Surface sulfonates lock serum albumin into a "hard" corona.

The composition of the layer of proteins adsorbed to macro- or microscopic surfaces of synthetic origin influences the response of living systems to these materials. This adsorbed layer of proteins usually comprises a "soft" coating or corona of labile or exchangeable adsorbed proteins on top of a more tenaciously held "hard" corona in contact with the surface. Here, we link the dependence of cell adhesion on a 20 nm film of polyelectrolyte complex to the "hardness" of the initial corona using albumin, the most prevalent protein in serum. The ease with which albumin can be lost depends on the surface functional group - carboxylate or sulfonate, in particular aromatic sulfonate. Carboxylate permits easier loss of albumin, which presumably allows the subsequent adsorption of proteins such as fibronectin, required for cell adhesion. Sulfonate holds on to albumin more strongly, producing a persistent hard corona likely to remain biocompatible. The mechanism is thought to be related to the higher energy of interaction between sulfonate and amine than between carboxylate and amine, and provides insight on possible reasons why so-called "tissue culture plastic" works so well for in vitro cell culture.

Stressful Surfaces: Cell Metabolism on a Poorly Adhesive Substrate.

The adhesion and proliferation of cells are exquisitely sensitive to the nature of the surface to which they attach. Aside from cell counting, cell "health" on surfaces is typically established by measuring the metabolic rate with dyes that participate in the metabolic pathway or using "live/dead" assays with combinations of membrane permeable/impermeable dyes. The binary information gleaned from these tests-whether cells are attached or not, and whether they are living or dead-provides an incomplete picture of cell health. In the present work, proliferation rates and net metabolism of 3T3 fibroblasts seeded on "biocompatible" ultrathin polyelectrolyte multilayer films and on control tissue culture plastic were compared. Cells adhered to, and proliferated on, both surfaces, which were shown to be nontoxic according to live/dead assays. However, adhesion was poorer on the multilayer surface, illustrated by diffuse organization of the actin cytoskeleton and less-developed focal adhesions. Proliferation was also slower on the multilayer. When normalized for the total number of cells, it was shown that cells on multilayers experienced a five-day burst of metabolic stress, after which the metabolic rate approached that of the control surface. This initial state of high stress has not been reported or appreciated in studies of cell growth on multilayers, although the observation period for this system is usually a few days.

Cell Adhesion and Proliferation on the "Living" Surface of a Polyelectrolyte Multilayer.

The adhesion of living eukaryotic cells to a substrate, one of the most complex problems in surface science, requires adsorption of extracellular proteins such as fibronectin. Thin films of polyelectrolyte complex made layer-by-layer (polyelectrolyte multilayers or PEMUs) offer a high degree of control of surface charge and composition-interconnected and essential variables for protein adhesion. Fibroblasts grown on multilayers of poly(styrenesulfonate), PSS, and poly(diallyldimethylammonium), PDADMA, with increasing thickness exhibit good adhesion until the 12th layer of polyelectrolyte has been added, whereupon there is a sudden transition to nonadhesive behavior. This sharp change is due to the migration of excess positive charge to the surface-a previously unrecognized property of PEMUs. Precise radiotracer assays of adsorbed (125)I-albumin show how protein adsorption is related to multilayer surface charge. With more negative surface charge density from the sulfonates of PSS, more albumin adsorbs to the surface. However, a loosely held or "soft corona" exchanges with serum protein under the Vroman effect, which is correlated with poor cell adhesion. A comprehensive view of cell adhesion highlights the central role of robust protein adhesion, which is required before any secondary effects of matrix stiffness on cell fate can come into play.

Quasi-Spherical Cell Clusters Induced by a Polyelectrolyte Multilayer.

Fibroblasts cultured on polyelectrolyte multilayers, PEMUs, made from poly(diallyldimethylammonium), PDADMA, and poly(styrene sulfonate), PSS, showed a variety of attachment modes, depending on the charge of the last layer and deposition conditions. PEMUs terminated with PDADMA (cationic) were cytotoxic when built in 1.0 M NaCl but cytophilic when built in 0.15 M NaCl. Cells adhered poorly to all PSS-capped (anionic) films. PEMUs built in 0.15 M NaCl but terminated with a layer of PSS in 1.0 M NaCl induced most cells to form spherical clusters after about 48 h of culture. These clusters still interrogated the surface, and when they were replated on control tissue culture plastic, cells emerged with close to 100% viability. Differences between the various surfaces were probed in an effort to identify the mechanism responsible for this unusual behavior, which did not follow accepted correlations between substrate stiffness and cell adhesion. No significant differences in roughness or wetting were observed between cluster-inducing PSS-capped multilayers and those that did not produce clusters. When the surface charge was assayed with radiolabeled ions a strong increase in negative surface charge was revealed. Viewing the multilayer as a zwitterionic solid and comparing its surface charge density to that of a cell membrane yields similarities that suggest a mechanism for preventing protein adhesion to the surface, a necessary step in the integrin-mediated mechanotransduction properties of a cell.

COMPORTAMIENTO DEL CATALIZADOR CoMo/[1]-Al2O3 MODIFICADO CON BORO Y POTASIO EN LAS REACCIONES SIMULTÁNEAS DE HIDROGENACIÓN DE OLEFINAS E HIDRODESULFURACIÓN DE 2-METILTIOFENO / THE PERFORMANCE OF CoMo/[1]-Al2O3 CATALYST MODIFIED WITH BORON AND POTASSIUM IN SIMULTANEUOUS REACTIONS OF OLEFINS HYDROGENATION AND 2-METHYLTHIOPHENE HYDRODESULFURIZATION / COMPORTAMENTO DO CATALISADOR CoMo/[1]-Al2O3 MODIFICADO COM BORO E POTÁSSIO NAS REAÇÕES SIMULTÂNEAS DO HIDROGENAÇÃO DE OLEFINAS E HIDRODESULFONAÇÃO DO 2 METILTIOFENO

Las características ácido-base de la [1]-alúmina, soporte de catalizadores sulfurados CoMo/[1]-Al2O3, fueron modificadas por la adición de boro (2, 3, 5, y 8% B2O3) y potasio (1, 3 y 5%K). Estos catalizadores fueron ensayados en reacciones simultáneas de hidrogenación (HID) de olefinas (2,4,4 trimetil-1-penteno y 2,4,4 trimetil- 2-penteno; 3:1) e hidrodesulfuración (HDS) de 2-metiltiofeno, simulando el hidrotratamiento (HDT) de una nafta de craqueo o de FCC (Fluidized Catalytic Cracking). Los resultados mostraron que aunque los cambios inducidos en las características ácido-base del soporte de alúmina disminuyeron la actividad para la HID y la HDS de los catalizadores, también dieron lugar a cambios apreciables en la selectividad hacia la HDS de tiofenos relativa a la HID de olefinas, lo que los convierte en buenos candidatos para el HDT de la nafta de FCC. Por otro lado, el craqueo, la alquilación y la isomerización de metilos se favorecieron cuando se hicieron las modificaciones con B, lo mismo que la migración del doble enlace de las olefinas ramificadas cuando se hizo la modificación con K. Los resultados de este trabajo permiten concluir que la actividad y selectividad del catalizador CoMo/[1]-Al2O3 para la HDS de la nafta de FCC pueden ser controladas por una adecuada modificación de las propiedades ácido-base del soporte de alúmina mediante la adición de pequeñas cantidades de B y de K.

The acid-base characteristics of the [1]-alumina support of sulfide CoMo catalysts were modified by boron (2, 3, 5, y 8% B2O3) and potassium (1, 3 y 5% K) addition. The catalysts were tested in simultaneous olefins (2,4,4-trimethyl-1-pentene y 2,4,4 trimethyl-2-pentene) hydrogenation (HYD) and 2-methyl-thiophene hydrodesulfurization (HDS) reactions, simulating the hydrotreating (HDT) of FCC (Fluidized Catalytic Cracking) naphtha. Results show that acid-base changes induced by B and K addition lead to an activity decrease in both HYD and HDS reactions, but, by contrast, an interesting increase in selectivity toward HDS over olefins HYD was also observed. These characteristics make these modified catalysts good candidates to be used in the naphtha HDT process. On the other hand, addition of B promotes cracking and methyl isomerization reactions, while, modification with K favors the double bond migration in branched olefins. The results of this work allow us to conclude that the activity and selectivity of CoMo/[1]-alumina catalysts for the HDS of FCC naphtha can be controlled by a proper modification of the acid-base properties of the alumina support by the addition of small quantities of B and K.

As características ácido-base da [1]-alumina, suporte de catalisadores sulfurados CoMo/[1]-Al2O3, foram modificadas pela adição de boro (2, 3, 5 e 8% B203) e potássio (1, 3 e 5% K). Estes catalisadores foram ensaiados em reações simultâneas de hidrogenação (HID) de olefinas (2,4,4 trimetil- 1-penteno y 2,4,4 trimetil-2-penteno; 3:1) e hidrosulfuração (HDS) de 2-metiltiofeno, simulando o hidrotratamento (HDT) de uma nafta de craqueo ou de FCC (Fluidized Catalytic Cracking). Os resultados mostraram que embora as mudanças induzidas nas características ácido - base do suporte de alumina, diminuíram a atividade para a HIDe a HDS dos catalisadores, também deram lugar a mudanças apreciáveis na seletividade pela HDS de tiofenos relativa à HID de olefinas, o que os converte em bons candidatos para o HDT da nafta de FCC. Por outro lado, o craqueo, a alquilação e a isomerização de metilos foram favorecidos quando foram feitas as modificações com B, o mesmo que a migração do enlace duplo das olefinas ramificadas quando se fez amodificação com K. Os resultados deste trabalho permitem concluir que a atividade e seletividade do catalisador CoMo/[1]-Al2O3 para a HDS da nafta de FCC, pode ser controlada por uma adequada modificação das propriedades ácido-base do suporte de alumina perante a adição de pequenas quantidades de B e de K.