Changyou Gao

Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering

Porous scaffolds for skin tissue engineering were fabricated by freeze-drying the mixture of collagen and chitosan solutions. Glutaraldehyde (GA) was used to treat the scaffolds to improve their biostability. Confocal laser scanning microscopy observation confirmed the even distribution of these two constituent materials in the scaffold. The GA concentrations have a slight effect on the cross-section morphology and the swelling ratios of the cross-linked scaffolds. The collagenase digestion test proved that the presence of chitosan can obviously improve the biostability of the collagen/chitosan scaffold under the GA treatment, where chitosan might function as a cross-linking bridge. A detail investigation found that a steady increase of the biostability of the collagen/chitosan scaffold was achieved when GA concentration was lower than 0.1%, then was less influenced at a still higher GA concentration up to 0.25%. In vitro culture of human dermal fibroblasts proved that the GA-treated scaffold could retain the original good cytocompatibility of collagen to effectively accelerate cell infiltration and proliferation. In vivo animal tests further revealed that the scaffold could sufficiently support and accelerate the fibroblasts infiltration from the surrounding tissue. Immunohistochemistry analysis of the scaffold embedded for 28 days indicated that the biodegradation of the 0.25% GA-treated scaffold is a long-term process. All these results suggest that collagen/chitosan scaffold cross-linked by GA is a potential candidate for dermal equivalent with enhanced biostability and good biocompatibility.

Pillar[6]arene/Paraquat Molecular Recognition in Water: High Binding Strength, pH-Responsiveness, and Application in Controllable Self-Assembly, Controlled Release, and Treatment of Paraquat Poisoning

The complexation between a water-soluble pillar[6]arene (WP6) and paraquat (G1) in water was investigated. They could form a stable 1:1 [2]pseudorotaxane with an extremely high association constant of (1.02 ± 0.10) × 10(8) M(-1) mainly driven by electrostatic interactions, hydrophobic interactions, and π-π stacking interactions. This molecular recognition has not only high binding strength but also pH-responsiveness. The threading and dethreading processes of this [2]pseudorotaxane could be reversibly controlled by changing the solution pH. This novel recognition motif was further used to control the aggregation of a complex between WP6 and an amphiphilic paraquat derivative (G2) in water. The reversible transformations between micelles based on G2 and vesicles based on WP6⊃G2 were realized by adjusting the solution pH due to the pH-responsiveness of WP6. The controlled release of water-soluble dye molecules from the vesicles could be achieved by the collapse of the vesicles into the micelles upon changing the solution pH to acidity. Additionally, the high binding affinity between WP6 and paraquat could be utilized to efficiently reduce the toxicity of paraquat. After the formation of a stable host-guest complex between WP6 and paraquat, less opportunity was available for paraquat to interact with the reducing agents in the cell, which made the generation of its radical cation more difficult, resulting in the efficient reduction of paraquat toxicity.

Layer-by-layer assembly of microcapsules and their biomedical applications

Nanoengineered multifunctional capsules with tailored structures and properties are of particular interest due to their multifunctions and potential applications as new colloidal structures in diverse fields. Among the available fabrication methods, the layer-by-layer (LbL) assembly of multilayer films onto colloidal particles followed by selective template removal has attracted extensive attention due to its advantages of precise control over the size, shape, composition, wall thickness and functions of the obtained capsules. The past decade has witnessed a rapid increase of research concerning the new fabrication strategies, functionalization and applications of this kind of capsules, particularly in the biomedical fields such as drug delivery, biosensors and bioreactors. In this critical review, the very recent progress of the multilayer capsules is summarized. First, the advances in assembly of capsules by the LbL technique are introduced with focus on tailoring the properties of hydrogen-bonded multilayer capsules by cross-linking, and fabrication of capsules based on covalent bonding and bio-specific interactions. Then the fabrication strategies which can speed up capsule fabrication are reviewed. In the following sections, the multi-compartmental capsules and the capsules that can transform their shape under stimulus are presented. Finally, the biomedical applications of multilayer capsules with particular emphasis on drug carriers, biosensors and bioreactors are described (306 references).

A Sugar-Functionalized Amphiphilic Pillar[5]arene: Synthesis, Self-Assembly in Water, and Application in Bacterial Cell Agglutination

A novel sugar-functionalized amphiphilic pillar[5]arene containing galactose groups as the hydrophlic part and alkyl chains as the hydrophobic part was designed and synthesized. It self-assembles in water to produce nanotubes as confirmed by TEM, SEM, and fluorescence microscopy. These nanotubes, showing low toxicity to both cancer and normal cells, can be utilized as excellent cell glues to agglutinate E. coli. The existence of galactoses on these nanotubes provides multivalent ligands that have high affinity for carbohydrate receptors on E. coli.

Surface modification of polycaprolactone with poly(methacrylic acid) and gelatin covalent immobilization for promoting its cytocompatibility.

Polycaprolactone (PCL) membrane was modified by grafting copolymerization of methacrylic acid (MAA) initiated under UV light. The covalent immobilization of gelatin on PCL-g-PMAA surface was consequently performed by using condensing agent, 1-ethyl-3-(3-dimethylamino propyl) carbodiimide hydrochloride. The occurrence of grafting copolymerization of PMAA and further immobilization of gelatin was confirmed by ATR-FTIR and X-ray photoelectron spectroscopy characterizations. The existence of carboxyl groups grafted on PCL surface was verified quantitatively by absorbance spectroscopy where rhodamine 6G was employed to react with carboxyl groups to generate an absorbance at 512 nm. The endothelial cell culture proved that the PCL membrane slightly modified with suitable amount of PMAA or gelatin had better cytocompatibility than control PCL or PCL membrane heavily modified with PMAA or gelatin.

Endothelium regeneration on luminal surface of polyurethane vascular scaffold modified with diamine and covalently grafted with gelatin.

Using the recently developed surface modification technique, free amino groups have been introduced onto polyester-type polyurethane (PU) scaffolds. The introduction of these free amino groups increases the surface energy and provides a convenient way to further immobilize bioactive species such as gelatin, collagen or chitosan, etc. on the scaffold surface by employing glutaraldehyde as a coupling agent. These modifications are advantageous to enhance cell-material interaction. The culture of human umbilical vein endothelial cells (HUVECs) in vitro proved that the cell proliferation ratio of both the aminolyzed and the biomacromolecules-immobilized PU membranes was improved greatly comparing with the control PU. Scanning electron microscopy and confocal laser scanning microscopy observations displayed that the gelatin-immobilized PU vascular scaffold had formed a monolayer of endothelial intima on its luminal surface after HUVECs were cultured for 6 d. Therefore, the aminolysis and the following biomacromolecule immobilization is a promising way to enhance the cell-PU interaction that can accelerate the endothelium regeneration, which is crucial for blood vessel tissue engineering.

Multilayer microcapsules with tailored structures for bio-related applications

The technique for fabrication of hollow multilayer microcapsules is initially established by layer-by-layer (LbL) assembly of oppositely charged polyelectrolytes onto sacrificial colloidal particles followed by core removal. It has the advantages of precise control over the size, shape, composition, wall thickness and functions of the obtained hollow microcapsules. More recently, the technique has been developed fast from the fabrication and basic property investigation to functionalization and applications, in particular in bio-related fields such as controlled drug delivery, biosensors and bioreactors. In this article, we shall review recent advances in property manipulation of electrostatic and hydrogen bonded microcapsules by chemical crosslinking, and microcapsules assembled by other driving forces such as covalent interaction, base pair interaction, host–guest interaction and van der Waals interaction. Then the possible applications of LbL microcapsules in bio-related fields are summarized with particular emphasis on very recent achievements in terms of loading and release, surface modification for stealth function or targeting and use as biosensors and bioreactors.

Immobilization of Biomacromolecules onto Aminolyzed Poly(L-lactic acid) toward Acceleration of Endothelium Regeneration

By reaction of poly(L-lactic acid) (PLLA) membrane with 1,6-hexanediamine, free amino groups were introduced onto a PLLA surface, through which biocompatible macromolecules such as gelatin, chitosan, or collagen were covalently immobilized by employing glutaraldehyde as a coupling agent. The existence of free amino groups on the aminolyzed PLLA surface was verified quantitatively by the ninhydrin analysis method, which revealed that surface NH(2) density increased with 1,6-hexanediamine concentration or aminolyzing time. Scanning force microscopy measurements detected an increase in surface roughness after aminolysis. The culture of human umbilical vein endothelial cells (HUVECs) in vitro proved that the cell proliferation rate and cell activity of both aminolyzed and biomacromolecule-immobilized PLLAs were improved compared with control PLLA. Scanning electron microscopy observation showed more spreading and flat cell morphology after HUVECs were cultured for 4 days on either aminolyzed or biomacromolecule-immobilized PLLA membranes. Confluent cell layers were observed on the modified PLLA. Measurement of von Willebrand factor secreted by these HUVECs confirmed that endothelium function was maintained. Therefore, aminolysis and biomacromolecule immobilization are promising ways to accelerate endothelium regeneration, which is crucial for blood vessel tissue engineering.

Gelatin/chitosan/hyaluronan scaffold integrated with PLGA microspheres for cartilage tissue engineering.

Poly(lactide-co-glycotide) (PLGA) microspheres integrated into gelatin/chitosan/hyaluronan scaffolds were fabricated by freeze-drying and crosslinking with 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide. The effects of the microspheres on porosity, density, compressive modulus, phosphate-buffered saline uptake ratio and weight loss of the scaffolds were evaluated. Generally, a scaffold with a higher PLGA content had a lower porosity and weight loss, and a medium uptake ratio, but a larger apparent density and compressive modulus. When the PLGA content was lower than 50%, the PLGA-integrated scaffolds had a similar pore size (approximately 200microm) as that of the control, and as much as 90% of their porosity could be preserved. In vitro chondrocyte culture in the 50% PLGA-integrated scaffold demonstrated that the cells could proliferate and secrete extracellular matrix at the same level as in the control gelatin/chitosan/hyaluronan scaffold.

Biological cells as templates for hollow microcapsules.

Microcapsules in the micrometer size range with walls of nanometer thickness are of both scientific and technological interest, since they can be employed as micro- and nano-containers. Liposomes represent one example, yet their general use is hampered due to limited stability and a low permeability for polar molecules. Microcapsules formed from polyelectrolytes offer some improvement, since they are permeable to small polar molecules and resistant to chemical and physical influences. Both types of closed films are, however, limited by their spherical shape which precludes producing capsules with anisotropic properties. Biological cells possess a wide variety of shapes and sizes, and, thus, using them as templates would allow the production of capsules with a wide range of morphologies. In the present study, human red blood cells (RBC) as well as Escherichia coli bacteria were used; these cells were fixed by glutardialdehyde prior to layer-by-layer (LbL) adsorption of polyelectrolytes. The growth of the layers was verified by electrophoresis and flow cytometry, with morphology investigated by atomic force and electron microscopy; the dissolution process of the biological template was followed by confocal laser scanning microscopy. The resulting microcapsules are exact copies of the biological template, exhibit elastic properties, and have permeabilities which can be controlled by experimental parameters; this method for microcapsule fabrication, thus, offers an important new approach for this area of biotechnology.Microcapsules in the micrometer size range with walls of nanometer thickness are of both scientific and technological interest, since they can be employed as micro- and nano-containers. Liposomes represent one example, yet their general use is hampered due to limited stability and a low permeability for polar molecules. Microcapsules formed from polyelectrolytes offer some improvement, since they are permeable to small polar molecules and resistant to chemical and physical influences. Both types of closed films are, however, limited by their spherical shape which precludes producing capsules with anisotropic properties. Biological cells possess a wide variety of shapes and sizes, and, thus, using them as templates would allow the production of capsules with a wide range of morphologies. In the present study, human red blood cells (RBC) as well as Escherichia coli bacteria were used; these cells were fixed by glutardialdehyde prior to layer-by-layer (LbL) adsorption of polyelectrolytes. The growth of the layers was verified by electrophoresis and flow cytometry, with morphology investigated by atomic force and electron microscopy; the dissolution process of the biological template was followed by confocal laser scanning microscopy. The resulting microcapsules are exact copies of the biological template, exhibit elastic properties, and have permeabilities which can be controlled by experimental parameters; this method for microcapsule fabrication, thus, offers an important new approach for this area of biotechnology.