Preparation and Characterization of Chitosan Composite Membranes Crosslinked by Carboxyl-capped Poly（ethylene glycol）

In this study a series of chemically crosslinked chitosan/poly（ethylene glycol） （CS/PEG） composite membranes were prepared with PEG as a crosslinking reagent other than an additional blend. First, carboxyl-eapped poly（ethylene glycol） （HOOC-PEG-COOH） was synthesized. Dense CS/PEG composite membranes were then prepared by casting/evaporation of CS and HOOC-PEG-COOH mixture in acetic acid solution. Chitosan was chemically crosslinked due to the amidation between the carboxyl in HOOC-PEG-COOH and the amino in chitosan under heating, as confirmed by FTIR analysis. The hydrophilicity, water-resistance and mechanical properties of pure and crosslinked chitosan membranes were characterized, respectively. The results of water contact angle and water absorption showed that the hydrophilicity of chitosan membranes could be significantly improved, while no significant difference of weight loss between pure chitosan membranes and crosslinked ones was detected, indicating that composite membranes with amidation crosslinking possess excellent water resistanance ability. Moreover, the tensile strength of chitosan membranes could be significantly enhanced with the addition of certain amount of HOOC-PEG-COOH crosslinker, while the elongation at break didn＇t degrade at the same time. Additionally, the results of swelling behaviors in water at different pH suggested that the composite membranes were pH sensitive.     

壳聚糖-纤维素硫酸钠聚电解质复合物膜对药物表观渗透系数的测试

药物渗透系数是考察复合物膜的药物释放性能的重要参数. 本文以溶解性不同的两种药物扑热息痛和5-氨基水杨酸(5-ASA)为模型药物研究了其在壳聚糖-纤维素硫酸钠聚电解质复合物膜中的渗透性能. 结果表明：壳聚糖-纤维素硫酸钠聚电解质复合物膜的渗透性能与其溶胀性能密切相关;复合物膜中壳聚糖和纤维素硫酸钠的配比、相对分子量和pH值对膜的渗透性能和溶胀性能影响显著,以扑热息痛作为模型药物研究了壳聚糖-纤维素硫酸钠聚电解质复合物膜在模拟胃肠液中对药物的渗透性能. 通过调整该复合物膜的配方,可以使该膜分别实现胃、小肠和结肠定位释药的目的.     

Study of Protein Adsorption/Adhesion Behaviors on Solid Beads Surface with Different Surface Properties

Three kinds of chitosan (CS) derivatives have been prepared via polyethyleneimine (PEI) or polyethylene glycol (PEG) together with PEI modified chitosan. FTIR, x-ray photoelectron spectroscopy (XPS), and thermal gravimetric analysis (TGA)/differential thermal analysis (DTA) proved the successful linkage, and XPS and zeta potential analysis show that these derivatives beads possess different surface nature. The PEI modified chitosan derivative is made in acid solution (A-PEI-CS) and the one added with additional PEG (PEG–CS-PEI) has showed higher swelling degree than their counterpart derivative prepared at basic condition (B-PEI-CS). Although the nitrogen content of A-PEI-CS is higher than that of B-PEI-CS, due to the better surface hydrophilicity, the B-PEI-CS beads are more capable of adsorbing bovine serum albumin (BSA) protein. Electrostatic attraction facilitates protein adsorption in a narrow acidic pH range while hydrophilicity hinders protein adhering to the beads’ surface at any pH.     

Novel nylon-supported organic-inorganic hybrid membrane with hierarchical pores as a potential immobilized metal affinity adsorbent

Chitosan-based porous organic-inorganic hybrid membranes supported by microfiltration nylon membranes were prepared, in which gamma-glycidoxypropyltrimethoxysilane (GPTMS) was used as an inorganic source as well as crosslinking reagent. Polyethylene glycol (PEG) with different molecular weight and content was used as imprinting molecule for morphology control. In situ crosslinking of chitosan and simultaneous polymerization of GPTMS in PEG template environment endowed the hybrid membrane with specific characteristics. Distinct hybrid effect between chitosan (CS) and GPTMS was revealed by shifting in X-ray diffraction (XRD) pattern, decomposition in simultaneous thermogravimetry and differential scanning calorimetry (TG/DSC) testing. As manifested by scanning electron microscopy (SEM), the molecular weight and content of PEG had remarkable effect on the resulting surface morphology of the hybrid membrane and a given surface morphology could be obtained by extracting of the imprinted PEG molecular. Among three types of porogen used: PEG 400, PEG 4000 and PEG 20000, only PEG 20000 could result in a porous surface. Moreover, a special porous surface with three-dimensional (3D) hierarchical structure-in-structure pore fashion was obtained when content of PEG 20000 was controlled at 15%. Experimental results also showed that the hybrid membrane had low swelling ratio and high stability in acidic solution. After conveniently coordinated with copper ions, the porous metal chelating hybrid membrane could effectively adsorb the model protein, bovine serum albumin (BSA). As expected, the hybrid membrane imprinted with 15% PEG 20000 had remarkably high copper ion binding and BSA adsorption capacity, which might result from the large surface area, high ligand density and suitable interconnected 3D hierarchical porous surface.     

Macroporous chitosan layer coated on non-porous silica gel as a support for metal chelate affinity chromatographic adsorbent

A new immobilized metal affinity chromatography (IMAC) matrix was prepared by coordinating Cu2+ with cross-linked chitosan coated on non-porous silica gel (Cu-CTS-SiO2). Macroporous structure could be formed on the coated layer by imprinting polyethylene glycol (PEG) in chitosan film. The surface morphology changes on Cu-CTS-SiO2 bead prepared in different condition were confirmed by scanning electron microscopy (SEM). Effects of chitosan and PEG content in coating solution, the molecular mass of PEG on the surface macropore formation and adsorption capacity of bovine serum albumin (BSA) were investigated. Results indicated that coating solution with 2% chitosan and 10% PEG 20000 was optimal. Batch experiments were also conducted for elucidating the optimal pH, the adsorption isotherm and adsorption kinetics of BSA. Adsorption isotherm of trypsin on the same adsorbent was also performed. Results showed that the support itself had low non-specific interaction with both BSA and trypsin. The maximum adsorption capacity for BSA and trypsin on the prepared IMAC adsorbent could reach 192 mg and 5000 IU, respectively calculated by every gram of chitosan. The binding and eluting condition for BSA were tested on column filled with the adsorbent. Crude BSA sample could be purified on the IMAC column.     

N-(2-磺酸基苯甲基)壳聚糖的合成、表征及其水凝胶的pH敏感性

通过两步反应合成了水溶性的N-(2-磺酸基苯甲基)壳聚糖(SBCS), 用IR, 1H NMR和UV-Vis谱对产物的结构进行了表征. 用胶体滴定法测定了N上2-磺酸基苯甲基的取代度. 以戊二醛为交联剂制备了N-(2-磺酸基苯甲基)壳聚糖水凝胶(SBCSG), 考察了凝胶在不同pH值缓冲溶液中的溶胀行为. 实验结果表明, SBCSG溶胀度随着凝胶交联度的增大而减小. 在碱性介质中SBCSG的溶胀度显著增大, 而在酸性介质中溶胀度显著减小, 在pH= 5.0缓冲液中的溶胀度达到最小值. SBCSG在碱性介质中的溶胀度随着侧链N上2-磺酸基苯甲基取代度增大而增大. 在pH=7.4的人工肠液和pH=1.0的人工胃液中SBCSG的溶胀-收缩具有可逆性, 显示出良好的pH敏感性. 有望作为pH敏感口服结肠定位给药系统药物载体.     

（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖/聚乙二醇互穿网络凝胶的制备及其释药作用

将壳聚糖改性为（2-羟基-3-丁氧基）丙基 羟丙基壳聚糖（2-H-3-B-P-HPCS）,并以（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖和聚乙二醇（PEG）为原料制备（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖/聚乙二醇互穿网络凝胶,研究了（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖浓度、聚乙二醇的用量、交联剂戊二醛用量、反应温度对该凝胶溶胀性能的影响。 通过红外光谱分析和扫描电子显微镜的方法比较了壳聚糖、（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖和（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖/聚乙二醇互穿网络凝胶结构和形态上的不同。 以阿昔洛韦为模型药物研究了其释药性能。 结果表明,该凝胶均具有良好的溶胀性、pH敏感性和药物缓释作用,有望用作新型的药物载体。     

壳聚糖-聚丙烯酸复合水凝胶的药物控制释放行为

以壳聚糖(Cs)和丙烯酸(AA)为原料,利用自由基聚合法制备了具有孔洞结构的复合水凝胶Cs-PAA,并研究了AA的量、交联剂的量、聚合温度和AA的中和度对水凝胶溶胀度的影响以及复合水凝胶对烟酸的控制释放。 结果表明,Cs-PAA复合水凝胶具有良好的pH值、离子强度敏感性,且溶胀度最高达1228 g/g,其在pH=686的缓冲溶液中的烟酸累积释放率明显大于其在pH=1.80的缓冲溶液,因此Cs-PAA水凝胶可作为肠口服药物的载体。     

聚乙烯醇/羧甲基壳聚糖共混水凝胶的辐射合成及性能

采用电子加速器辐照法制备了聚乙烯醇(PVA)/羧甲基壳聚糖(CMCH)共混水凝胶;研究了PVA与CMCH的配比、辐照剂量、温度以及pH值对PVA/CMCH共混水凝胶性能的影响.实验发现,PVA与CMCH在辐照剂量为40 kGy、配比为w(PVA)/w(CMCH)=5/1的条件下可得到强度较好的PVA/CMCH共混水凝胶,该水凝胶具有一定的温度和pH敏感性:在5~20℃时具有较高的溶胀率,温度在20℃以上溶胀率较低;水凝胶在pH<4.0和pH>6.0时溶胀率均较大,而当pH为4.0~6.0时溶胀率较小.     

Preparation and characterization of collagen/chitosan poly (ethylene glycol)/nanohydroxyapatite composite scaffolds

Porous three‐dimensional collagen/chitosan scaffolds combined with poly (ethylene glycol) (PEG) and hydroxyapatite were obtained through a freeze‐drying method. Physical cross‐linking was examined by dehydrothermal treatment. The prepared materials were characterized by different analyses, eg, scanning electron microscopy (SEM), measurements of porosity and swelling, mechanical properties, and resistance to enzymatic degradation. The porosity of scaffolds and their swelling ratio decreased with the addition of hydroxyapatite. Moreover, after exposure to collagenase, the collagen/chitosan matrices containing PEG showed much faster degradation rate than matrices with the addition of hydroxyapatite. The results indicated that the addition of hydroxyapatite led to improvement of stiffness. The highest degree of porosity and swelling were demonstrated by collagen/chitosan/PEG matrices without hydroxyapatite.     

Chitosan nanocapsules: Effect of chitosan molecular weight and acetylation degree on electrokinetic behaviour and colloidal stability

In recent years, chitosan nanocapsules have shown promising results as carriers for oral drug or peptide delivery. The success in their applicability strongly depends on the stability of these colloidal systems passing through the digestive tract. In gastric fluids, clear stability comes from the high surface charge density of the chitosan shell, which is completely charged at acidic pH values. However, in the intestinal fluid (where the pH is almost neutral) the effective charge of these nanocapsules approaches zero, and the electrostatic forces cannot provide any stabilization. Despite the lack of surface charge, chitosan nanocapsules remain stable in simulated intestinal fluids. Recently, we have demonstrated that this anomalous stability (at zero charge) is owed to short-range repulsive forces that appear between hydrophilic particles when immersed in saline media. The present work examines the influence of the chitosan hydrophobicity, as well as molecular weight, in the stability of different chitosan nanocapsules. A study has been made of the size, polydispersity, electrophoretic mobility, and colloidal stability of eight core-shell nanocapsule systems, in which the chitosan-shell properties have been modified using low-molecular-weight (LMW) and high-molecular-weight (HMW) chitosan chains having different degrees of acetylation (DA). With regard to the stability mediated by repulsive hydration forces, the LMW chitosan provided the best results. In addition, contrary to initial expectations, greater stability (also mediated by hydration forces) was found in the samples formed with chitosan chains of high DA values (i.e. with less hydrophilic chitosan). Finally, a theoretical treatment was also tested to quantify the hydrophilicity of the chitosan shells.     

pH-sensitive polyelectrolyte complex gel microspheres composed of chitosan/sodium tripolyphosphate/dextran sulfate: swelling kinetics and drug delivery properties

Porous chitosan (CS) polyelectrolyte complex (PEC) hydrogel microspheres were prepared via either wet phase-inversion or ionotropic crosslinking with sodium tripolyphosphate (Na+ - TPP) and dextran sulfate (DS). The resulting microspheres were characterized using scanning electron microscopy (SEM) and elemental analysis (EA). The controlled release behavior of ibuprofen (IBU) from these microspheres was investigated. The PEC microspheres were about 700-950 microm in diameter with large pores and open porous structure. The CS/TPP/DS microspheres resisted hydrolysis in strong acid and biodegradation in enzymatic surroundings. The swelling kinetics for CS microspheres was close to Fickian diffusion, whereas those for CS/TPP and CS/TPP/DS were non-Fickian. Furthermore, the equilibrium water content (EWC) and water diffusion coefficient (D) increased with the pH of the media. The release profiles of IBU from CS/TPP/DS microspheres were slow in simulated gastric fluid (SGF, pH 1.4) over 3 h, but nearly all of the initial drug content was released in simulated intestinal fluid (SIF, pH 6.8) within 6 h after changing media. Overall the results demonstrated that CS/TPP/DS microspheres could successfully deliver a hydrophobic drug to the intestine without losing the drug in the stomach, and hence could be potential candidates as an orally administered drug delivery system.     

Antibacterial activity of pH‐sensitive genipin cross‐linked chitosan/poly(ethylene glycol)/silver nanocomposites

Novel nanocomposites consisting of genipin cross‐linked chitosan (GC), poly(ethylene glycol) (PEG), and silver nanoparticles were prepared for such biomedical applications as the wound‐healing materials. Various amounts of silver nanoparticles were dispersed in the GC/PEG hydrogel matrix without severe aggregation. The effects of composition and silver nanoparticles on the physico‐chemical properties of samples were evaluated by infrared analysis, contact angle measurements, and swelling tests. The GC/PEG/Ag nanocomposite showed a pH‐sensitive swelling behavior. The surface hydrophilicity of GC/PEG/Ag nanocomposites was improved with the increase of silver nanoparticle content. L929 cell attachment was improved in the presence of silver nanoparticles. The antimicrobial function was assessed for the GC/PEG/Ag nanocomposites containing the silver content over 100 ppm. The silver nanoparticles had the dual functions of reinforcing structural stability and enhancing antimicrobial activity of GC/PEG/Ag nanocomposites. Copyright © 2010 John Wiley & Sons, Ltd.     

Degradation of PEG and non-PEG alginate-chitosan microcapsules in different pH environments

Bioencapsulation allows the protection of biologically active substances or cells from the biological environment. As such, bioencapsulation is often used for the delivery of drugs, growth factors and therapeutically useful cells. Depending on the site of implantation, the biocapsules are subjected to different pH environments, which will affect the degradation properties, mechanical properties and swelling behaviour of the biocapsules. As such, the encapsulation material plays an important role in the long term stability and performance of the biocapsules in vivo. In this study, five types of encapsulation materials were investigated: (i) alginate (A), (ii) alginate-chitosan (AC), (iii) alginate-chitosan-alginate (ACA), (iv) alginate-chitosan-polyethylene glycol (PEG) (ACP) and (v) alginate-chitosan-polyethylene glycol (PEG)-alginate (ACPA). Degradation studies were carried out by immersing the microcapsules in solutions of different pH values to investigate the role of the material as well as the number of encapsulation layers in maintaining the stability of the microcapsules in the different pH environments. Compression testing indicated that even with the presence of PEG on the surface membrane, there was not much difference in mechanical strength between ACA and ACPA microcapsules. However, the use of PEG did affect the weight change of the ACPA microcapsules when immersed in water and three different pH solutions. For the swelling test, the ACPA microcapsules showed a lower water uptake than ACA microcapsules. For degradation, the presence of PEG led to a lower increase in weight change compared to non-PEG chitosan microcapsules. Hence, the study revealed that PEG influenced the integrity of the surface membrane and not the mechanical strength of the microcapsules. With the inclusion of PEG, the interpenetrating network on the surface membrane would be further reinforced. As such, the addition of PEG to the alginate-chitosan microcapsules led to protection against an acidic environment, whilst the number of coating layers only influences the swelling properties and not the degradation and Young’s modulus of the microcapsules.     

Formulation characterization and in vitro evaluation of acacia gum–calcium alginate beads for oral drug delivery systems

In the present work, new matrix bead formulations based on linear and branched polysaccharides have been developed using an ionic gelation technique, and their potential use as oral drug carriers has been evaluated. Using calcium chloride as a cross‐linking agent and sodium diclofenac (SD), as a model drug, acacia gum–calcium alginate matrix beads were formulated. The response surface methodology based on 3² factorial design was used as a statistical method to evaluate and optimize the effects of the biopolymers‐blend ratio and the concentration of calcium chloride on the particle size (mm), density (g/cm³), drug encapsulation efficiency (%), and the cumulative drug release after 8 hours (R_8h,%). The optimized beads with the highest drug encapsulation efficiency were examined for a drug‐excipients compatibility by powder X‐ray diffraction, differential scanning calorimetry, thermo‐gravimetric analysis, and Fourier transform‐infrared spectroscopy analyses. The swelling and degradation of the matrix beads were found to be influenced by the pH of medium. Higher degrees of swelling were observed in intestinal pH than in stomach pH. Accordingly, the drug release study showed that the amount of SD released from the acacia gum–calcium alginate beads was higher in intestinal pH than in stomach pH. Therefore, the in vitro drug release from the SD‐loaded beads appears to follow the controlled‐release (Hixson‐Crowell) pattern involving a case‐2 transport mechanism operated by swelling and relaxation of the polymeric blend matrix.     

壳聚糖-丝心蛋白合金膜的研究(Ⅱ)──半互穿聚合物网络型膜对pH和离子的敏感性

研究了壳聚糖-丝心蛋白半互穿聚合物网络型膜对pH值和离子的敏感性,发现该膜在pH<3.3时溶胀度剧烈增加;在pH=2时,是否出现溶胀极大值与膜组分的含量及交联剂的含量有关;膜在不同pH值溶液中可交替溶胀和收缩,且这种溶胀-收缩行为重复可逆。同时,该膜在不同离子溶液中的溶胀度亦不同;在相同离子强度的溶液中,此膜在3价离子中的溶胀度最大。     

肽类药物口服剂型材料及控制释放性能研究Ⅰ.壳聚糖─海藻酸盐微囊对胰岛素的控释作用

应用壳聚糖-海藻酸盐微囊技术制备了一系列胰岛素微囊，并研究了不同反应条件如海藻酸钠浓度、壳聚糖浓度、壳聚糖分子量及壳聚糖溶液pH值对微囊的胰岛素包封率及其释放性能的影响。结果表明，海藻酸钠浓度越高，微囊对胰岛素的包封率越高，在模拟小肠液中释放速率越低；壳聚糖浓度越大，微囊的胰岛素包封率及其在模拟胃液中释放率越高，在模拟肠液中释放达最大值所需时间越长；而随壳聚糖分子量减小，微囊在胃液中释放率增高；壳聚糖溶液pH值的变化对微囊的胰岛素包封率未造成明显影响。     

Preparation and release profiles of pH/temperature-responsive carboxymethyl chitosan/P(2-(dimethylamino) ethyl methacrylate) semi-IPN amphoteric hydrogel

Thermo- and pH-responsive semi-IPN polyampholyte hydrogels were prepared by using carboxymethyl chitosan and P(2-(dimethylamino) ethyl methacrylate) with N N'-Methylenebisacrylamide (BIS) as crosslinking agent. It was found that the semi-IPN hydrogel shrunk most at the isoelectric point (IEP) and swelled when pH deviated from the IEP. Its swelling ratio dramatically decreased between 30 and 50 °C at pH 6.8 buffer solution. It also showed good reversibility. The UV results showed that when the pH values of drug release medium were 3.7, 6.8, and 9 at 25 °C, the cumulative release rates reached 83.1, 51.5, and 72.2%, respectively. The release rate of coenzyme A (CoA) was higher at 50 °C than 37 and 25 °C at pH 6.8 solution. The release rate decreased with increasing the content of carboxymethyl chitosan at 25 °C in pH 6.8 solution. The results showed that semi-IPN hydrogel seems to be of great promise in pH/temperature drug delivery systems.     

Morphology and temperature responsiveness–swelling relationship of poly(N‐isopropylamide–chitosan) copolymers and their application to drug release

Poly [N‐isopropylacrylamide (NIPAAm)–chitosan] crosslinked copolymer particles were synthesized by soapless emulsion copolymerization of NIPAAm and chitosan. An anionic initiator [ammonium persulfate (APS)] and a cationic initiator [2,2′‐azobis(2‐methylpropionamidine)dihydrochloride (AIBA)] were used to initiate the reaction of copolymerization. The chitosan–NIPAAm copolymer synthesized by using APS as the initiator showed a homogeneous morphology and exhibited the characteristic of a lower critical solution temperature (LCST). The copolymer synthesized by using AIBA as an initiator showed a core–shell morphology, and the characteristic of LCST was insignificant. The LCST of the chitosan–NIPAAm copolymer depended on the morphology of the copolymer particles. In addition, the chitosan–NIPAAm copolymer particles were processed to form copolymer disks. Then, the effect of various variables such as the chitosan/NIPAAm weight ratio, the concentration of crosslinking agent, and the pH values on the swelling ratio of chitosan–NIPAAm copolymer disks were investigated. Furthermore, caffeine was used as the model drug to study the characteristics of drug loading of the chitosan–NIPAAm copolymer disks. Variables such as the chitosan/NIPAAm weight ratio and the concentration of the crosslinking agent significantly influenced the behavior of caffeine loading. Two factors (pore size and swelling ratio) affected the behavior of caffeine release from the chitosan–NIPAAm copolymer disks. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 3029–3037, 2004     

Synthesis and characterization of a pH responsive and mucoadhesive drug delivery system for the controlled release application of anti-cancerous drug

In this work a novel pH sensitive composite, polyacrylamide grafted succinyl chitosan intercalated bentonite (AAm-g-NB/SC) was prepared as a drug carrier system for the controlled delivery of paclitaxel. Characterization of the drug delivery system was carried out using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), thermal analysis etc. The equilibrium swelling behaviour of the composite was studied and the result showed a maximum at pH 7.4. The in vitro drug release study of paclitaxel indicated that about 15.6% of drug release was found to be occurred at pH 1.2 within 16 h, whereas about 82.5% of drug release was occurred at the intestinal pH condition of 7.4. In vitro biocompatibility study was performed and the result showed good biocompatibility of the composite in the concentration range 6.25–100 µg/mL. The cytotoxicity assay was carried out in cancerous cell line of Human colorectal Adenocarcinoma. Mucous glycoprotein assay study showed that the drug delivery system having good apparent adhering property towards mucin. The investigation indicated that paclitaxel, an anticancer drug can be successfully entrapped in the AAm-g-NB/SC composite for the controlled and targeted delivery for colorectal cancer therapy.