploited to expand the utilization of these systems in drug delivery applications.     

Liposome: classification,preparation, and applications

Liposomes, sphere-shaped vesicles consisting of one or more phospholipid bilayers, were first described in the mid-60s. Today, they are a very useful reproduction, reagent, and tool in various scientific disciplines, including mathematics and theoretical physics, biophysics, chemistry, colloid science, biochemistry, and biology. Since then, liposomes have made their way to the market. Among several talented new drug delivery systems, liposomes characterize an advanced technology to deliver active molecules to the site of action, and at present, several formulations are in clinical use. Research on liposome technology has progressed from conventional vesicles to ‘second-generation liposomes’, in which long-circulating liposomes are obtained by modulating the lipid composition, size, and charge of the vesicle. Liposomes with modified surfaces have also been developed using several molecules, such as glycolipids or sialic acid. This paper summarizes exclusively scalable techniques and focuses on strengths, respectively, limitations in respect to industrial applicability and regulatory requirements concerning liposomal drug formulations based on FDA and EMEA documents.     

Synthesis and characterization of chitosan derivatives carrying galactose residues

A series of water‐soluble chitosan derivatives, carrying galactose residues, were synthesized by using an alternative method in which the galactose groups were introduced into amino groups of the derivatives. First, hydroxyethyl chitosan (HECS) and hydroxypropyl chitosan (HPCS) were synthesized under alkaline conditions by using chitosan and propylene or chitosan and ClCH₂CH₂OH as the starting materials, respectively. Then lactobionic acid was added into the systems so as to form galactosylated HECS (Gal‐HECS) and galactosylated HPCS (Gal‐HPCS) with substitution degrees of 53 and 47%, respectively. Lactosaminated HPCS (Lac‐HPCS) and Lactosaminated HECS (Lac‐HECS) were obtained with substitution degrees of 42 and 38%, respectively, by the reductive amination of the mixtures of lactose and HECS or lactose and HPCS with potassium borohydride present in the reaction. The chemical structures of new chitosan derivatives were characterized by FTIR, ¹H NMR, ¹³C NMR, and elemental analysis. Some physical properties were also analyzed by wide angle X‐ray diffraction (WAXD) and differential scanning calorimetry (DSC). The novel chitosan derivatives carrying galactose residues may be used as additives for hepatic targeting delivery. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 2161–2167, 2005     

Synthesis of a novel polymeric surfactant by reductive N-alkylation of chitosan with 3-O-dodecyl-d-glucose

A novel chitosan-based polymeric surfactant, DG-chitosan, was prepared via reductive N-alkylation of chitosan with 3-O-dodecyl-d-glucose in acetate buffer (pH 4.3, 0.1 M)-methanol in the presence of sodium cyanoborohydride (NaBH₃CN). DG-chitosan was swelling in water, partly dissolvable in pyridine and DMF, and completely soluble in 0.1% aqueous acetic acid. ¹H and ¹³C NMR spectroscopic analyses in 2% acetic acid-d₄-methanol-d₄ together with elemental analysis showed the degree of substitution was 27%. Formation of polymeric micelles was observed by use of pyrene as a fluorescent probe, and the critical aggregation concentration (CAC) of DG-chitosan was marked equal to 28.1 mg/L.     

Synthesis,characterization, and drug‐release behavior of amphiphilic quaternary ammonium chitosan derivatives

A new type of amphiphilic quaternary ammonium chitosan derivative, 2‐N‐carboxymethyl‐6‐O‐diethylaminoethyl chitosan (DEAE–CMC), was synthesized through a two‐step Schiff base reaction process and applied to drug delivery. In the first step, benzaldehyde was used as a protective agent for the incorporation of diethylaminoethyl groups to form the intermediate (6‐O‐diethylaminoethyl chitosan). On the other hand, NaBH₄ was used as a reducing agent to reduce the Schiff base, which was generated by glyoxylic acid, for the further incorporation of carboxymethyl groups to produce DEAE–CMC. The structure, thermal properties, surface morphology, and diameter distribution of the resulting chitosan graft copolymers were characterized by Fourier transform infrared spectroscopy, ¹H‐NMR, thermogravimetric analysis, differential scanning calorimetry, X‐ray powder diffraction, scanning electron microscopy, and laser particle size analysis. Benefiting from the amphiphilic structure, DEAE–CMC was able to be formed into microspheres in aqueous solution with an average diameter of 4.52 ± 1.21 μm. An in vitro evaluation of these microspheres demonstrated their efficient controlled release behavior of a drug. The accumulated release ratio of vitamin B₁₂ loaded DEAE–CMC microspheres were up to 93%, and the duration was up to 15 h. The grafted polymers of DEAE–CMC were found to be blood‐compatible, and no cytotoxic effect was shown in human SiHa cells in an MTT [3‐(4, 5‐dimethyl‐thiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide] cytotoxicity assay. These results indicate that the DEAE–CMC microspheres could be used as safe, promising drug‐delivery systems. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39890.     

苯硼酸及其衍生物在医药与化工领域的应用研究进展

苯硼酸及其衍生物可以与多羟基化合物（多糖、糖脂、糖蛋白及核苷酸等物质）形成可逆络合物,因此可用于这些物质的识别、分离与检测。综述了苯硼酸及其衍生物在自律式胰岛素给药系统、组织工程、生物物质分离系统以及传感器方面的应用研究进展,展望了苯硼酸及其衍生物在医药与化工领域的广阔前景。     

Carbon nanotubes: properties,synthesis, purification,and medical applications

Current discoveries of different forms of carbon nanostructures have motivated research on their applications in various fields. They hold promise for applications in medicine, gene, and drug delivery areas. Many different production methods for carbon nanotubes (CNTs) have been introduced; functionalization, filling, doping, and chemical modification have been achieved, and characterization, separation, and manipulation of individual CNTs are now possible. Parameters such as structure, surface area, surface charge, size distribution, surface chemistry, and agglomeration state as well as purity of the samples have considerable impact on the reactivity of carbon nanotubes. Otherwise, the strength and flexibility of carbon nanotubes make them of potential use in controlling other nanoscale structures, which suggests they will have a significant role in nanotechnology engineering.     

Chitin and chitosan in selected biomedical applications

Chitin (CT), the well-known natural biopolymer and chitosan (CS) (bio-based or “artificial polymer”) are non-toxic, biodegradable and biocompatible in nature. The advantages of these biomaterials are such that, they can be easily processed into different forms such as membranes, sponges, gels, scaffolds, microparticles, nanoparticles and nanofibers for a variety of biomedical applications such as drug delivery, gene therapy, tissue engineering and wound healing. Present review focuses on the diverse applications of CT and CS membranes and scaffolds for drug delivery, tissue engineering and targeted regenerative medicine. The chitinous scaffolds of marine sponges’ origin are discussed here for the first time. These CT based scaffolds obtained from Porifera possess remarkable and unique properties such as hydration, interconnected channels and diverse structural architecture. This review will provide a brief overview of CT and CS membranes and scaffolds toward different kinds of delivery applications such as anticancer drug delivery, osteogenic drug delivery, and growth factor delivery, because of their inimitable release behavior, degradation profile, mucoadhesive nature, etc. The review also provides an overview of the key features of CT and CS membranes and scaffolds such as their biodegradability, cytocompatibility and mechanical properties toward applications in tissue engineering and wound healing.     

Preparation and evaluation of thermo-reversible copolymer hydrogels containing chitosan and hyaluronic acid as injectable cell carriers

Poly(N-isopropylacrylamide) end-capped with a carboxyl group (PNIPAM-COOH) was grafted to chitosan for synthesizing thermo-reversible chitosan-g-poly(N-isopropylacrylamide) (CPN), which was further grafted with hyaluronic acid (HA) to form hyaluronic acid-g-CPN (HA-CPN). PNIPAM-COOH, CPN and HA-CPN formed injectable free-flowing aqueous solutions and exhibited reversible sol-to-gel phase transition (above 5% polymer concentration) at 30 °C. Chemical properties and temperature-dependent physical properties of the polymer hydrogels, such as rheological behavior, phase transition kinetics, and water content were characterized in detail. The mechanical stiffness of hydrogels increased with the presence of chitosan in the copolymer, but decreased after conjugation with HA. Chitosan and HA grafting also endowed higher water content and resistance to volume contraction during phase change of the copolymer solution. In vitro cell culture experiments with chondrocytes and meniscus cells in HA-CPN hydrogel showed beneficial effects on the cell phenotypic morphology, proliferation, and differentiation. Progressive tissue formation was demonstrated by monotonic increases in extracellular matrix contents and mechanical properties.     

Synthesis and characterization of a novel derivative of chitosan

The graft-copolymer of chitosan with 4-(6-methacryloxyhexyloxy)-4′-nitrobiphenyl I was synthesized by radical polymerization. Graft-copolymerization was carried out under homogeneous conditions with AIBN as initiator and 2 wt% acetic acid as solvent. Homopolymerization of I was also carried out to obtain material for comparative analysis. The resulting chitosan graft-copolymers were isolated, purified and characterized with FTIR, NMR, DSC, and TGA. Films were then prepared and characterized by X-ray diffraction, wave guide coupling and infrared dichroism techniques.     

Synthesis and self‐assembly study of zwitterionic amphiphilic derivatives of chitosan

New zwitterionic derivatives of chitosan (CH) were synthesized through the Michael addition reaction of 1‐(3‐sulfopropyl)‐2‐vinylpyridine hydroxide (SPP) with primary amines of deacetylated CHs (with weight‐average molecular weights of 46 and 216 kDa) to obtain SPP‐substituted CHs. The hydrophilic derivatives were subsequently modified with 2.1, 4.6, and 9.7% of dodecyl groups [degree of substitution by dodecyl groups (DS_Dod)]. The SPP‐substituted CH derivatives were characterized by ¹H‐NMR, Fourier transform infrared spectroscopy, and gel permeation chromatography. Aqueous solutions of SPP‐substituted CH samples remained clear, independently of the pH (3.0 < pH < 12.0). The self‐association study of the amphiphilic derivatives was performed in aqueous buffered solution at pH 5.0 and 7.4, and the critical aggregation concentration values varied from 5.6 × 10^?3 to 0.02 g/L. The measurements of dynamic light scattering and ζ potentials showed that the self‐assembly behavior was dependent on the pH and DS_Dod. At pH 7.4, the measured ζ potentials were near zero, and colloidal stability was provided by the hydrated zwitterionic shell of the aggregates. Transmission electron microscopy revealed spherelike microsized particles of broad distribution. The amphiphilic SPP‐substituted CH samples were shown to be nontoxic with a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay performed with HeLa cells. The remarkable water solubility and nontoxicity displayed by the new SPP‐substituted CH derivatives showed promising properties for the design of CH‐based biomaterials and nanoparticles. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 44176.     

pH-responsive micelles based on (PCL)2(PDEA-b-PPEGMA)2 miktoarm polymer: controlled synthesis,characterization, and application as anticancer drug carrier

Amphiphilic A₂(BC)₂ miktoarm star polymers [poly(ϵ-caprolactone)]₂-[poly(2-(diethylamino)ethyl methacrylate)-b- poly(poly(ethylene glycol) methyl ether methacrylate)]₂ [(PCL)₂(PDEA-b-PPEGMA)₂] were developed by a combination of ring opening polymerization (ROP) and continuous activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP). The critical micelle concentration (CMC) values were extremely low (0.0024 to 0.0043 mg/mL), depending on the architecture of the polymers. The self-assembled empty and doxorubicin (DOX)-loaded micelles were spherical in morphologies, and the average sizes were about 63 and 110 nm. The release of DOX at pH 5.0 was much faster than that at pH 6.5 and pH 7.4. Moreover, DOX-loaded micelles could effectively inhibit the growth of cancer cells HepG2 with IC₅₀ of 2.0 μg/mL. Intracellular uptake demonstrated that DOX was delivered into the cells effectively after the cells were incubated with DOX-loaded micelles. Therefore, the pH-sensitive (PCL)₂(PDEA-b-PPEGMA)₂ micelles could be a prospective candidate as anticancer drug carrier for hydrophobic drugs with sustained release behavior.     

Synthesis,characterization, and in vitro cytotoxicity of chitosan hydrogels containing nanogold

Hydrogels due to their unique properties are characterized by a wide range of applications. In presented research a series of hydrogel based on chitosan and modified with nanogold have been prepared by photopolymerization. Based on the research it can be concluded that presented hydrogels are interesting materials that can play a key role in medicine. They are characterized by biocompatibility to simulated physiological fluids and do not show any cytotoxicity to the dermal cells. Furthermore, they are characterized by relatively high sorption capacity. These results show the possibility of the potential use of these hydrogels for biomedical purposes.     

Alginate: properties and biomedical applications

Alginate is a biomaterial that has found numerous applications in biomedical science and engineering due to its favorable properties, including biocompatibility and ease of gelation. Alginate hydrogels have been particularly attractive in wound healing, drug delivery, and tissue engineering applications to date, as these gels retain structural similarity to the extracellular matrices in tissues and can be manipulated to play several critical roles. This review will provide a comprehensive overview of general properties of alginate and its hydrogels, their biomedical applications, and suggest new perspectives for future studies with these polymers.     

Amphiphilic chitosan graft copolymer via combination of ROP, ATRP and click chemistry: Synthesis, self-assembly, thermosensitivity, fluorescence, and controlled drug release

Novel amphiphilic chitosan-g-poly(ε-caprolactone)-(g-poly(2-(2-methoxyethoxy)ethyl methacrylate)-co-oligo(ethylene glycol) methacrylate) (CS-g-PCL(-g-P(MEO₂MA-co-OEGMA))) copolymers with double side chains of PCL and P(MEO₂MA-co-OEGMA) were synthesized via combination of ring-opening polymerization (ROP), atom transfer radical polymerization (ATRP) and click chemistry. The molar ratio of PCL and P(MEO₂MA-co-OEGMA) was varied through variation of the feed ratio and the coupling efficiency of click chemistry is comparatively high. The graft copolymers can assemble into spherical micelles. The micelles show thermosensitive properties and the lower critical solution temperatures (LCSTs) were influenced by CS chains and the ratio of PCL and P(MEO₂MA-co-OEGMA) side chains. Moreover, the micelles can reversibly swell and shrink in response to the change of temperatures. Furthermore, the micelles present obvious fluorescence and the fluorescent intensity can be adjusted by altering the temperatures. The investigation of doxorubicin release from the micelles indicated that the release rate of the drug could be effectively controlled by altering the temperatures.