Synthesis and Properties of Dendronized Chitosan

Summary: Chitosan films and microspheres were prepared and their surfaces were functionalized with first generation dendritic molecules. The films were modified by Weisocyanate dendron, while Behera's and bis Behera's amine dendrons were used to modify the microspheres. Prior to dendronization films were prepared by blending chitosan with 18% of polyvinyl pyrrolidone (PVP), and casting the resulting mixture. The degree of dendronization reached was 28%. The microspheres were prepared by coacervation/precipitation, after which the surfaces were activated with either epychlorohydrine (ECH) or 1,4-butanediol diglycidyl ether (BDGE). The oxirane groups were utilized to form covalent bonds between chitosan and dendrons. The degree of dendronization yielded with Behera's amine was 60% for both activating agents. When bis Behera's amine was used, the dendronization reached values of 15 and 21% when ECH or BDGE were used, respectively. The dendronized products were characterized through spectroscopic and microscopic studies and by determination of swelling indexes. Only one of the surfaces was dendronized in every film, which therefore presented a hydrophobic and a hydrophilic surface. Since these films maintain the properties of chitosan, they offer interesting potential as dressings for exuding wounds. The different surfaces make the microspheres potentially applicable as carriers for delivery and controlled release of drugs.     

Solution-based assembly of conductive gold film on flexible polymer substrates

Conductive films of gold were assembled on flexible polymer substrates such as Kapton and polyethylene using a solution-based process. The polymer substrates were modified by using argon plasma and subsequent coupling of silanes with amino- or mercapto- terminal groups. These modified surfaces were examined by X-ray photoelectron spectroscopy and contact angle measurements. Colloidal gold was assembled onto the silane-modified surface from solution. The gold particles are attached to the surface by covalent interactions with the thiol or amine group. Formation of a conductive film is achieved by increasing the coverage of gold by using a "seeding" method to increase the size of the attached gold particles. Field emission scanning electron microscopy was used to follow the growth of the film. The surface resistance of the films, measured using a four-point probe, was about 1 Omega/sq.     

Preparation and Characterization of Titania-grafted Poly（styrene-divinybenzene） Nanocomposite Microspheres

Titania-grafted poly（styrene-divinylbenzene）（TiO2/PSt-DVB） nanocomposite microspheres were prepared by an open-ring reaction and radical grafting copolymerization method. The TiO2 nanoparticles were first modified by attachment of epoxy groups to their surfaces to provide reactive groups that could covalently bond to the polymer （PSt-DVB） microspheres. The nanocomposite obtained was characterized by FTIR, SEM, XRD, and TGA analyses as well as UV-Vis spectrophotometry. The results indicated that the TiO2 nanoparticles were uniformly grafted onto the surface of the polymer microsphere producing grain sizes of about 5―10 μm. The modified TiO2 showed better UV absorbing property than the unmodified form, and the nanocomposite also retained the same UV absorbing property as the free modified TiO2 nanoparticle.     

Protein surface patterning using nanoscale PEG hydrogels

We have used focused electron-beam cross-linking to create nanosized hydrogels and thus present a new method with which to bring the attractive biocompatibility associated with macroscopic hydrogels into the submicron length-scale regime. Using amine-terminated poly(ethylene glycol) thin films on silicon substrates, we generate nanohydrogels with lateral dimensions of order 200 nm which can swell by a factor of at least five, depending on the radiative dose. With the focused electron beam, high-density arrays of such nanohydrogels can be flexibly patterned onto silicon surfaces. Significantly, the amine groups remain functional after e-beam exposure, and we show that they can be used to covalently bind proteins and other molecules. We use bovine serum albumin to amplify the number of amine groups, and we further demonstrate that different proteins can be covalently bound to different hydrogel pads on the same substrate to create multifunctional surfaces useful in emerging bio/proteomic and sensor technologies.     

Surface modification. II. Functionalization of solid surfaces with vinylic monomers

A new method to functionalize surfaces of solid substrates such as glass, silicon crystals, and silica microspheres with appropriate vinylic monomers, i.e., methyl vinyl ketone, methyl acrylate, methacrolein, and acrolein, is described. The surface modification process was performed through the following sequence of reactions: (a) derivatization of the surfaces with to-nitrile groups by interacting the substrates with SiCl₃(CH₂)₃CN; (b) subsequent reduction of the a)-nitrile groups with diborane to w-amine groups; (c) binding of the vinylic monomers to the surfaces via the to-amine groups. pK_1/2 of the surface primary amine groups, as determined by contact angle titration, was found to be 2–4 units lower than the pK_1/2 values of primary amine analogous in solution. Methyl vinyl ketone and methyl acrylate were covalently bound to the amine surfaces only under basic conditions via the Michael addition reaction. Methacrolein and acrolein were covalently bound to the amine surfaces under both acidic and basic conditions via two major reactions: the Michael addition reaction and Schiff base bond formation. The concentration of the aldehyde groups of the surfaces obtained by the reaction with methacrolein and acrolein was significantly higher than that obtained using the common, published method in which glutaraldehyde interacts with the amine surfaces.     

壳聚糖膜固定生物活性短肽RGDS的研究

为改善壳聚糖对细胞的特异性吸附,采用水溶性碳二亚胺将生物活性短肽精氨酸-甘氨酸-天冬氨酸-丝氨酸(RGDS)固定到壳聚糖膜的表面,采用X射线光电子能谱检测固定多肽前后的壳聚糖膜表面,发现反应后壳聚糖膜表面氮元素含量增大,Nls和Cls曲线拟合谱中酰胺键增多,表明RGDS短肽已固定到壳聚糖膜的表面;人角膜缘上皮细胞体外培养实验表明,固定RGDS后壳聚糖膜的细胞黏附率有了明显提高,固定RGDS后的壳聚糖膜在角膜组织工程支架等方面有更好的应用潜力。     

Covalent VEGF protein immobilization on resorbable polymeric surfaces

Vascular endothelial growth factor type protein (VEGF), a potent angiogenic effector molecule, was successfully covalently immobilized onto the surfaces of the resorbable polymers poly(L‐lactic acid) (PLLA) and poly(ε‐caprolactone) (PCL) through a three‐step strategy. The surfaces were first covalently grafted with poly(acrylic acid) using non‐destructive and solvent free vapor‐phase grafting. A diamine spacer was coupled to the carboxylic acid pendant groups on the graft chains using EDC/NHS chemistry and VEGF was finally covalently attached to the amine linkers. The chemistry and topography of the modified substrates were quantitatively and qualitatively verified with XPS, ATR‐FTIR, UV–VIS, SEM, and ELISA. Copyright © 2010 John Wiley & Sons, Ltd.     

Crosslinked Polyacrolein Microspheres with High Loading of Aldehyde Groups for Use as Scavenger Resins in Organic Synthesis

Summary: Suspension polymerization yielded microspheres (40–50 μm) of polyacrolein. Smooth and rugged surfaces can be created by varying the polymerization procedure. We have shown that the polyacrolein resins with a high loading of aldehyde groups serve as effective scavengers for primary amines and may be used to remove compounds bearing amino groups in the combinatorial synthesis of compound libraries. Copolymerization with styrene can help to separate the adjacent aldehyde groups, thus making the functional groups more available in organic reactions. The polyacrolein resins in the aldehyde form or after appropriate chemical modifications may also be useful as support materials in solid‐phase synthesis.

The SEM image of macroporous polyacrolein microspheres with toluene as porogen prepared by free radical polymerization.     

Chitosan modified poly(L-lactide) microspheres as cell microcarriers for cartilage tissue engineering

The surfaces of poly(l-lactide) (PLLA) microspheres were modified by chitosan via a method of hydrolysis and grafting-coating to improve their compatibility to chondrocytes. The PLLA microspheres with a diameter of 74-150mum were fabricated by an oil/water emulsion solvent evaporation method, followed by hydrolysis in alkaline solution to produce a larger number of carboxyl groups. Using water-soluble carbodiimide as a coupling reagent, chitosan was covalently grafted onto the microspheres. Due to the physical entanglement and insolubility at neutral pH, unbonded chitosan molecules were stably remained to yield a large amount of coated chitosan. Biological performance of the control PLLA and the chitosan-coated PLLA microspheres were assessed by in vitro culture of rabbit auricular chondrocytes. After 24h and 7d culture, the chitosan-coated PLLA microspheres, especially the ones with larger chitosan amount, exhibited stronger ability to promote cell attachment and proliferation, and maintain the secretion function of the chondrocytes. Therefore, the chitosan-coated PLLA microspheres can be potentially used as the injectable cell microcarriers for chondrogenesis in cartilage tissue engineering.     

Vibrational spectroscopy of dimethylchlorooctadecylsilane covalently bonded to ultrathin silica films immobilized on Ag surfaces

FTIR and Raman spectroscopies have been used to characterize the structure and conformational order of dimethylchlorooctadecylsilane (DOS) covalently bonded to ultrathin silica films supported on Ag substrates. Ultrathin silica films of ca. 30 A thickness prepared from sol-gel methods are immobilized on Ag surfaces modified with a self-assembled monolayer of (3-mercaptopropyl)trimethoxysilane (3MPT). This layered structure provides a unique opportunity for acquiring complementary spectral data from both FTIR and Raman spectroscopies, which are useful in elucidating alkylsilane conformation pertaining to stationary phases for reversed-phase liquid chromatography (RPLC). Characterization of octadecyltrichlorosilane (OTS) layers on thin silica films of ca. 800 A thickness on 3MPT-modified Ag surfaces has been reported previously. Differences between the ultrathin silica films used in this study and the thin silica films used in this previous study are considered. The results from both FTIR and Raman spectroscopy presented here suggest that bonded DOS alkyl chains are in a disordered, liquid-like state with close to monolayer surface coverage.     

Synthesis and characterization of highly ordered mesoporous thin films with -COOH terminated pore surfaces

Highly ordered mesoporous inorganic-organic hybrid thin films with covalently bonded carboxylic acid (-COOH) terminal groups on the pore surfaces were synthesized by evaporation induced self-assembly of tetraethoxysilane, organosilanes, and a nonionic surfactant followed by acid hydrolysis and characterized using transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, surface acoustic wave (SAW) based N2 sorption, and thermogravimetric analysis (TGA) techniques.     

Interaction of antigen and antibody on core-shell polymeric microspheres

Monodispersed microspheres with polystyrene as the core and poly(acrylamide-co-N-acryloxysuccinirnide) as the shell were synthesized by a two-step surfactant-free emulsion copolymerization.The core-shell morphology of the microspheres was shown by scanning electron microscopy and transmission electron microscopy.Rabbit immunoglobulin G (as antigen) was covalently coupled onto the microspheres by the reaction between succinimide-activated ester groups on the shell of the microspheres and amino groups of the antigen molecules.The size of particles was characterized by dynamic light scattering technique and was found to vary upon bioconjugation and interaction with proteins.The binding process was shown to be specific to goat anti-rabbit immunoglobulin G(as antibody) and reversible upon the addition of free antigen into the system.     

Vibrational spectroscopy of dimethylchlorooctadecylsilane covalently bonded to ultrathin silica films immobilized on Ag surfaces

FTIR and Raman spectroscopies have been used to characterize the structure and conformational order of dimethylchlorooctadecylsilane (DOS) covalently bonded to ultrathin silica films supported on Ag substrates. Ultrathin silica films of ca. 30 Å thickness prepared from sol-gel methods are immobilized on Ag surfaces modified with a self-assembled monolayer of (3-mercaptopropyl)trimethoxysilane (3MPT). This layered structure provides a unique opportunity for acquiring complementary spectral data from both FTIR and Raman spectroscopies, which are useful in elucidating alkylsilane conformation pertaining to stationary phases for reversed-phase liquid chromatography (RPLC). Characterization of octadecyltrichlorosilane (OTS) layers on thin silica films of ca. 800 Å thickness on 3MPT-modified Ag surfaces has been reported previously. Differences between the ultrathin silica films used in this study and the thin silica films used in this previous study are considered. The results from both FTIR and Raman spectroscopy presented here suggest that bonded DOS alkyl chains are in a disordered, liquid-like state with close to monolayer surface coverage.     

Surface modification of polymers. V. Biomaterial applications

Polyethylene films were surface grafted with glycidyl methacrylate (GMA) by UV irradiating the film for 5 min together with benzophenone. Poly(ethylene glycol) (PEG) was attached to the grafted surface through reaction with the epoxy groups. This yielded a surface which consisted of 95% PEG as measured with ESCA. The adsorption of human transferrin onto this film was significantly reduced as compared with a pure polyethylene film. Heparin was also reacted with a GMA grafted PE surface. ESCA showed that heparin was grafted to the surface, and in vitro blood clotting tests on the heparinized PE surface showed a reduced thrombus formation. GMA grafted polystyrene wells were reacted with carbohydrazide, to the formed carbohydrazide surface a rabbit antibody raised against mouse urinary protein (RaMUP) was covalently coupled. The RaMUP coupled surfaces was used in the detection of mouse urinary protein (MUP) at low concentrations (ca. 1 ng/mL) with an ELISA technique.     

Simple and versatile methods for the fabrication of arrays of live mammalian cells

Single-step methods for the generation of patterned surfaces on hydrogels are presented. Poly(vinyl alcohol) films covalently bonded on glass cover slips and commercially available hydrogel-coated polystyrene plates were used as cell-repellent surfaces. Cell-adhesive domains were created by spotting dilute solutions of sodium hypochlorite onto the surfaces. Alternatively, domains supporting cell attachment were created by exposure to UV light from a xenon excimer lamp, employing a contact mask. Rat skeletal myoblast cells, HEK 293 human embryonic kidney cells and Caco-2 colon carcinoma cells adhered and spread exclusively on modified areas. The surfaces are durable for weeks under cell culture conditions and re-usable after removal of the cells by trypsin treatment. Arrays of adhesive spots seeded with cells at a low density permitted dynamic monitoring of cell proliferation. Selected colonies can be harvested from the surfaces by means of local trypsination. Thus, these techniques may provide useful tools for the isolation of clonal cell populations. Additionally, we demonstrate the possibility of surface-mediated gene delivery from the micro patterns. We show that DNA, complexed with a lipid reagent, can be adsorbed on modified poly(vinyl alcohol) coatings, resulting in spatially controlled adhesion and reverse transfection of HEK 293 cells.     

Graft polymerization of styrene initiated by covalently bonded peroxide groups on silica

The graft polymerization of styrene initiated by immobilized peroxide groups was investigated. Three different types of modification reactions were used to introduce peroxide groups which are directly attached onto the surface of two different silica supports. Silanol groups were chlorinated using thionyl chloride or tetrachlorosilane. In another reaction pathway 1,3,5-benzenetricarbonyl chloride enabled the introduction of free acid chloride residues bonded onto the surface of silica. tert-Butyl hydroperoxide (TBHP) was used to transform the chlorosilyl and the acid chloride groups into peroxide residues. In a further reaction step the covalently bonded peroxides initiated the polymerization of styrene to form grafted polystyrene directly attached onto the silica support. Solid-state 13C CP/MAS NMR spectroscopy, and thermogravimetric and scanning electron microscope measurements enabled a clear structure and property elucidation of the different bonded phases. The highest amount of grafted polystyrene was achieved employing the acid chloride synthesis pathway with silica-gel, whereas modification of spherical silica only led to minor amounts of grafted polymer. The results contribute to the evolving need to understand particle surface modifications and may have positive impact on development of new HPLC stationary phases for improved elutant resolution.     

载入树状分子内部的纳米金修饰玻碳电极的制备及其对亚硝酸根的电化学测定

建立了一种简单、可靠的基于载入树状分子内部的纳米金修饰电极用于亚硝酸根测定的电化学分析方法.将壳聚糖(Chit)修饰在玻碳电极表面,在偶联活化剂碳二亚胺存在的条件下,4.5代羧基末端的树状分子(Dendrimer)通过其外围的羧基与壳聚糖的氨基形成酰胺键而连接在电极表面.Au(Ⅲ)通过与树状分子内部氮的配位作用被结合在...     

Electrode surface modification by a spirobifluorene derivative. An XPS and electrochemical investigation

Ordered thin layers of a spirobifluorene derivative containing an amino group were formed by grafting them onto a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid (11-MUA) on gold. Either physical (H-bonding) or chemical bonding (activated by EDCl) was investigated. X-ray photoelectron spectroscopy and electrochemical impedance spectroscopy confirmed that both methods can be used to effectively graft 2-amino-9,9'-spirobifluorene molecules onto the SAM surface, giving high surface coverages, with a significantly higher packing in the case of chemisorbed films. EIS measurements also showed that the covalently bonded spirobifluorene SAMs act as an effective barrier to both ion penetration and heterogeneous electron transfer.     

Characterization of PET films modified by tetraethylenepentamine (TTEPA)

Tetraethylenepentamine, a long‐chained multifunctional amine, was used to attach nitrogen (N) groups to the surface of a polyethylene terephthalate film. The N content of the modified films was determined by X‐ray photoelectron spectroscopic analysis as well as titration measurements. The physical properties of the modified films were studied using weight‐loss measurements, X‐ray diffraction, and environmental scanning electron microscopy. Contact‐angle measurements were used to establish the changes in wettability of the modified films. The applicability of the surface‐tension‐component theory and the equation‐of‐state approach to this system is briefly discussed. © 2001 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 39: 623–633, 2001     

The pellicular monolith: pore-surface functionalization and surface-phase construction in macroporous polymeric materials

We report synthesis and characterization of a macroporous polymeric material containing a covalently immobilized pore-surface phase of well-defined thickness, gel-phase porosity and organic functional group content. The pore surfaces of otherwise inert macroporous (32 μm mean pore size) ultrahigh-molecular-weight polyethylene (UHMWPE) are aminated throughout using a low-pressure flowing-discharge process to enable covalent immobilization of lightly cross-linked polymer colloid particles on all pore surfaces in the monolith. Solvent swelling and chemical derivitization of the covalently immobilized polymer colloid particles produce a pore-surface gel phase of well-defined thickness, organic amine content, and gel-phase porosity. The low degree of cross-linking in the polymer colloid particles prevents dissolution of the immobilized colloid in good solvents and enables the formation of pore-surface gel phases having high gel porosity on swelling in good solvents. The pore-surface amination introduced by the flowing discharge process varies by less than 17% through 5-mm thickness of the macroporous UHMWPE material. The properties of the pore-surface gel phase also vary by less than 17% through the cross section. The pore-surface immobilized polymer colloid particles swell by a factor of 10 in water and tetrahydrofuran after derivitization with polyethylene glycol. Received: 20 November 1998 Accepted in revised form: 21 January 1999