Chitosan-Based Dressing Materials for Problematic Wound Management.

Wound healing is a complex mechanism involving a variety of factors and is a representative process of tissue growth and regeneration in our body. Surface-based interactions between the dressing material and the wound may significantly influence the healing phase. Advances in understanding the mechanism of wound healing have led to the development of numerous dressing materials that can accelerate the healing process. However, these materials have a passive role in wound healing. It is therefore necessary to develop novel wound dressing materials, especially effective for clinically problematic wounds. Chitosan-based dressing materials are considered suitable for clinically problematic wounds as they exhibit several characteristic features, such as facilitating hemostasis, enhanced wound healing during the inflammatory and proliferative phases, antimicrobial effect, etc. Here, we review the current status of clinically available dressing materials and studies on the biological characteristics of chitosan, and discuss the potential applications of chitosan in multi-functional dressing materials for accelarated wound healing.

Hydrolyzed hexagonal boron nitride/polymer nanocomposites for transparent gas barrier film.

A flexible thin gas barrier film formed by layer-by-layer (LBL) assembly has been studied. We propose for the first time that hexagonal boron nitride (h-BN) can be used in LBL assembly. When dispersed in water through sonication-assisted hydrolysis, h-BN develops hydroxyl groups that electrostatically couple with the cationic polymer polydiallyldimethylammonium chloride (PDDA). This process produces hydroxyl-functional h-BN/PDDA nanocomposites. The nanocomposites exhibit well exfoliated and highly ordered h-BN nanosheets, which results in an extremely high visual clarity, with an average transmittance of 99% in the visible spectrum. Moreover, well aligned nanocomposites extend gas diffusion path that reduce water vapor transmission rate to 1.3 × 10-2 g m-2 d-1. The simple and fast LBL process demonstrated here can be applied in many gas barrier applications.

Self-assembled nanocomplex between polymerized phenylboronic acid and doxorubicin for efficient tumor-targeted chemotherapy.

Since the discovery that nano-scaled particulates can easily be incorporated into tumors via the enhanced permeability and retention (EPR) effect, such nanostructures have been exploited as therapeutic small molecule delivery systems. However, the convoluted synthetic process of conventional nanostructures has impeded their feasibility and reproducibility in clinical applications. Herein, we report an easily prepared formulation of self-assembled nanostructures for systemic delivery of the anti-cancer drug doxorubicin (DOX). Phenylboronic acid (PBA) was grafted onto the polymeric backbone of poly(maleic anhydride). pPBA-DOX nanocomplexes were prepared by simple mixing, on the basis of the strong interaction between the 1,3-diol of DOX and the PBA moiety on pPBA. Three nanocomplexes (1, 2, 4) were designed on the basis of [PBA]:[DOX] molar ratios of 1:1, 2:1, and 4:1, respectively, to investigate the function of the residual PBA moiety as a targeting ligand. An acid-labile drug release profile was observed, owing to the intrinsic properties of the phenylboronic ester. Moreover, the tumor-targeting ability of the nanocomplexes was demonstrated, both in vitro by confocal microscopy and in vivo by fluorescence imaging, to be driven by an inherent property of the residual PBA. Ligand competition assays with free PBA pre-treatment demonstrated the targeting effect of the residual PBA from the nanocomplexes 2 and 4. Finally, the nanocomplexes 2 and 4, compared with the free DOX, exhibited significantly greater anti-cancer effects in vitro and even in vivo. Our pPBA-DOX nanocomplex enables a new paradigm for self-assembled nanostructures with potential biomedical applications.

Clinical effect of a polysaccharide-rich extract of Acanthopanax senticosus on alcohol hangover.

The present study aimed to examine the effects polysaccharide-rich extract of Acanthopanax senticosus (PEA) on blood alcohol concentration (BAC) and hangover as well as blood lab parameters. A randomized, placebo-controlled, double-blind crossover trial was conducted. The PEA was orally administered before and after consuming alcohol 1.75 g/kg of pure alcohol. After alcohol consumption, BAC was measured for evaluation of alcohol pharmacokinetics. In the second day morning, subjects were asked to complete the Acute Hangover Scale (AHS) questionnarie. BAC results showed little difference between placebo and PEA groups, indicating that PEA does not have an effect on the pharmacokinetics of alcohol. However, several AHS items (i.e., tired, headache, dizziness, stomachache and nausea) and AHS total score were significantly improved by PEA. Blood lab parameters were significantly altered by alcohol in the placebo group. The alteration by alcohol of glucose and C-reactive protein (CRP) level was significantly attenuated by PEA. Therefore, PEA may have potential to reduce the severity of the alcohol hangover by inhibiting the alcohol-induced hypoglycemia and inflammatory response.

Phage lambda capsids as tunable display nanoparticles.

Nanoparticle technologies provide a powerful tool for the development of reagents for use in both therapeutic and diagnostic, or "theragnostic" biomedical applications. Two broad classes of particles are under development, viral and synthetic systems, each with their respective strengths and limitations. Here we adapt the phage lambda system to construct modular "designer" nanoparticles that blend these two approaches. We have constructed a variety of modified "decoration" proteins that allow site-specific modification of the shell with both protein and nonproteinaceous ligands including small molecules, carbohydrates, and synthetic display ligands. We show that the chimeric proteins can be used to simultaneously decorate the shell in a tunable surface density to afford particles that are physically homogeneous and that can be manufactured to display a variety of ligands in a defined composition. These designer nanoparticles set the stage for development of lambda as a theragnostic nanoparticle system.

Pathogen-derived oligosaccharides improve innate immune response to intracellular parasite infection.

Pathogen glycolipids, including Leishmania spp. lipophosphoglycan (LPG) and Mycobacterium tuberculosis mannosylated lipoarabinomannan (ManLAM), modulate essential interactions with host phagocytic cells. Polysaccharide and lipid components promote immunomodulation. Owing to the stereochemistry required to synthesize oligosaccharides, the roles for oligosaccharides in the pathogenesis of infectious diseases have remained largely unknown. Recent advances in carbohydrate chemistry allowed us to synthesize pathogen surface oligosaccharides to discern their immune response-altering activities. Trimannose cap carbohydrates from ManLAM and LPG altered the production of proinflammatory cytokines via a toll-like receptor (TLR2)-mediated mechanism in vitro and in vivo. In vivo treatment with trimannose led to increased Th1-polarizing, IL-12p40-producing cells from the draining lymph nodes of treated Leishmania major-infected mice compared with cells from untreated infected mice. Trimannose treatment increased the production of other Th1 proinflammatory cytokines (ie, interferon-γ, IL-6, and tumor necrosis factor-α) critical for a productive immune response to either pathogen. This significant difference in cytokine production between trimannose cap sugar-treated and control groups was not observed in draining lymph node cells from TLR2(-/-) mice. Type of inflammation and rate of bead entry into macrophages and dendritic cells were different for trimannose-coated beads compared with control oligosaccharide-coated beads, indicating selective lectin receptor/oligosaccharide interactions mediating cell entry and cytokine production. These novel findings may prompt the development of targeted oligosaccharide adjuvants against chronic infections.

Fuzzy nanoassembly of polyelectrolyte and layered clay multicomposite toward a reliable gas barrier.

Flexible and transparent gas-diffusion barriers have played an important role in recent years. The present study describes a flexible barrier film with a tailored architecture of cationic polyelectrolytes and clay/polymer nanoassemblies. Highly oriented and well-aligned barrier films were achieved by the consecutive absorption of flexible cationic polymer and anionic montmorillonite platelets. The experimental results showed that the layer-by-layer deposition of oppositely charged thin films containing self-assembled poly(vinyl alcohol) and montmorillonites improved their gas barrier characteristics based on the Ca degradation test, enhancing their optical transparency. This nanostructure, fabricated using a solution process, is useful in many applications, for example, flexible and moisture-free organic electronics. This simple and fast method is suitable for the mass coating of large surface areas, as required in industry.

Mannose-functionalized "pathogen-like" polyanhydride nanoparticles target C-type lectin receptors on dendritic cells.

Targeting pathogen recognition receptors on dendritic cells (DCs) offers the advantage of triggering specific signaling pathways to induce a tailored and robust immune response. In this work, we describe a novel approach to targeted antigen delivery by decorating the surface of polyanhydride nanoparticles with specific carbohydrates to provide "pathogen-like" properties that ensure nanoparticles engage C-type lectin receptors on DCs. The surface of polyanhydride nanoparticles was functionalized by covalent linkage of dimannose and lactose residues using an amine-carboxylic acid coupling reaction. Coculture of functionalized nanoparticles with bone marrow-derived DCs significantly increased cell surface expression of MHC II, the T cell costimulatory molecules CD86 and CD40, the C-type lectin receptor CIRE and the mannose receptor CD206 over the nonfunctionalized nanoparticles. Both nonfunctionalized and functionalized nanoparticles were efficiently internalized by DCs, indicating that internalization of functionalized nanoparticles was necessary but not sufficient to activate DCs. Blocking the mannose and CIRE receptors prior to the addition of functionalized nanoparticles to the culture inhibited the increased surface expression of MHC II, CD40 and CD86. Together, these data indicate that engagement of CIRE and the mannose receptor is a key mechanism by which functionalized nanoparticles activate DCs. These studies provide valuable insights into the rational design of targeted nanovaccine platforms to induce robust immune responses and improve vaccine efficacy.

Synthesis of multivalent tuberculosis and Leishmania-associated capping carbohydrates reveals structure-dependent responses allowing immune evasion.

Mycobacterium tuberculosis and the protozoan parasites of the genus Leishmania are intracellular pathogens that can survive in macrophages--the very white blood cells of the immune system responsible for engulfing and ultimately clearing foreign invaders. The ability of these pathogens to hide within immune cells has made the design of effective therapies, including vaccines, to control tuberculosis and leishmaniasis particularly challenging. Herein we present the synthesis and development of carbohydrate-based probes to demonstrate that changes in pathogen-associated surface oligosaccharides are sufficient to alter cellular immune responses and thereby let a pathogen hide from immune surveillance.

Highly conformal SiO2/Al2O3 nanolaminate gas-diffusion barriers for large-area flexible electronics applications.

The present study demonstrates a flexible gas-diffusion barrier film, containing an SiO(2)/Al(2)O(3) nanolaminate on a plastic substrate. Highly uniform and conformal coatings can be made by alternating the exposure of a flexible polyethersulfone surface to vapors of SiO(2) and Al(2)O(3), at nanoscale thickness cycles via RF-magnetron sputtering deposition. The calcium degradation test indicates that 24 cycles of a 10/10 nm inorganic bilayer, top-coated by UV-cured resin, greatly enhance the barrier performance, with a permeation rate of 3.79 × 10(-5) g m(-2) day(-1) based on the change in the ohmic behavior of the calcium sensor at 20 °C and 50% relative humidity. Also, the permeation rate for 30 cycles of an 8/8 nm inorganic bilayer coated with UV resin was beyond the limited measurable range of the Ca test at 60 °C and 95% relative humidity. It has been found that such laminate films can effectively suppress the void defects of a single inorganic layer, and are significantly less sensitive against moisture permeation. This nanostructure, fabricated by an RF-sputtering process at room temperature, is verified as being useful for highly water-sensitive organic electronics fabricated on plastic substrates.

Porous calcium phosphate-collagen composite microspheres for effective growth factor delivery and bone tissue regeneration.

Microspheres are beneficial for filling defects of various shapes and provide a large surface area for cell attachment. Porous microspheres have attracted particular attention because they can deliver cells and bioactive molecules such as growth factors. In this study, BCP-collagen composite microspheres were developed for growth factor delivery in bone regeneration. Firstly, porous biphasic calcium phosphate (BCP) microspheres were fabricated by applying a water-in-oil emulsion technique using camphene as a pore generator. Then, porous BCP-collagen composite microspheres were fabricated by repetitively dip coating the microspheres in a collagen solution to effectively deliver growth factor to bone defects. Characterization of the microspheres and in vitro studies were conducted to investigate the effect of collagen infiltration on bone regeneration. In addition, in vitro evaluation demonstrated the sustained bone morphogenetic protein-2 (BMP-2) delivery of the microspheres and the effect of cell differentiation, and in vivo assessment with rabbits revealed that the microspheres filled the defect well and that bone could be regenerated through the microspheres. Moreover, the composite system was more effective for bone regeneration than the bare BCP microspheres because of the drug retention of collagen. These findings indicate that the porous microspheres are effective for tissue regeneration by continuous growth factor delivery.

Nanoparticle-based computing architecture for nanoparticle neural networks.

The lack of a scalable nanoparticle-based computing architecture severely limits the potential and use of nanoparticles for manipulating and processing information with molecular computing schemes. Inspired by the von Neumann architecture (VNA), in which multiple programs can be operated without restructuring the computer, we realized the nanoparticle-based VNA (NVNA) on a lipid chip for multiple executions of arbitrary molecular logic operations in the single chip without refabrication. In this system, nanoparticles on a lipid chip function as the hardware that features memory, processors, and output units, and DNA strands are used as the software to provide molecular instructions for the facile programming of logic circuits. NVNA enables a group of nanoparticles to form a feed-forward neural network, a perceptron, which implements functionally complete Boolean logic operations, and provides a programmable, resettable, scalable computing architecture and circuit board to form nanoparticle neural networks and make logical decisions.

In-vitro blood and vascular compatibility of sirolimus-eluting organic/inorganic hybrid stent coatings.

The surface characteristics of coronary stents play a pivotal role in inhibiting in-stent restenosis and late-stent thrombosis. In this study, a sol-gel-derived silica xerogel-chitosan hybrid coating was applied to Co-Cr stent and was reported, for the first time, as a biocompatible drug delivery tool in vascular stent application. A dense and uniform chitosan-silica xerogel hybrid coating (<1-µm thick) was applied on bare Co-Cr material. Sirolimus was well incorporated into the hybrid coatings without re-crystallization. The chitosan-silica hybrid coating with 30 wt% silica xerogel showed better mechanical stability and good adhesive strength without any cracking or delamination. The chitosan-silica hybrid coated Co-Cr surface exhibited significantly improved wettability and corrosion resistance compared to the chitosan coated Co-Cr surface. In addition, the hybrid coating layer enabled efficient loading of sirolimus, owing to the unique mesoporous structure of silica xerogel, which further allowed the sustained release of sirolimus over 3 weeks. In-vitro tests with human umbilical cord vein endothelial cells and blood platelets confirmed that the chitosan-silica hybrid coating had excellent cytocompatibility and hemocompatibilty. Thus, this study demonstrated that the chitosan-silica hybrid material is a promising material for coating coronary stents, with minimal risk of in-stent restenosis and thrombogenicity.

Use of thioglycerol on porous polyurethane as an effective theranostic capping agent for bone tissue engineering.

Thiolated biodegradable polyurethane (TG-DPU) was synthesized using a one-pot reaction with thioglycerol adopted as a functionalized chain extender. After characterization of the chemical structure of TG-DPU using proton nuclear magnetic resonance spectroscopy, bone morphogenetic protein (BMP-2) was loaded in the TG-DPU under oxidative conditions to form disulfides between the free thiol of TG-DPU and BMP-2. The interaction between TG-DPU and BMP-2, so-called bioconjugates, was investigated using X-ray photoelectron spectroscopy analysis; the appearance of disulfide (S-S) linkage indicated the formation of a polymer/growth factor conjugate system. The covalently linked bioconjugates provided stability with minimal loss during the drug delivery with prolonged release performance in in vitro release tests. The effects of the drugs delivered by TG-DPU were also confirmed by in vitro alkaline phosphatase tests using pre-osteoblasts and in vivo bone regeneration tests. The drugs effectively induced cell differentiation and promoted mature bone recovery.

Reduced fibrous capsule formation at nano-engineered silicone surfaces via tantalum ion implantation.

Although the design of more biocompatible polymeric implants has been studied for decades, their intended functionality continues to be impaired by the response of the host tissue to foreign bodies at the tissue-implant interface. In particular, the formation and contracture of fibrous capsules prevent the intimate integration of an implant with surrounding tissues, which leads to structural deformation of the implants and persistent discomfort and pain. We report a new surface nano-engineered silicone implant that reduces fibrous capsule formation and improves the biocompatibility of it via sputtering-based plasma immersion ion implantation (S-PIII). This technique can introduce biologically compatible tantalum (Ta) on the silicone surface to produce a Ta-implanted skin layer (<60 nm thick) as well as generate either smooth (Smooth/Ta silicone) or nano-textured (Nano/Ta silicone) surface morphologies. The biologically inert chemical structure and strong hydrophobic surface characteristics of bare silicone are substantially ameliorated after Ta ion implantation. In particular, the Nano/Ta silicone implant's combination of surface nano-texturing as a physical cue and the Ta-implanted layer as a chemical cue was found to be very effective at achieving outstanding hydrophilicity and fibroblast affinity compared to the bare and Smooth/Ta silicone implants. In a mouse in vivo study conducted for 8 weeks, the Nano/Ta silicone implant inhibited fibrous capsule formation and contracture on its surface better than the bare silicone based on an analysis of the number of macrophages, myofibroblast differentiation and activation, collagen density, and thickness of fibrous capsules.

Fluorine-ion-releasing injectable alginate nanocomposite hydrogel for enhanced bioactivity and antibacterial property.

The creation of a moist environment and promotion of cell proliferation and migration together with antibacterial property are critical to the wound-healing process. Alginate (Alg) is an excellent candidate for injectable wound dressing materials because it can form a gel in a mild environment. Taking advantage of its gelation property, an injectable nano composite hydrogel containing nano-sized (about 90 nm) calcium fluoride (CaF2) particles was developed using in-situ precipitation process. The amount of released fluorine (F-) ion from the nanocomposite hydrogel increased with increasing CaF2 content inside the composite hydrogel and the ions stimulated both the proliferation and migration of fibroblast cells in vitro. The antibacterial property of the composite hydrogel against E. coli and S. aureus was confirmed through colony formation test where the number of bacterial colonies significantly decreased compared to Alg hydrogel. The in vivo results based on a full-thickness wound model showed that the nanocomposite hydrogel effectively enhanced the deposition of the extracellular matrix compared to that of the Alg hydrogel. This study demonstrates the potential of this nanocomposite hydrogel as a bioactive injectable wound-dressing material with the ability to inhibit bacterial growth and stimulate cell proliferation and migration for accelerated wound healing.

Enhanced Osseointegration Ability of Poly(lactic acid) via Tantalum Sputtering-Based Plasma Immersion Ion Implantation.

Poly(lactic acid) (PLA) is the most utilized biodegradable polymer in orthopedic implant applications because of its ability to replace regenerated bone tissue via continuous degradation over time. However, the poor osteoblast affinity for PLA results in a high risk of early implant failure, and this issue remains one of the most difficult challenges with this technology. In this study, we demonstrate the use of a new technique in which plasma immersion ion implantation (PIII) is combined with a conventional DC magnetron sputtering. This technique, referred to as sputtering-based PIII (S-PIII), makes it possible to produce a tantalum (Ta)-implanted PLA surface within 30 s without any tangible degradation or deformation of the PLA substrate. Compared to a Ta-coated PLA surface, the Ta-implanted PLA showed twice the surface roughness and substantially enhanced adhesion stability in dry and wet conditions. The strong hydrophobic surface properties and biologically relatively inert chemical structure of PLA were ameliorated by Ta S-PIII treatment, which produced a moderate hydrophilic surface and enhanced cell-material interactions. Furthermore, in an in vivo evaluation in a rabbit distal femur implantation model, Ta-implanted PLA demonstrated significantly enhanced osseointegration and osteogenesis compared with bare PLA. These results indicate that the Ta-implanted PLA has great potential for orthopedic implant applications.

Acceleration of the healing process of full-thickness wounds using hydrophilic chitosan-silica hybrid sponge in a porcine model.

In this study, we evaluated the surface characterization of a novel chitosan-silica hybridized membrane and highlighted the substantial role of silica in the wound environment. The chemical coupling of chitosan and silica resulted in a more condensed network compared with pure chitosan, which was eventually able to stably maintain its framework, particularly in the wet state. In addition, we closely observed the wound-healing process along with the surface interaction between chitosan-silica and the wound site using large-surface-area wounds in a porcine model. Our evidence indicates that chitosan-silica exerts a synergetic effect of both materials to promote a remarkable wound-healing process. In particular, the silica in chitosan-silica accelerated wound closure including wound contraction, and re-epithelialization via enhancement of cell recruitment, epidermal maturity, neovascularization, and granulation tissue formation compared with pure chitosan and other commercial dressing materials. This advanced wound dressing material may lead to effective treatment for problematic cutaneous wounds and can be further applied for human skin regeneration.

Effective Wound Healing by Antibacterial and Bioactive Calcium-Fluoride-Containing Composite Hydrogel Dressings Prepared Using in Situ Precipitation.

In this study, we report the development of a hyaluronic acid (HA)-based composite hydrogel containing calcium fluoride (CaF2) with good biocompatibility and antibacterial properties for multifunctional wound dressing applications. CaF2 was newly selected for incorporation within HA because it can release both Ca2+ and F- ions, which are well-known ions for affecting cell proliferation and inhibiting bacterial growth, respectively. In particular, an in situ precipitation process enables easy control over the released amount of F- ions by simply adjusting the precursor solutions (calcium chloride (CaCl2) and ammonium fluoride (NH4F)) used for the CaF2 precipitation. CaF2 particles were uniformly embedded within a HA-based pure hydrogel using an in situ precipitation process. Through variation of the CaCl2 and NH4F concentrations used in the precipitation as well as the precipitation time, composite hydrogels with different ion-release profiles were obtained. By controlling the precipitation time, especially for 10 min and after 30 min, large differences in the ion-release profiles as a function of CaF2 concentration were observed. A shorter precipitation time resulted in faster release of fluoride, whereas for the 30 min and 1 h samples, sustained ion release was achieved. Colony tests and live/dead assays using Escherichia coli and Staphylococcus aureus revealed a lower density of bacteria on the CaF2 composite hydrogels than on the pure hydrogel for both strains. In addition, improved cellular responses such as cell attachment and proliferation were also observed for the CaF2 composite hydrogels compared to those for the pure hydrogel. Furthermore, the composite hydrogels exhibited excellent wound healing efficiency, as evidenced by an in vitro cell migration assay. Finally, monitoring of the wound closure changes using a full-thickness wound in a rat model revealed the accelerated wound healing capability of the CaF2 composite hydrogels compared with that of the pure hydrogel. Based on our findings, these CaF2 composite hydrogels show great potential for application as advanced hydrogel wound dressings with antibacterial properties and accelerated wound-healing capabilities.

Biomimetic Coating of Hydroxyapatite on Glycerol Phosphate-Conjugated Polyurethane via Mineralization.

In this study, glycerol phosphate was introduced into polyurethane (PU) to promote the coating stability of hydroxyapatite (HA) during its mineralization on the PU surface. Glycerol phosphate was successfully conjugated with the PU chain during polymerization. Phosphate groups in glycerol phosphate accelerated the nucleation of HA under calcium phosphate ion-rich conditions (concentrated simulated body fluid), resulting in the enhancement of structural stability. The robust interface between HA and PU also improved mechanical properties. Hydrophilic phosphate groups and bioactive HA improved in vitro cellular responses in terms of the attachment and proliferation of L929 fibroblasts and MC3T3-E1 preosteoblasts. Thus, the highly elastic and bioactive PU-gp-HA could be a promising candidate for tissue engineering applications that experience frequent deformation, including diverse cartilage replacements.