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Superior colliculus stimulation enhances neocortical serotonin release and electrocorticographic activation in the urethane-anesthetized rat.

Recent evidence indicates that the superior colliculus (SC), in addition to its functions in sensory detection, also participates in controlling the generalized activation state of the forebrain, as measured by the electroencephalogram (EEG) or electrocorticogram (ECoG). The mechanisms by which the SC modulates forebrain activation are not well understood. By using in vivo microdialysis, we examined the role of serotonin release as a mechanism by which the SC can control neocortical activity in the urethane-anesthetized rat. Electrical 100 Hz stimulation of the SC increased frontal cortex serotonin output to 116, 118, and 140% of baseline levels for stimulation intensities of 0.5, 0.75, and 1.0 mA, respectively. Further, 75% of extracellularly recorded single (putative serotonergic) dorsal raphe neurons increased their discharge rate in response to 100 Hz stimulation of the SC. Stimulation of the SC also suppressed frontal cortex low frequency (1-6 Hz) synchronized ECoG activity, replacing it with high-frequency desynchronization. This activation response was resistant to cholinergic-muscarinic receptor antagonists (atropine, 50 mg/kg; scopolamine, 2 mg/kg), but was reduced or abolished by systemic treatment with the serotonergic receptor antagonists ketanserin (10 mg/kg) or methiothepin (5 mg/kg). These data suggest that efferents from the SC, possibly by an excitatory action on serotonergic dorsal raphe cells, produce an enhanced release of serotonin and ECoG activation in the neocortex. The stimulation of cortical serotonin output may constitute a mechanism by which the SC acts on the forebrain to increase cortical excitability in response to sensory stimuli processed by SC neurons.

Animals↗

Surface coating of stearyl poly(ethylene oxide) coupling-polymer on polyurethane guiding catheters with poly(ether urethane) film-building additive for biomedical applications.

Three types of stearyl poly(ethylene oxide) (SPEO) with Mn of 2,300, 6,000 and 12,000 were synthesized; accordingly, three types of amphiphilic coupling-polymer SPEO-MDI-SPEO (MSPEO) were prepared by the reactions with 4,4'-methylene diphenyl diisocyanate (MDI). As the surface-modifying additives (SMA), MSPEOs were coated onto the outer wall of the medical guiding catheters. Due to the lack of stability, when coated, MSPEO blended with the film building agent (FBA), poly(ether urethane) (PEL). The process of coating was performed with a lifter. With invariable speed, the PU guiding catheter was vertically dipped into the coating mixture of SMA-MSPEO and FBA-PEL. The surface analysis was carried out by ATR-FTIR and contact angle measurements. It was proved that the surface enrichment of PEO on water interface was much higher than that on air interface. Three kinds of static clotting time tests, PRT, PT and TT, as well as the static platelet adhesion experiment were performed. The results indicated that the coated surface could resist the blood coagulation effectively. In order to test the blood compatibility of the coated catheters under a shear of blood flow, the dynamic experiment was performed with a closed-loop tubular system under a shear rate of 1,500 s(-1). The blood regular testing was carried out on the samples taken out at six different times (0, 5,10, 20, 30 and 60 min). The results were ideal. Finally, the SMA-MSPEO was proved to be non-acute-toxic by LD50 test.

Blood Coagulation↗

In vivo biostability of a poly(carbonate-urea)urethane graft.

In peripheral and coronary bypass surgery, the patency of prosthetic grafts is inferior to autologous vein, mainly due to intimal hyperplasia caused in part by compliance mismatch between rigid graft and elastic host artery. We have developed a compliant poly(carbonate-urea)urethane vascular graft "MyoLink" which was biostable in vitro degradation studies. To further investigate the biostability of this material, we report a long-term in vivo study on 8 beagle dogs (15+/-3 kg) implanted with this graft (ID 5mm) in the aorta-iliac position; three grafts were harvested at 18 months to assess short-term biodegradation, with one animal having died from an unrelated infection. The 4 remaining grafts were harvested at 36 months for analysis by: (1) histology, (2) compliance measurements and (3) environmental scanning electron microscopy (ESEM); gel permeation chromatography (GPC); attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) and radial tensile strength analysis. There was no infection or inflammation of the grafts or surrounding tissues. Histological analysis showed a well-developed neointima but only at the distal anastomosis. There were no significant differences in compliance pre- and post-implantation and no evidence of material curvature, radial expansion or chemical breakdown, ESEM and GPC showed no signs of degradation. Peak height analysis with ATR-FTIR of the 1740 cm(-1) (C=O of carbonate) and 1253 cm(-1) bands (C-O-C of CO-O-C) showed a loss of carbonate carbonyl but was not statistically significant. Radial tensile strength remained within batch release specifications.This polyurethane graft retains its compliance post-implantation, whilst exhibiting only a minor hydrolysis of the amorphous segment, confirming its biostability in vivo up to 3 years.

Anastomosis, Surgical↗

Adhesion, apoptosis and cytokine release of human mononuclear cells cultured on degradable poly(urethane urea), polystyrene and titanium in vitro.

Early interactions between materials and mononuclear cells may influence the viability and secretory response of the cells. Such effects may in turn influence the subsequent inflammatory and repair phases around the materials. In the present study, it was examined if mononuclear cells cultured in vitro either unstimulated or stimulated with lipopolysaccharide (LPS) (10ng/ml) revealed differences regarding cell viability and apoptosis. A major interest was to study the influence of different material properties on the parameters of the inflammatory response upon cell adhesion to materials with widely different surface chemical properties but similar surface topography: degradable poly(urethane urea) (PUUR), cell culture treated polystyrene (PS) surfaces, and commercially pure (c.p.) titanium (Ti). Finally, the secretion of the proinflammatory tumor necrosis factor-a (TNF-alpha) and the downregulating interleukin-10 (IL-10) cytokines was examined in the supernatants from 24h mononuclear cell cultures. No differences in cell viability as measured by lactate dehydrogenas (LDH) were observed between the three materials. The number of material-surface adherent cells was higher on PUUR than the more hydrophilic PS and Ti as judged by quantification of material surface-associated DNA, light microscopic morphological examination of DAPI-stained cells and SEM. LPS increased the number of adherent cells, irrespective of the type of material. The lowest number of apoptotic (annexin-V) and necrotic (propidium iodide) mononuclear cells was detected on PUUR. LPS decreased the number of both apoptotic and necrotic cells, irrespective of material. Low TNF-alpha levels were detected in unstimulated conditions, irrespective of material types. A significantly lower amount of TNF-alpha was found with unstimulated cells on PUUR than on Ti. A significantly higher IL-10 level was detected in unstimulated Ti cultures compared with PUUR and PS. Secretion of IL-10 was predominantly stimulated by LPS on PUUR and Ti. The data indicate that material-related differences are expressed in differences in cell adherence, apoptosis and cytokine secretion. Further, degradable PUUR has equal or less cell-activating properties than Ti and PS under in vitro conditions.

Absorbable Implants↗

Biodegradable poly(ether ester urethane)urea elastomers based on poly(ether ester) triblock copolymers and putrescine: synthesis, characterization and cytocompatibility.

Polymers with elastomeric mechanical properties, tunable biodegradation properties and cytocompatibility would be desirable for numerous biomedical applications. Toward this end a series of biodegradable poly(ether ester urethane)urea elastomers (PEEUUs) based on poly(ether ester) triblock copolymers were synthesized and characterized. Poly(ether ester) triblock copolymers were synthesized by ring-opening polymerization of epsilon-caprolactone with polyethylene glycol (PEG). PEEUUs were synthesized from these triblock copolymers and butyl diisocyanate, with putrescine as a chain extender. PEEUUs exhibited low glass transition temperatures and possessed tensile strengths ranging from 8 to 20MPa and breaking strains from 325% to 560%. Increasing PEG length or decreasing poly(caprolactone) length in the triblock segment increased PEEUU water absorption and biodegradation rate. Human umbilical vein endothelial cells cultured in a medium supplemented with PEEUU biodegradation solution suggested a lack of degradation product cytotoxicity. Endothelial cell adhesion to PEEUUs was less than 60% of tissue culture polystyrene and was inversely related to PEEUU hydrophilicity. Surface modification of PEEUUs with ammonia gas radio-frequency glow discharge and subsequent immobilization of the cell adhesion peptide Arg-Gly-Asp-Ser increased endothelial adhesion to a level equivalent to tissue culture polystyrene. These biodegradable PEEUUs thus possessed properties that would be amenable to applications where high strength and flexibility would be desirable and exhibited the potential for tuning with appropriate triblock segment selection and surface modification.

Absorbable Implants↗

Characterization of the structures of poly(urea-urethane) microcapsules.

A series of poly(urea-urethane) microcapsules containing phthalate derivatives as a core material were prepared by an interfacial polymerization process in order to investigate the structural formation mechanism. Scanning electron microscopy (SEM) analysis for the cross sections of microcapsules revealed the systematic formations of porous structures followed by the formation of core/shell structures. Critical values of the core oil content for the formation of porous and core/shell structures were obtained from SEM results and the critical values were found to be proportional to the solubility parameters of core materials. Dynamic mechanical measurements indicated an amorphous structure of wall membrane and the glass transition temperature was found to decrease with increasing the core oil content suggesting a plasticizing effect. The surface amount of the core oils absorbed in the wall membrane was estimated using time of flight secondary ion spectroscopy analysis and found to increase with increasing the oil content before reaching constant. This tendency was interpreted in terms of the structural formation of the microcapsules. The results obtained in the present investigation were reasonably understood on the basis of swelling theory of wall membrane and the Flory-Huggins interaction parameters of the systems were discussed.

Algorithms↗

Effects of a benzodioxan derivative MKC-242 on the firing activity of the dorsal hippocampus CA1 pyramidal neurons in awake and urethane- anaesthetized rats: in vivo electrophysiological evidence for a 5-HT 1A agonistic property.

The aim of the present in vivo study was to determine whether a benzodioxan derivative MKC-242, (S)-5-[3-[(1,4-benzodioxan-2-ylmethyl)amino]propoxy]-1,3-benzodioxole hydrochloride, possesses an agonistic activity at postsynaptic serotonin (5-HT)1A receptors in the rat hippocampus when administered systemically. We examined the effects of acute administrations of MKC-242 on the firing activities of dorsal hippocampus CA1 pyramidal neurons. In quiet awake rats, s.c. administrations of MKC-242 significantly decreased the spontaneous firing activity in a dose-dependent manner at doses of 0.3-6 mg/kg. In urethane-anaesthetized rats, i.v. injections of MKC-242, at cumulative doses of 0.3-3 mg/kg, also significantly and dose-dependently inhibited the firing activity induced by microiontophoretically applied quisqualate. These decreasing effects were antagonized by the selective 5-HT1A antagonists WAY-100135 (5 mg/kg, s.c.) and WAY-100635 (0.2 mg/kg, s.c. to the awake rats and 0.4 mg/kg, i.v. to the anaesthetized rats), thereby confirming that MKC- 242 decreased the firing activities by stimulating 5-HT1A receptors. The selective depletion of 5-HT produced by the 3-d administration of the 5-HT synthesis inhibitor, parachlorophenylalanine (500 mg/kg.d, i.p.), did not affect the decreasing effect of MKC-242 in the awake animals, indicating that postsynaptic 5-HT1A receptors mediated the decreasing effect. The present results provided the first in vivo electrophysiological evidence that MKC-242, when systemically administered, exerts a 5-HT1A agonistic action at the postsynaptic level.

Journal Article↗

Use of iron monocarboxylates in the two-step preparation of poly(ester-urethane)s.

Five different iron monocarboxylates were used as catalysts in the two-step preparation route of lactic acid based poly(ester-urethane)s (PEU). In the first step, a hydroxyl-terminated poly(lactic acid) prepolymer was prepared, which in the second step was linked with 1,6-hexamethylene diisocyanate. The resulting polymers were characterized by titration, size exclusion chromatography, differential scanning calorimetry, and nuclear magnetic resonance spectroscopy, and the mechanical properties were tested as well. Iron monocarboxylates proved to be efficient catalysts in the preparation of a hydroxyl-terminated prepolymer (lowest acid number obtained: 0.08). The same catalyst systems proved also to be highly efficient in the linking step yielding a high molar mass PEU. Semicrystalline PEUs could be prepared at 160 and 180 degrees C by using the iron acetate of different oxidation state. PEU prepared at 200 degrees C was amorphous, which could be related to racemization during the polycondensation. By using the fluorinated iron acetate amorphous PEUs was prepared at all reaction temperatures. The molar mass of the prepolymers and the PEUs increased as a function of the polycondensation temperature for all catalysts used. The highest weight-average molar masses (M(w)) were obtained by using the fluorinated iron acetate.

Calorimetry, Differential Scanning↗

Synthesis and properties of degradable poly(urethane urea)s to be used for ligament reconstructions.

In the present study we describe the synthesis, wet spinning, mechanical testing, and degradation of poly(urethane urea)s (PUURs) intended for clinical use in anterior cruciate ligament (ACL) reconstruction. The effects of soft segment chemical composition and molar mass and the kind of diamine chain extender on the material properties were investigated. It was found that the fibers made of PUUR with polycaprolactone diol (PCL530) as soft segment and MDI/1,3-DAP as hard segment (PCL530-3) have high tensile strength and high modulus and when degraded keep their tensile strength for the time demanded for the application. In conclusion, from a chemical and mechanical point of view PUUR fibers of PCL530-3, ARTELON, are suitable for designing a degradable ACL device.

Biocompatible Materials↗

Sonogashira cross-couplings of ynamides. Syntheses of urethane- and sulfonamide-terminated conjugated phenylacetylenic systems.

[reaction: see text] The first successful Sonogashira coupling of ynamides with aryl and vinyl iodides is described here. This study resolves the problem of the competing pathway involving homocoupling of ynamides and provides a practical entry to novel urethane- or sulfonamide-terminated conjugated acetylenic systems. An interesting tandem hydrohalogenation and Sonogashira coupling was also observed to give an en-ynamide.

Journal Article↗

A novel urethane containing copolymer as a surface modification additive for blood contact materials.

Surface modification to develop a biomolecules-presenting surface is of interest, both from a scientific and an industrial point of view. In this research, a penta-block-coupling polymer of warfarin-PEO-MDI-PEO-warfarin was specially designed as the surface modifying additive (SMA). The warfarin-modified polyurethane surfaces were then prepared by dip-coating method. Attenuated total reflection fourier transform infra-red (ATR-FTIR) spectra revealed that the urethane segments in the SMA could penetrate into the hard block of segmented polyurethane (SPU) via intermolecular hydrogen bonds. The X-ray photoelectron spectroscopy (XPS) results indicated that the intermolecular hydrogen bonds were strong enough to form stable warfarin-PEO composite surfaces in an aqueous environment. Fibrinogen and albumin adsorption onto unmodified and SMA-modified SPU was investigated by the (125)I-labeled method. The surface for attracting and reversibly binding albumin, which was proved to suppress the platelet adhesion and prolong the clotting time, has been developed by the simple coating of the novel SMA in SPU.

Journal Article↗

Decomposition analysis of binary polyfunctional urethane monomer mixtures.

Thermal decomposition in three binary monomer mixture systems containing an experimentally synthesized monomer (EXP3) and a commercial polyfunctional urethane monomer (U-4TXA) was examined in terms of differential thermal analysis (DTA) and simultaneous thermogravimetry (TG) analysis when a sample was heated to 800 degrees C. The binary EXP3/U-4TXA monomer mixtures were visible light-cured (VLC) resins which included CQ (camphorquinone=0.5 wt%) and DMAEMA (dimethylaminoethyl methacrylate=0.5 wt%) as the photoinitiator. Their DTA curves showed that thermal decomposition initiated at around 300 degrees C and finished at around 500 degrees C. TG curves showed complete weight loss at 800 degrees C during thermally induced decomposition: the decomposition reaction at weight losses of 5 to 50% was calculated from the slope of the (1/T) versus log10(Hr) diagram, because the exothermic decomposition temperature shown on DTA curves increased with increasing heating rate from 2 to 20 degrees C/min. Thermal decomposition analysis showed that a decomposition with weight change had exothermic heats ranging from 0.38 to 1.07 kJ/g for the EXP U1 mixture, 1.06 to 1.76 kJ/g for EXP U2 and 1.74 to 2.02 kJ/g for EXP U3. Activation energy ranged from 1.42 to 1.89 kJ/mol at weight losses of 5 to 50% in the three binary EXP3/U-4TXA monomer mixture systems.

Journal Article↗

The effects of different additives on the dielectric relaxation and the dynamic mechanical properties of urethane dimethacrylate.

The polymer-filler interaction of a dental composite was examined by dielectric measurements to determine how a non-compatible inorganic phase modifies the molecular behaviour of the polymer chains and how modification of this phase by silanation can affect these molecular behaviours. Urethane dimethacrylate (UDMA) monomer and zirconia-silica (ZS) powder were used as organic and inorganic phases, respectively. 3-Acryloxypropyltrimethoxysilane (MAPM) and 4-aminobutyltriethoxysilane (ABTE) were used as silanating coupling agents. The components of the composite were mixed in different ratios after treating the filler with from 0 to 30 times the minimum uniform coverage with the silane, then the composite was made into thin films. The dielectric spectra of five replicas for each filler-monomer ratio were measured, and three replicas were measured for each silane and for each amount of silane used to treat the filler. Increasing the filler concentration in the composites decreased the intensity of Tan(delta)epsilon for the alpha- and beta-relaxations, where the beta-relaxations also became broader and the alpha-relaxations were totally obstructed. The alpha-relaxations were shifted to higher temperatures, while the beta-relaxations were shifted to lower temperatures. Filler treatment with small amounts of MAPM shifted the alpha-relaxations to higher temperatures; they were shifted back to lower temperatures when the filler was treated with large amounts of silane. Filler treatment with large amounts of ABTE (30-fold) caused an extra peak to emerge in the high-temperature region. It can be concluded that increasing filler concentration restricts the mobility of the main chains and decreases the thickness of the surface layer, while allowing more movement of the local chains. Filler treatment with MAPM was shown to be a compatible coupling agent with the ZS and the UDMA systems. Such compatibility was observed through the effectiveness of the chemical linkage of the silane to the filler and the polymer without forming a separate inter-phase. Filler treatment with ABTE was shown to be an incompatible coupling agent for the ZS-filled UDMA system. The incompatibility was observed by the occurrence of a new peak that characterizes an inter-phase.

Acrylates↗

The effects of moisture on the dielectric relaxation of urethane dimethacrylate polymer and composites.

Dielectric characterization evaluated the influence of sorbed moisture on urethane dimethacrylate (UDMA) polymer matrix and unsilanated and silanated zirconia-silica (ZS)-filled UDMA composites. The moisture content (MC) of samples tested ranged from 0 to 7.32% with maximum values for UDMA, silanated and unsilanated ZS-filled UDMA were 3.72, 6.0 and 7.32%, respectively. Dielectric spectra were measured from 0.053 to 100 kHz and from -180 to 180;C. Small MC stiffened the polymers, which was shown by a shift of beta-peaks to higher temperatures, a decrease in the half width of bands and an increase in the change of activation energies. Higher MC resulted in water plasticizing the polymer by shifting the beta-peaks to lower temperatures. Maximum water sorption restricted the motion of the polymer chains and the b-peaks shifted to higher temperatures. Further, increasing MC shifted the a-relaxation to lower temperatures in both UDMA and composites. However, in composites with unsilanated ZS, a new relaxation occurred at high temperatures and low frequencies which was attributed to Maxwell-Wagner-Sillars (MWS) polarization. Thus, in UDMA, water occurs as fragments that form non-freezing clusters. However, in unsilanated ZS-UDMA composites, bulk water can exist at the polymer-filler interface and form disordered structures of ice at low temperatures which can evaporate at high temperatures. Silanated ZS-UDMA composites function similarly to UDMA at low MC but are more plasticized at high temperatures.

Absorption↗

Optimal endothelialisation of a new compliant poly(carbonate-urea)urethane vascular graft with effect of physiological shear stress.

OBJECTIVE: to define the optimal seeding conditions of a new stress free poly(carbonate-urea)urethane (CPU) graft with compliance similar to that of human artery with honeycomb structure engineered during the manufacturing process to enhance adhesion and growth of endothelial cells. METHODS: (111)Indium-oxine radiolabeled human umbilical vein endothelial cells (HUVEC) were seeded onto CPU grafts at (a) concentrations from 2-24x10(5)cells/cm(2)and (b) incubated for 0.5, 1, 2, 4 and 6 h. Following incubation, graft segments were subjected to three washing/gamma counting procedures and scanning electron microscopy (SEM). Cell viability was measured using a modified Alamar blue(TM)assay. To test physiological retention a pulsatile flow phantom was used to subject optimally seeded (16x10(5), 4 h) CPU grafts to arterial shear stress for 6 h with real time acquisition of scintigraphic images of seeded grafts using a nuclear medicine gamma camera system. RESULTS: the seeding efficiency of 54+/-13% post three washes was achieved using 16x10(5)cells/cm(2). Similarly in SEM micrographs a seeding density of 16x10(5)cells/cm(2)resulted in a confluent monolayer. Seeded CPU segments incubated for 4 h exhibited significantly higher resistance to wash-off than segments incubated for 30 min (p <0.05). Exposure of seeded grafts to pulsatile shear stress resulted in some cell loss with 67+/-3% of cells adherent following 6 h of perfusion with ongoing metabolic activity. Thus, optimal conditions were 16x10(5)cells/cm(2)at 4 h. CONCLUSIONS: the optimal seeding conditions have been defined for "tissue-engineered" vascular graft which allow complete endothelialisation and high cell-to-substrate strength that resists hydrodynamic stress.

Biocompatible Materials↗

An assessment of covalent grafting of RGD peptides to the surface of a compliant poly(carbonate-urea)urethane vascular conduit versus conventional biological coatings: its role in enhancing cellular retention.

The aim of sodding prosthetic grafts with endothelial cells (EC) is to establish a functioning antithrombogenic monolayer of EC. Application of basement membrane proteins improves EC adherence on ePTFE grafts. Their addition to a biodurable compliant poly(carbonate-urea)urethane graft (CPU) was studied with respect to EC adherence. Preclot, fibronectin, gelatin, and collagen were coated onto CPU. RGD peptide, heparin, and both RGD and heparin were chemically bonded to CPU. Human umbilical vein EC (HUVEC) labeled with 111-Indium oxine were sodded (1.8 x 10(6) EC/cm(2)) onto native and the modified CPU. The grafts were washed after 90 min and EC retention determined. The experiments were repeated six times. EC retention on native CPU was 1.0 +/- 0.2 x 10(5) EC/cm(2). The application of preclot, fibronectin, gelatin, and collagen did not improve EC retention, which was 0.8 +/- 0.1, 0.4 +/- 0.1, 0.3 +/- 0.08, and 0.5 +/- 0.2 x 10(5) EC/cm(2), respectively. Bonding RGD, heparin, and both RGD and heparin significantly improved EC retention to 1.9 +/- 0.6, 1.7 +/- 0.5, and 2.6 +/- 0.6 x 10(5) EC/cm(2), respectively (p < 0.01). Bonding of RGD, heparin, and both RGD and heparin accelerates and enhances EC retention onto CPU. Simple coating of basement membrane proteins confers no advantage over native CPU.

Coated Materials, Biocompatible↗

Cardiovascular responses to medullary microinjections of opiate agonists in urethane-anesthetized rats.

Pressor (VLPA) and depressor (VLDA) areas of the ventrolateral medulla were identified by microinjections of L-glutamate in urethane-anesthetized rats. Cardiovascular effects of opiate agonists microinjected into the same sites were then studied. Agents used to stimulate mu, delta, sigma, kappa, and beta-endorphin (epsilon) receptors were morphiceptin, D-Ala2-D-Leu5-enkephalin, N-allyl-normetazocine, dynorphin, and beta-endorphin, respectively. Opiate receptor stimulation in VLPA decreased blood pressure (BP) and heart rate (HR), while in VLDA it increased BP and HR. Thus, it is the site of injection rather than the type of opiate receptor that determines cardiovascular responses. Naloxone, an opiate antagonist, reversed and prevented these responses. Abolition of cardiovascular responses by spinal transection at the C1 level indicated that the sympathetic nervous system mediated these responses. The following mechanism is proposed for these actions of opiates: Cell bodies in VLPA, but not in VLDA, project to the intermediolateral cell column of the spinal cord. Opiates inhibit VLPA and lower BP and HR by decreasing sympathetic outflow. Opiate-induced inhibition of VLDA, which has an inhibitory effect on VLPA, results in an increase in BP and HR.

Animals↗