Correction to: Reply to the letter: N-Butyl Cyanoacrylate-Lipiodol Mixture for Endovascular Purpose: Polymerization Kinetics Differences Between In Vitro and In Vivo Experiments.

We would like to correct one of the references that was listed incorrectly in our article.

Is the Cessation of Blood Flow Faster than the Polymerization of an n-Butyl Cyanoacrylate-Lipiodol Mixture? An In Vitro Phantom Study.

PURPOSE: To compare the polymerization time of n-butyl cyanoacrylate (NBCA) and lipiodol mixture in a static model and a pulsating flow model simulating embolization procedure of small caliber arteries. MATERIALS AND METHODS: The polymerization time of NBCA-lipiodol mixture was measured by the morphological changes of a glue droplet in a petri dish. For the flow model, we used a 2-mm-inner-diameter polyvinyl alcohol tube connected to a pulsation pump. Bovine serum was supplied from the pump and circulated into the system at 30 ml/min and 60 bpm. A 0.64-mm-inner-diameter silicon microcatheter was inserted into this system, and then, 0.5 ml of glue was injected into the tube. The flow cessation time was defined as the time it took to stop the serum draining from the end of the tube. Six samples of 100, 66, 50, 40, 33, and 20 vol% NBCA were assessed. RESULTS: The median polymerization times for each concentration were 0.12, 3.72, 12.30, 27.41, 57.68, and 63.67 s, respectively. The median flow cessation times were 0.28, 0.78, 1.43, 3.75, 4.50, and 9.29 s, respectively. The flow cessation time was significantly shorter than the polymerization time for all samples except for 100 vol% cyanoacrylate (p < 0.05). CONCLUSION: The flow cessation time of cyanoacrylate glue was significantly shorter than the polymerization time in an in vitro experiment. The injected glue possibly stops the blood flow before the completion of polymerization in the vascular system.

Surface plasmon field-enhanced fluorescence spectroscopy apparatus with a convergent optical system for point-of-care testing.

Surface plasmon field-enhanced fluorescence spectroscopy (SPFS) is a promising methodology for point-of-care (POC) testing. The SPFS devices that have been reported are equipped with an angle rotating stage to adjust the surface plasmon resonance (SPR) angle. In a clinical setting, however, the SPR angle determination is a tedious and time-consuming process. In this study, we employed an SPFS instrument with a convergent optical system that allows the omission of this procedure. We demonstrated that this instrumentation allowed the sensitive determination of low concentrations of α-fetoprotein in serum and reduced the variation effect caused by the protein concentrations in samples. The SPFS with a convergent optical system is suitable for POC testing.

Effect of dielectric spacer thickness on signal intensity of surface plasmon field-enhanced fluorescence spectroscopy.

Surface plasmon field-enhanced fluorescence spectroscopy (SPFS) combines enhanced field platform and fluorescence detection. Its advantages are the strong intensity of the electromagnetic field and the high signal/noise (S/N) ratio due to the localized evanescent field at the water/metal interface. However, the energy transfer from the fluorophore to the metal surface diminishes the fluorescence intensity, and this reduces the sensitivity. In this study, we tested whether polystyrene (PSt) could act as a dielectric layer to suppress the energy transfer from the fluorophore to the metal surface. We hypothesized that this would improve the sensitivity of SPFS-based immunoassays. We used α-fetoprotein (AFP) as a model tumor biomarker in the sandwich-type immunoassay. We determined the relationship between fluorescent signal intensity and PSt layer thickness and compared this to theoretical predictions. We found that the fluorescence signal increased by optimally controlling the thickness of the PSt layer. Our results indicated that the SPFS-based immunoassay is a promising clinical diagnostic tool for quantitatively determining the concentrations of low-level biomarkers in blood samples.

Surface plasmon resonance in monitoring of complement activation on biomaterials.

When artificial materials come into contact with blood, various biological responses are induced. For successful development of biomaterials used in biomedical devices that will be exposed to blood, understanding and control of these interactions are essential. Surface plasmon resonance (SPR) spectroscopy is one of the surface-sensitive optical methods to monitor biological interactions. SPR enables real-time and in situ analysis of interfacial events associated with biomaterials research. In this review, we describe an SPR biosensor and its application to monitor complement activation onto biomaterials surface. We also discuss the effect of surface properties of the material on complement activation.

Coils coated with the cyclic peptide SEK-1005 accelerate intra-aneurysmal organization.

BACKGROUND: Although the treatment of intracranial aneurysms with detachable coils is now widely accepted, the problem of coil compaction and recanalization remains to be solved. OBJECTIVE: To prevent recanalization by inducing intra-aneurysmal organization through prepared platinum coils coated with a novel cyclic peptide, SEK-1005, which can accelerate wound healing. METHODS: Using a rat aneurysm model, we examined the tissue response to these coils. An SEK-1005-coated coil (SC) or unmodified coil was inserted into the ligated external carotid artery (ECA) sac of rats. The sacs were removed on day 14 or 42 after coil insertion and subjected to conventional and immunohistochemical examination. We evaluated the tissue response in the ECA sacs and compared the percentage of organized areas in the ECA sacs of rats with SCs and unmodified coils. RESULTS: In SC rats, tissue organization was accelerated and the proliferation of α-smooth muscle actin- and vimentin-positive cells was promoted. On days 14 and 42, tissue organization was significantly greater in the ECA sacs of rats with SCs. CONCLUSION: SCs accelerated intra-aneurysmal organization in our rat aneurysm model suggesting that platinum coils coated with the novel cyclic peptide SEK-1005 may prevent recanalization and improve the clinical outcome in patients treated by coil embolization.

Simple immersion of filter devices into an urokinase solution prevents fibrin net formation during carotid artery stenting.

Slow-flow phenomenon is frequently observed during carotid artery stenting (CAS) with a filter embolic protection device. It results in technical difficulties and can lead to adverse neurological events. Flow impairment is thought to be caused by plaque entrapped by the filter and/or blood coagulation on the filter. Characteristics of heparin- or urokinase-treated polyurethanes were analyzed by surface plasmon resonance, and the fibrinolytic activity of the urokinase-treated filter of Angioguard XP was estimated by the fibrin plate assay. A filter membrane of Angioguard XP protection device was treated with a heparin or urokinase solution. In clinical studies, six and nine patients were treated by CAS using Angioguard XP modified with heparin and urokinase, respectively. Filter membranes were examined by scanning electron microscopy (SEM). From in vitro studies, it appeared that urokinase adsorbed and remained on the Angioguard XP filter, and its fibrinolytic activity was demonstrated even after washing with saline; heparin, however, was easily washed out from the surface. From clinical study, some filter pores were obstructed in all six patients in the heparin group and in three patients in the urokinase group. Fibrin net was found on the filter in five of six patients in the heparin group and in one of nine patients in the urokinase group. Treatment of an Angioguard XP filter with a urokinase solution is effective in preventing pore occlusion and may reduce occurrence of the slow-flow phenomenon.

Effect of swelling of poly(vinyl alcohol) layers on complement activation.

Polymers carrying hydroxyl groups have the potential ability to activate the complement system when in contact with blood. However, the effects of their surface structure on complement activation are still not fully understood. In this study, we examined complement activation by poly(vinyl alcohol) (PVA) layers formed on a gold surface modified with aldehyde groups. The complement system was strongly activated by a PVA surface with a dry thickness of 2.9 nm, while it was poorly activated by a PVA surface with a dry thickness of 7.4 nm. Annealing of the latter for 2 h at 150 degrees C converted the surface into a complement activating surface. The difference in complement activation between PVA layers was associated with the water content of PVA layers. These results suggest that complement activation by hydrated polymers highly depends on the water content of the polymer layers.

Effects of hydrophobicity and electrostatic charge on complement activation by amino groups.

Some studies have demonstrated that amino groups, acting as nucleophiles, are potent activators of the complement system, but others not. To clarify these contradictory results, we examined complement activation on two series of NH(2)/CH(3) and NH(2)/COOH mixed self-assembled monolayers (SAMs). NH(2)/CH(3) mixed SAMs were not potent activators of the complement system regardless of the ratio of NH(2)/CH(3) in mixed SAMs. Numerous serum proteins, such as albumin, were adsorbed onto those SAMs and formed a protein layer which inhibited access of C3b to amino groups. In contrast, much C3b and/or C3bBb were deposited on NH(2)/COOH mixed SAMs with approximately 50-60% NH(2) density on the surface and SC5b-9 was found in serum exposed to this SAM, indicating activation of the complement system. These results suggest that C3b can easily access nucleophilic NH(2) groups because of the decrease in electrostatic interaction between negatively charged proteins and the NH(2) SAM surface.

Complement activation on degraded polyethylene glycol-covered surface.

Surface modification with polyethylene glycol (PEG) has been employed in the development of biomaterials to reduce unfavorable reactions. However, unanticipated body reactions have been reported, with activation of the complement system being suggested as having involvement in these responses. In this study, we prepared a PEG-modified surface on a gold surface using a monolayer of alpha-mercaptoethyl-omega-methoxy-polyoxyethylene. We observed neither protein adsorption nor activation of the complement system on the PEG-modified surface just after preparation. Storage of the PEG-modified surface in a desiccator under ambient light for several days or following ultraviolet irradiation, reflection-adsorption (FTIR-RAS) and X-ray photo spectrometry revealed deterioration of the PEG layer, which became a strong activator of the complement system through the alternative pathway.

Development of nanofiber-covered stents using electrospinning: in vitro and acute phase in vivo experiments.

There are some technical difficulties in treating for a broad necked aneurysm and a higher incidence of recurrence. Because of these drawbacks, more innovative techniques for superior endovascular reconstructive treatment are required. We developed a novel covered stent employing electrospinning to deposit fine polyurethane (PU) fibers onto stents. An in vitro water leak test was designed and applied prior to animal testing to estimate the performance of covered stents and to determine the appropriate amount of PU fibers on a stent. Two tenths of a milligram of PU fibers proved to be sufficient to prevent water leakage. Then, the efficacy of the covered stents to that of bare stents was compared using 10 rabbits in which model aneurysms had been formed at the right common carotid artery by the elastase method. Angiographic evaluation on day 1 posttreatment (acute phase) revealed complete occlusion of the aneurysms and the patency of the parent arteries in animals treated with covered stents. At 10 days poststenting (subacute phase), the aneurysm neck was completely covered with neointimal layer as shown by scanning electron microscopic examination. The PU-covered stent holds promise as a device for treating cerebral aneurysms.

Complement activation by polymers carrying hydroxyl groups.

Hydrogels of polymers carrying surface hydroxyl groups strongly activate the complement system through the alternative pathway, although it has also been reported that solutions of polymers do not. To address these curious, inconsistent results, we examined the effect of polymer states, either immobilized on a surface or soluble in serum, on the complement activation using a surface plasmon resonance apparatus and enzyme-linked immunosorbent assay. We clearly showed that dextran- and poly(vinyl alcohol)-immobilized surfaces strongly activated the complement system but that soluble polymers could not, even when the amounts of the soluble polymers added to serum were 4-2000 times higher than those on the polymer-immobilized surfaces.

Complement activation on surfaces carrying amino groups.

The complement system is strongly activated by surfaces carrying nucleophilic groups, such as hydroxyl (OH) groups, and triggered by deposition of complement protein fragment, C3b. Surfaces carrying amino groups, the other representative nucleophilic group, are expected to be potential activators of the complement system through the alternative pathway. Few studies thus far have examined the potential of artificial materials carrying amino groups in activating the complement system. In this study, we employed a self-assembled monolayer (SAM) of 11-amino-1-undecanethiol (NH2-SAM) and a polyethyleneimine (PEI)-coated surface as model surfaces to study interactions between amino groups and serum complement pathway. SAMs of 11-mercaptoundecanol (OH-SAM) and 1-dodecanethiol (CH3-SAM) were used as control surfaces, respectively. Although much protein was adsorbed from serum solutions on the two types of amino surfaces, amounts of C3b deposition were much less than those observed on OH-SAM. Amounts of C3a released on the amino surfaces were same levels as that of CH3-SAM, but significantly smaller than that on OH-SAM. These facts suggest that the nucleophilic amino groups on NH2-SAM and PEI-coated surfaces do not directly activate the alternative pathway, but the protein adsorbed layers formed on amino surfaces activate it, but to an extent much smaller than that on OH-SAM. In addition, we found no deposition of C1q molecules on the amino surfaces, suggesting that these surfaces fail to activate the classical pathway. However, more careful studies are needed to conclude it, because it is known that C1q is only transiently detected at typical classical activation interfaces.

Complement activation on surfaces modified with ethylene glycol units.

Poly(ethylene glycol) (PEG) has been widely used for the preparation of biomedical devices and drug carriers to reduce the interaction of proteins with artificial materials. However, unanticipated body reactions such as hypersensitivity reactions caused by PEG-modified surfaces have been reported. Body reactions to PEG-modified materials still remain unclear. In this study, complement activation on surfaces modified with ethylene glycol units was examined. Two surfaces modified with tri(ethylene glycol)-terminated alkanethiol (HS-TEGOH) and methoxy-terminated PEG-thiol (HS-mPEG) were employed as model surfaces. Complement activation was assessed by the binding of an antibody against complement C3b after exposure to diluted human serum using a surface plasmon resonance (SPR) apparatus. Strong complement activation was observed on HS-TEGOH but not on HS-mPEG surfaces. The terminal hydroxyl group of HS-TEGOH is involved in complement activation. Although the HS-mPEG surface just after preparation did not induce complement activation, it became a stronger activator with storage period under room light at room temperature. Complement activation on the HS-mPEG surface was further accelerated by irradiation with UV light. These results suggest that functional groups which can activate the complement system are introduced onto the HS-mPEG surface by oxidation during long-term storage and UV irradiation.

Antibody arrays for quantitative immunophenotyping.

Detection of multiple surface antigens expressed on living cell is an important step for cell processing and clinical diagnosis. Here we describe the preparation of antibody arrays that allow parallel detection of multiple surface antigens through affinity binding of living cells. An antibody array was fabricated by photo-assisted patterning of an alkanethiol monolayer formed on a gold-coated glass plate and subsequent immobilization of antibodies specific for cell surface antigens in an array format. We demonstrate here that rapid phenotyping can be performed on the array for both adhesion-dependent and non-dependent cells by direct cell binding assays. The density of bound cells on each antibody spot was in accordance with their contents in an original suspension. This result suggests the feasibility of the array-based method for quantitative assessment of multiple antigen expression. These findings will serve to extend the range of fundamental and clinical applications of antibody arrays.

Immobilization of basic fibroblast growth factor on a platinum microcoil to enhance tissue organization in intracranial aneurysms.

OBJECT: To enhance tissue organization in an aneurysm lumen, the authors prepared a platinum microcoil carrying basic fibroblast growth factor (bFGF) and analyzed its effectiveness in the treatment of aneurysms. METHODS: Ultrathin multiorganic layers were assembled on a platinum coil through successive deposition of cationic polyethylenimine and anionic heparin, and then bFGF was immobilized through an affinity interaction with heparin. The bFGF was effectively immobilized on the surface of the platinum coil without deterioration of the coil's mechanical properties. Coil embolization of aneurysms constructed using a canine common carotid artery was performed via the endovascular approach. The aneurysms together with parent arteries were harvested 2 weeks after coil embolization. Platinum coils unmodified, coated with heparin, or immobilized with heparin and bFGF were examined. The percentage of occlusion at the aneurysm orifice in animals treated with bFGF-immobilized coils (92.99+/-7.94%) was significantly greater than that in animals treated with heparin-coated coils (57.26+/-10.76%) or unmodified coils (52.86+/-8.54%). The histological score of the aneurysms treated with bFGF-immobilized coils was also significantly greater than the scores in the control group. CONCLUSIONS: These results indicated that bFGF-immobilized microcoils may be beneficial in the obliteration of aneurysms.

Vascular endothelial growth factor immobilized on platinum microcoils for the treatment of intracranial aneurysms: experimental rat model study.

Platinum microcoils coated with immobilized recombinant human vascular endothelial growth factor (rhVEGF) were prepared and the effectiveness for the embolization of aneurysms was investigated using a rat model. Platinum coils were prepared by successive deposition of cationic polyethyleneimine and anionic heparin, and VEGF was immobilized through affinity interaction with heparin. Unmodified, heparin-coated, or rhVEGF-immobilized platinum coil segments were inserted into the ligated external carotid arteries at the bifurcation of the common carotid artery (CCA) of adult female rats. The bifurcation segments of the CCA were harvested 2 weeks after the coil placement. rhVEGF-immobilized coils showed significantly greater endothelial formation at the aneurysm orifice and cell infiltration in the aneurysm body compared with the unmodified and heparin-coated coils. The percentage of sac occlusion was significantly greater in the rhVEGF-immobilized group (77.53 +/- 27.58%) than in the heparin-coated group (44.81 +/- 38.30%) and unmodified group (34.99 +/- 28.15%). Scanning electron microscopy showed a tendency for more fibrotic and cellular collections on the coil surface and more tissue mass filling in the coil lumen in the rhVEGF-immobilized group. Platinum microcoils coated with immobilized rhVEGF may be effective for the obliteration of aneurysms.

Deposition of complement protein C3b on mixed self-assembled monolayers carrying surface hydroxyl and methyl groups studied by surface plasmon resonance.

Since complement activation is recognized as a common response of the host defense system when an artificial medical device is applied to a patient, great effort has been devoted to studies on the interaction of the complement system with artificial materials. However, some uncertainties remain, partially because of the lack of well characterized surfaces and suitable analytic methods for study of the surface phenomena that occur on artificial materials under physiologic conditions. In this study, we employed self-assembled monolayers (SAMs) and the surface plasmon resonance (SPR) technique to study interactions of the serum complement with well characterized surfaces. Self-assembled monolayers carrying various concentrations of hydroxyl groups were prepared using 11-mercapto-1-undecanol (C11-OH) and one of n-nonanethiol, n-dodecanethiol, and n-hexadecanethiol. The amount of NHS deposition on the SAMs increased with increasing C11-OH content of the SAMs, and the amount of anti-C3b antibody immobilization formed on the NHS deposition layers increased with increasing C11-OH content of the SAMs. These results clearly demonstrate that a large amount of C3b, produced through the activation of the complement system, binds covalently to and is adsorbed by hydroxyl-group-rich surfaces. The combination of SAMs and the SPR technique is suitable for studying the interaction of the complement system with solid surfaces, and the results should give basic information needed for a rational design of biocompatible surfaces on synthetic materials.