n-o-dimethacryloylhydroxylamine and ethylene-dimethacrylate

Molecularly imprinted polymers (MIPs) against fructosyl valine (Fru-Val), the N-terminal constituent of hemoglobin A1c beta-chains, were prepared by cross-linking of beta-D-Fru-Val-O-bis(4-vinylphenylboronate) with an excess of ethylene glycol dimethacrylate (EDMA) or trimethylolpropane trimethacrylate (TRIM). Control MIPs were prepared in analogy by cross-linking the corresponding vinylphenylboronate esters of fructose and pinacol. After template extraction batch rebinding studies were performed using different pH values and buffer compositions. The Fru-Val imprinted TRIM cross-linked polymer binds about 1.4 times more Fru-Val than the fructose imprinted polymer and 2.7 times more Fru-Val than pinacol imprinted polymer. The highest imprinting effect was obtained in 100 mM sodium carbonate/10% methanol (pH 11.4). The TRIM cross-linked Fru-Val imprinted polymer showed a better specificity than the EDMA cross-linked polymer. The binding of valine was very low. Thermo gravimetric analysis indicated that the generated Fru-Val imprinted polymer has high thermo stability. No change in binding was observed after incubation of the polymers in buffer at 80 degrees C for 36 h. Since the functional group of the polymers (phenyl boronic acid) targets the sugar part of Fru-Val the imprint technique used should also be applicable for the development of MIPs against other glycated amino acids and peptides.

Topics: Glycated Hemoglobin; Humans; Hydroxylamines; Methacrylates; Polymers; Temperature; Valine

Poly(2-hydroxyethyl methacrylate) (PHEMA) crosslinked with ethylene dimethacrylate (EDMA) or N,O-dimethacryloylhydroxylamine (DMHA) was obtained in the form of slabs by bulk radical polymerization. Two porosity-inducing methods were investigated, phase separation using a low-molecular-weight porogen and a salt-leaching technique using NaCl and saccharose. Compared with the phase separation, the salt-leaching created open porous structures with voids of the size and shape of crystallites. To address its potentials in the context of stem cell therapies, undifferentiated mouse embryonic stem cells D3 (ES D3 cells) were seeded on the slabs and analyzed for the ability to grow on different types of non-degradable and/or degradable porous PHEMA hydrogels. The cells were able to proliferate only on PHEMA crosslinked with EDMA or 2 wt% DMHA. In order to assess the effect of gelatin, which is routinely used for ES cell cultures, PHEMA slabs were soaked in gelatin solutions and compared the number of cells on gelatin-treated and untreated slabs 4 days after cell seeding. Surprisingly, the number of cells was only slightly higher on gelatin-treated slabs.

Topics: Animals; Biocompatible Materials; Biodegradation, Environmental; Cell Culture Techniques; Cells, Cultured; Cross-Linking Reagents; Cyclohexanols; Embryo, Mammalian; Hydrogen-Ion Concentration; Hydrolysis; Hydroxylamines; Indicators and Reagents; Methacrylates; Mice; Microscopy, Electron; Microscopy, Electron, Scanning; Nitrogen; Polyhydroxyethyl Methacrylate; Polymers; Polymethacrylic Acids; Sodium Chloride; Stem Cells; Sucrose; Time Factors