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A system to impose prescribed homogenous strains on cultured cells.

There is presently significant interest in cellular responses to physical forces, and numerous devices have been developed to apply stretch to cultured cells. Many of the early devices were limited by the heterogeneity of deformation of cells in different locations and by the high degree of anisotropy at a particular location. We have therefore developed a system to impose cyclic, large-strain, homogeneous stretch on a multiwell surface-treated silicone elastomer substrate plated with pulmonary epithelial cells. The pneumatically driven mechanism consists of four plates each with a clamp to fix one edge of the cruciform elastomer substrate. Four linear bearings set at predetermined angles between the plates ensure a constant ratio of principal strains throughout the stretch cycle. We present the design of the device and membrane shape, the surface modifications of the membrane to promote cell adhesion, predicted and experimental measurements of the strain field, and new data using cultured airway epithelial cells. We present for the first time the relationship between the magnitude of cyclic mechanical strain and the extent of wound closure and cell spreading.

Algorithms↗

Solid supports for microarray immunoassays.

Stimulated by the achievements of the first phase in genomics and the resulting need of assigning functions to the acquired sequence information, novel formats of immunoassays are being developed for high-throughput multi-analyte studies. In principle, they are similar in nature to the microarray assays already established at the level of nucleic acids. However, the biochemical diversity and the sheer number of proteins are such that an equivalent analysis is much more complex and thus difficult to accomplish. The wide range of protein concentration complicates matters further. Performing microarray immunoassays already represents a challenge at the level of preparing a working chip surface. Arrays have been produced on filter supports, in microtiter plate wells and on glass slides, the last two usually coated with one-, two- or three-dimensionally structured surface modifications. The usefulness and suitability of all these support media for the construction and application of antibody microarrays are reviewed in this manuscript in terms of the different kinds of immunoassay and the various detection procedures. Additionally, the employment of microarrays containing alternative sensor molecules is discussed in this context. The sensitivity of microspot immunoassays predicted by the current analyte theory is not yet a reality, indicating the extent of both the technology's potential and the size of the task still ahead.

Acrylic Resins↗

[Capsular sack implantation of heparin-modified posterior chamber lenses].

In a prospective study, heparin surface modified posterior chamber lenses were implanted into the capsular bag in 50 patients. The exact positioning of the biconvex, one-piece lens in the bag is the prerequisite for good centration and for an optimal result. The hydrophilisation of the originally hydrophobic PMMA material is achieved by heparin surface modification and results in less inflammation. The aim of this study was to investigate the postoperative behavior of the HSM posterior chamber lens implanted in the bag, with special consideration given to intraocular inflammation. In the postoperative follow-up time of one to seven months, hundred percent of the patients with physiological fundus reached a postoperative visual acuity of 0.7 or better.

Aged↗

Adhesion and proliferation of human endothelial cells on photochemically modified polytetrafluoroethylene.

We studied the adhesion and proliferation of human endothelial cells on photochemically modified polytetrafluoroethylene samples. The polymer surfaces were modified by exposure to the ultraviolet light of a Xe(2)(*)-excimer lamp at a wavelength of 172 nm in an ammonia atmosphere. Treatment times were between 10 and 20 min. The endothelial cell density was determined 1, 3 and 8 days after seeding by image analysis. Surface modification of the samples resulted in a significant increase in the number of adhering cells and in the formation of a confluent cell layer after 3-8 days. The results were comparable than those obtained on polystyrene Petri dishes, which are used as standard substrates in cell cultivation. Thus modified PTFE appears to be a promising material for the fabrication of artificial vascular prostheses coated with endothelial cells.

Biocompatible Materials↗

Preparation of multilayered gold-silica-polystyrene core-shell particles by seeded polymerization.

A preparation method for multilayered gold-silica-polystyrene core-shell composite particles is proposed. The gold-silica core-shell particles of 192-nm-sized, synthesized by coating the 18-nm-sized gold particles with silica by a seeded growth technique, were used as cores for succeeding polystyrene coating. After surface modification of gold-silica composite particles by methacryloxypropyltrimethoxysilane (MPTMS), polymerizations of styrene (0.16-0.4 M) were conducted with 8 x 10(-3) M of potassium persulfate initiator in the presence of 1 x 10(-3) M of sodium p-styrenesulfonate anionic monomer. Multilayered core-shell gold-silica-polystyrene particles that contained a single core could be obtained. The coefficient of variation of size distribution (CV) of the composite particles was less than 7%, and polystyrene shell thickness was in a range of 193 to 281 nm.

Gold↗

Effect of surface composition on triolein hydrolysis in phospholipid vesicles and microemulsions by a purified acid lipase.

Sonicated dispersions of egg yolk phosphatidylcholine and triolein as vesicles and microemulsions have been used as substrates for the assay of a purified acid lipase. Previous studies have also shown that triolein localized in the surface phase of emulsions is the preferred substrate. In this study, we examined enzyme activity following several surface modifications using both vesicles and microemulsions. When the acidic phospholipids phosphatidylserine and phosphatidic acid were incorporated into both vesicles and microemulsions at up to 10 mol % of the total phospholipid, a dose-dependent reduction in the apparent Km was observed. Using the vesicles as substrate, a dose-dependent decrease in Vmax was also observed. Agarose gel electrophoresis was used to verify suspected changes in net particle charge. Analogous inclusion of phosphatidylethanolamine, sphingomyelin, or cholesterol did not affect kinetic parameters. Addition of oleic acid to sonication mixtures produced vesicles with a decreased apparent Km and Vmax, but triolein hydrolysis in microemulsions was not significantly altered. Triolein-containing vesicles prepared by using dimyristoyl- or dipalmitoylphosphatidylcholine were hydrolyzed maximally at the gel liquid-crystalline transition temperatures of the appropriate phospholipid. Differential scanning calorimetry was used to verify the temperatures of transition in these vesicles. The results indicate that acid lipase activity is influenced by the charge or physical state of the surface phase of model substrates and suggest that degradation of core components of naturally occurring substrates such as lipoprotein may be influenced by chemical changes on the surface of these particles.

Animals↗

Phospholipid impregnation of abdominal rubber drains: resistance to bacterial adherence but no effect on drain-induced bacterial translocation.

In order to evaluate the effect of surface modification of biomaterials on bacterial adherence and bacterial translocation after intraperitoneal biomaterial implantation, phosphatidylcholine- or phosphatidylinositol-impregnated rubber drain pieces, which had been intraperitoneally implanted in the rat for 2 and 7 days, or unimplanted, were incubated in vitro with 3H-labelled Escherichia coli and Enterobacter cloacae. As compared with unimpregnated pieces, the adherence of bacteria significantly decreased to phosphatidylcholine- and phosphatidylinositol-impregnated rubber drain pieces that were either unimplanted or implanted for 2 days, but not for 7 days. The supplementation of albumin in the medium reduced the adherence of bacteria to the unimplanted, unimpregnated drain pieces, but did not further decrease adherence of bacteria to the unimplanted, phospholipid-impregnated brain pieces. Bacterial growth was inhibited after incubation in nutrient broth supplemented with phospholipids. The incidence of enteric bacterial translocation induced by intraperitoneal drain implantation did not differ between phospholipid-impregnated and unimpregnated drain pieces. Scanning electron microscopy revealed a large amount of biofilm and fibrous deposition on the surface of the implanted, phospholipid-impregnated rubber drain pieces. Thus, phospholipid impregnation of rubber drains reduces bacterial adherence and inhibits bacterial growth, without influencing the incidence of bacterial translocation.

Abdomen↗

Clay-based polymer nanocomposites: research and commercial development.

This paper reviews the recent research and development of clay-based polymer nanocomposites. Clay minerals, due to their unique layered structure, rich intercalation chemistry and availability at low cost, are promising nanoparticle reinforcements for polymers to manufacture low-cost, lightweight and high performance nanocomposites. We introduce briefly the structure, properties and surface modification of clay minerals, followed by the processing and characterization techniques of polymer nanocomposites. The enhanced and novel properties of such nanocomposites are then discussed, including mechanical, thermal, barrier, electrical conductivity, biodegradability among others. In addition, their available commercial and potential applications in automotive, packaging, coating and pigment, electrical materials, and in particular biomedical fields are highlighted. Finally, the challenges for the future are discussed in terms of processing, characterization and the mechanisms governing the behaviour of these advanced materials.

Aluminum Silicates↗

Edge transfer lithography using alkanethiol inks.

Edge lithographic patterning techniques are based on the utilization of the edges of micrometer-sized template features for the reproduction of submicrometer structures. Edge transfer lithography (ETL) permits local surface modification in a single step by depositing self-assembled monolayers onto a metal substrate selectively along the feature edges of an elastomeric stamp. In this report two stamp designs are described that now allow for the use of alkanethiol inks in ETL and their use as etch resists to reproduce submicrometer structures in gold. Anisotropically modified stamps are shown to combine the potential for very high-resolution patterning with the versatility and simplicity of microcontact printing.

Alkanes↗

The comparison of provisional luting agents and abutment surface roughness on the retention of provisional implant-supported crowns.

STATEMENT OF PROBLEM: In immediate implant loading, it is important to keep provisional restorations in place during early-phase healing. Current luting agents for provisional restorations may provide inadequate retention, creating a clinical challenge. PURPOSE: This study compared the retention of provisional autopolymerizing acrylic resin implant-supported single restorations with combinations of different implant abutment surface conditions and provisional luting agents. MATERIAL AND METHODS: Thirty solid titanium implant abutments (ITI), 4 mm high, were divided into 3 groups. Ten abutments were unaltered, 10 were airborne-particle abraded with 50-microm aluminum oxide, and 10 were roughened with a medium-roughness diamond rotary cutting instrument. Thirty implant analogs (ITI) were mounted in autopolymerizing acrylic resin blocks. A solid titanium implant abutment was placed in each implant analog and torqued to 35 N.cm. After fabrication of 4 provisional acrylic resin crowns for each abutment, provisional luting agents TempBond, TempBond NE, Life, and Zone were used to secure the provisional crowns to the respective abutments. All specimens were luted with one of the provisional luting agents for a given test. After ultrasonic cleaning of the abutments, another provisional crown was luted with another luting agent. All specimens were stored in 100% humidity environment for 1 day at 37 degrees C prior to testing. Each provisional acrylic resin crown was pulled from the abutment with a 500-kg load cell in a universal testing machine at a crosshead speed of 0.5 cm/minute, and tensile strength was recorded (N). Data were analyzed using analysis of variance (ANOVA) and the Scheffe test (alpha=.05). RESULTS: Tensile strength was significantly higher for Life and TempBond NE than for TempBond and Zone, regardless of the surface conditions (P=.0001). The result of the 2-way ANOVA indicated that a significant interaction existed between the provisional luting agents and surface conditions (P=.0039). TempBond NE showed significantly higher tensile strength when combined with airborne-particle-abraded surfaces compared to other combinations (P=.011). However, no difference was found in tensile strength of Life and Zone between different combinations. The tensile strength of TempBond was lower when used with the unaltered surface compared to other combinations (P=.001). CONCLUSION: Surface modification of an implant abutment by airborne-particle abrasion or diamond rotary cutting instrument did not improve retention of a provisional acrylic crown when Life or Zone was used as the luting agent. Airborne-particle abrasion may be an effective method to increase retention of a provisional acrylic crown when TempBond NE is used.

Acrylic Resins↗

Properties of phospholipid monolayer deposited on a fluorinated polyurethane.

A simple procedure for surface modification of polyurethane by the Langmuir-Blodgett (LB) method using the amphiphile 1,2-distearoyl-sn-glycero-3-phosphocholine dihydrate (DSPC) was developed. The polyurethane containing the fluorinated moiety was prepared via the perfluoro-containing chain extender 2,2,3,3-tetrafluoro-1,4-butanediol. The fluorinated polyurethane (FPU) films were prepared by spin coating and dipping methods. The spin-coated FPU films exhibited hydrophobic characteristic and, thus, enhanced the transferability of DSPC. Held at constant pressure of 45 mN/m, the DSPC monolayer was transferred successfully to FPU films with a near-unity transfer ratio. The in vitro platelet adhesion assay revealed that the FPU modified with DSPC monolayer was more platelet compatible than the unmodified FPU substrates with no pseudopods and flattening of adherent platelets as well as lower platelet adhesion density. Moreover, the DSPC monolayer remained intact after platelet adhesion testing. In addition, the platelet compatibility of the unmodified FPU was affected by the film preparation methods. This might be attributed to the distinctive surface micromorphology formed. This simple DSPC deposition scheme by a LB technique would be very useful to further enhance the platelet compatibility of hydrophobic substrate and can be utilized for biomedical application in which the flow shear rate is not too high.

Biocompatible Materials↗

Biomimetic polymer/apatite composite scaffolds for mineralized tissue engineering.

The material surface must be considered in the design of scaffolds for bone tissue engineering so that it supports bone cells adhesion, proliferation and differentiation. A biomimetic approach has been developed as a 3D surface modification technique to grow partially carbonated hydroxyapatite (the bonelike mineral) in prefabricated, porous, polymer scaffolds using a simulated body fluid in our lab. For the rational design of scaffolding materials and optimization of the biomimetic process, this work focused on various materials and processing parameters in relation to apatite formation on 3D polymer scaffolds. The apatite nucleation and growth in the internal pores of poly(L-lactide) and poly(D,L-lactide) scaffolds were significantly faster than in those of poly(lactide-co-glycolide) scaffolds in simulated body fluids. The apatite distribution was significantly more uniform in the poly(L-lactide) scaffolds than in the poly(lactide-co-glycolide) scaffolds. After incubation in a simulated body fluid for 30 d, the mass of poly(L-lactide) scaffolds increased approximately 40%, whereas the mass of the poly(lactide-co-glycolide) scaffolds increased by about 15% (see Figure). A higher ionic concentration and higher pH value of the simulated body fluid enhanced apatite formation. The effects of surface functional groups on apatite nucleation and growth were found to be more complex in 3D scaffolds than on 2D films. Surprisingly enough, it was found that carboxyl groups significantly reduced the apatite formation, especially on the internal pore surfaces of 3D scaffolds. These findings are critically important in the rational selection of materials and surface design of 3D scaffolds for mineralized tissue engineering and may contribute to the understanding of biomineralization as well.SEM micrograph of a poly(L-lactide) scaffold.

Apatites↗

A new strategy toward improving immunoprotection in cell therapy for diabetes mellitus: long-functioning PEGylated islets in vivo.

Surface modification of islets using poly(ethylene glycol) (PEG) has been studied toward preventing immune responses of host for successful islet transplantation. In this study, we assessed the functionality of PEGylated islets and immune responses of host, as well as the synergistic effects of using PEGylation simultaneously with cyclosporine A (CsA) to prevent immune reactions. The average period of time for which PEGylated islets functioned normally following islets transplantation was 14 days, whereas it was only 5 days for unmodified islets. Host's immune cells did not eliminate the PEGylated islets but were observed to gather around the islets. On the other hand, unmodified islets were completely eliminated. When a low dose of CsA (3 mg/kg/day) was intravenously administered at the same time, the PEGylated islets showed stable activity and survived for 100 days before, that is, the PEGylated islets secreted insulin and were preserved from the immune cells. However, unmodified islets survived for 14 days on the average, even when CsA was administered. Consequently, it can be suggested that the PEGylation of islet surfaces can be an effective approach toward reducing the immunogenecity of islets, thus improving the functionality and survival time of transplanted islets, especially when CsA was simultaneously administered.

Animals↗

Electropolymerization of a phenol-modified peptide for use in receptor-ligand interactions studied by surface plasmon resonance.

The combination of surface plasmon resonance (SPR) with an electrochemical method for surface modification is presented. The SLP1 sequence of the sodium channel protein of rat cardiac muscle cells was N-terminally modified with an electropolymerizable group and immobilized on a gold-coated glass slide by oxidative polymerization. The resulting peptide-functionalized substrate was incubated with a polyclonal-specific anti-SLP1 serum. Growth of the peptide layer and the immunological reaction between ligand and receptor were detected on-line by SPR. The applicability of this approach for the rapid and selective analysis of receptor-ligand interactions is demonstrated.

Amino Acid Sequence↗

Poly(ethylene oxide)-modified poly(beta-amino ester) nanoparticles as a pH-sensitive system for tumor-targeted delivery of hydrophobic drugs: part 2. In vivo distribution and tumor localization studies.

PURPOSE: This study was carried out to determine the biodistribution profiles and tumor localization potential of poly(ethylene oxide) (PEO)-modified poly(beta-amino ester) (PbAE) as a novel, pH-sensitive biodegradable polymeric nanoparticulate system for tumor-targeted drug delivery. METHODS: The biodistribution studies of PEO-modified PbAE and PEO-modified poly(epsilon-caprolactone) (PCL), a non-pH-sensitive polymer, nanoparticle systems were carried out in normal mice using 111indium-oxine [111In] as a lipophilic radiolabel encapsulated within the polymeric matrix, and the distribution of the nanoparticles was studied in plasma and all the vital organs following intravenous administration. Solid tumors were developed on nude mice using human ovarian carcinoma xenograft (SKOV-3) and the change in concentrations of tritium [3H]-labeled paclitaxel encapsulated in polymeric nanoparticles was examined in blood, tumor mass, and liver. RESULTS: Study in normal mice with a gamma-emitting isotope [111In] provided a thorough biodistribution analysis of the PEO-modified nanoparticulate carrier systems, whereas 3H-paclitaxel was useful to understand the change in concentration and tumor localization of anticancer compound directly in major sites of distribution. Both PEO-PbAE and PEO-PCL nanoparticles showed long systemic circulating properties by virtue of surface modification with PEO-containing triblock block copolymer (Pluronic stabilizer. Although the PCL nanoparticles showed higher uptake by the reticuloendothelial system, the PbAE nanoparticles effectively delivered the encapsulated payload into the tumor mass. CONCLUSIONS: PEO-modified PbAE nanoparticles showed considerable passive tumor targeting potential in early stages of biodistribution via the enhanced permeation and retention (EPR) mechanism. This prompts a detailed biodistribution profiling of the nanocarrier for prolonged periods to provide conclusive evidence for superiority of the delivery system.

Amino Acids↗

The complement- but not mannose receptor-mediated phagocytosis is involved in the hepatic uptake of cetylmannoside-modified liposomes in situ.

In the elimination of injected liposomes in vivo, it is considered that several serum components play an important role on hepatic uptake of them. This study was conducted to clarify the hepatic uptake mechanism of cetylmannoside (Man)-modified multilamellar vesicles (Man-MLV) using perfused rat liver. In the presence of serum, Man-MLV was taken up by the liver depending on the serum concentration, and it showed an approximately two-fold higher accumulation than MLV without any surface modifications (PC-MLV). These hepatic uptakes of liposomes were obviously inhibited by preheating the serum at 56 degrees C for thirty minutes or by the treatment with anti-rat C3 antiserum. Further, SDS-PAGE followed by immunoblot analysis showed the deposition of iC3b on the opsonized Man-MLV. These results obtained in the present study suggested that hepatic uptake of Man-MLV was mainly mediated by complement receptor rather than mannose receptor on Kupffer cells in vivo.

Animals↗

Topographic control of lipid-raft reconstitution in model membranes.

Liquid-ordered (L(O)) domains reconstituted in model membranes have provided a useful platform for in vitro studies of the lipid-raft model, in which signalling membrane molecules are thought to be compartmentalized in sphingolipid- and cholesterol-rich domains. These in vitro studies, however, have relied on an uncontrolled phase-separation process that gives a random distribution of L(O) domains. Obviously, a precise control of the size and spatial distribution of the L(O) domains would enable a more systematic large-scale in vitro study of the lipid-raft model. The prerequisite for such capability would be the generation of a well-defined energy landscape for reconstituting the L(O) domain without disrupting the two-dimensional (2D) fluidity of the model membrane. Here we report controlling the reconstitution of the L(O) domains in a spatially selective manner by predefining a landscape of energy barriers using topographic surface modifications. We show that the selective reconstitution spontaneously arises from the 2D brownian motion of nanoscale L(O) domains and signalling molecules captured in these nanodomains, which in turn produce a prescribed, concentrated downstream biochemical process. Our approach opens up the possibility of engineering model biological membranes by taking advantage of the intrinsic 2D fluidity. Moreover, our results indicate that the topographic configuration of cellular membranes could be an important machinery for controlling the lipid raft in vivo.

Biophysics↗

Synthesis of amphiphilic superparamagnetic ferrite/block copolymer hollow submicrospheres.

Self-assembly can be a powerful, but simple, synthetic method for the fabrication and surface modification of nanometer- to micrometer-sized hollow spheres. Here we report a facile route for preparation of submicrometer ferrite hollow spheres which are amphiphilic and superparamagnetic. This unique approach involves the formation of ferrite nanocrystals and the simultaneous self-assembly of nanocrystals and block copolymer PEO-PPO-PEO into hollow spheres. Furthermore, this approach is general for the preparation of a series of ferrite hollow spheres, including Fe3O4, Co1-xFe2+xO4, and Mn1-xFe2+xO4. Unlike conventional hollow spheres, which are either hydrophobic or hydrophilic, the products we obtained exhibit excellent dispersibility in both polar and nonpolar solvents.

Ferric Compounds↗