Antibacterial and swelling properties of N-isopropyl acrylamide grafted and collagen/chitosan-immobilized polypropylene nonwoven fabrics

The different molar ratios of collagen/chitosan were used to be immobilized on polypropylene nonwoven fabrics grafted with N-isopropyl acrylamide (abbr. PP-g-NIPAAm-i-Col/Chi). For a controlled immobilizing time and NIPAAm concentration, the collagen/chitosan immobilized values and the antibacterial properties of PP-g-NIPAAm-i-Col/Chi increased with increasing amount of chitosan in the mixture of collagen/chitosan. The crosslinking reaction between the grafted polyNIPAAm and collagen/chitosan molecules was clearly confirmed by the examination of the spectra of the surface reflection infrared spectroscopy (IR). The values of water absorption and water diffusion coefficient of PP-g-NIPAAm-i-Col/Chi decreased with increase of the chitosan in the mixture of collagen/chitosan and the value of immobilized collagen/chitosan at the same pH value of buffering water. The PP-g-NIPAAm-i-Col/Chi have excellent water absorption, water permeability, and antibacterial properties and would be suitable for the healing of wounded skin area.     

Enhanced biocompatibility and antibacterial property of polyurethane materials modified with citric acid and chitosan

Citric acid (CA) and chitosan (CS) were covalently immobilized on polyurethane (PU) materials to improve the biocompatibility and antibacterial property. The polyurethane pre-polymer with isocyanate group was synthesized by one pot method, and then grafted with citric acid, followed by blending with polyethersulfone (PES) to prepare the blend membrane by phase-inversion method so that chitosan can be grafted from the membrane via esterification and acylation reactions eventually. The native and modified membranes were characterized by attenuated total reflectance-Fourier transform infrared spectroscope, X-ray photoelectron spectroscopy, scanning electron microscopy, water contact angle measurement, and tensile strength test. Protein adsorption, platelet adhesion, hemolysis assay, activated partial thromboplastin time, prothrombin time, thrombin time, and adsorption of Ca²⁺ were executed to evaluate the blood compatibility of the membranes decorated by CA and CS. Particularly, the antibacterial activities on the modified membranes were evaluated based on a vitro antibacterial test. It could be concluded that the modified membrane had good anticoagulant property and antibacterial property.     

Microbiological interaction and diffusion properties of hydrophilic and hydrophobic PU membranes

In this study, the diffusion characteristics of Diclofenac-Na through hydrophobic and hydrophilic polyurethane (PU) based membranes are investigated. Hydrophilic polymers are obtained via graft copolymerization of PU with acrylic acid (AA) and crotonic acid (CA). The membranes are prepared by solvent-casting method and characterized by FTIR spectra and SEM analysis. The diffusion measurements are performed using a diffusion cell during 10 h at 37 degrees C. Permeability coefficients calculated from diffusion experiments are found to be approximately three times higher in hydrophilic membrane than hydrophobic PU membrane. The adherence of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) onto the membranes at 37 degrees C were also investigated. It is determined that the microorganisms didn't adhere onto the membranes compared by control suspension.     

Chitosan/poly(vinyl alcohol) blending hydrogel coating improves the surface characteristics of segmented polyurethane urethral catheters

Segmented polyurethane (SPU) is commonly used to manufacture urethral catheters. Surface modifications for SPU catheters are needed to reduce friction and protein adsorption, in order to minimize catheter-related complications, including urethral trauma, encrustation, catheter obstruction, bacterial colonization, and infection. In this study, a four-step surface modification method was developed to create a thin lubricious layer of chitosan/poly(vinyl alcohol) (PVA) hydrogel on the SPU catheter. Modification steps included oxidation of the SPU surface, functionalities modification, carbodiimide reaction and coupling, and hydrogel crosslinking. The success of each modification step was confirmed by Fourier transform infrared spectroscopy. Measurement of the water contact angle revealed that hydrogel coating created a highly hydrophilic surface and atomic force microscope analyses demonstrated that the surface was slippery. Protein absorption of the SPU catheter was significantly reduced by coating hydrogel. Chitosan in the hydrogel could provide antimicrobial activity, and the hydrogel coating SPU samples showed significant antibacterial effects in this study. In summary, the four-step modification method developed in this study provided a simple and effective way to coat the surface of SPU catheters with a chitosan/PVA blending hydrogel that could help to minimize the risk of complications related to the use of urethral catheters.     

Synthesis and characterization of thermosensitive chitosan copolymer as a novel biomaterial

Novel water-soluble thermosensitive chitosan copolymers were prepared by graft polymerization of N-isopropylacrylamide (NIPAAm) onto chitosan using cerium ammonium nitrate (CAN) as an initiator. The physicochemical properties of the resulting chitosan-g-NIPAAm copolymers were characterized by Fourier transform infrared (FT-IR) spectroscopy, 1H-nuclear magnetic resonance, X-ray diffraction measurement, thermogravimetric analysis (TGA) and solubility test. Sol-gel transition behavior was investigated by the cloud point measurement of the chitosan-g-NIPAAm aqueous solution. The gelling temperature was examined using the vial inversion method. The percentage of grafting (%) and efficiency of grafting (%) were investigated according to concentrations of monomer and initiator. The maximum grafted chitosan copolymer was obtained with 0.4 M NIPAAm and 6 x 10(-3) M CAN. Water-soluble chitosan-g-NIPAAm copolymers were prepared successfully and they formed thermally reversible hydrogel, which exhibits a lower critical solution temperature (LCST) around 32 degrees C in aqueous solutions. A preliminary in vitro cell study showed nontoxic and biocompatible properties. These results suggest that chitosan-g-NIPAAm copolymer could be very useful in biomedical and pharmaceutical applications as an injectable material for cell and drug delivery.     

新型纳米银/聚氨酯胆道支架表面抗菌涂层的体外抑菌试验

背景：胆管支架已广泛应用于胆管各种良恶性狭窄的姑息治疗,临床短期疗效显著,但长期疗效由于支架再狭窄而受到限制。 目的：研制新型纳米银/聚氨酯胆管内支架复合抗菌涂层并检测其体外抗菌性能。 方法：原位还原法用纳米银、聚氨酯制备纳米银/聚氨酯抗菌材料,采用抑菌环试验法分别测定实验组(纳米银/聚氨酯)、阳性对照(硝酸银纸片)和阴性对照(普通聚氨酯材料)3组对金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌、肺炎克雷伯杆菌和肠球菌的抑菌环直径并记录分析数据。 结果与结论：纳米银/聚氨酯在体外对金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌、肺炎克雷伯杆菌和肠球菌均有抑制效果,与硝酸银纸片相比,抑菌效果差异无显著性意义。结果显示纳米银/聚氨酯复合物具有良好的抗菌效果,且具有较好的稳定性。     

Preparation and surface properties of PEO-sulfonate grafted polyurethanes for enhanced blood compatibility

In order to improve the thromboresistance of the commercial polyurethane(PU), its surface modification was accomplished by three new different methods and their surface characteristics were investigated using ATR-FTIR, ESCA, SEM, and dynamic contact angle measurements. Sulfonations using propane sultone were performed directly onto PU or onto hydrophobic dodecanediol (DDO) grafted PU or onto hydrophilic poly(ethyleneoxide) (PEO) grafted PU. ESCA data coincided well with ATR-IR results, as more 0 at. % for PEO grafted PUs and the presence of S for the sulfonated PUs were revealed. At SEM observation the surfaces of PU-DDO and PU-PEO were relatively smooth, whereas all the sulfonated PU surfaces showed excellent smoothness and homogeneity. The hydrophilicity of the surfaces was considerably increased after PEO grafting or sulfonation. In addition, all the sulfonated PU surfaces, particularly PU-PEO-SO₃, which has further hydrophilicity, exhibited complete wetting behavior due to the negatively charged SO₃ groups.     

Poly(methyl methacrylate)-grafted chitosan microspheres for controlled release of ampicillin

Microspheres of 50-500 microm diameter were prepared from a blend of chitosan and chitosan-g-PMMA. Environmental scanning electron microscopic and SEM studies revealed that the microspheres are porous and the pores extend toward the inner core of the microspheres. The microspheres were also found to be hemocompatible and cytocompatible. A model drug ampicillin was used to evaluate the drug loading capacity and the controlled release properties of the microspheres. The system maintained a sustained release of ampicillin for a period of more than 8 days. The drug-loaded chitosan/chitosan-g-PMMA microspheres exhibited higher antibacterial activity for both the gram positive (ATCC 25923 S. aureus) and gram negative (ATCC 25922 E. coli) bacteria than the drug-loaded virgin chitosan microspheres. The percentage release and bioactivity of ampicillin was found to be higher for the chitosan/chitosan-g-PMMA microspheres than the virgin chitosan microspheres. Potential applications such as oral drug delivery, wound dressings, tissue engineering, and so forth, are envisaged from these microspheres.     

Covalently crosslinked chitosan hydrogel: properties of in vitro degradation and chondrocyte encapsulation

In vitro degradation and chondrocyte-encapsulation of chitosan hydrogel made of crosslinkable and water-soluble chitosan derivative (CML) at neutral pH and body temperature were studied with respect to weight loss, cytoviability, DNA content and cell morphology. In vitro degradation of the chitosan hydrogels was sensitive to their crosslinking degree and existence of lysozyme in the solution. Chitosan hydrogel (Gel-I5) fabricated from 1% CML and 5mM ammonium persulfate (APS)/N,N,N',N'-tetramethylethylenediamine (TMEDA) displayed no degradation in phosphate buffered saline (PBS) after 18d, but degraded completely at 8d in 1mg/ml lysozyme/PBS. The chitosan hydrogel fabricated from 10mM APS/TMEDA was non-degradable even in lysozyme/PBS solution after 18d. The hydrogel loaded with chondrocytes in cell culture medium, however, was susceptible to degradation during the in vitro culture. In vitro culture of the encapsulated chondrocytes in the chitosan hydrogel demonstrated that the cells retained round shaped morphology and could survive through a 12d-culture period, although the DNA assay detected an overall reduction of the cell number. These features provide a great opportunity to use the chitosan hydrogel as an injectable scaffold in tissue engineering and orthopaedics.     

Protein adsorption,fibroblast activity and antibacterial properties of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) grafted with chitosan and chitooligosaccharide after immobilized with hyaluronic acid

Poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) membrane was treated with ozone and grafted with acrylic acid. The resulting membranes were further grafted with chitosan (CS) or chitooligosaccharide (COS) via esterification. Afterward hyaluronic acid (HA) was immobilized onto CS- or COS-grafting membranes. The antibacterial activity of CS and COS against Staphylococus aureus, Escherichia coli, and Pseudomonas aeruginosa was preserved after HA immobilization. Among them, CS-grafted PHBV membrane showed higher antibacterial activity than COS-grafted PHBV membrane. In addition, after CS- or COS-grafting, the L929 fibroblasts attachment and protein adsorption were improved, while the cell number was decrease. After immobilizing HA, the cell proliferation was promoted, the protein adsorption was decreased, and the cell attachment was slightly lower than CS- or COS-grafting PHBV.     

In vitro antibacterial and cytotoxicity assay of multilayered polyelectrolyte-functionalized stainless steel

Infection of implanted materials by bacteria constitutes one of the most serious complications following prosthetic and implant surgery. In the present study, a new strategy for confering stainless steel with antibacterial property via the alternate deposition of quaternized polyethylenimine (PEI) or quaternized polyethylenimine-silver complex and poly(acrylic acid) (PAA) was investigated. The success of the deposition of the polyelectrolyte multilayers (PEM) and its chemical nature was investigated by static water contact angle and X-ray photoelectron spectroscopy (XPS), respectively. The antibacterial activity was assessed using Escherichia coli (E. coli, a gram-negative bacterium) and Staphylococcus aureus (S. aureus, a gram-positive bacterium). The inhibition of E. coli and S aureus growth on the surface of functionalized films was clearly shown using the LIVE/DEAD Baclight bacterial viability kits and fluorescence microscopy. The cytotoxicity of the PEM to mammalian cells, evaluated by the MTT assay, was shown to be minimal and long-term antibacterial efficacy can be maintained. These results indicate new possibilities for the use of such easily built and functionalized architectures for the functionalization of surfaces of implanted medical devices.     

Characterization and behavior of anesthetic bioactive textile complex in vitro condition

In this study, a bioactive complex containing nonwoven textile material (polypropilene (PP)/viscose), chitosan hydrogel, and lidocaine hydrochloride, was designed. The purpose of such biomedical textile was in the treatment of painful sites. Mercury intrusion porosimetry was used in order to estimate the influence of medical impregnation on porous structure of nonwoven material. It was estimated that more than 97% of pores in untreated nonwoven sample were larger than 15 μm. Anesthetic treatment of nonwoven reduced total pore volume of ultramacropores and macropores, while total pore volume of mesopores slightly increased. Lidocaine hydrochloride release from the anesthetic/chitosan hydrogel/nonwoven complex was measured in vitro by Franz diffusion cell technique. Mathematical model was developed to estimate the release of the lidocaine from obtained bioactive textile material. The diffusive transport of lidocaine hydrochloride through three connected layers, i.e., polymer hydrogel, membrane, and solution is modeled based on Fick's second law. Taking all the relevant conditions, regarding this experiment, into consideration, the coefficient of lidocaine diffusion through the polymer hydrogel, as well as the concentration ratio parameter were determined by the mathematical model.     

A mechanistic study of the antibacterial effect of silver ions on Escherichia coli and Staphylococcus aureus

To investigate the mechanism of inhibition of silver ions on microorganisms, two strains of bacteria, namely Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus), were treated with AgNO(3) and studied using combined electron microscopy and X-ray microanalysis. Similar morphological changes occurred in both E. coli and S. aureus cells after Ag(+) treatment. The cytoplasm membrane detached from the cell wall. A remarkable electron-light region appeared in the center of the cells, which contained condensed deoxyribonucleic acid (DNA) molecules. There are many small electron-dense granules either surrounding the cell wall or depositing inside the cells. The existence of elements of silver and sulfur in the electron-dense granules and cytoplasm detected by X-ray microanalysis suggested the antibacterial mechanism of silver: DNA lost its replication ability and the protein became inactivated after Ag(+) treatment. The slighter morphological changes of S. aureus compared with E. coli recommended a defense system of S. aureus against the inhibitory effects of Ag(+) ions.     

Studies of the antimicrobial activity of ovotransferrin

The antibacterial activity of ovotransferrin (conalbumin) against different bacterial species was studied in vitro. The most sensitive species were Pseudomonas sp., E. coli, S. mutans; and the most resistant ones S. aureus, Proteus sp., Klebsiella. The bacteriostatic activity of conalbumin in various conditions was also tested. The presence of bicarbonate ions always increased the activity of conalbumin, while an antagonistic effect of citrate was observed in bacteria with a receptor for the iron-citrate complex. Experiments with conalbumin covalently linked to Sepharose 4B indicated that its antibacterial activity may not be due simply to the removal of iron from the medium, but probably involves other metals and an interaction with the bacterial surface. The in vitro studies carried out with conalbumin and lactoferrin demonstrated that the two proteins produced a similar inhibition of growth of E. coli and S. mutans. The in vivo studies showed that the protective effect of conalbumin in newborn guinea pigs with E. coli by gastrointestinal route was similar to that naturally provided by the lactoferrin present in milk.     

Preparation and antibacterial properties of O-carboxymethyl chitosan/lincomycin hydrogels

In this study, O-carboxymethyl chitosan (O-CMCS) was synthesized from chitosan and monochloroacetic acid. Then O-CMCS hydrogel was prepared by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) in which the lincomycin was packaged. The Fourier transform infrared spectrum and scanning electron microscopy were adopted to characterize the structure and morphology of the product. The influences of dosage of EDC/NHS and concentration of O-CMCS on the swelling properties of the hydrogels were investigated. The hydrogels performed good swelling capacities and obvious pH-sensitive properties. The antibacterial activities of the hydrogels were tested against Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus). Compared with pure O-CMCS hydrogels, the antibacterial activities of O-CMCS/lincomycin hydrogels were significantly improved with the increase in the concentration of lincomycin against E. coli and S. aureus. With the increase in dosage of crosslinking agent or concentration of O-CMCS, the antibacterial activities both decreased gradually against the two bacteria. O-CMCS/lincomycin hydrogel was expected to be used for antibacterial material in view of its significant antibacterial activities.     

Synergistic actions of antibacterial neutrophil defensins and cathelicidins

OBJECTIVE: Activated neutrophils extracellularly release antibacterial defensins and cathelicidins from the granules. In this study, to elucidate the interactions between defensins and cathelicidins in the extracellular environment, we evaluated the individual and synergistic actions of defensins and cathelicidins in the presence of physiological concentration of NaCl (150 mM). MATERIALS AND METHODS: Antibacterial activities against Escherichia coli and Staphylococcus aureus were assessed using human and guinea pig defensins and cathelicidins. Furthermore, the effect of defensins and cathelicidins on membrane permeabilization was examined using E. coli ML-35p, as a target organism. RESULTS: In the absence of NaCl, human defensin (HNP-1) and guinea pig defensins (GNCPs) exhibited the antibacterial activities in a dose-dependent manner (0.1-10 microg/ml); however, their activities were completely lost in the presence of 150 mM NaCl. In contrast, the antibacterial activities of human cathelicidin (CAP18/LL-37) and guinea pig cathelicidin (CAP11) were resistant to NaCl. Interestingly, HNP-1 and GNCPs synergized with CAP18/LL-37 and CAP11 to enhance the antibacterial activities against E. coli and S. aureus in the presence of 150 mM NaCl (p<0.05). Similarly, HNP-1 and GNCPs were synergistic with CAP18/LL-37 and CAP11 to potentiate the outer and inner membrane permeabilization of E. coli ML-35p (p<0.05). CONCLUSIONS: Together these observations indicate that when extracellularly released from neutrophils, defensins cannot function as antibacterial molecules by themselves, but can synergistically work with cathelicidins to exert the antibacterial activity in the extracellular milieu by augmenting the membrane permeabilization of target cells.     

Water absorbing and antibacterial properties of N-isopropyl acrylamide grafted and collagen/chitosan immobilized polypropylene nonwoven fabric and its application on wound healing enhancement

A durable sandwich wound dressing system with high liquid absorbing, biocompatibility, and antibacterial properties was designed. Various solution weight ratios of collagen to chitosan were used to immobilize on the polypropylene nonwoven fabric, which were pregrafted with acrylic acid (AA) or N-isopropyl acrylamide (NIPAAm) to construct a durable sandwich wound dressing membrane with high water absorbing, easy removal, and antibacterial activity. Swelling properties and antibacterial activity of the membranes were measured, and wound healing enhancement by skin full-thickness excision on animal model was examined. The results indicated that NIPAAm-grafted and collagen/chitosan-immobilized polypropylene nonwoven fabric (PP-NIPAAm-collagen-chitosan) showed a better healing effect than AA-grafted and collagen/chitosan-immobilized polypropylene nonwoven fabric (PP-AA-collagen-chitosan). The wound treated with PP-NIPAAm-collagen-chitosan demonstrated the excellent remodeling effect in histological examination with respect to the construction of vein, epidermis, and dermis at 21 days after skin injury. The values of water uptake and water diffusion coefficient for PP-NIPAAm-collagen-chitosan were higher than that for PP-AA-collagen-chitosan under a given solution weight ratio of collagen/chitosan. Both PP-NIPAAm-collagen-chitosan and PP-AA-collagen-chitosan demonstrated antibacterial activity.     

Preparation and characterization of alpha-amylase-immobilized thermal-responsive composite hydrogel membranes.

Composite hydrogel membranes of crosslinked poly(N-isopropylacrylamide-co-N-acryloxysuccinimide-co-2-hydroxyet hyl methacrylate) [P(NIPAAm-NAS-HEMA)] with starch, as a macropore forming agent, on nonwoven polyester was prepared. The membranes could swell and de-swell around the characteristic lower critical solution temperature (LCST) of poly(N-isopropylacrylamide) (PNIPAAm). It was demonstrated that the presence of macropores in the membranes could improve the immobilization efficiency as well as lead to a short responding time upon temperature change across the LCST. Immobilized alpha-amylase could retain as high as 33% of the activity of the free enzyme with a loading level of 0.60-0.65 mg/cm2 when the membrane preparation and enzyme immobilization conditions were optimized. The half time (T0.5) for the swelling or de-swelling response of the gel phase within the membranes was less than 2 min, and the 90% time (T0.9) was less than 6 min. The permeability for maltose through the membranes could change as much as 4.9-fold when the temperature was raised above or reduced below the LCST.     

Chemical, biological and microbiological evaluation of cyclodextrin finished polyamide inguinal meshes

This study describes the use of cyclodextrins (CDs) as a finishing agent of polyamide (PA) fibers used in order to obtain inguinal meshes with improved antibiotic delivery properties. The finishing process involved polymerization between citric acid and CDs, which yielded a cross-linked polymer that physically adhered to the surface of PA fibers. This permanent functionalization was characterized by evaluating the damping property with a polar liquid (glycerol) via the drop contact angle method for various rates of modification of the fabrics. The biological and microbiological effects of the PA, which were functionalized with hydroxypropylated derivate of gamma-CD (HP-gamma-CDs) and charged with ciprofloxacin (CFX), were evaluated by cell culture assays. We observed a good adhesion and proliferation of fibroblastic cells (NIH3T3) after 3 and 6 days and no detectable toxicity of the modified substrate. The in vitro antibacterial activity of the HP-gamma-CD grafted PA fabrics charged with CFX against the bacteria Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli was greatly superior to that of the virgin sample within a 24h batch experiment in human blood plasma medium. In conclusion, these results from our study offer an insight into the efficient performance of CDs as drug delivery systems for multiple applications in the fields of biomaterials and medical textiles.     

纳米载银磷酸锆抗菌聚氨酯的抗菌性能****

背景：聚氨酯材料具有优异的物理和化学性能,良好的生物相容性和抗凝血性能,且易加工成形,但聚氨酯制造的人工器官容易受到细菌等微生物的入侵。 目的：观察纳米载银无机抗菌剂对聚氨酯抗菌性能的影响。 方法：将纳米载银无机抗菌剂RHA-2,按0%(空白对照组),0.5%,1%,1.5%,2%,2.5%,5%比例添加到聚氨酯中。采用薄膜密着法检测抗菌聚氨酯对金黄色葡萄球菌、大肠杆菌的抑菌作用,并分析比较抗菌剂添加比例与聚氨酯抗菌性能的相关性。 结果与结论：添加纳米载银无机抗菌剂的聚氨酯对金黄色葡萄球菌、大肠杆菌具有良好的抑菌作用。抗菌剂添加比例0.5%~5%组对金黄色葡萄球菌的抑菌率分别为80.23%,91.32%,95.23%,99.19%,99.87%,99.93%,对大肠杆菌的抑菌率分别为76.70%,86.96%,92.92%,95.43%,99.34%,99.87%,显示抗菌性能随抗菌剂添加比例的上升而明显提高。表明纳米载银无机抗菌剂的添加赋予了聚氨酯优异的抗菌性能,且从抗菌角度出发,推荐纳米载银无机抗菌剂在聚氨酯中的添加比例不应低于1.5%。