Comparison of Gelatin/Polylysine- and Silk Fibroin/SDF-1α-Coated Mesenchymal Stem Cell-Seeded Intracranial Stents

Endothelialization of the aneurysmal neck is essential for aneurysm healing after endovascular treatment. Mesenchymal stem cell (MSC)-seeded stents can promote aneurysm repair. The biological effects of coated and uncoated nitinol intracranial stents seeded with MSCs on vascular cells and macrophage proliferation and inflammation are investigated. Two stent coatings that exert pro-aggregation effects on MSCs via different mechanisms are examined: gelatin/polylysine (G/PLL), which enhances cell adhesion, and silk fibroin/SDF-1α (SF/SDF-1α), which enhances chemotaxis. The aim is to explore the feasibility of MSC-seeded coated stents in the treatment of intracranial aneurysms. The G/PLL coating provides the highest cytocompatibility and blood compatibility substrate for MSCs and vascular cells and promotes cell adhesion and proliferation. Moreover, it enhances MSC secretion and regulation of vascular cell and macrophage proliferation and chemotaxis. Although the SF/SDF-1α coating promotes MSC secretion and vascular cell chemotaxis, it induces a greater degree of macrophage proliferation, chemotaxis, and secretion of pro-inflammatory factors. MSC-seeded stents coated with G/PLL may benefit stent surface endothelialization and reduce the inflammatory response after endovascular treatment of intracranial aneurysm. These effects may improve aneurysm healing and increase the cure rate.     

Chitosan Based Scaffolds by Lyophilization and sc.CO2 Assisted Methods for Tissue Engineering Applications

Porous chitosan scaffolds with possible tissue engineering applications were synthesized by using lyophilization and supercritical carbon dioxide (sc.CO₂) drying technique. 1% Chitosan (CS) solution in aq. acetic acid was treated with 37% formaldehyde solution; the resulting hydrogels were subjected to solvent-exchange prior to the final treatment procedures. Their morphology, pore structure, and physical properties were characterized by Fourier transform infrared spectroscopy (FTIR), thermal analysis, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and the specific surface areas and porosities of scaffolds were determined by using N₂ adsorption. The sc.CO₂ treated scaffolds showed a much greater surface area in comparison to the lyophilized one. Hence, sc.CO₂ treated scaffolds is better for cell proliferation. We further investigated the bioactivity of sc.CO₂ treated scaffolds using simulated body fluid (SBF). The sc.CO₂ assisted chitosan scaffold prepared by using green chemistry approach is highly pure and from a hygienic point of view, it is an ideal material for tissue engineering applications.     

Preparation of glycerol plasticized chitosan films using AlCl3·6H2O as the solvent: optical,crystalline, mechanical and barrier properties

Chitosan films plasticized with the complex of AlCl₃·6H₂O and glycerol were prepared by using AlCl₃·6H₂O as the solvent. The effect of AlCl₃·6H₂O and glycerol complex with Chitosan was studied by SEM, XRD, TGA, and tensile testing. The complex would increase the water resistance and destroy the crystals of chitosan film. The TGA results proved that the chitosan film plasticized with the complex showed higher thermal stability at the temperature below 200?°C than pure chitosan film. With the addition of the complex of AlCl₃·6H₂O and glycerol, the tensile strength of chitosan film decreased and the elongation at break increased.     

Inhibition of corneal neovascularization by rapamycin

The purpose of this study was to determine whether rapamycin could inhibit corneal angiogenesis induced by basic fibroblast growth factor (bFGF). Using human dermal microvascular endothelial cells (HDMECs), we examined the effect of rapamycin on cell proliferation and migration, and the expression of vascular endothelial growth factor (VEGF). The rabbit's eye was implanted intrastromally into the superior cornea with pellet containing bFGF for the control group and pellet containing bFGF and rapamycin for the rapamycin group. Biomicrographically, corneal angiogenesis was evaluated for 10 days after pellet implantation. The neovascularized cornea also was examined histologically. bFGF induced corneal neovascularization was significantly reduced by treatment with rapamycin. Using in vitro model, rapamycin strongly inhibited bFGF induced proliferation, migration, and VEGF secretion of HDMECs. We could observe that the bFGF induced corneal angiogenesis was inhibited by rapamycin in a micropocket rabbit model. The score of neovascularization was significantly decreased in the rapamycin group than in the control group at 10 days after pellet implantation. Histologically, the cornea of rapamycin group also showed much less new vessels than that of control group. Collectively, rapamycin appears to inhibit bFGF induced angiogenesis in a rabbit corneal micropocket assay and may have therapeutic potential as an antiangiogenic agent.     

Cold atmospheric plasma modification of curcumin loaded in tri‐phosphate chitosan nanoparticles enhanced breast cancer cells apoptosis

The anti‐cancer mechanisms of curcumin have been reported to include suppressions of angiogenesis and tumor proliferation. The main goal of this research is to increase the solubility of curcumin by cold atmospheric plasma (CAP) and assess the effects of modified curcumin by charging with tri‐polyphosphate chitosan nanoparticles for MCF‐7, MDA‐MB‐231 breast cancer cells, and human fibroblast cells. Curcumin modification was done by CAP and its solubility was evaluated by spectrophotometry. After loading modified curcumin into nano‐chitosan‐TPP, nanocurcumin was characterized by scanning electron microscopy. Cellular viability and apoptosis of treated cells were assessed by MTT and Annexin V. The changes of messenger RNA expression of TP5353 and VEGF genes were analyzed by real‐time PCR. CAP was able to transform the curcumin to possess hydrophilic characteristics after 90 seconds. The mean diameter of Curcumin loaded chitosannanoparticles (NPs) were determined as 48 nm. MTT results showed that the IC₅₀ of nano Cur‐chitosan‐TPP was effectively decreased compared to free curcumin in MCF‐7 (15 μg/mL at 72 hours vs 20 μg/mL at 48 hours). Additionally, nano Cur‐chitosan‐TPP had no significant effect on normal cells (Human dermal fibroblas: HDF), whereas it also decreased the viability of triple negative breast cancer cell line (MDA‐MB‐231). Real‐time PCR results showed that expression level of TP53 gene was upregulated (P = .000), whereas VEGF gene downregulated (P = .000) in treated MCF‐7 cells. Curcumin loaded chitosan nanoparticles have led to an induction of apoptosis (79.93%) and cell cycle arrest (at S and G2M). Modified‐curcumin‐tri‐polyphosphate chitosan nanoparticles using CAP can be considered as a proper candidate for breast cancer treatment.     

Core–shell structure PEO/CS nanofibers based on electric field induced phase separation via electrospinning and its application

Core–shell structured PEO‐chitosan nanofibers have been produced from electric field inducing phase separation. Chitosan, a positive charged polymer, was dissolved in 50 wt % aqueous acetic acid and the amino group on polycation would protonize, which would endow chitosan electrical properties. Chitosan molecules would move along the direction of the electric field under the electrostatic force and formed the shell layer of nanofibers. Preparation process of core – shell structure is quite simple and efficient without any post‐treatment. The core–shell structure and existence of chitosan on the shell layer were confirmed before and after post‐treatment by TEM and further supported by SEM, FTIR, XRD, DSC, and XPS studies. Blending ratio of PEO and chitosan, molecular weight of chitosan for the mobility of chitosan are thought to be the key influence factors on formation of core–shell structure. Drug release studies show that the prepared core–shell structure nanofibers has a potential application in the biomedical fields involving drug delivery. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2015 , 53, 2298–2311     

Hydrolysis of mixed monomolecular films of tricaprylin/dilauroylphosphatidylcholine by lipase and phospholipase A₂

The potential of chitosan, a fishery waste-based material, as a soil amendment to clean-up metal contaminated soil was investigated. Chitosan was treated using glutaraldehyde (GLA), epichlorohydrin (ECH) and ethylene glycol diglycidyl ether (EGDE) as cross-linking reagents to enhance its chemical stability in acidic media and to improve its physical properties. Cross-linking treatment had significant effects on chitosan surface area, pore diameter, surface morphology and crystallinity. Interaction with Ag(I), Pb(II) and Cu(II) decreased the crystallinity of the materials and changed their surface morphology significantly. FTIR analysis confirmed that N and O atoms served as binding sites for the metal ions. Chitosan and treated chitosans were able to bind metal ions, even in the presence of K⁺, Cl^? and NO₃^?, which are dominant ions in soil. Therefore, remediation of metal contaminated soil using chitosan and cross-linked treated chitosans as soil amendments is feasible.     

角膜内皮细胞载体膜片的性质研究及移植试验

利用溶剂浇铸/颗粒沥滤技术制备出壳聚糖和硫酸软骨素共混膜, 研究了共混膜的透光性、 表面结构、 红外图谱、 X-射线衍射图谱以及生物相容性和生物降解性. 结果表明, 共混膜透明度高, 结构均匀, 孔隙大小合适, 壳聚糖分子与硫酸软骨素达到分子水平分散, 且分子间存在较强的相互作用, 共混膜具有良好的生物相容性和生物降解性. 以共混膜为载体培养兔角膜内皮细胞, 结果表明, 共混膜非常适合角膜细胞贴附生长, 10 d内长成良好单层细胞. 将载有角膜内皮细胞的膜片植入内皮缺损兔眼中, 在56 d内术眼基本保持透明, 之后角膜水肿, 逐渐模糊并出现新生血管, 在230 d时术眼新生血管基本消失, 角膜恢复透明.     

Cellulose-based materials as scaffolds for tissue engineering

Two types of cellulose-based materials, 6-carboxycellulose with 2.1 or 6.6 wt% of –COOH groups, were prepared and tested for potential use in tissue engineering. The materials were functionalized with arginine, i.e. an amino acid with a basic side chain, or with chitosan, in order to balance the relatively acid character of oxidized cellulose molecules, and were seeded with vascular smooth muscle cells (VSMC). The cell adhesion and growth were then evaluated directly on the materials, and also on the underlying polystyrene culture dishes. Of these two types of studied materials, 6-carboxycellulose with 2.1 wt% of –COOH groups was more appropriate for cell colonization. The cells on this material achieved an elongated shape, while they were spherical in shape on the other materials. The number of cells and the concentration (per mg of protein) of contractile proteins alpha-actin and SM1 and SM2 myosins, i.e. markers of the phenotypic maturation of VSMC, were also significantly higher on this material. Functionalization of the material with arginine and chitosan further improved the phenotypic maturation of VSMC. Chitosan also improved the adhesion and growth of these cells. In comparison with the control polystyrene dishes, the proliferation of cells on our cellulose-based materials was relatively low. This suggests that these materials can be used in applications where high proliferation activity of cells is not desirable, e.g. proliferation of VSMC on vascular prostheses. Alternatively, the cell proliferation might be enhanced by another more efficient modification, which would require further research.     

Bioactive Properties of Chitin/Chitosan—Calcium Phosphate Composite Materials

Calcium phosphate coating over phosphorylated derivatives of chitin/chitosan material was produced by a process based on phosphorylation, Ca(OH)₂ treatment and SBF (simulated body fluid solution) immersion. Chitin/chitosan phosphorylated using urea and H₃PO₄ and then soaked in saturated Ca(OH)₂ solution at ambient temperature, which lead to the formation of thin coatings formed by partial hydrolysis of the PO₄ functionalities, were found to stimulate the growth of a calcium phosphate coating on their surfaces after soaking in 1.5 × SBF solution for as little as one day. The Ca(OH)₂ treatment facilitates the formation of a calcium phosphate precursor over the phosphorylated chitin/chitosan, which in turn encourages the growth of a calcium deficient apatite coating over the surface upon immersion in SBF solution. The bio-compatibility of calcium phosphate compound—chitin/chitosan composite materials was evaluated by cell culture test using L-929 cells. The initial anchoring ratio and the adhesive strength of L-929 cells for composites was higher than that for the polystyrene disk (LUX, control). The results of in-vitro evaluation suggested that the calcium phosphate—chitin/chitosan composite materials were suitable for cell carrier materials.     

Effect of molecular weight and degree of deacetylation on controlled release of isoniazid from chitosan microspheres

Glutaraldehyde cross‐linked chitosan microspheres for controlled release of isoniazid were prepared using chitosan of different molecular weights (MWs) and degrees of deacetylation (DDAs). Chitosan microspheres were characterized for their size, hydrophobocity, degree of swelling and loading of isoniazid. Hydrophobicity of chitosan microspheres increased on increasing the degree of cross‐linking and MW of chitosan. Chitosan microspheres with high degree of deacetylation (DDA) (75 wt%), high MW chitosan (2227 kg mol^?1), and with 12 wt% concentration of glutaraldehyde showed optimum loading and release of isoniazid. The isoniazid from chitosan microspheres was released in two steps, i.e. burst (%R_B) and controlled (%R_C) steps. The microspheres with low MW chitosan (260 kg mol^?1) and low DDA (48 wt%) showed prominent burst release of isoniazid, but microspheres with high MW chitosan (2227 kg mol^?1) and high DDA (75 wt%) have released more isoniazid in a controlled manner (60 wt%) at 37°C in a solution of pH 5.0 ± 0.1. The burst step of drug release (%R_B) has followed first order kinetics, whereas controlled step of drug release (%R_C) followed zero order kinetics. The burst step of drug release was Fickian and controlled step was non‐Fickian in nature. The diffusion constant (D) for isoniazid release was influenced by the properties of chitosan and degree of cross‐linking. Copyright © 2007 John Wiley & Sons, Ltd.     

Regioselective synthesis and biological activity of oxidized chitosan derivatives

Oxidized chitosan derivatives with various degrees of oxidation (DS, 0.1–1.0) were prepared by the treatment of chitosan with CrO₃/aq HClO₄ or by the oxidation of ­3‐O‐ and N‐protected chitosan with 30% aq H₂O₂/Na₂WO₄ followed by 3‐O‐ and N‐deprotection. The oxidized products were then N‐acetylated with Ac₂O in order to improve their water‐solubility. Although the oxidized chitosan derivative of DS 0.28 and the degree of N‐acetylation of chitosan (DA) 38% was insoluble in the pH 3–8 region, that of DS 0.26 and DA 76% was soluble in the neutral pH range. The newly‐prepared acetylated and oxidized chitosan derivatives were found to suppress the chemiluminescence response of inflammatory cells such as canine polymorphonuclear cells (PMNs). Analysis by the surface plasmon resonance method revealed that the bind and release behavior of PMNs to acetylated oxidized chitosan derivatives was similar to that against carboxymethylated chitosan derivatives. The amount of water‐soluble chitosan derivative bound to cytokine IL‐8 was found to be affected by the structural and electronic features of the chitosan substituents in the chitosan chain. Copyright © 2003 John Wiley & Sons, Ltd.     

Synthesis and cell activity of novel galactosylated chitosan as a gene carrier

It is important for gene carrier to transport DNA into target cells. Although viral vectors are very efficient gene-transfer vehicles, significant drawbacks limit their applications. Chitosan (CS) has been researched widely as a non-viral vector. However, the low cell specificity and low transfection efficiency of chitosan need to be overcome. In order to conquer the drawback of chitosan, the present paper is concerned with the synthesis of novel galactosylated chitosan (GC) through etherization of chitosan and galactose in THF using BF₃·OEt₂ as promoter. The final product was characterized and confirmed by FT-IR and ¹H NMR. The degree of O-substitution (DS) of chitosan by galactose was measured to be 10.38% using anthrone-sulfuric acid colorimetric method. The mean particle diameter and average zeta potential of the GC/DNA complex were 350 nm and +22.1 mV, respectively. The GC/DNA nanoparticle was tested to transfect HEK293 cells, and the viability of HEK293 cells was not affected by the GC/DNA nanoparticle compared to that of the control.     

Chitosan/poly(tetrafluoroethylene) composite membranes using in pervaporation dehydration processes

Chitosan/PTFE composite membranes were prepared from casting a γ-(glycidyloxypropyl)trimethoxysilane (GPTMS)-containing chitosan solution on poly(styrene sulfuric acid) grafted expended poly(tetrafluoroethylene) film surface. The adhesion between the chitosan skin layer and the PTFE substrate was pretty good to warrant the high performance of chitosan/PTFE composite membranes using in pervaporation dehydration processes on isopropanol. The chitosan/PTFE membrane exhibited a permeation flux of 1730 g/m² h and a separation factor of 775 at 70 °C on pervaporation dehydration of a 70 wt% isopropanol aqueous solution. The membrane also survived after a long-term operation test in 45 days.     

Chitosan: An Attractive Biocompatible Polymer for Microencapsulation

Chitosan is a weak cationic polysaccharide composed essentially of β(1 → 4) linked glucosamine units together with some N‐acetylglucosamine units. It is obtained by extensive deacetylation of chitin, a polysaccharide common in nature. Chitosan is a biocompatible, biodegradable, and nontoxic natural polymer that exhibits excellent film‐forming ability. As a result of its cationic character, chitosan is able to react with polyanions giving rise to polyelectrolyte complexes. Therefore, because of these interesting properties, it has become the subject of numerous scientific reports and patents on the preparation of microspheres and microcapsules. The techniques employed to microencapsulate with chitosan include, among others, ionotropic gelation, spray drying, emulsion phase separation, simple and complex coacervation, and polymerization of a vinyl monomer in the presence of chitosan. The aim of this work is to review some of the more common techniques used and to put forward the results obtained by our research group in preparing chitosan‐based microcapsules: for taste masking and improving the stability of a nutritional oil, the sustained release of drugs, as well as the preparation of chitosan superparamagnetic microcapsules for the immobilization of enzymes.

Scanning electron micrograph of some superparamagnetic chitosan particles and magnetic hysteresis loop of the microparticles.     

Synthesis,NMR Study and Preliminary Sorption Properties of two N-Benzyl Sulfonated Chitosan Derivatives 1

Abstract

Two chitosan based sorbents for the uptake of metallic cations in acidic solutions were synthesized. Chitosan was reacted with 2-formylbenzene sodium sulfonate and 4-formyl-1,3-benzene sodium disulfonate in the presence of NaCNBH₃ to yield N-benzyl mono and disulfonate derivatives of chitosan. NMR spectra confirmed the presence of benzyl sulfonate groups which were subsequently used to determine the degree of substitution. The results of sorption experiments for mono and divalent metals at pH=2 are reported.

     

Modification of mechanical and thermal property of chitosan–starch blend films

Chitosan–starch blend films (thickness 0.2 mm) of different composition were prepared by casting and their mechanical properties were studied. To improve the properties of chitosan–starch films, glycerol and mustard oil of different composition were used. Chitosan–starch films, incorporated with glycerol and mustard oil, were further modified with monomer 2-hydroxyethyl methacrylate (HEMA) using gamma radiation. The modified films showed improvement in both tensile strength and elongation at break than the pure chitosan–starch films. Water uptake of the films reduced significantly than the pure chitosan–starch film. Thermo gravimetric analysis (TGA) and dynamic mechanical analysis (DMA) showed that the modified films experience less thermal degradation than the pure films. Scanning electron microscopy (SEM) and FTIR were used to investigate the morphology and molecular interaction of the blend film, respectively.     

Biomimicking polysaccharide nanofibers promote vascular phenotypes: a potential application for vascular tissue engineering

The potential of electrospun pullulan/dextran (P/D) nanofibers (average diameter = 323 nm) for vascular tissue engineering applications is explored. The mechanical properties of the nanofibers are of the same order of magnitude as that of human arteries (Young's modulus ≈0.88 MPa; tensile strength ≈0.35 MPa). It is demonstrated that the nanofiber topography enables cell adhesion and that the endothelial phenotype is maintained on the nanofibers. Moreover, P/D nanofibers support a stable confluent monolayer of endothelial cells over 14 d. SMCs seeded on nanofibers display similar levels of alpha smooth muscle actin and a lower proliferation rate than cells on 2D cultures. The observations suggest that nanofibers promote a shift to a quiescent contractile phenotype in SMCs.

     

Temperature Responsive Shape‐Memory Scaffolds with Circumferentially Aligned Nanofibers for Guiding Smooth Muscle Cell Behavior

Structural simulation of the smooth muscle layer plays an important role in tissue engineering of blood vessels for the replacement of damaged arteries. However, it is difficult to construct small‐diameter tubular scaffolds to homogenously locate and align smooth muscle cells (SMCs). In this work, novel temperature responsive shape‐memory scaffolds are designed for SMC culturing. The scaffolds are composed of an outer layer of poly(lactide–glycolide–trimethylene carbonate) (PLGATMC) for programming the deformation from planar to small‐diameter tubular shape and an inner layer of aligned nanofibrous membrane of poly(lactide–glycolide)/chitosan (PLGA/CS) to regulate cell adhesion, proliferation, and morphology. The SMC behaviors and functions are dependent on the PLGA/CS ratios of membranes, and the scaffold with PLGA/CS 7:3 membrane exhibits the most suitable ability to regulate SMC behavior. The PLGA/CS@PLGATMC scaffold can be deformed into a temporary planar at 20 °C for convenient seeding and attachment of SMCs and then immediately self‐rolled into 3D tube at 37 °C. The proposed strategy offers a practical approach for the development of small‐diameter vascular scaffolds from 2D planar into 3D tubular shape by self‐rolling.     

Crystal transition from hydrated chitosan and chitosan/monocarboxylic acid complex to anhydrous chitosan investigated by X‐ray diffraction

We have studied the crystal transition behaviors from hydrated chitosan to anhydrous chitosan by X‐ray diffraction analyses. Hydrated chitosan prepared by deacetylation of crustacean α‐chitin was subjected to the two conversion methods, hydrothermal treatment and high‐humidity treatment via chitosan/monocarboxylic acid complex. The transition by hydrothermal treatment progressed with increasing treatment temperature and time, and the rapid transition occurred above 200 °C. Chitosan/acetic acid complex and chitosan/formic acid complex were prepared by immersing hydrated chitosan in acid solution. The transition from chitosan/acetic acid complex to anhydrous chitosan in high relative humidity condition proceeded with increasing temperature and was complete at 80 °C for 1 h, whereas chitosan/formic acid complex did not convert to anhydrous chitosan under the same conditions. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2015 , 53, 1065–1069