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Results for “Polyhydroxyethyl Methacrylate”

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PubMed2000

Effect of an immobilization matrix and capsule membrane permeability on the viability of encapsulated HEK cells.

The effect of inclusion of an immobilization matrix and the capsule membrane permeability on the viability, metabolic activity, and proliferation of encapsulated HEK cells was investigated in vitro. In the absence of a matrix, a particular transfected HEK cell line formed a single aggregate in the core of the poly(hydroxyethyl methacrylate-co-methyl methacrylate) (HEMA-MMA) capsule, and the number of live cells decreased significantly with the passage of time. In contrast, co-encapsulation with a 1% (w/v) ultralow gelling temperature agarose matrix promoted the proliferation of the encapsulated cells. The initial number of approximately 200 live cells/capsule doubled 14 d after encapsulation and reached a plateau of approximately 500 live cells/capsule 28 d after encapsulation. The agarose matrix provided uniform distribution of the cells within the capsule core giving rise to multiple aggregates upon proliferation. Reduction of the polymer solution concentration, and hence the increase of the permeability of the capsule membrane, did not have an effect on the extent or rate of proliferation of cells co-encapsulated with agarose, and did not improve the viability of cells that were encapsulated without a matrix. These cells (transfected with the cDNA for human hepatic lipase) served as a model as part of a program evaluating the use of encapsulated cells for gene therapy.

Biocompatible Materials↗

PubMed1986

Suppression of platelet activity on microdomain surfaces of 2-hydroxyethyl methacrylate-polyether block copolymers.

Block copolymers constructed from chains of poly(2-hydroxyethyl methacrylate) (PHEMA) and either poly-ethyleneoxide (PEO) or poly-propyleneoxide (PPO) were synthesized. These block copolymers exhibited microdomain structure. Platelet adhesion on their surfaces was investigated by a column elution method to examine the effect of microdomain structure. The number of platelets adhered from whole blood was smaller for the block copolymer systems than for the homopolymers. Minimum points of platelet adhesion appeared at approximately 0.38 mol fraction of HEMA in the HEMA-PO system. Both block copolymer surfaces showed microdomains of alternate lamellar structure. Furthermore, the percent of platelets released from the column after incubation was investigated using PRP. In the case of homopolymers, released platelet percentages decreased with an increase of incubation time. Released platelet percentages from the block copolymers, however, were nearly constant with changing incubation time. These results show that HEMA-EO and HEMA-PO block copolymers had the ability to suppress both reversible and irreversible adhesion of platelets to their respective microdomain surfaces.

Animals↗

PubMed1993

Protein adsorption and platelet adhesion onto polyurethane grafted with methoxy-poly(ethylene glycol) methacrylate by plasma technique.

Graft polymerization of methoxy-poly(ethylene glycol) methacrylate, an ester of methacrylic acid and monomethoxy-poly(ethylene glycol) (PEO), was performed onto a polyetherurethane (PU) film and tube under different polymerization conditions by a plasma treatment technique. The surface of grafted PU film was characterized by staining with dye, x-ray photoelectron spectroscopy, contact angle, and zeta potential. All these measurements indicated that water-soluble chains were immobilized on the PU surface, their location being restricted to the film surface region. The PU surface showed reduced protein adsorption in vitro and reduced platelet adhesion in vitro and ex vivo. The optimum graft density suppressing the protein adsorption was as low as 5 micrograms cm(-2). When a small amount of dimethacrylate was added to the monomer solution for graft polymerization to introduce crosslinking in the grafted layer, protein adsorption was further slightly reduced. The extent of reduction in serum albumin adsorption was always less than that of gamma-globulin. Although platelet adhesion was largely reduced by the surface graft polymerization, a definite amount of protein was always adsorbed to the grafted surface.

Adsorption↗

PubMed1992

Reduced lymphocyte activation in space: role of cell-substratum interactions.

We investigated the effect of substratum adhesiveness on stimulated lymphocyte blastogenesis by reducing and blocking cell adhesion with poly (2-hydroxyethyl methacrylate) (poly-HEMA) in a simple on-ground system. Cells grown on medium-thick and thick poly-HEMA films were rounded in shape and displayed no signs of spreading. By contrast, on tissue culture plastic and very thin poly-HEMA films, they showed clear signs of spreading. The mitogenic response of lymphocytes grown on thick poly-HEMA films was reduced by up to 68% of the control (tissue culture plastic). Interferon-gamma production was near zero when the cells were grown on the least adhesive substratum. On uncoated plastic, activated lymphocytes subjected to high gravity (20g) exhibited an increased proliferation rate (40%) compared with 1g. By contrast, on poly-HEMA, high gravity did not improve lymphocyte responsiveness. These results show that activated lymphocytes need to anchor and spread prior to achieving an optimal proliferation response. We conclude that decreased lymphocyte adhesion could contribute to the depressed in vitro lymphocyte responsiveness found in the microgravity conditions of space flight.

Cell Adhesion↗

PubMed2004

Polymer-brush stationary phases for open-tubular capillary electrochromatography.

Synthesis of poly(2-hydroxyethyl methacrylate) (PHEMA) brushes from the inside of silica capillaries by surface-initiated atom transfer radical polymerization (ATRP) yields unique stationary phases for open-tubular capillary electrochromatography (OT-CEC). Although PHEMA brushes have only a small effect on the separation of a set of phenols and anilines, derivatization of PHEMA with ethylenediamine (en) allows baseline resolution of several anilines that co-elute from bare silica capillaries. Derivatization of PHEMA with octanoyl chloride (C8-PHEMA films) affords even better resolution in the separation of a series of phenols and anilines. Increasing the thickness of C8-PHEMA coatings by a factor of 2 enhances resolution for several solute pairs, presumably because of an increase in the effective stationary phase to mobile phase volume ratio. Thus, this work demonstrates that thick polymer brushes provide a tunable stationary phase with a much larger phase ratio than is available from monolayer wall coatings. Through appropriate choice of derivatizing reagents, these polymer brushes should allow separation of a wide range of neutral molecules as well as compounds with similar electrophoretic mobilities.

Chromatography, Micellar Electrokinetic Capillary↗

PubMed1997

Microcapsules as bio-organs for somatic gene therapy.

Current human gene therapy relies on genetic modification of the patient's own cells. An alternate non-autologous approach is to use universal cell lines engineered to secrete therapeutic products. Protection with immuno-isolation devices would allow the same recombinant cell line to be used for different patients, thus potentially lowering the cost of treatment. The feasibility of this idea has now been demonstrated in vitro and in vivo. Recombinant gene products with potential therapeutic applications (human growth hormone, factor IX, lysosomal enzymes, adenosine deaminase) have been expressed from genetically modified cells after encapsulation with alginate-poly-L-lysine-alginate or hydroxyethyl methacrylate-methyl methacrylate. We have also demonstrated the feasibility of this idea in vivo. After intraperitoneal implantation, genetically modified mouse Ltk- fibroblasts or C2C12 myoblasts encapsulated in alginate-poly-L-lysine-alginate could deliver recombinant gene products (human growth hormone, human factor IX) to the systemic circulation of mice. The clinical efficacy of this novel approach to gene therapy has now been shown in murine models of human diseases. In the Snell dwarf mice deficient in growth hormone production, implantation of encapsulated mouse myoblasts engineered to secrete mouse growth hormone resulted in increases in body weight, length and organ sizes, some to > 25% above those of the controls. In the Gus/Gus mice suffering from the lysosomal storage disease mucopolysaccharidosis type VII due to deficient beta-glucuronidase, implantation of encapsulated mouse fibroblasts engineered to secrete mouse beta-glucuronidase resulted in delivery of normal levels of the enzyme in the plasma and significant correction of the organ histopathology. Hence, delivery of recombinant gene products through bioartificial devices appears to be a promising strategy for the treatment of genetic diseases.

Alginates↗

PubMed1996

Poly (2-hydroxyethyl methacrylate) particles for management of hemorrhage of complicated origin: treatment of hemobilia.

Poly (2-hydroxyethyl methacrylate) (PHEMA) particles of cylindrical and spherical shape were developed as a preparation for tumor treatment or control of hemorrhage by blocking their blood supply. In this report, PHEMA particles were used for the management of hemobilia, that is, bleeding into biliary passages. The origin of hemobilia in 31 patients was localized by selective angiography. With the objective of prophylaxy of hemorrhage, selective embolization with PHEMA particles of the branches of the hepatic artery responsible for the supply of blood to the focus of damage was used in 18 patients. This low-trauma method allowed either a complete control of bleeding or, at least, intraoperative blood loss was reduced more than twice. Histological investigation of the occluded blood vessels showed that the thrombus was attached to the particles and was reinforced by the porous structure of the polymer. A hypercoagulation reaction was observed in the postembolization period. This allowed correction of the hypocoagulation in the hemostasis system.

Bile Ducts, Intrahepatic↗

PubMed2006

Gradient substrate assembly for quantifying cellular response to biomaterials.

Using quantitative fluorescence microscopy in conjunction with a method of gradient substrate assembly established in their group, the authors were able to introduce and measure reproducible changes in cellular morphology and cell density by manipulating polymer grafting density. The mechanism behind this change in cellular behavior was explained by a semiempirical, geometric model that describes the effect of the spatial distribution of the polymer on protein attachment. A 10-fold increase in graft density of poly(2-hydroxyethyl methacrylate) [PHEMA] along the surface of a gradient sample, preexposed to bovine fibronectin, caused a change in the size of fibroblasts on the surface (i.e., cell spreading) from (1238 +/- 704) to (377 +/- 216) microm(2). The results were in quantitative agreement with those obtained on three separate gradient samples. Both cellular response and fibronectin adsorption (as measured via ellipsometry) were found to vary sigmoidally with graft density of PHEMA, demonstrating the high degree of correlation between the two phenomena. A simple, rigid-disk model accounting for the surface coverage of PHEMA was able to predict the amount of adsorbed fibronectin with a correlation coefficient of 0.97. Maximal cell adhesion and cell spreading were found to occur at fibronectin surface densities of 50 and 100 ng/cm(2), respectively. The results demonstrate the role of gradient substrate assembly as a method for quantifying the relationship between protein and cellular response to technologically relevant polymeric materials.

Adsorption↗

PubMed1997

Detection and characterization of Sp1 binding activity in human chondrocytes and its alterations during chondrocyte dedifferentiation.

We have detected DNA binding activity for a synthetic oligonucleotide containing an Sp1 consensus sequence in nuclear extracts from human chondrocytes. Changes in the levels of Sp1 oligonucleotide binding activity were examined in nuclear extracts from freshly isolated human chondrocytes, from chondrocytes that had been cultured under conditions that allowed the maintenance of a chondrocyte-specific phenotype on plastic dishes coated with the hydrogel poly(2-hydroxyethyl methacrylate), and from chondrocytes induced to dedifferentiate into fibroblast-like cells by passage in monolayer culture on plastic substrata. It was observed that Sp1 binding was 2-3-fold greater in nuclear extracts from dedifferentiated chondrocytes than in nuclear extracts from either freshly isolated chondrocytes or from cells cultured in suspension. The Sp1 binding activity was specific, since it was competed by unlabeled Sp1 but not by AP1 or AP2. The addition of a polyclonal antibody against Sp1 to nuclear extracts from freshly isolated chondrocytes or to extracts isolated from chondrocytes cultured in monolayer decreased the binding of Sp1 by approximately 85%. However, when the same experiment was carried out with nuclear extracts prepared from cells cultured on poly(2-hydroxyethyl methacrylate)-coated plates, only a very slight inhibition of Sp1 binding was observed. When fragments of the COL2A1 promoter containing putative Sp1 binding sites amplified by polymerase chain reaction were examined, it was found that the amounts of DNA-protein complex formed with nuclear extracts from dedifferentiated chondrocytes were 2-3-fold greater than the amounts formed with nuclear extracts from freshly isolated chondrocytes or from cells cultured in suspension. Quantitation of DNA binding activity by titration experiments demonstrated that nuclear extracts from fibroblast-like cells contained approximately 2-fold greater Sp-1 specific binding activity than nuclear extracts from chondrocytes. The direct role of Sp1 in type II collagen gene transcription was demonstrated by co-transfection experiments of COL2A1 promoter-CAT constructs in Drosophila Schneider line L2 cells that lack Sp1 homologs. This is the first demonstration of Sp1 binding activity in human chondrocytes and of differences in Sp1 DNA binding activity between differentiated and dedifferentiated chondrocytes.

Animals↗

PubMed1992

In vitro cytotoxicity of melanized poly(2-hydroxyethyl methacrylate) hydrogels, a novel class of ocular biomaterials.

Due to their ability to absorb ultraviolet and visible radiation, we have proposed the melanized poly(2-hydroxyethyl methacrylate) hydrogels as biomaterials suitable for the manufacture of soft artificial intraocular lenses. Their biocompatibility has not been evaluated so far. In this study, poly(2-hydroxyethyl methacrylate) containing various amounts of adrenochrome-melanin were synthesized and the cytotoxicity of their aqueous extracts was assessed by using four in vitro testing techniques (trypan blue dye exclusion, inhibition of DNA synthesis, lactate dehydrogenase release, and inhibition of cell growth). Assays were based on incubation with human choroidal fibroblasts. By the first three techniques, no cytotoxicity was found in the extracts. The inhibition of cell growth test showed a slight cellular protein loss, however only in the extracts of polymers with high melanin content. The results suggest that the release of potentially toxic agents from melanized hydrogels into an aqueous medium is not significant. However, when an assay in collagen gel was carried out in the presence of specimens of melanized hydrogels, a toxic reaction was clearly revealed. This can be caused by a delayed release of toxic molecules from melanin, or by some other mechanism. The use of melanin-containing polymers as implant materials becomes questionable and further research is necessary.

Biocompatible Materials↗

PubMed1998

Effect of charged groups on the adsorption and penetration of proteins onto and into carboxymethylated poly(HEMA) hydrogels.

A range of carboxymethylated poly(hydroxyethyl methacrylate) (CM-PHEMA) hydrogels with varying degrees of carboxymethylation was synthesized for a systematic study of the effects of ionized groups ('charge') on the uptake by hydrogel matrices of the proteins, lysozyme and human serum albumin (HSA). Using a radiolabel-tracer technique, X-ray photoelectron spectroscopy, and laser scanning confocal microscopy, we attempted to differentiate between protein molecules that were irreversibly adsorbed onto the hydrogel surface and those that penetrated into the hydrogel matrix. The effective pore size of the CM-PHEMA hydrogels was modelled and compared with the known molecular dimensions of the two proteins. The effects of the presence of varying amounts of ionized groups in the hydrogel matrix differed for the two proteins. For lysozyme, increased uptake was observed at higher carboxymethylation; this is interpreted as resulting from a combination of electrostatic attraction and increasing ease of penetration of the protein into the more porous hydrogel matrix. For HSA, on the other hand, the uptake was primarily by surface adsorption, with little diffusive penetration into the matrix.

Adsorption↗

PubMed2000

Microencapsulation of normal and transfected L929 fibroblasts in a HEMA-MMA copolymer.

Mouse L929 fibroblasts transfected to express a secreted form of human alkaline phosphatase (SEAP) were encapsulated in approximately 400-microm poly(hydroxyethyl methacrylate-co-methyl methacrylate) (HEMA-MMA) microcapsules as a baseline for the use of genetically engineered cells in encapsulation therapy. Although incubation of microcapsules with serum-containing medium resulted in maintaining the number of live encapsulated cells with the passage of time, incubation in a serum-free medium resulted in a three-fold proliferation of the encapsulated cells within a 3-week observation period. Similar to the results for incubation with serum-containing medium, co-encapsulation with a bovine dermal type I collagen, i.e., the inclusion of a matrix in the core of the capsules, resulted in maintenance of the initial number of live cells with the passage of time. SEAP measurements indicated that the transfected cells not only continued to express the transgene product after encapsulation, but also adapted to the capsule microenvironment to secrete SEAP at progressively larger amounts with the passage of time. However, SEAP expression only occurred when the transfected cells (encapsulated or non-encapsulated) were cultivated in serum-containing medium.

Alkaline Phosphatase↗

PubMed1994

Interpenetrating polymer network (IPN) as a permanent joint between the elements of a new type of artificial cornea.

The combination at the interface between two chemically identical polymers was investigated by light and electron (scanning, transmission) microscopy. The polymers constitute elements of a new type of artificial cornea in which the peripheral skirt is made from spongy poly(2-hydroxyethyl methacrylate) (PHEMA) and the central optical zone from homogeneous, transparent PHEMA. Their two-phase combination along the boundary fulfill formally the requirements for an interpenetrating polymer network (IPN). The procedure for the manufacture of prosthesis was described in detail. Thin and ultrathin sections excised from the interface region were investigated using microscopic techniques. Light microscopy allowed the measurement of the diffusion path length of transparent PHEMA into sponge, which was approximately 0.5 mm. Transmission electron microscopy revealed a cellular-like morphology as well as larger segregated zones, which indicated network interpenetration on a molecular level and also a relatively poor miscibility of the two polymers despite their identical chemical structure. The latter was interpreted as a result of the submicroscopic restraints imposed by polymer I (sponge) upon polymer II. This study provides evidence that the interface combination of the prosthetic elements should be regarded as a gradient homo-IPN. This system offers a union between elements much stronger than those previously reported in artificial corneas.

Biocompatible Materials↗

PubMed2001

Capillary zone electrophoresis of proteins with poly(2-hydroxyethyl methacrylate)-coated capillaries: fundamental and applications.

Fused silica capillaries have been modified by atom-transfer radical polymerization (ATRP) to generate covalently bonded polymer films of 2-hydroxyethyl methacrylate. Because the kinetics of ATRP have mainly been investigated in bulk solutions, a GC experiment was set up to examine monomer conversion inside narrow-bore capillaries. It was shown that after 1 to 4 h the reaction was nearly complete. The coating process was further optimized by monitoring EOF, because low EOF indicates high surface coverage. To deal with the very low EOF values, a new approach was used to dramatically reduce the measurement time by overlaying hydrodynamic flow on the electroosmotic flow. The corresponding equations are derived separately in detail. Capillaries were then coated under optimum conditions with linear or cross-linked polymer films. The EOF was reduced over a wide range of pH values. A long-term reproducibility test with both types of functionalization showed that the efficiency of the linear polymer coating decreased significantly over time. With cross-linked films, however, the efficiency even increased. Relative standard deviations for protein migration times were also much lower in cross-linked coated capillaries. Highly efficient separations could be performed for basic and acidic proteins in acidic media, and for the latter even in basic media.

Electrophoresis, Capillary↗

PubMed1989

The viability of mononuclear phagocytes in vitro is diminished by the interaction of cells with serum proteins bound to the culture substratum.

The long-term viability of bone marrow-derived mononuclear phagocytes in vitro was inversely correlated with the capacity of the cells to attach to the culture substratum. Mononuclear phagocytes suspended in medium containing 10% fetal bovine serum and subcultured on substrata coated with a 0.1% solution of poly(2-hydroxyethyl methacrylate) formed a population of non-adherent or loosely attached cells that remained viable for a 7 day incubation period. In contrast, cells subcultured under otherwise identical conditions on substrata optimal for cell attachment exhibited a 40-fold decline in cell number during the same period of time. The survival of mononuclear phagocytes subcultured under conditions which promoted cell attachment was increased by reducing the concentration of serum in the medium. Thus, cells subcultured 7 days in the complete absence of serum exhibited only a two-fold decline in cell number. However, mononuclear phagocytes subcultured in the absence of serum exhibited a ten-fold decline in cell number when cultured on substrata coated with serum proteins. This decline was reversed by the addition of the mononuclear phagocyte-specific growth factor, colony stimulating factor-1 (CSF-1) to the medium. These results indicate that serum proteins bound to the culture substratum exert a significant influence on the viability of adherent mononuclear phagocytes in vitro and on the requirement of cells for CSF-1 in order to survive.

Animals↗

PubMed1999

Adhesion mechanisms of human lens epithelial cells on 4 intraocular lens materials.

PURPOSE: To evaluate lens epithelial cell (LEC) adhesion on different intraocular lens (IOL) materials with particular attention to the distribution of proteins located in the focal contacts. SETTING: Center of Biotechnological and Clinical Research in Ophthalmology, University of Bologna, Italy. METHODS: The IOL materials tested were poly(methyl methacrylate) (PMMA), heparin-surface-modified PMMA (HSM PMMA), polyHEMA, and silicone. Primary cultures of human LECs were established from human anterior capsules obtained during cataract surgery. The mean number of cells attached per square millimeter was calculated for each material after 24 and 72 hours. Transmission electron microscopy and immunocytochemical analysis were performed to detect the proteins actin, vinculin, and talin. RESULTS: Mean adhesiveness of human LECs increased over time with PMMA and decreased with the other materials. At 72 hours, mean LECs ranged from 54.8 cells/mm2 +/- 12.8 (SD) on PMMA to 2.1 +/- 0.7 cells/mm2 on silicone. The means for HSM PMMA and polyHEMA fell in between. The cytoskeletal proteins were arranged to produce focal contacts in only the LECs cultured on PMMA. The LECs cultured on polyHEMA, HSM PMMA, and silicone attached but failed to develop focal contacts or stress fibers. CONCLUSION: This study confirms the multifactorial pathogenesis of posterior capsule opacification and suggests its incidence will be reduced by improving surgical techniques and using IOL surfaces that discourage cell adhesion.

Actins↗

PubMed2005

Synthesis, structure, and physical properties of hybrid nanocomposites for solid-state dye lasers.

We report on the synthesis, structural characterization, physical properties, and lasing action of two organic dyes, Rhodamine 6G (Rh6G) and Pyrromethene 597 (PM597), incorporated into new hybrid organic-inorganic materials, where the organic component was either poly(2-hydroxyethyl-methacrylate) (PHEMA) or copolymers of HEMA with methyl methacrylate (MMA), and the inorganic counterpart consisted of silica derived from hydrolysis-condensation of methyltriethoxysilane (TRIEOS) in weight proportion of up to 30%. Lasing efficiencies of up 23% and high photostabilities, with no sign of degradation in the initial laser output after 100 000 pump pulses at 10 Hz, were demonstrated when pumping the samples transversely at 534 nm with 5.5 mJ/pulse. A direct relationship could be established between the structure of the hybrid materials, analyzed by solid-state NMR, and their laser behavior. An inorganic network dominated by di-/tri- substituted silicates in a proportion approximately 35:65, corresponding to samples of HEMA with 15 and 20 wt % proportion of TRIEOS, optimizes the lasing photostability. The thermal properties of these materials, together with the high homogeneity revealed by atomic force microscopy (AFM) images, even in compounds with high silica content, indicate their microstructure to be a continuous phase, corresponding to the polymer matrix, which "traps" the silica components at molecular level via covalent bonding, with few or no silica islands.

Chemical Phenomena↗

PubMed1980

On the anomalous sorption behaviour of chlorhexidine with poly(2-hydroxyethyl methacrylate).

General observations of solute: plastic interactions would suggest that the sorption of the cationic preservative, chlorhexidine, with poly (2-hydroxyethyl methacrylate) (PHEMA) is not only unexpected but also atypical in its properties. The polymer and preservative were investigated independently and the sorption of PHEMA with simple solutes, namely benzoic acid, benzocaine and aniline found to exhibit conventional properties. The single exception was the uptake of small amounts of benzocaine and aniline cations at acidic pH's. In order to elucidate the sorption characteristics of chlorhexidine a series of N1-(p-chlorophenyl)-N5-alkylbiguanide acetates were synthesized. The anomalous sorption behaviour observed between PHEMA and the bis biguanide, chlorhexidine, was also found to be characteristic of these monofunctional biguanide derivatives. The extent of the interactions increased with increasing alkyl chain length (R = methyl to n-octyl), this being interpreted in terms of an interaction binding mechanism via the biguanide functional group stabilized by van der Waal's forces between the alkyl chain and the polymer backbone. Atypical sorption behaviour was observed for simple organic cations, biguanide and N2-phenylbiguanide acetate with PHEMA, a possible inference being that this is a general characteristic of all cationic sorption with PHEMA.

Adsorption↗