Surface immobilization of galactose onto aliphatic biodegradable polymers for hepatocyte culture

A novel surface modification method of biodegradable polymers was investigated for inducing the attachment of specific cells onto the polymer surface via ligand-receptor interactions. Galactose, a targeting ligand specific to asialoglycoprotein receptors present on cell membrane of hepatocytes, was introduced on the surface of poly(D,L-lactic-co-glycolic acid) (PLGA) films. A terminal end group of carboxylic acid in PLGA was activated by dicyclohexylcarbodiimide and N-hydroxysuccinimide for the direct conjugation of lactose by reductive amination reaction. Di-block copolymers of PLGA-b-poly(ethylene glycol) (PEG) having a free terminal amine group were also synthesized and used for the conjugation of galactose for the introduction of a PEG spacer between PLGA and galactose. The presence of galactose moieties on the blend film surface was characterized by measuring water contact angle and X-ray photon spectroscopy, and the amount of galactose was indirectly determined by a specific lectin-binding assay. With increasing the galactose concentration on the blend film surface, the initial attachment as well as the cell viability of hepatocyates concomitantly increased. The introduction of PEG spacer reduced the cell attachment and viability. Albumin secretion rate from hepatocytes was enhanced for galactose modified surfaces, whereas it was reduced for the surfaces not having galactose moieties.     

Surface esterification of corn starch films: Reaction with dodecenyl succinic anhydride

Surface of corn starch films was modified through esterification using dodecenyl succinic anhydride (DDSA) as reactant. The effects of reaction temperature and time, concentration of alkaline aqueous solution for activating starch, concentration of ethanol diluted DDSA, and time of alkaline aqueous solution treatment on the physical properties related to material’s hydrophilic nature, such as moisture absorption and surface water contact angle, were investigated. It was found that the surface esterification modification significantly reduced the moisture sensitivity and surface hydrophilic character of starch film. Due to the hydroxyl groups in the film surface layer were reacted with DDSA, the equilibrium moisture content of the starch film under 95% RH declined 22% and the surface water contact angle increased up to 82% after surface esterification modification.     

转化人胚肌腱细胞力学特性研究

探讨转化人胚腱细胞９Ｔｒａｎｓｆｏｒｍｅｄ ｈｕｍａｎ ｅｍｂｒｙｏｎｉｃ ｔｅｎｄｏｎ ｅｅｌｌｓ,ＴＨＥＴＣｓ）力学特性,以阐明在应变条件下进行细胞－支架材料复合培养时腱铁力学相容性。取人腱细胞（Ｈｕｍａｎ ｅｍｂｒｙｏｎｉｃ ｔｅｎｄｏｎ ｃｅｌｌｓ,ＨＥＴＣｓ）和ＴＨＥＴＣｓ,采用微吸管吸吮实验技术,测量单个细胞的粘生及其对聚乳酸（Ｐｏｌｙｌａｃｔｉｃ ａｃｉｄ,ＰＬＡ）和聚乳酸羟基乙酸共     

PHBHHx膜表面亲水改性及其与神经干细胞生物相容性

对羟基丁酸-羟基己酸共聚酯(PHBHHx)膜进行表面改性,研究神经干细胞(NSCs)在改性后的PHBHHx膜表面的贴附、增殖及分化情况,为开发新型脑组织工程支架材料奠定基础。采用溶剂挥发法制备PHBHHx膜,扫描电镜观察其表面性状;分别通过脂肪酶处理,NaOH处理的方法对PHBHHx膜进行表面改性,测量接触角以检测膜表面亲水性。分离培养孕14.5 d大鼠胚胎大脑皮质NSCs,接种在表面改性后的PHBHHx膜表面进行体外培养,扫描电镜观察膜表面细胞形态,MTT法检测细胞活力,免疫细胞化学染色观察NSCs存活和分化情况。结果显示,与未处理的PHBHHx膜相比,脂肪酶、NaOH处理能够显著提高PHBHHx膜表面亲水性,增加NSCs在PHBHHx膜表面贴附数量;NSCs在改性后的PHBHHx膜表面能够良好地存活并分化为神经元和胶质细胞。结果提示PHBHHx膜表面碱处理通过提高材料表面亲水性和粗糙程度,增加其与NSCs的生物相容性,改性后的PHBHHx材料是一种非常有潜力的新型脑组织工程支架材料,有望在NSCs移植修复脑损伤中发挥作用。     

氨基修饰的纳米纤维对人和大鼠 MSCs增殖分化的影响

目的:探讨氨基修饰后的静电纺丝纳米纤维对大鼠和人骨髓来源的间充质干细胞(Rat and human bone marrow mesenchymal stem cells, r MSCs and hMSCs)增殖及成骨分化的影响。方法:采用静电纺丝法制备聚乳酸-羟基乙酸共聚物(poly(lactic-co-glycolic acid),PLGA)纳米纤维,用氨气等离子体处理其表面来接枝氨基;通过测量PLGA纳米纤维(NF)及氨基修饰后的纳米纤维(NF-NH_2)接触角来证明修饰效果;将r MSCs和hMSCs分别接种于NF和NF-NH_2,用CCK-8试剂盒检测接种后1, 3 (4), 7天的细胞增殖;接种后的21天,用茜素红S染色(ARS)法检测细胞成骨分化情况。结果:氨气等离子体处理后纳米纤维接触角从81.28±0.33降低至53.99±0.79,说明氨基修饰后的PLGA NF亲水性增加;CCK-8结果显示氨基修饰增加了r MSCs的黏附,接种24 h后r MSCs在NF和NF-NH_2上的检测吸光值分别为0.096±0.011和0.175±0.014(P0.001),而对hMSCs黏附和增殖没有影响,接种24 h后hMSCs在NF和NF-NH_2上的检测吸光值分别为0.237±0.004和0.238±0.006(P0.05);ARS染色结果显示氨基修饰后r MSCs成骨分化增多(在NF和NF-NH_2表面ARS染色区域比例分别13.147±3.223%和36.677±5.230%),而hMSCs在修饰前后的纳米纤维上均有表达(修饰前后ARS染色比例分别为50.283±2.942%和38.254±3.272%)。结论:氨基修饰的NF可以促进大鼠来源的MSCs黏附增殖以及成骨分化,而对人骨髓来源的MSCs没有显著影响,这提示我们MSCs的增殖分化行为可能具有种属依赖性。     

Surface modification of chitosan films. Effects of hydrophobicity on protein adsorption

The surface of chitosan films was modified using acid chloride and acid anhydrides. Chemical composition at the film surface was analyzed by attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). ATR-FTIR data verified that the substitution took place at the amino groups of chitosan, thus forming amide linkages, and the modification proceeded to the depth at least 1 microm. Choices of molecules substituted at the amino groups of the glucosamine units did affect the hydrophobicity of the film surface, as indicated by air-water contact angle analysis. The surface became more hydrophobic than that of non-modified film when a stearoyl group (C(17)H(35)CO-) was attached to the films. The reaction of chitosan films with succinic anhydride or phthalic anhydride, however, produced more hydrophilic films. Selected modified films were subjected to protein adsorption study. The amount of protein adsorbed, determined by bicinchoninic acid (BCA) assay, related to the types of attached molecules. The improved surface hydrophobicity affected by the stearoyl groups promoted protein adsorption. In contrast, selective adsorption behavior was observed in the case of the chitosan films modified with anhydride derivatives. Lysozyme adsorption was enhanced by H-bonding and charge attraction with the hydrophilic surface. While the amount of albumin adsorbed was decreased possibly due to negative charges that gave rise to repulsion between the modified surface and albumin. This study has demonstrated that it is conceivable to fine-tune surface properties which influence its response to bio-macromolecules by heterogeneous chemical modification.     

Controllable surface modification of poly(lactic-co-glycolic acid) (PLGA) by hydrolysis or aminolysis I: physical, chemical, and theoretical aspects

While biodegradable, biocompatible polyesters such as poly (lactic-co-glycolic acid) (PLGA) are popular materials for the manufacture of tissue engineering scaffolds, their surface properties are not particularly suitable for directed tissue growth. Although a number of approaches to chemically modify the PLGA surface have been reported, their applicability to soft tissue scaffolds, which combine large volumes, complex shapes, and extremely fine structures, is questionable. In this paper, we describe two wet-chemical methods, base hydrolysis and aminolysis, to introduce useful levels of carboxylic acid or primary and secondary amine groups, respectively, onto the surface of PLGA with minimal degradation. The effects of temperature, concentration, pH, and solvent type on the kinetics of these reactions are studied by following changes in the wettability of the PLGA using contact angle measurements. In addition, the treated surfaces are studied using X-ray photoelectron spectroscopy (XPS) to determine the effect on the surface chemical structure. Furthermore, we show using XPS analysis that these carboxyl and amine groups are readily activated to allow the covalent attachment of biological macromolecules.     

Platelet adhesion to polyurethane blended with polytetramethylene oxide

The surface and blood compatibility characteristics of Pellethane polyurethane blended with 1% or 5% (w/w) polytetramethylene oxide (PTMO) were evaluated. Analysis by X-ray photoelectron spectroscopy indicated that blending of PTMO caused an increased amount of amide wax, a processing agent present in Pellethane, to be expressed on the surface of the blended films in vacuo. Dynamic contact angle measurements in water, however, showed that PTMO was preferentially expressed on the blend film surfaces in water. The two lower molecular weight species, PTMO and amide wax, were thus capable of reorienting, depending on the environmental conditions. An in vitro assay of platelet adherence and thrombosis showed that polyurethane blended with 5% PTMO had about two-thirds fewer adherent platelets compared to unblended polyurethane and that a blend containing 1% PTMO was intermediate in platelet adherence. Measurements of albumin adsorption from binary solution with fibrinogen indicated that PTMO blends did not preferentially adsorb albumin compared to unblended polyurethane. (c) 1996 John Wiley & Sons, Inc.     

Biomimetic Hydrophobic Surfaces with Low or High Adhesion Based on Poly(vinyl alcohol) and SiO2 Nanoparticles

Superhydrophobic surfaces are often found in nature,such as plant leaves and insect wings.Inspired by superhydrophobic phenomenon of the rose petals and the lotus leaves,biomimetic hydrophobic surfaces with high or low adhesion were prepared with a facile drop-coating approach in this paper.Poly(vinyl alcohol) (PVA) was used as adhesive and SiO2 nanoparticles were used to fabricate surface micro-structure.Stearic acid or dodecafluoroheptyl-propyl-trimethoxysilane (DFTMS) were used as low surface energy materials to modify the prepared PVA/SiO2 coating surfaces.The effects of size of SiO2 nanoparticles,concentration of SiO2 nanoparticle suspensions and the modifications on the wettability of the surface were investigated.The morphology of the PVA/SiO2 coating surfaces was observed by using scanning electron microscope.Water contact angle of the obtained superhydrophilic surface could reach to 3°.Stearic acid modified PVA/SiO2 coating surfaces showed hydrophobicity with high adhesion.By mixing the SiO2 nanoparticles with sizes of 40 nm and 200 nm and modifying with DFTMS,water contact angle of the obtained coating surface could be up to 155° and slide angle was only 5°.This work provides a facile and useful method to control surface wettability through changing the roughness and chemical composition of a surface.     

Surface modification of polyhydroxyalkanoate films and their interaction with human fibroblasts

A poly(3-hydroxybutylate-co-hydroxyvalerate) (PHA) film containing 34 mol.% 3-hydroxyvalerate (Biopol D600P) was prepared by the solvent cast method using a 10 wt.% chloroform solution of PHA. The PHA film was exposed to an oxygen plasma glow discharge to produce peroxides on its surfaces. These peroxides were then used as catalysts for the polymerization of acrylic acid (AA) in order to prepare carboxyl group-introduced PHA (PHA-C). Insulin-immobilized PHA was prepared using the coupling reaction of PU-C with insulin. The surface-modified PHAs were then characterized by attenuated total reflection Fourier transform infrared spectroscopy, electron spectroscopy for chemical analysis, and a contact angle goniometer. The amounts of insulin directly coupled to the carboxyl groups on PHA-C and coupled to the terminus amino groups of the grafted polyethylene oxide were 2.9 and 0.8 microg cm(-2), respectively. The PHA water contact angle (75 degrees ) decreased with AA grafting (33 degrees ) and insulin immobilization (31 degrees ), thereby exhibiting the increased hydrophilicity of the modified PHAs. When compared with PHA and PHA-C, the proliferation of human fibroblasts in the presence of serum was significantly accelerated on the insulin-immobilized PHAs.     

Bioactive thin film of acidic fibroblast growth factor fabricated by layer-by-layer assembly

A new class of bioactive thin films using growth factors as building blocks has been fabricated via layer-by-layer assembly (LBL) technique. Acid fibroblast growth factor (aFGF) in the presence of heparin was used as negatively charged polyelectrolytes, while poly(ethyleneimine) (PEI) was chosen as a positively charged counterpart. The self-deposition process and surface morphology of the resultant multilayers were monitored and detected by UV-vis absorbance spectra, advanced contact angle measurements, and scanning force microscopy (SFM) observations. Cell culture was performed to assess the efficiency of the growth factors. The fibroblasts proliferated faster on the surface assembled with five bilayers of (aFGF/heparin)/PEI with apparent higher cytoviability than on those surfaces modified by one bilayer of (aFGF/heparin)/PEI, five bilayers of aFGF/PEI, or five bilayers of heparin/PEI, and tissue culture polystyrene. Enhanced secretion of collagen type I and interleukin 6 (IL-6) by the fibroblasts seeded on the five bilayers of (aFGF/heparin)/PEI was also verified by immunohistochemical examination. The bioactivity of the (aFGF/heparin)/PEI multilayers could be largely preserved when stored at -20 degrees C.     

The effect of titanium with heparin/BMP-2 complex for improving osteoblast activity

The objective of this study was to investigate the enhanced osteoblast activity of MG-63 cells cultured on titanium (Ti) with a heparin/BMP-2 (Hep/BMP-2) complex. The Ti substrates were initially modified by chemical grafting poly-l-lysine (PLL) using condensing agent, followed by immobilizing the heparin/BMP-2 complex to the PLL-grafted Ti substrate via electrostatic interactions. The surface modification of Ti substrates with PLL and/or Hep/BMP-2 complex were confirmed with scanning electron microscopy, contact angle measurements, and X-ray photoelectron spectroscopy. Immobilized BMP-2 was released from the Hep/BMP-2/Ti substrate in a sustained manner. In vitro studies revealed that osteoblasts grown on Hep/BMP-2/Ti substrate increased ALP activity, calcium deposition, ALP and osteocalcin levels as compared to those grown on pristine Ti or PLL-Ti. These results indicated that heparin/BMP-2 complex immobilized Ti substrate can be useful to effectively improve osteoblast activity.     

Interrelationship between water film thicknesses and contact angles and a model for CO2 adhesion

Thermodynamics of contact angle phenomena is strongly affected by the presence of thin liquid films. However, at present, studies for CO₂/brine/mineral systems only consider the films apart from contact angles. In this paper, molecular dynamics (MD) simulations have been performed to simultaneously investigate the interrelationship between water film thicknesses and water contact angles. Two types of contact angles were considered namely Young’s contact angle (no water film is present) and contact angle with film (a stable film is present). The results showed that as Young’s contact angle increased, film thickness decreased which leading to increasing of contact angle with film. The effects of CO₂-mineral pre-contact have also been investigated and it has been found that on mediate hydrophilic surfaces (Q³), water films were present when CO₂ droplets were placed above the surfaces, however, water films were absent when CO₂ droplets directly contact with the surfaces. This phenomenon implies that water films on mineral surfaces have a possibility to rupture and a film rupture mechanism for CO₂ adhesion on hydrated mineral surfaces was proposed. These results may provide new information on interactions among CO₂, water/brine and mineral to better understand the behaviour of CO₂ during geologic sequestration.     

Amorphous characteristics of an ultrathin cellulose film

Swelling behavior and rearrangements of an amorphous ultrathin cellulose film (20 nm thickness) exposed to water and subsequently dried were investigated with grazing incidence X-ray diffraction, neutron reflectivity, atomic force microscopy, and surface energy calculations obtained from contact angle measurements. The film swelled excessively in water, doubling its thickness, but shrunk back to the original thickness upon water removal. Crystallinity (or amorphousness) and morphology remained relatively unchanged after the wetting/drying cycle, but surface free energy increased considerably (ca. 15%) due to an increase in its polar component, that is, the hydrophilicity of the film, indicating that rearrangements occurred during the film's exposure to water. Furthermore, stability of the films in aqueous NaOH solution was investigated with quartz crystal microbalance with dissipation monitoring. The films were stable at 0.0001 M NaOH but already 0.001 M NaOH partially dissolved the film. The surprising susceptibility to dissolve in dilute NaOH was hypothetically attributed to the lack of hierarchical morphology in the amorphous film.     

Preparation and characterization of monolayer and multilayer Langmuir-Blodgett films of a series of 6-O-alkylcelluloses

Monolayer and multilayer Langmuir-Blodgett (LB) films of 6-O-alkylcelluloses with various chain lengths were prepared and studied. The surface pressure (pi)-area (A) isotherms of 6-O-alkylcelluloses exhibited characteristic behaviors depending on the length of the alkyl chain and temperature. 6-O-Stearylcellulose on the subphase formed a homogeneous monolayer at 10 mN m(-1). By transfer ratio, FT-IR, and contact angle measurements, it was proved that the monolayer of 6-O-stearylcellulose on the water surface was transferred successfully onto a substrate by a vertical dipping method to form a Z-type LB film. The transmission and reflection absorption IR spectrum indicated that the hydrocarbon chains had all-trans rotamers and were oriented nearly perpendicular to the surface in the film. AFM section analysis revealed that the thickness per layer was calculated to be 2.35 nm. These results suggested that the hydrocarbon chains were inclined at an angle of about 25.3 degrees to have high packing density in the alkyl region.     

A novel strategy to obtain a hyaluronan monolayer on solid substrates

The aim of this study was to find a novel simple method to obtain polysaccharide ultrathin layers on solid substrates to investigate the interaction between the surface and the biological environment. A Hyaluronan (Hyal) monolayer with a well-defined chemistry was obtained by exploiting the capability of organosilanes to spontaneously adhere onto glass surfaces. A silane alkylic chain was conjugated with Hyal, and the derivatized polysaccharide was allowed to spontaneously adhere onto a glass surface. The elemental analysis of the modified polysaccharide demonstrated that one out of five disaccharide units was conjugated with the alkyl silane chain, corresponding to a substitution degree of the carboxylate groups of approximately 20%. The film of the modified polysaccharide was characterized by means of X-ray photoelectron spectroscopy (XPS), water contact angle, and atomic force microscopy (AFM) measurements. XPS analysis demonstrated that we obtained a Hyal layer with a thickness of about 2.0 nm corresponding to a Hyal monolayer. The Hyal-coated surfaces appeared to be rather smooth and highly hydrophilic and showed significant resistance to nonspecific cell adhesion.     

Studies of chitosan: II. Preparation and characterization of chitosan/poly(vinyl alcohol)/gelatin ternary blend films

Chitosan/poly(vinyl alcohol)/gelatin (CS/PVA/GA) ternary blend films were prepared by solution blending method in this study. The thermal properties of the CS/PVA/GA ternary blend films were examined by differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The melting point of the CS/PVA/GA ternary blend film was increased when the amount of GA in the blend film was increased based upon the DSC thermal analysis. Results of X-ray diffraction (XRD) analyses indicated that the intensity of diffraction peak at 19 degrees of PVA became lower and broader with increasing the amount of GA in the CS/PVA/GA ternary blend film. Although CS, PVA, and GA are hydrophilic biodegradable polymers, the results of water contact angle measurements are still as high as 83 degrees, 68 degrees, and 66 degrees, respectively. A minimum water contact angle (56 degrees) was observed when the ternary blend film contains 50 wt.% GA (i.e. GA5). This behavior is primarily due to the reorientation of polar functional groups toward to the top surface of CS/PVA/GA ternary blend films.     

Cellulose microfibrils grafted with PBA via surface-initiated atom transfer radical polymerization for biocomposite reinforcement

Immobilizing poly(butyl acrylate) (PBA) on cellulose microfibrils (CMFs) by atom transfer radical polymerization (ATRP) of butyl acrylate (BA) on the surface of 2-bromoisobutyryl-functionalized CMF generated highly hydrophobic microfibrils (CMF-PBA) with a hard core and a soft-shell structure. TGA and static water contact angle results suggested that the surfaces of the modified CMF samples were not completely covered by PBA chains until the molecular weight of grafts became sufficiently long. The GPC results indicated that the grafts with low molecular weight showed controlled/"living" characteristics of the surface-initiated ATRP; however, there existed more side reactions with the increase in molecular weights. Biocomposites consisting of polypropylene (PP) and CMF-PBA samples exhibited significantly improved compatibility, interface adhesion, and mechanical properties with the increase in PBA graft length. The findings confirmed that the longer grafts facilitated the better entanglement of PBA grafts with PP macromolecules and thus further improved the mechanical properties.     

Superhydrophobicity of Bionic Alumina Surfaces Fabricated by Hard Anodizing

Bionic alumina samples were fabricated on convex dome type aluminum alloy substrate using hard anodizing technique.The convex domes on the bionic sample were fabricated by compression molding under a compressive stress of 92.5 MPa.The water contact angles of the as-anodized bionic samples were measured using a contact angle meter (JC2000A) with the 3 μL water drop at room temperature.The measurement of the wetting property showed that the water contact angle of the unmodified as-anodized bionic alumina samples increases from 90° to 137° with the anodizing time.The increase in water contract angle with anodizing time arises from the gradual formation of hierarchical structure or composite structure.The structure is composed of the micro-scaled alumina columns and pores.The height of columns and the depth of pores depend on the anodizing time.The water contact angle increases significantly from 96° to 152° when the samples were modified with self-assembled monolayer of octadecanethiol (ODT),showing a change in the wettability from hydrophobicity to super-hydrophobicity.This improvement in the wetting property is attributed to the decrease in the surface energy caused by the chemical modification.