Papers by Keyword: Protein Adsorption

Abstract: This paper introduces a new method of surface modification by self-assembled monolayer (SAM) and polymer monolayer grafting. Since most of the glass surfaces lack the reactive functional group, an activation process with 3-(trimethoxysilyl)propyl methacrylate(TPM) is used in our experiment to generate the vinyl reactive sites on the substrate surface for further graft polymerization. The TPM saline layer acts as the “anchor” part to link the functional part onto the surface of substrate. The paper summarizes the surface modifications by the polymerizations of PEGMA, AA(Acrylic acid) and NVP(Nitrogen-vinyl-2-pyrrolidone) respectively and their applications for protein adsorption and cell adhesion through a series of measurements. In previous research, AA and NVP had also been adopted for surface treatment and had achieved good results. The substrate can be glass, alumina, silicon, metals or stainless steel. We choose glass as our substrate during the experiment.

Abstract: Real time adsorption behaviors of six proteins with different isoelectric points on hydroxyapatite (HAp) nanocrystal surfaces have been investigated by using HAp sensors for quartz crystal microbalance with dissipation technique (QCM-D). The dissipation (D)–frequency (f) plots clearly showed that the different types of protein adsorption behaviors; the D-f plots of acidic proteins lie on one straight line with a constant slope under all initial protein concentrations, while those of neutral and basic proteins lie on two straight lines with different slopes. The acidic proteins formed a monolayer, while the neutral and basic proteins could cause conformational changes with the adsorbed amount of proteins. The QCM-D technique with novel HAp nanocrystal sensor is useful for the liquid phase changes of proteins on the surface.

Abstract: The protein adsorption behavior was investigated for highly (001) oriented hydroxypatatite coatings (HACs). Highly (001) oriented (HO-) HACs and HAC with low orientation (LO-HAC) were prepared on titanium (Ti) substrates through a radio-frequency thermal plasma spraying method. Sintered HA pellets (S-HA) was also prepared as a control. The solution of 22 'g/100 'l PBS of the fluorescein-isothiocyanate (FITC) labeled bovine serum albumin (FITC-BSA), immunoglobulin G (FITC-IgG) and cytochrom c (FITC-CCC) was separately dropped on the surface of HACs and was incubated for 30 min. After the incubation, such HACs were washed with the PBS additionally supplemented with 125mM NaCl and observed using a fluorescence microscope. Fluorescence microscopic examination indicates that FITC labeled proteins somewhat adsorbed on the HACs, while proteins adsorbed little on S-HA surface. In particular, it can be seen that FITC-CCC adsorbed more prominently on the HO-HAC.

Abstract: The adsorption properties of metal ions containing hydroxyapatite (Ca10(PO4)6(OH)2, HAp) were elucidated for the development of protein drug carrier. As-prepared metal ion containing HAp nanocrystal showed plate-like morphology with 10-20nm in length and 5-10nm in width. The metal ion containing HAp microparticles had higher specific surface area than the HAp microparticle. The adsorption amount per unit area of HAp showed higher than those of metal ion containing HAp. The adsorption behavior followed the Langmuir curves for each protein, indicating the monolayer adsorption. The loaded amount of proteins could be one of the most important properties for the application of drug delivery carrier.

Abstract: Micrometer-sized hydroxyapatite (HA) with rod-shape was prepared to investigate protein adsorption onto its crystal face, through hydrothermal processing. Adsorption of Bovine serum albumin (BSA) and Lysozyme from chicken egg white (LSZ) on rod-shaped HA crystals with approximately 50 μm in length and 1 μm thickness was examined in phosphate buffer solution. The amount of adsorbed BSA was decreased with increasing phosphate concentration in the examined solution. Adsorption of LSZ was increased with increasing phosphate concentration in the buffer solution up to 50 mmol/L (mM), while it was remarkably decreased when the phosphate concentration reached approximately 100 mM. These phenomena lead to understanding of chromatographic characters of HA for separation of acidic and basic proteins.

Abstract: Nano-crystalline Mg-containing hydroxyapatite (Mg·HAp) were prepared by a wet chemical method, for which selective adsorption of proteins was examined, taking bovine serum albumin (BSA) and a pathogenic protein β2-microglobulin (β2-MG) as the model proteins. Increase in the Mg content led to smaller crystallites and larger specific surface area (SA) of Mg·HAps as well as zeta potential, while the amount both of BSA and β2-MG adsorption on Mg·HAp particles. It is thus concluded that the adsorption of BSA and β2-MG on Mg•HAp was associated with surface charges.

Abstract: The aim of this study was to set up a SPR sensor and to investigate protein adsorption onto biomaterial surface using it. Methods: The adsorption of human albumin (Alb), fibrinogen (Fib) and immunoglobulin G (IgG) onto polyurethane (PU) H50-50 and Au surface were measured using this SPR sensor. Results: The results of protein adsorption showed that the amount of Alb, Fib and IgG adsorbed on PU (H50-50) are smaller than Au, and on these two material surfaces, the amount sequence of adsorbed protein is: IgG> Fib > Alb. Both the Alb/Fib ratio and Alb/IgG ratio on PU (H50-50) surface are higher than Au. Conclusion: The antithrombogenicity of PU (H50-50) is better than Au.

Abstract: The effects of various processing parameters of electro spinning such as concentration, applied voltage, distance from needle to collector on the morphology(especially fiber diameter) of nanofiber matrix was investigated in this study. We found out the concentration of polymer solution was the key parameter to control the fiber diameter. Such a difference on surface morphology of electro spun nanofiber matrix under various processing parameters will show the different behaviors on protein adsorption on the surface contacting with body fluid. Consequently, this results different cell motions on the matrix used on tissue engineering. To evaluate the adsorption of proteins on the surface of sheet type nanofiber matrix, matrix obtained by electro spinning were immersed in FITC labeled proteins solutions. And then, we confirmed adsorption of proteins using laser scanning confocal microscopy (LSCM). The quantitative analysis of adsorbed proteins was also investigated by UV spectroscopy.

Abstract: The contact of a cell on the biomaterial’s surface is mediated by its adhesion components. The topography of titanium surfaces influences these adhesion components of osteoblasts, e.g. the integrins, the adapter proteins and the actin cytoskeleton. In our current experiments we were interested in why osteoblasts were strongly aligned to the grooves of a structured pure titanium surface (grade 2). The titanium was characterized by EIS to get insights in the electro-chemically active surface. We used MG-63 human bone cells, cultured in DMEM with 10% FCS at 37°C. For protein adsorption the titanium discs were incubated for 24h with complete medium containing soluble fibronectin at 37°C. Interestingly, only in the grooves cells adhered and were aligned and this is not dependent on the gravitation. The cell adhesion seems to depend on the protein adsorption of fibronectin which we could find to be adsorbed exclusively in the valleys. We speculate that there are local differences in electro-chemical characteristics of this structured titanium surface.

Abstract: The aim of present work was to study the interaction between human plasma protein-albumin (Alb) and immunoglobulin G (IgG) and the surfaces of two kinds of diamond-like carbon (DLC-A and DLC-B) and titanium (Ti) film. Fourier transform infrared spectroscopy (FTIR) was used to perform both quantity investigation and secondary structure analysis of above two proteins adsorbed on material surfaces. A modified Coomassie brilliant blue (CBB) protein assay was also used to study the amount of adsorbed proteins. The result of FTIR quantitative evaluation shows that the ratio of adsorbed Alb to IgG (RA/I) on three kinds of material surface has an order: DLC-A > DLC-B > Ti, which is coincide with the result from CBB protein assay. The result of secondary structure analysis shows that the conformation of Alb and IgG changes in a largest degree after adsorbed on Ti and a smallest degree on DLC-A surface. Both the results indicate that the anti-thrombogenicity of DLC-A seems to be the best and Ti is the worst.