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Surface modifications at the periosseous region of chick osteoclast as revealed by freeze-substitution.

Improved preservation of osteoclast fine structure can be achieved by quick freezing, freeze-substitution, or detergent extraction. With such techniques the ruffled border mainly contains a disorganized, interconnected meshwork of microfilaments (5-7 nm in diameter), whereas in the clear zone a few ordered arrays of intermediate-type filaments (10-12 nm in diameter) are detectable among the network of microfilaments. In well-frozen samples, well-preserved matrix may have occluded the cytoskeleton; detergent extraction permits visualization of the cytoskeletal components. In fresh-frozen cells an extracellular fuzzy coat overlays the ruffled border. At the site of attachment of the clear zone to the bone surface, extracellular cementing material is detected only after quick freezing. The superiority of quick freezing to preserve ultrastructure is shown in various cytoplasmic organelles. Most vesicles and vacuoles found close to the ruffled border seemed not to make contact with the extracellular matrix. Anhydrous procedures using quick freezing and freeze-substitution stabilize bone mineral in some vacuoles and in the channels of the ruffled border.

Animals↗

Surface modification of oxidic nanoparticles using 3-methacryloxypropyltrimethoxysilane.

Tin oxide, antimony-doped tin oxide (ATO), and silica nanosized particles in aqueous dispersion were reacted with various amounts of 3-methacryloxypropyltrimethoxysilane (MPS). The kinetics were followed by 29Si NMR and the products were analyzed by FTIR and 29Si NMR. The kinetic experiments on ATO and silica revealed that the hydrolysis is the rate-determining step in these reactions. The reaction of MPS with the particles is favored over the homocondensation of MPS. Quantitative analysis using FTIR revealed that the amount of MPS grafted onto the tin oxide and silica particles is limited to the amount needed to fill one monolayer. For ATO the maximum amount of grafted MPS was only 50-70% of the amount that is needed for a closed monolayer. The MPS molecules are for the most part oriented parallel to the oxide surface, and a hydrogen bond between the MPS-carbonyl and the oxide is formed.

Journal Article↗

Surface modification without desorption: recycling of Cl on Si(100)-(2 x 1).

We demonstrate chlorine-induced modification of Si(100)-(2 x 1) under conditions where Cl is recycled rather than desorbed as SiCl2. A dimer with 2 Cl atoms, 2SiCl, converts to SiCl2+Si, allowing the bare Si atom to escape onto the terrace. At temperatures below the desorption threshold, the SiCl2 unit decays through Cl diffusion, allowing the second Si atom to escape. The result is a dimer vacancy, terrace regrowth structures, and Cl that is able to participate in another pitting event. Access to this unexpected roughening pathway is controlled by the Cl concentration and temperature. This previously overlooked process represents an important component of Si(100) surface processing.

Journal Article↗

Novel core(polyester)-shell(polysaccharide) nanoparticles: protein loading and surface modification with lectins.

This study describes new lectin-decorated or protein-loaded nanoparticles with a hydrophobic poly(epsilon-caprolactone) (PCL) core and a hydrophilic dextran (Dex) corona. In this view, a family of block Dex-PCLn copolymers was first synthesized, consisting of a Dex backbone to which n preformed PCL blocks were grafted. The ability of these new copolymers to form nanoparticles was evaluated in comparison with a series of PCL homopolymers of various molecular weights (2000, 10,000 and 40,000 g/mole). Two different nanoparticle preparation methods have been developed and tested for their efficacy to incorporate proteins. For this, three proteins were used: a model protein, bovine serum albumin (BSA), a lectin from leaves of Bauhinia monandra (BmoLL) and Lens culinaris (LC) lectin. All these proteins were successfully incorporated in nanoparticles with a mean diameter around 200 nm. Lectins could also be adsorbed onto the surface of Dex-PCLn nanoparticles. Surface-bound BmoLL conserved its hemagglutinating activity, suggesting the possible application of this type of surface-modified nanoparticles for targeted oral administration. Caco-2 cellular viability was higher than 70% when put in contact with Dex-PCLn nanoparticles, even at concentrations as high as 660 microg/ml.

Animals↗

Surface-modification of polystyrene-microtitre plates via grafting of glycidylmethacrylate and coating of poly-glycidylmethacrylate.

Photografting of glycidylmethacrylate (GMA) onto commercially available polystyrene-microtitre plates was carried out in methanol as well as butanol with benzophenone (BP) as photoinitiator. Alternatively, prepolymers of polyglycidylmethacrylate (PGMA) were synthesized in methanol with azo-iso-butyrodinitrile (AIBN) as initiator and dip-coated onto polystyrene-microtitre plates. Both modification methods were tested in order to reach a high binding capacity for proteins. Peroxidase was used as model protein. In addition, the immobilization of myelin basic protein (MBP) to epoxy-modified microtitre plates is shown and a MBP-based ELISA has been developed.

Biocompatible Materials↗

The inhibitory mechanism of gap junctional intercellular communication induced by polyethylene and the restorative effects by surface modification with various proteins.

Gap junctional intercellular communication (GJIC) is a function that plays an important role in maintaining cell and tissue homeostasis and in regulating cell growth, development, and differentiation. Change in this function of V79 fibroblasts cultured on polyethylene films modified with albumin or collagen was estimated using fluorescence redistribution after photobleaching (FRAP) analysis. The GJIC function of V79 cells on nontreated polyethylene was strongly inhibited in comparison with those on a glass coverslip. When the cells were culture on collagen-immobilized polyethylene film, this function was recovered to about 70% of the cells cultured on the coverslip. However, albumin immobilization did not recover the function as much as collagen immobilization. Western blotting analysis and immunostaining of connexin 43, which is a major protein constituting gap junctional channel of these cells, revealed its abnormal expression and distribution in the cells on nontreated polyethylene, whereas its almost normal distribution was observed in the cells on collagen-immobilized polyethylene. This abnormal expression and distribution of connexin 43 induced by the surface of polyethylene may be ascribed to a strong inhibition of GJIC of V79 fibroblasts.

Albumins↗

Surface modification of titanium based biomaterials by ion beam.

Ion beam enhanced deposition (IBED) was adopted to synthesize biocompatible titanium oxide film. Structure characteristics of titanium oxide film were investigated by RBS, AES, and XRD. The blood compatibility of the titanium oxide film was studied by measurements of blood clotting time and platelet adhesion. The results show that the anticoagulation property of titanium oxide film is improved significantly. The mechanism of anticoagulation of the titanium oxide film was discussed.

Animals↗

Surface modification of proteins. Activation of monomethoxy-polyethylene glycols by phenylchloroformates and modification of ribonuclease and superoxide dismutase.

A single-step method of activation of monomethoxypolyethylene glycols suitable for its binding to polypeptides and proteins is proposed. Based on the reaction with 2,4,5-trichlorophenylchloroformate or p-nitrophenylchloroformate, it gives reactive PEG-phenylcarbonate derivatives. The PEG intermediate is stable on storage, the activating group is easily quantified,and the reaction with amino acid and proteins proceeds rapidly at pH near neutrality. The PEG derivatization of enzymes with this procedure is less inactivating than those previously reported. Ribonuclease and superoxide dismutase were modified and the effect of (a) bound polymer on clearance time in rats, (b) antibody recognition, and (c) on the enzymatic activity toward low and high molecular weight substrates were studied.

Amino Acids↗

Gelatin nanoparticles by two step desolvation--a new preparation method, surface modifications and cell uptake.

A new two-step desolvation method for manufacturing gelatin nanoparticles was developed. After the first desolvation step, the low molecular gelatin fractions present in the supernatant were removed by decanting. The high molecular fractions present in the sediment were redesolved and then desolvated again at pH 2.5 in the second step. The resulting particles can then be easily purified by centrifugation and redispersion. The different fractions obtained during the process were analysed by gel permeation chromatography (GPC). Based on these results, it can be concluded that the molecular weight of gelatin has a decisive influence on the stability of the manufactured gelatin nanoparticles. In addition, two fluorescent dyes (Texasred and fluoresceinamine) were coupled to the nanoparticles for cell uptake studies. The fluorescent nanoparticles showed a high uptake into monocytes/macrophages.

Cells, Cultured↗

Novel surface-modification techniques for polymer-based separation media. Stimulus-responsive phenomena based on double polymeric selectors.

A pair of polymeric selectors potentially responding to stimulation was introduced onto monosized porous polymer particles to be evaluated as a packing material for HPLC. Possible complexes formed between polyacrylamide (PAAm) and poly(methacrylic acid) (PMAA) were utilized as stimulus responsive polymeric selectors. Uniformly sized base polymer particle was prepared by multi-step swelling and polymerization method, while the introduction of PAAm and PMAA was done by newly invented modification technique. In this technique, a solvent in which both acrylamide (AAm) and methacrylic acid (MAA) monomers are soluble, but PAAm and PMAA are insoluble, was utilized as a modification medium. The polymer particle doubly modified with PAAm and PMAA was utilized as packing material for HPLC and the stimulus responses were evaluated by changing temperature or pH to check change of the slope of a Van't Hoff plot. By using water as a mobile phase, the expected inflection point of the Van't Hoff plot was observed at upper critical solution temperature (UCST) of the polymer complexes and the temperature responsive ability was observed. Moreover, pH responsive ability was studied by using buffer of either pH 4 or 10 as mobile phase. Slope of the plot was changed in buffer of pH 4, but no change of slope was observed in the buffer of pH 10.

Chromatography, High Pressure Liquid↗

Surface modifications evoked by estradiol and diethylstilbestrol in isolated endometrial cells: evidence from lectin probes and extracellular release of lysosomal protease.

Endometrial cells were isolated from the uteri of ovariectomized rats. The inhibitory effect of alpha-methyl-D-mannoside on fluorescein-labeled concanavalin A (Con A) binding to these cells indicates that they possess specific binding sites for Con A. The lectin also mediates adsorption of homologous erythrocytes to these cells. Both Con A binding and Con A-mediated hemadsorption to endometrial cells are depressed at 4 C compared with these functions in cells maintained at 22 C. Gross elevations in lectin-mediated hemadsorption to endometrial cells are evident following prior exposure to 1 X 10(-9)M concentrations of diethylstilbestrol (DES) or estradiol-17beta, but not to the physiologically inactive 17 alpha-epimer, at 22 C. The enhancement of hemagglutinability cannot be attributed to a corresponding increase in lectin binding at 22 C. Although estrogen treatment elicited significant increments in Con A binding as early as 5 min after addition of estrogen to cell suspensions, the increment in agglutination attributable to hormone treatment consistently ranged from 1.5-3 times greater than the increase in lectin binding. These estrogenic effects were reduced by incubation of the endometrial cells at 4 C or when cortisol, 3 X 10(-6)M, was present with estradiol-17beta. In parallel experiments, treatment with DES and estradiol-17beta, but not estradiol-17 alpha, also enhanced the release of cathepsin B 1 and acid phosphatase from uterine segments into the particle-free extracellular media in which the tissues had been incubated for 30-60 min. The marked increment in the extracellular activity of the lysosomal hydrolases induced by estrogen treatment was suppressed in cells incubated at 4 C or when cortisol was present concomitantly. These and related data suggest the hypothesis that acute increments in lysosomal hydrolase activity may contribute to cell surface alterations which have been described in both normal and aberrant processes of cell growth.

Acid Phosphatase↗

Lymphatic uptake and biodistribution of liposomes after subcutaneous injection: III. Influence of surface modification with poly(ethyleneglycol).

PURPOSE: The aim of the present paper was to assess the effect of inclusion of distearoylphosphatidylethanolamine-poly(ethyleneglycol) (DSPE-PEG) into liposomal bilayers on the lymphatic uptake and lymph node localization of liposomes after subcutaneous administration. METHODS: [3H]-Cholesteryloleylether labeled liposomes of various composition and sizes were injected s.c. into the dorsal side of the foot of rats. At several time-points after injection, blood levels of liposomes were determined. Lymphatic uptake from the s.c. site of injection and lymph node localization in regional lymph nodes were determined at the end of the 52 h observation period. RESULTS: The results demonstrate that inclusion of DSPE-PEG into several types of liposomes has only a modest effect on lymphatic uptake. Also lymph node localization is only slightly affected by PEG-mediated steric stabilization. CONCLUSIONS: Factors other than the presence of a steric barrier are more important in determining lymphatic uptake from the s.c. injection site. The observation that lymph node localization was only slightly affected by PEG-coating strongly suggests that macrophage uptake is not the only important mechanism of lymph node localization of s.c. administered liposomes.

Animals↗

Irradiation effects in MALDI, ablation, ion production, and surface modifications. Part II. 2,5-dihydroxybenzoic acid monocrystals.

Irradiation effects at low and high laser fluence on 2,5-dihydroxybenzoic acid large crystals were investigated. Contrary to what was observed for matrices as cinnamic acid derivatives, no chemical degradation of matrix is evidenced and continuous ablation as well as ion production resulted of extended irradiation in all the fluence range corresponding to classical matrix-assisted laser desorption /ionization. Ripples are formed on the base of the crater for a limited number of laser shots under moderate fluence. For extended irradiation, conical shape craters are formed with the axis of the crater oriented along the incident direction of the laser beam. A study of the craters showed that ablation through the ablated volume slowly varied with the laser fluence when a strong increase of ion production (matrix and analyte) was recorded. Ablation volume was found to vary non-linearly with the number of laser shots. On a same spot, the ablated volume and the ion production were measured as a function of the laser energy. With an increasing laser energy (or fluence), the ablated volume slowly increases when the ion production strongly increases. This gives evidence of a decoupling between ablation and ionization. Interaction of the plume with the incoming beam is thus probable.

Crystallization↗

Surface modification of extrasynovial tendon by chemically modified hyaluronic acid coating.

We investigated a method for chemically binding hyaluronic acid (HA) to extrasynovial tendon and the effect of chemically modified HA on the gliding resistance of tendon. Canine peroneus longus (PL) tendons were immersed into one of three different solutions (saline, 1% HA, or 1% chemically modified HA) for 2 h. The gliding resistance of treated PL tendons was measured at 1, 5, 10, 20, 50, and 100 cycles in a saline bath. After treatment with unmodified HA and chemically modified HA, the gliding resistance of the PL tendons decreased significantly compared with the saline-treated tendons (p < 0.05), and this effect of the two HA treatments persisted through 10 cycles. For cycles 20-100, the gliding resistance of PL tendons treated with chemically modified HA remained significantly lower than that of tendons treated either with saline or unmodified HA (p < 0.01). The effect of paratenon removal on gliding resistance was neither statistically significant for repetitions beyond 10 cycles, nor was it an independent predictor of gliding resistance, as the effect (higher resistance after paratenon removal) was mainly seen in the saline-treated tendons. Chemically modified HA-treated extrasynovial tendons may improve gliding of tendon graft and reduce adhesion postoperatively, compared with traditional grafts.

Animals↗

A new porous surface modification technology for peritoneal dialysis catheters as an exit-site cuff to reduce exit-site infections.

Catheter exit-site infection continues to be a more common morbid event in patients undergoing peritoneal dialysis. Previous attempts to place a biointegration material at the next site have failed to reduce infection rates. This study reports the use of an innovative microporous silicone material placed as a cuff around the catheter at the exit site. The porous material has a pore-sized distribution that stimulates and facilitates capillary ingrowth into the pores. This capillary ingrowth prevents scar tissue formation, increases blood supply, and theoretically improves the immunological competence of the tissue at the vulnerable exit site. Twenty-five test catheters (12 using standard exit-site creation and 13 using the Moncrief-Popovich implantation technique) were implanted in a canine model. The exit-site infection rate in a canine model without the microporous material was 100% at 2 months. The corresponding results with the microporous material was 40% at 2 months. The majority of the test catheters showed progressive drying and healing at the exit site. Sixty percent of the catheters healed quickly and remained infection-free. Biointegration of the microporous material at the exit-site was demonstrated. Several exit site infections with the test catheters treated only with local therapy (without systemic or topical antibiotics) demonstrated progressive healing and secondary adequate biointegration. Because of these encouraging results, human studies were initiated, with the first human implant occurring in August, 1994. A 10-patient project is planned for the next year.

Animals↗

Amphiphilic poly-N-vinylpyrrolidones: synthesis, properties and liposome surface modification.

Certain amphiphilic water-soluble polymers including amphiphilic derivatives of polyvinyl pyrrolidone (PVP) were found to be efficient steric protectors for liposomes in vivo. In this study, we have tried to develop synthetic pathways for preparing amphiphilic PVP and to investigate the influence of the hydrophilic/hydrophobic blocks on some properties of resulting polymers and polymer-coated liposomes. To prepare amphiphilic PVP with the end stearyl (S) or palmityl (P) residues, amino- and carboxy-terminated PVP derivatives were first synthesized by the free-radical polymerization of vinyl pyrrolidone in the presence of amino- or carboxy-mercaptans as chain transfer agents, and then modified by interaction of amino-PVP with stearoyl chloride or palmitoyl chloride, or by dicyclohexyl carbodiimide coupling of stearylamine with carboxy-PVP. ESR-spectra of the hydrophobic spin-probe, nitroxyl radical N-oxyl-2-hexyl-2-(10-methoxycarbonyl)decyl-4,4'-dimethyl oxazoline, in the presence of amphiphilic PVP demonstrated good accessibility of terminal P- and S-groups for the interaction with other hydrophobic ligands. Spontaneous micellization and low CMC values (in a low micromolar range) were found for amphiphilic PVP derivatives using the pyrene method. In general, S-PVP forms more stable micelles than P-PVP (at similar MW, CMC values for S-PVP are lower than for P-PVP). It was found that amphiphilic PVP incorporated into negatively charged liposomes effectively prevents polycation(poly-ethylpyridinium-4-vinylchloride)-induced liposome aggregation, completely abolishing it at ca. 10 mol% polymer content in liposomes. Additionally, the liposome-incorporated PVP prevents the fluorescence quenching of the membrane-incorporated hydrophobic fluorescent label [N-(4-fluoresceinthiocarbamoyl)dipalmitoyl-PE] by the free polycation. PVP-modified liposomes were loaded with a self-quenching concentration of carboxyfluorescein, and their destabilization in the presence of mouse serum was investigated following the release of free dye. Amphiphilic PVP with MW between 1,500 and 8,000 provides good steric protection for liposomes. The degree of this protection depends on both polymer concentration and molecular size of the PVP block.

Animals↗

Polymeric surface modifications of tantalum stents.

PURPOSE: To compare two kinds of polymer-coated tantalum stents with bare tantalum stents (control) to determine if the coatings can improve thromboresistance. METHODS: Twenty-seven Fontaine-Dake stents were balloon expanded in three 8-mm x 80-cm polytetrafluoroethylene (PTFE) grafts; 9 stents were bare tantalum (T); 9 were coated with polyetherurethane (PL); and 9 were coated with parylene (PA). There were 9 stents placed in each graft as follows: 3 tantalum, 3 polyetherurethane, and 3 parylene. In swine whose platelets had been radiolabeled with indium 111, the ends of each stented graft were connected to 14F femoral and venous sheaths to create an ex vivo fistula. Each graft was exposed to blood for 30, 60, and 120 minutes. At the end of each test period, the stented grafts were disconnected from the sheaths, flushed with saline until clear, and then flushed with formalin. The stents were removed from the grafts, and a radionuclide well counter recorded radionuclide counts from each stent type at each period of blood contact. These values were converted to platelet density per 1000 microns 2. Stents were then photographed and scanned with electron microscopy (EM) for qualitative analysis. Possible significant differences in platelet adhesion with the three types of stents (both between stent groups and within stent groups) were examined using a two-tailed Student's t-test. RESULTS: There were significantly fewer platelets adsorbed on PA versus T at all time periods (p < 0.005); on PL versus T at 60 and 120 minutes (p < 0.005); and on PA versus PL at 30 and 120 minutes (p < 0.0005). There was no significant difference in platelet density within each stent group (p = 0.1). Mean platelet density (number of platelets per 1000 microns 2 +/- SD) was as follows: at 30 minutes: T = 1891 +/- 965; PL = 373 +/- 193; and PA = 27 +/- 3; at 60 minutes: T = 6226 +/- 1621; PL = 1573 +/- 793; and PA = 1185 +/- 710; at 120 minutes: T = 5307 +/- 591; PL = 3164 +/- 318; and PA = 180 +/- 100. Gross inspection of the 120-minute groups demonstrated focal areas of thrombus on T, less on PL, and none on PA. Scanning EM demonstrated extensive platelet accumulation covering T at all time periods, less on PL, and even less on PA.

Animals↗