Chitosan-dextran sulphate nanocapsule drug delivery system as an effective therapeutic against intraphagosomal pathogen Salmonella.

OBJECTIVES: The ability to target conventional drugs efficiently inside cells to kill intraphagosomal bacteria has been a major hurdle in treatment of infective diseases. We aimed to develop an efficient drug delivery system for combating infection caused by Salmonella, a well-known intracellular and intraphagosomal pathogen. Chitosan-dextran sulphate (CD) nanocapsules were assessed for their efficiency in delivering drugs against Salmonella. METHODS: The CD nanocapsules were prepared using the layer-by-layer method and loaded with ciprofloxacin or ceftriaxone. Antibiotic-loaded nanocapsules were analysed in vitro for their ability to enter epithelial and macrophage cells to kill Salmonella. In vivo pharmacokinetics and organ distribution studies were performed to check the efficiency of the delivery system. The in vivo antibacterial activity of free antibiotic and antibiotic loaded into nanocapsules was tested in a murine salmonellosis model. RESULTS: In vitro and in vivo experiments showed that this delivery system can be used effectively to clear Salmonella infection. CD nanocapsules were successfully employed for efficient targeting and killing of the intracellular pathogen at a dosage significantly lower than that of the free antibiotic. The increased retention time of ciprofloxacin in the blood and organs when it was delivered by CD nanocapsules compared with the conventional routes of administration may be the reason underlying the requirement for a reduced dosage and frequency of antibiotic administration. CONCLUSIONS: CD nanocapsules can be used as an efficient drug delivery system to treat intraphagosomal pathogens, especially Salmonella infection. This delivery system might be used effectively for other vacuolar pathogens including Mycobacteria, Brucella and Legionella.

Design of Tailor-Made Biopolymer-Based Capsules for Biological Application by Combining Porous Particles and Polysaccharide Assembly.

Various approaches have been described in the literature to demonstrate the possibility of designing biopolymer particles with well-defined characteristics, such as size, chemical composition or mechanical properties. From a biological point of view, the properties of particle have been related to their biodistribution and bioavailability. Among the reported core-shell nanoparticles, biopolymer-based capsules can be used as a versatile platform for drug delivery purposes. Among the known biopolymers, the present review focuses on polysaccharide-based capsules. We only report on biopolyelectrolyte capsules fabricated by combining porous particles as a template and using the layer-by-layer technique. The review focuses on the major steps of the capsule design, i.e., the fabrication and subsequent use of the sacrificial porous template, multilayer coating with polysaccharides, the removal of the porous template to obtain the capsules, capsule characterisation and the application of capsules in the biomedical field. In the last part, selected examples are presented to evidence the major benefits of using polysaccharide-based capsules for biological purposes.

Impact of Macrophages on the Interaction of Cetuximab-Functionalized Polyelectrolyte Capsules with EGFR-Expressing Cancer Cells.

Polyelectrolyte capsules (PCs) are a promising tool for anticancer drug delivery and tumor targeting. Surface functionalization of PCs with antibodies is widely used for providing their specific interactions with cancer cells. The efficiency of PC-based targeted delivery systems can be affected by the cellular heterogeneity of the tumor, particularly by the presence of tumor-associated macrophages. We used human epidermoid carcinoma cells and macrophages derived from human leukemia monocytic cells in either monoculture or coculture to analyze the targeting capacity and internalization efficiency of PCs with a mean size of 1.03 ± 0.11 µm. The PCs were functionalized with the monoclonal antibody cetuximab targeting the human epidermal growth factor receptor (EGFR). We have shown that surface functionalization of the PCs with cetuximab ensures a specific interaction with EGFR-expressing cancer cells and promotes capsule internalization. In monoculture, the macrophages derived from human leukemia monocytic cells have been found to internalize both nonfunctionalized PCs and cetuximab-functionalized PCs (Cet-PCs) more intensely compared to epidermoid carcinoma cells. The internalization of Cet-PCs by cancer cells is mediated by lipid rafts of the cell membrane, whereas the PC internalization by macrophages is only slightly influenced by lipid rafts. Experiments with a coculture of human epidermoid carcinoma cells and macrophages derived from human leukemia monocytic cells have shown that Cet-PCs preferentially interact with cancer cells, which are subsequently attacked by macrophages. These data can be used to further improve the strategy of PC functionalization for targeted delivery, with the cellular heterogeneity of the tumor microenvironment taken into consideration.

Encapsulation of Enzymes in Porous Capsules via Particle Templating.

The entrapment of enzymes in capsules is a smart strategy to concentrate them in confined spaces and control their exposure to outside environments. Enzymes can be caged in the interior of capsules during their formation (preloading) or postloaded within prefabricated and permeable hollow shells. On the other hand, enzymes can also be deposited within the shell or on the surface of the capsules. Each of these strategies has intrinsic limitations, and a common enemy is the undesired desorption of enzymes.Here, we describe the formation of enzyme-loaded polymeric capsules prepared with the Layer-by-Layer method and the template-assisted entrapment of enzymes through coprecipitation (preloading) within calcium carbonate particles, as an example of an efficient preloading strategy, and draw attention at the key parameters that influence this immobilization method.

Biodegradable Alginate Polyelectrolyte Capsules As Plausible Biocompatible Delivery Carriers.

Polyelectrolyte capsules made of different biodegradable and nonbiodegradable polymers can be designed as systems for effective encapsulation and delivery of compounds. The objective of this work was to synthesize biocompatible and biodegradable capsules (<1 µm) by the layer-by-layer (LbL) approach using alginate (ALGI) and poly-l-arginine (PARG) polyelectrolytes with a pH-sensitive outer layer of EUDRAGIT L 100 (EuL) polymer. Those capsules were loaded with curcumin as a model therapeutic drug, which possesses antioxidant, anti-inflammatory, and anticancer activity. Encapsulation of drugs inside capsules protects its therapeutic activity and increases its bioavailability. We report the capsule stability, loading efficiency, drug release, as well as capsule degradation studies as a function of pH. Furthermore, in vitro biocompatibility studies of capsules including cell viability and uptake studies were performed using HeLa cells. The here synthesized capsules exhibited good reproducibility, spherical shape, and high monodispersibility. The capsules showed good loading efficiency and drug release profile dependent upon pH environment. The in vitro studies indicate that the capsules exhibited acceptable biocompatibility and are highly internalized by cells. Our study thus suggests that alginate LbL capsules could be used as an efficient drug carrier with effective encapsulation and successful in vitro release of cargo in the cell.

Intracellular redox induced drug release in cancerous and mesenchymal stem cells.

In this report, we investigated intracellular redox induced drug release in cancerous cells and human mesenchymal stem cells (MSCs) as an example of healthy cells using redox-responsive microcapsules with covalently bonded anti-cancer drug (doxorubicin) via the amine-reactive cross-linker, 3,3'-dithiobis(sulfosuccinimidyl propionate) containing disulfide bond. Such rationally designed capsules with incorporated redox-sensitive cross-linker are capable of controllable Dox release in the presence of glutathione (GSH) due to a thiol-cleavable disulfide bonds. The treatment of human MSCs and human cervical cancer cell line (HeLa) with Dox-conjugated capsules showed that the Dox release was observed only when capsules incubated with HeLa cells which can be induced by high GSH level in cancerous (HeLa) cells. Moreover, the results of cell viability indicated that Dox-conjugated capsules are more effective when inducing cell death of HeLa than free Dox improving the anti-tumor efficacy of chemotherapeutic drug and simultaneously they possess lower cytotoxicity against MSCs compared to cancerous cells. Such properties are important in design of smart drug carriers for efficient cancer therapy.

Highly active antibody-modified magnetic polyelectrolyte capsules.

Polyelectrolyte hollow capsules are versatile platforms typically used for encapsulation of a wide variety of macromolecules in their cavity. The polymer shell of these capsules as composed by alternating layers of oppositely charged polyelectrolytes also allows for adding additional functionalities. The properties of the shell can be for example engineered by trapping different nanoparticles in-between the shell layers and/or by attaching bioactive molecules such as antibodies to the outermost layer. Herein, iron oxide NPs were inmobilized into the shell of polyelectrolyte capsules and the outermost layer of the shell was covalently modified with anti peroxidase antibodies. These capsules act as prototype model system, aiming to obtain a microstructure with the potential capability to specifically recognize and separate macromolecules. Due to the magnetic nanoparticles in the capsule shell, the capsules together with the attached target might be extracted by magnetic field gradients. Here we verified this approach by extracting horseradish peroxidase from a solution through magnetic separation with capsules bearing antibodies against horseradish peroxidase. The bioactivity of the capsules and the high degree of specific antibody functionalization were confirmed and quantified through an enzymatic reaction mediated by the extracted horseradish peroxidase.

Mesenchymal Stem Cell Magnetization: Magnetic Multilayer Microcapsule Uptake, Toxicity, Impact on Functional Properties, and Perspectives for Magnetic Delivery.

Mesenchymal stem cells (MSCs) are widely used in cell therapy due to their convenience, multiline differentiation potential, reproducible protocols, and biological properties. The potential of MSCs to impregnate magnetic microcapsules and their possible influence on cell function and ability to response to magnetic field have been explored. Interestingly, the cells suspended in media show much higher ability in internalization of microcapsules, then MSCs adhere into the surface. There is no significant effect of microcapsules on cell toxicity compared with other cell line-capsule internalization reported in literature. Due to internalization of magnetic capsules by the cells, such cell engineering platform is responsive to external magnetic field, which allows to manipulate MSC migration. Magnetically sorted MSCs are capable to differentiation as confirmed by their conversion to adipogenic and osteogenic cells using standard protocols. There is a minor effect of capsule internalization on cell adhesion, though MSCs are still able to form spheroid made by dozen of thousand MSCs. This work demonstrates the potential of use of microcapsule impregnated MSCs to carry internalized micron-sized vesicles and being navigated with external magnetic signaling.

Functionalized biocompatible polyelectrolyte multilayers for drug delivery: in situ investigation of mechanical properties by dissipative quartz crystal microbalance.

Nanostructured polymeric capsules have been applied in different fields, and specifically are regarded as promising for smart drug delivery applications. The physical-chemical and mechanical properties, and thus the permeability of the polyelectrolyte multilayer shell, play an important role in efficient delivery. Quartz crystal microbalance working in liquid has been used for the characterization of the buildup process and of the viscoelastic properties of biocompatible multilayers and of their functionalization by S-layer proteins. Optical and scanning electron microscopy have been used for the morphological characterization of nanostructured capsules obtained at physiological conditions by the assembly of the characterized multilayers onto spherical cores and by their subsequent removal. The proposed functionalized biocompatible capsules can be regarded as promising candidates for smart drug delivery applications.

Polyelectrolyte based molecular carriers: the role of self-assembled proteins in permeability properties.

Polyelectrolyte capsules are seen as promising nanotechnology based drug delivery systems. In previous works, we have demonstrated the possibility to fabricate bio-activated surface layer containing capsules with improved features in terms of biocompatibility. In this study, we have characterized the permeability properties of such capsules towards low and high molecular weight molecules, including proteins. The results indicated that the presence of the surface layer strongly affects the permeability properties of the capsules in terms of loading capacity which was found to be higher compared to that of plain capsules. These properties make such systems interesting candidates as drug delivery platforms.