No abstract

No abstract

Diblock copolymer grafts covalently attached to surfaces have attracted considerable attention because of their special structure and novel properties. In this work, poly(N-isopropylacrylamide)-block-polystyrene (PNIPAAm-b-PS) brushes were prepared via surface-initiated consecutive atom-transfer radical polymerization on initiator-immobilized silicon. Because of the inherent thermosensitivity of PNIPAAm and the hydrophobicity difference between the two blocks, the modified surfaces were responsive to both temperature and solvent. Moreover, the diblock copolymer brushes exhibited both resistance to nonspecific protein adsorption and unique cell interaction properties. They showed strong protein resistance in both phosphate-buffered saline and blood plasma. In particular, fibrinogen adsorption from plasma at either room temperature or body temperature was less than 8 ng/cm(2), suggesting that the surfaces might possess good blood compatibility. In addition, the adhesion and detachment of L929 cells could be "tuned", and the ability to control the detachment of cells thermally was restored by block polymerization of hydrophobic, cell-adhesive PS onto a thicker PNIPAAm layer. In addition to providing a simple and effective design for advanced cell-culture surfaces, these results suggest new biomedical applications for PNIPAAm.

No abstract

Well-controlled poly(N-vinylpyrrolidone) (PVP)-grafted silicon surfaces were prepared by surface-initiated atom transfer radical polymerization (SI-ATRP) with 1,4-dioxane/water mixtures as solvents and CuCl/5,7,7,12,14,14-hexamethyl-1,4,8,11-tetraazacyclotetradecane (Me6TATD) as a catalyst. The thickness of the PVP layer on the surface increased with reaction time, suggesting that the ATRP grafting of N-vinylpyrrolidone (NVP) from the silicon surfaces was a well-controlled process. The water contact angle and X-ray photoelectron spectroscopy (XPS) were used to characterize the modified surfaces. The protein adsorption property of the PVP-grafted surfaces was evaluated using a radiolabeling method. Compared with unmodified silicon surfaces, a Si-PVP60 surface with a PVP thickness of 15.06 nm reduced the level of adsorption of fibrinogen, human serum albumin (HSA), and lysozyme by 75, 93, and 81%, respectively. Moreover, the level of fibrinogen adsorption decreases gradually with an increase in PVP thickness. However, no significant difference in fibrinogen adsorption was found when the PVP layer was thicker than the critical thickness of 13.45 nm.

A weak polyelectrolyte coating, carboxylated poly(oligo(ethylene glycol)methacrylate-co-2-hydroxyethylmethacrylate), was prepared via surface initiated polymerization (SIP) from initiators immobilized to gold surface through the Au-S bonds. When dry thickness increased up to 75 nm, this polyelectrolyte coating was pulled off the Au substrate by simply exposing to phosphate buffer saline (PBS, pH = 7.4, [Na(+)] = 150 mM). This covalent bond breaking (CBB) behavior was monitored in situ using a quartz crystal microbalance (QCM) and CBB was associated with the swelling of the anchored polyelectrolyte chains.

Bioactive films were produced by conjugation of a short peptide onto modified cellulose nanofibrils (CNF). Specifically, a hydrophilic copolymer, poly(2-aminoethyl methacrylate hydrochloride-co-2-hydroxyethylmethacrylate) (poly(AMA-co-HEMA)), was grafted via surface initiated polymerization from an initiator coupled to CNF. The poly(AMA-co-HEMA) was used as a spacer and support layer for immobilization of the peptide, acetylated-HWRGWVA, which has specific affinity with human immunoglobulin G (hIgG). Two methods for peptide grafting were compared: modification of CNF in aqueous suspension followed by assembly into a bioactive film and peptide grafting on a preformed CNF film. The CNF-based networks were examined on solid supports via atomic force microscopy (AFM) and extreme resolution imaging with ultralow electron landing energies (scanning low energy electron microscopy). The specific binding capability of hIgG and nonspecific protein resistance of the resultant peptide-modified CNF were evaluated by using quartz crystal microgravimetry (QCM). The effects of initiator concentration and thickness of poly(AMA-co-HEMA) layer on hIgG adsorption were investigated in the developed systems, which exhibited high signal-to-noise response.

Although polymeric membranes are widely used in the purification of protein pharmaceuticals, interactions between biomolecules and membrane surfaces can lead to reduced membrane performance and damage to the product. In this study, single-molecule fluorescence microscopy provided direct observation of bovine serum albumin (BSA) and human monoclonal antibody (IgG) dynamics at the interface between aqueous buffer and polymeric membrane materials including regenerated cellulose and unmodified poly(ether sulfone) (PES) blended with either polyvinylpyrrolidone (PVP), polyvinyl acetate-co-polyvinylpyrrolidone (PVAc-PVP), or polyethylene glycol methacrylate (PEGM) before casting. These polymer surfaces were compared with model surfaces composed of hydrophilic bare fused silica and hydrophobic trimethylsilane-coated fused silica. At extremely dilute protein concentrations (10(-3)-10(-7) mg/mL), protein surface exchange was highly dynamic with protein monomers desorbing from the surface within ∼1 s after adsorption. Protein oligomers (e.g., nonspecific dimers, trimers, or larger aggregates), although less common, remained on the surface for 5 times longer than monomers. Using newly developed super-resolution methods, we could localize adsorption sites with ∼50 nm resolution and quantify the spatial heterogeneity of the various surfaces. On a small anomalous subset of the adsorption sites, proteins adsorbed preferentially and tended to reside for significantly longer times (i.e., on "strong" sites). Proteins resided for shorter times overall on surfaces that were more homogeneous and exhibited fewer strong sites (e.g., PVAc-PVP/PES). We propose that strong surface sites may nucleate protein aggregation, initiated preferentially by protein oligomers, and accelerate ultrafiltration membrane fouling. At high protein concentrations (0.3-1.0 mg/mL), fewer strong adsorption sites were observed, and surface residence times were reduced. This suggests that at high concentrations, adsorbed proteins block strong sites from further protein adsorption. Importantly, this demonstrates that strong binding sites can be modified by changing solution conditions. Membrane surfaces are intrinsically heterogeneous; by employing single-molecule techniques, we have provided a new framework for understanding protein interactions with such surfaces.