Synthesis of photoreactive poly(ethylene glycol) and its application to the prevention of surface‐induced platelet activation

Phosphorylcholine groups attached to polymer surfaces are known to improve hemocompatibility. A photochemical method is presented to couple phosphorylcholine-containing aryl azides to poly(etherurethane) surfaces (PEUs). Two aryl azides that consist of a photoactivatable 4-azidobenzoyl group, a short spacer chain, and a phosphorylcholine endgroup were synthesized. The two compounds differ only in the type of spacer used: triethylene glycol for compound 1 and hexanediol for compound 2. These compounds were physically adsorbed to PEU surfaces. Upon UV irradiation, reactive intermediates are formed that react with nucleophilic groups on the polymer surface. The modified surfaces showed decreased underwater contact angles, indicating that hydrophilic phosphorylcholine groups are present at the surface. ESCA measurements showed the presence of phosphorus and positively charged nitrogen atoms in the outermost polymer layers (analyzed depth about 50 A), which is a strong indication of the presence of phosphorylcholine groups. Hemocompatibility in vitro was tested with thrombin generation assays and platelet adhesion tests. In thrombin generation assays the clotting time of platelet-rich plasma in contact with the polymer surface is determined. Clotting times were clearly prolonged for the modified surfaces. Surfaces modified with compound 2 showed slightly higher clotting times than those modified with compound 1. Repeated surface modification with compound 2 further increased the clotting time. For the tested surfaces an increase in the clotting time corresponds to an increase in the concentration of phosphorylcholine groups at the surface (as measured by ESCA and contact angle). Platelet adhesion studies with scanning electron microscopy demonstrated that fewer platelets (showing less activation) adhered to the modified surfaces than to the unmodified polyurethane.

“…(ethylene glycol) 27 and heparin. 28 polyethylene 5.7 min, polypropylene 7.3 min, polyureAryl azides containing phosphorylcholine endthane 8.0 min, and polyvinylchloride-T 9.9 min.…”

Section: Surface Analysisunclassified

A photochemical method for the surface modification of poly(etherurethanes) with phosphorylcholine-containing compounds to improve hemocompatibility

Heiden

Goebbels

Pijpers

et al. 1997

“…[14][15][16][17] Numerous investigators have found that foreign surfaces rapidly adsorb plasma proteins when exposed to blood, 18 so it is essential to have an understanding of protein-polymer interaction for the development of antithrombogenic material. Hydrogel grafted surfaces by ␥-irradiation method and the protein interaction and morphology of platelets onto the surfaces have been studied by Sharma et al 19,20 Thermoplastic elastomers are triblock copolymers, the central portion being elastomeric with short, glassy blocks on either side.…”

Section: Introductionmentioning

confidence: 99%

Preparation and characterization of heparin-containing SBS-g-DMAEMA copolymer membrane

Yang¹,

Jong²,

Hsu³

et al. 1998

The grafting of dimethyl amino ethyl methacrylate (DMAEMA) onto styrene-butadiene-styrene triblock copolymer (SBS) membrane was subsequently conducted by UV-radiation induced graft copolymerization without degassing to obtain the SBS-g-DMAEMA copolymer membrane. The substituted amino groups on the SBS-g-DMAEMA graft copolymer membrane were quaternized with iodomethane, and then the membrane was treated with heparin to prepare the heparin-containing SBS-g-DMAEMA copolymer membrane (SBS-g-DMAEMA-HEP). The graft copolymer membrane (SBS-g-DMAEMA) and the heparin-containing SBS-g-DMAEMA copolymer membrane (SBS-g-DMAEMA-HEP) were characterized by FTIR spectroscopy. The heparin content was determined by toluidine blue heparin assay. Contact angle, water content, and protein adsorption of fibrinogen and albumin experiments were also performed to evaluate the effect of graft amount and heparin content on the biocompatibility of SBS-g-DMAEMA and SBS-g-DMAEMA-HEP graft copolymer membranes. By using Kaelble's equation, the surface tension of SBS-g-DMAEMA and SBS-g-DMAEMA-HEP were determined. It was found that with increasing grafting amount and the heparin content, the surface tension and water content of SBS-g-DMAEMA membrane increased, whereas the contact angle decreased. The amount of the adsorption of albumin and fibrinogen decreased with increasing graft amount and heparin content. However, there was a minimum for adsorption of proteins in the SBS-g-DMAEMA and SBS-g-DMAEMA-HEP membranes.

“…1,2 On the contrary, highly hydrophilic molecules, such as 2-hydroxymethylmethacrylate and polyethyleneglycol, have been grafted onto surfaces in order to inhibit cell adhesion. 3,4 However, these surfaces are monofunctional and irresponsive to stimuli.…”

Section: Introductionmentioning

confidence: 99%

Signal transduction and cytoskeletal reorganization are required for cell detachment from cell culture surfaces grafted with a temperature-responsive polymer

Yamato

Okuhara

Karikusa

et al. 1999

135

105

We have developed a new cell culture substrate grafted with a temperature-responsive polymer, poly(N-isopropylacrylamide) (PIPAAm) using an electron beam irradiation method. These surfaces are hydrophobic in culture at 37 degrees C due to the hydration/dehydration changes intrinsic to PIPAAm at 32 degrees C, and they become highly hydrophilic below 32 degrees C. At 37 degrees C grafted and ungrafted surfaces showed no difference with regard to attachment, spreading, growth, confluent cell density, and morphology of bovine aortic endothelial cells. Stress fibers, peripheral bands, and focal contacts were established in similar ways. After the medium temperature was decreased to 20 degrees C, spread cells lost their flattened morphology, acquiring a rounded cell appearance similar to that of cells immediately after plating. After mild agitation cells floated free from the dish surface without trypsin treatment. Neither cell morphological changes nor cell detachment occurred on ungrafted surfaces. An ATP synthesis inhibitor, sodium azide, and a tyrosine kinase inhibitor, genistein, suppressed cell morphological changes and cell detachment while a protein synthesis inhibitor, cycloheximide, slightly enhanced cell detachment. An actin filament stabilizer, phalloidin, and its depolymerizer, cytochalasin D, also inhibited cell detachment. These findings suggest that cell detachment on grafted surfaces is mediated by intracellular signal transduction and reorganization of the cytoskeleton. While trypsinization causes damage to the cell membrane surface and extracellular matrix proteins, this alternative low temperature treatment is exceptionally noninvasive. The temperature-responsive cell culture surface also should prove useful for investigating the molecular machinery involved in cell-surface detachment.