Zhengke Wang

Gelation process visualized by aggregation-induced emission fluorogens

Alkaline-urea aqueous solvent system provides a novel and important approach for the utilization of polysaccharide. As one of the most important polysaccharide, chitosan can be well dissolved in this solvent system, and the resultant hydrogel material possesses unique and excellent properties. Thus the sound understanding of the gelation process is fundamentally important. However, current study of the gelation process is still limited due to the absence of direct observation and the lack of attention on the entire process. Here we show the entire gelation process of chitosan LiOH-urea aqueous system by aggregation-induced emission fluorescent imaging. Accompanied by other pseudo in situ investigations, we propose the mechanism of gelation process, focusing on the formation of junction points including hydrogen bonds and crystalline.

Orientation in multi-layer chitosan hydrogel: morphology, mechanism, and design principle.

Hydrogels with organized structure have attracted remarkable attentions for bio-related applications. Among the preparation of hierarchical hydrogel materials, fabrication of hydrogel with multi-layers is an important branch. Although the generation mechanism of layers had been fully discussed, sub-layer structure was not sufficiently studied. In this research, multi-layered chitosan hydrogel with oriented structure was constructed, and the formation mechanism of orientation was proposed, based on gelation behavior and entanglement of polymer chains in the hydrogel-solution system. Employing the layered-oriented characteristic, chitosan hydrogel materials with various shapes and structure can be designed and fabricated.

Fabrication of Chitosan Nanoparticles with Aggregation-Induced Emission Characteristics and Their Applications in Long-Term Live Cell Imaging

Chitosan with tetraphenylethene pendants (TPE-CS) are synthesized by reaction between amine and isothiocyanate groups of chitosan and tetraphenylethene (TPE), respectively. Nanoparticles of TPE-CS (TPE-CS NPs) are fabricated by ionic gelation method. The NPs are uniform in size, spherical in shape, monodispersed, and positive in surface charge. The suspension of TPE-CS NPs emits strong blue fluorescence under photoexcitation due to the aggregation-induced emission characteristics of the TPE moieties. The NPs can be internalized into cytoplasm through endocytosis pathway and retain inside the live cells to image the cells. Cytotoxicity assay reveals that TPE-CS NPs are cytocompatible and thus can be used for long-term live cell imaging.

Using absorbable chitosan hemostatic sponges as a promising surgical dressing

As absorbable hemostatic dressings, chitosan with a deacetylation degree of 40% (CS-40) and 73% (CS-73) have been fabricated into sponges via a modified method. The hemostatic, biocompatible and biodegradable properties were evaluated through in vivo assays. In a hepatic hemorrhage model, the chitosan sponges, with excellent blood compatibility, achieved less blood loss than the gelation sponge (GS). In addition, CS-40 showed better hemostatic capability and biodegradability than CS-73. After implantation, a histological analysis indicated that CS-40 exhibited the best biodegradability, tissue regeneration and least tissue adhesion. By contrasting CS-40 and CS-73, the deacetylation degree is confirmed to be a key factor for the hemostatic effect, biodegradability, biocompatibility and tissue regeneration. Our overall results demonstrated the potential application of CS-40 for use in absorbable hemostatic dressings.

Preparation and characterization of bionic bone structure chitosan/hydroxyapatite scaffold for bone tissue engineering

Three-dimensional oriented chitosan (CS)/hydroxyapatite (HA) scaffolds were prepared via in situ precipitation method in this research. Scanning electron microscopy (SEM) images indicated that the scaffolds with acicular nano-HA had the spoke-like, multilayer and porous structure. The SEM of osteoblasts which were polygonal or spindle-shaped on the composite scaffolds after seven-day cell culture showed that the cells grew, adhered, and spread well. The results of X-ray powder diffractometer and Fourier transform infrared spectrometer showed that the mineral particles deposited in the scaffold had phase structure similar to natural bone and confirmed that particles were exactly HA. In vitro biocompatibility evaluation indicated the composite scaffolds showed a higher degree of proliferation of MC3T3-E1 cell compared with the pure CS scaffolds and the CS/HA10 scaffold was the highest one. The CS/HA scaffold also had a higher ratio of adhesion and alkaline phosphate activity value of osteoblasts compared with the pure CS scaffold, and the ratio increased with the increase of HA content. The ALP activity value of composite scaffolds was at least six times of the pure CS scaffolds. The results suggested that the composite scaffolds possessed good biocompatibility. The compressive strength of CS/HA15 increased by 33.07% compared with the pure CS scaffold. This novel porous scaffold with three-dimensional oriented structure might have a potential application in bone tissue engineering.

A novel chitosan-based sponge coated with self-assembled thrombin/tannic acid multilayer films as a hemostatic dressing

In order to prepare a novel hemostatic dressing for uncontrolled hemorrhage, a porous chitosan sponge was coated with self-assembled (thrombin/tannic acid)n films, which were based on hydrogen bonding interactions between thrombin and tannic acid at physiologic pH. According to the whole blood clotting test, the coated chitosan sponges showed a significantly high rate of blood clotting due to the addition of thrombin. On the other hand, the storable half-life of immobilized thrombin is extended to 66.9 days at room temperature, which is 8.5 times longer than unfixed thrombin. It is because of the immobilization effect of, not only the porous structure of chitosan sponge but also the interactions between thrombin and tannic acid. In addition, the tannic acid has similar antibacterial effect to chitosan. Therefore, it is an excellent combination of chitosan, thrombin and tannic acid. Besides, all of materials in this research have been approved by the United States Food and Drug Administration (FDA). So the chitosan-based sponge is a promising candidate dressing for uncontrolled hemorrhage due to its storable, bio-safe and highly effective hemostatic properties.

Ferroferric oxide/chitosan scaffolds with three-dimensional oriented structure

A facile approach to construct ferroferric oxide/chitosan composite scaffolds with three-dimensional oriented structure has been explored in this research. Chitosan and ferroferric oxide are co-precipitated by using an in situ precipitation method, and then lyophilized to get the composite scaffolds. XRD indicated that Fe3O4 was generated during the gel formation process, and increasing the content of magnetic particles could destruct the crystal structure of chitosan. When the content of magnetic particles is lower than 10%, the layer-by-layer structure and wheel spoke structure are coexisting in the scaffolds. Increasing the content of magnetic particles, just layer-by-layer structure could be observed in the scaffolds. Ferroferric oxide particles were uniformly distributed in the matrix, the size of which was about 0.48 μm in diameter, 2 μm in length. Porosity of magnetic chitosan composite scaffolds is about 90%. When the ratio of ferroferric oxide to chitosan is 5/100, the compressive strength of the material is 0.4367 MPa, which is much higher than that of pure chitosan scaffolds, indicating that the layer-by-layer and wheel spokes complex structure is beneficial for the improvement of the mechanical properties of chitosan scaffolds. However, increasing the content of ferroferric oxide, the compressive strength of scaffolds decreased, because of the decreasing of chitosan crystallization and aggregation of magnetic particles as stress centralized body. Another reason is that the layer-by-layer and wheel spokes complex structure makes bigger contributions for the compressive strength than the layer-by-layer structure does. Three-dimensional ferroferric oxide/chitosan scaffolds could be used as hyperthermia generator system, improving the local circulation of blood, promoting the aggradation of calcium salt and stimulating bone tissue regeneration.

Monitoring layer-by-layer self-assembly process of natural polyelectrolytes by fluorescent bioconjugate with aggregation-induced emission characteristic

A novel chitosan-based fluorescent bioconjugate (TPE-CS) with aggregation-induced emission (AIE) characteristic is synthesized and used as a fluorescent probe for monitoring layer-by-layer self-assembly process of natural polyelectrolytes. QCM results and contact angle measurement indicate that this AIE active TPE-CS bioconjugate can be assembled with alginate (ALG) through layer-by-layer deposition. Ellipsometry and fluorescence (FL) spectroscopy show an exponential growth of the TPE-CS/ALG multilayer films. Moreover, the exponential relationship between the FL intensity and the number of bilayers, which is in accordance with the thickness variation of multilayer films, provides solid evidence for its capacity to monitor the layer-by-layer self-assembly process.

Catechol-Functional Chitosan/Silver Nanoparticle Composite as a Highly Effective Antibacterial Agent with Species-Specific Mechanisms

In this study, silver nanoparticles (Ag NPs) coated with catechol-conjugated chitosan (CSS) were prepared using green methods. Interestingly, we uncovered that CSS-coated Ag NPs (CSS-Ag NPs) exhibited a higher toxicity against gram-negative Escherichia coli (E. coli) bacteria than against gram-positive Staphylococcus aureus (S. aureus) bacteria. The differences revealed that the CSS-Ag NPs killed gram bacteria with distinct, species-specific mechanisms. The aim of this study is to further investigate these underlying mechanisms through a series of analyses. The ultrastructure and morphology of the bacteria before and after treatment with CSS-Ag NPs were observed. The results demonstrated the CSS-Ag NPs killed gram-positive bacteria through a disorganization of the cell wall and leakage of cytoplasmic content. In contrast, the primary mechanism of action on gram-negative bacteria was a change in membrane permeability, induced by adsorption of CSS-Ag NPs. The species-specific mechanisms are caused by structural differences in the cell walls of gram bacteria. Gram-positive bacteria are protected from CSS-Ag NPs by a thicker cell wall, while gram-negatives are more easily killed due to an interaction between a special outer membrane and the nanoparticles. Our study offers an in-depth understanding of the antibacterial behaviors of CSS-Ag NPs and provides insights into ultimately optimizing the design of Ag NPs for treatment of bacterial infections.

Fluorescent detection of Cu(II) by chitosan-based AIE bioconjugate

Detection of Cu(II) is very important in disease diagnose, biological system detection and environmental monitoring. Previously, we found that the product TPE-CS prepared by attaching the chromophores of tetraphenylethylene (TPE) to the chitosan (CS) chains showed excellent fluorescent properties. In this study, we tried to use TPE-CS for detecting Cu(II) because of the stable complexation of CS with heavy metals and the luminosity mechanism of the Restriction of Intramolecular Rotations (RIR) for aggregation-induced emission (AIE)-active materials. The fluorescence intensity changed when TPE-CS was contacted with different metal ions, to be specific, no change for Na+, slightly increase for Hg2+, Pb2+, Zn2+, Cd2+, Fe2+, Fe3+ due to the RIR caused by the complexation between CS and metal ions. However, for Cu2+, an obvious fluorescence decrease was observed because of the Photoinduced-Electron-Transfer (PET). Moreover, we found that the quenched FL intensity of TPE-CS was proportional to the concentration of Cu(II) in the range of 5 μmol/L to 100 μmol/L, which provided a new way to quantitatively detect Cu(II). Besides, TPE-CS has excellent water-solubility as well as absorbability (the percentage of removal, R = 84%), which is an excellent detection probe and remover for Cu(II).