Size and density of polystyrene particles measure by ultracentrifugation.
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Highly selective separation of yttrium (and lanthanides) is of interest for the design of radiopharmaceuticals, and an efficient method based on the ion-imprinting concept is proposed here. The synthesis and structural, thermodynamic and photophysical characterization of complexes of trivalent yttrium and lanthanides with two new vinyl derivatives of dipicolinic acid, HL1 and L2, are described. The feasibility of using ion-imprinted resins for yttrium and lanthanide separation is demonstrated. The resins were obtained by copolymerization with styrene and divinylbenzene and subsequent acid treatment to remove the metal ion. High-resolution Eu luminescence experiments revealed that the geometry of the complexation sites is well preserved in the imprinted polymers. The ion-imprinted polymer based on HL1 proved to be particularly well adapted for yttrium extraction, having a sizeable capacity (8.9+/-0.2 mg g(-1) resin) and a fast rate of extraction (t(1/2)=1.7 min). In addition, lighter and heavier lanthanide ions are separated. Finally, the resin displays high selectivity for yttrium and lanthanide cations against alkali and alkaline earth metals. For instance, in a typical experiment, 10 mg of yttrium was extracted from 5 g of milk ash sample by 2 g of the resin. The good separation properties displayed by the resin based on HL1 open interesting perspectives for the production of highly pure (90)Y and radiolanthanides for medical applications, and for trace analysis of these radiochemicals in food and in the environment.
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RINm5F cells (an insulin-secreting cell line) were cultured on PSSO3Na microbeads under static conditions. The cell growth rate was either identical to that of cells grown on plastic wells or slower, depending on the initial cell concentration. With both supports, it was similarly influenced by the fetal calf serum concentration in the culture medium, and protein content per cell was identical. However, no spreading was observed when cells were cultured on microbeads. RINm5F cells cultured on plastic wells responded to arginine + theophylline and to leucine + theophylline by a significant increase in insulin secretion. By contrast, in cells cultured on PSSO3Na microbeads, the increase in this secretion was only slight or nil. All these abnormalities were reversible. Thus, when cells cultured on microbeads were detached and seeded on plastic wells, normal spreading and insulin secretion were observed. Lastly, PSSO3Na beads had an acute suppressive effect on insulin secretion by cells cultured on plastic wells. This study provides an example of cell-biomaterial interaction in which cell growth is possible, but with altered cell function.
The surface characteristics of intravenously administered particulate drug carriers decisively influence the protein adsorption that is regarded as a key factor for the in vivo fate of the carriers. Latex nanoparticles were synthesized to study the influence of different basic and acidic functional groups on particulate surfaces on the protein adsorption from human serum. The protein mass adsorbed to the particles was assessed by BCA protein assay, the protein adsorption patterns were analyzed by two-dimensional electrophoresis. Considerable differences in the protein adsorption with regard to preferential adsorbed proteins were detectable for the different functional groups. Possible correlations between the surface characteristics and the protein adsorption are shown and discussed. The knowledge concerning the interactions of proteins and nanoparticles can be used for a rational development of particulate drug carriers and can also be useful for an optimized design of medical devices, e.g., hemodialysis membranes or implants.
Extracellular matrix (ECM) protein adsorption and organization serves as a critical first step in the development and organization of tissues. Advances in tissue engineering, therefore, will depend on the ability to control the rate and pattern of ECM formation. Fibronectin is a prominent component of the ECM, which undergoes fibrillogenesis in the presence of cells. Using sulfonated polysyrene surfaces, we showed that fibronectin undergoes a transition from monolayer to multilayer adsorption at calculated surface charge densities above 0.03 Coulombs (C)/m(2). At charge densities above approximately 0.08 C/m(2), distinct fibronectin fibrillar networks are observed to form with a fibril morphology similar to those observed to form in situ on cell surfaces. This self-organization process is time dependent, with the fibrils achieving dimensions of 30-40 microm in length and 1 microm in height after 72 h of incubation. We suggest that the polarization of charge domains on the polyampholytic fibronectin molecules near high charge density surfaces is sufficient to initiate the multilayer adsorption and the organization of these fibrillar structures. These results suggest that the nonlinear dependence of adsorption on surface charge density may play an important role in the self-organization of many matrix components.
Adhesion of baby hamster kidney fibroblasts (BHK cells) to a Falcon tissue culture flask was measured under various physiological conditions. While 75-80% of the fibroblasts adhere at temperatures from 19-50 degrees, cellular adhesion decreased dramatically below 19 degrees. Less than 10% of the cells adhere to the substratum even after prolonged incubations at temperatures of 8 degrees or below. This lack of adhesion at low temperatures cannot be overcome by the application of increased gravitational force to the cells. No correlation exists between cellular ATP concentrations or respiration rates and the rate of cell adhesion to the substratum. One millimolar Na F and 1 mM 2,4 dinitrophenol together lower cellular ATP concentration by 95% but adhesion is reduced by only 50%. NaN3 and KCN greatly lower cellular ATP concentrations without a corresponding inhibition of adhesion. Inhibition of cellular respiration by these compounds occurs at lower concentrations than does the inhibition of adhesion. Two micrograms/milliliters of cytochalasin B inhibits adhesion by 90%, 0.1 mM vinblastine sulphate or colchicine by less than 50% and 50 microgram/ml colcemid by less than 30%. Fixing the cells with formaldehyde, hardening their membranes with ZnCl2 or treating the cells with toluene, all cause an inhibition in adhesion. Again, application of increased gravitational force cannot overcome these latter inhibitions of BHK cell adhesion to the surface of the flasks.
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The delayed ion extraction method has been used to study characteristics of the initial velocity distributions of positive and negative ions produced simultaneously by laser desorption/ionization (LDI) from non-impacted single aerosol polymeric particles, using a bipolar time-of-flight (TOF) instrument (LAMPAS 2). Due to the geometry of the setup and the characteristics of the ablation process, only the projections of the velocities on the axis of the mass spectrometer can be directly studied. Additionally, since the mean initial velocity under these conditions should be close to zero, it was necessary to extend the method by taking into account higher order contributions of the velocity distribution. Theoretical expressions for these higher order terms are presented and discussed. The bipolar characteristics of the instrument permit evaluation and treatment of a possible instrumental artifact caused by small inclinations of the ionizing laser with respect to the ideal incidence direction. Results of a number of experiments are presented and discussed in relation to the theoretical expressions presented, and to possible ablation scenarios. Evidence pointing out that, under our experimental conditions, only partial ablation of the latex particles occurs was obtained. The variance of the distribution of the projection of the initial velocities can be directly estimated from these results. By assuming that the total initial velocities of the ions are developed completely according to a single-temperature adiabatic expansion mechanism, temperatures of approximately 50 K/Da can be assigned to the ion clouds from the variance estimations. If a two-temperature model is used, a radial temperature of about 100 K/Da results. These values are in reasonable agreement with results for polymer ablation from the literature.
The application of shadow nanosphere lithography for the preparation of large-area, two-dimensional, metallic nanostructures of different shape is described. Through changing the mask morphology by temperature processing and varying the evaporation conditions, particles with morphologies such as rings, rods, and dots have been produced. This process allows outstanding control of the size and morphology of the particles. The efficient technique is shown to scale down the size of metallic nanoparticles from 200 to 30 nm, while preserving the original nanosphere spacing and order. The 150-nm-diameter Fe rings produced by this method show ferromagnetic behavior, which was predicted by theoretical simulation. All the experimental results were confirmed by computer simulations, which also showed the possibility of creating periodic arrays of any other geometrical shape.
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Immobilization of proteins on microplate wells by simple adsorption (e.g., for ELISA) is convenient, but it can be inefficient, especially if proteins are hydrophilic or small in size. This problem was alleviated by the use of polyvinylbenzyl lactonoylamide (PVLA). PVLA is strongly adsorbed to the hydrophobic well surface, and its lactonamide part can be oxidized with periodate to generate aldehydo groups. Proteins are then immobilized covalently to the aldehydo groups by reductive amination under mild conditions. Using this method, henceforth termed the PVLA method, alkaline phosphatase (AP) was immobilized to microplates six- to sevenfold greater than by simple adsorption (as measured by activity). Similarly, the activity of immobilized mannose-binding protein A (MBP-A) was 4- to 8-fold higher by the PVLA method than by simple adsorption. The PVLA-coated plates needed as little as 200 ng of MBP-A per well to have a sufficient amount of MBP-A immobilized for the measurement of binding of 125I-labeled mannosylated bovine serum albumin (125I-Man-BSA), but unmodified plates required as much as 20 micrograms/well MBP-A to obtain the same response. Recommended conditions for the PVLA method are 40 microliters of 2 mg/ml of PVLA for coating, 1 mM NaIO4 for the generation of the aldehydo groups, and a 2-h reductive amination at 37 degrees C between pH 8 and 9 for the protein ligation.