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Dipole-like neuronal sources of theta rhythm in dorsal hippocampus, dentate gyrus and cingulate cortex of the urethane-anesthetized rat.

Spatial distribution of theta activity was investigated in the dorsal hippocampal formation and overlying neocortex of the urethane-anesthetized rat. Laminar phase profiles from semi-microelectrode penetartions showed approximately 180 degrees phase shifts combined with small amplitude values in stratum radiatum of CA1, instratum moleculare of the dentate gyrus and in layer V/VI of the cingulate cortex at theta peak frequency. Evidence has been presented that layers of neurons in CA1, in the dorsal granular layer and in the cingulate cortex are the sources of dipole-like theta field potentials. A strong linear relationship between the neuronal theta sources in hippocampal CA1, dentate area and cingulate cortex was found.

Animals↗

Urethane-lined silicone facial prostheses.

After testing a variety of materials to line the tissue-fitting surface of silicone prostheses in an attempt to render them adheophilic, tear-resistant, wettable, smooth, and resistant to fungal growth, a vacuum adaptable thermoplastic prefabricated thin urethane film was found to be most effective. A technique is described for fabricating a silicone prosthesis with such a lining.

Adhesiveness↗

Effect of kainic acid and NMDA on the pattern electroretinogram, the scotopic threshold response, the oscillatory potentials and the electroretinogram in the urethane anaesthetized cat.

Kainic acid (KA, 12.5-100 nmol) or N-methyl-D-aspartate (NMDA 25-250 nmol) was injected into the vitreous of one eye of urethane anaesthetized cats. Pattern electroretinograms (PERGs) were recorded to transient contrast reversing bars. Scotopic luminance electroretinograms (ERGs) were recorded to blue flashes. All doses of KA reduced the oscillatory potentials (OPs), PERG and focal ERG (FERG). At 50 nmol KA, the b-wave and scoptic threshold response (STR) were normal. At 100 nmol KA, the STR was absent and the b-wave reduced by over 50%. OPs and STRs were reduced in all NMDA injected eyes. NMDA at 25 nmol enhanced the FERG, PERG, and b-wave and high doses (above 150 nmol) reduced them. Light microscopic examination of retinas showed 25 nmol KA only damaged dendrites of ganglion cells. NMDA damage was slight with < 200 nmol. These data show that the cat PERG has a proximal component which is very sensitive to low doses of KA; the PERG and FERG are very similar; the STR and PERG are generated by different structures and that the OPs and the FERG and PERG are all generated close to the ganglion cell layer, proximal to the STR.

Animals↗

Grafting reactions and heparin adsorption of poly(amidoamine)-grafted poly(urethane amide)s.

Differently terminated poly(amidoamine) (PAA) oligomers were grafted on the surface of poly(ether urethane amide)s (PEUAm), with fumaric or maleic acid moieties. The grafting reaction was Michael-type addition of amino groups to activated double bonds in the PEUAm backbone. PAAs having primary amino, or secondary amino end-groups were directly grafted on the surface of PEUAm sheets. For vinyl-terminated chains an alpha, omega amino-polyether spacer was introduced initially, following the same addition mechanism. Ungrafted and grafted materials were characterized, besides other analytical techniques, by ATR FT-IR spectroscopy. The heparin adsorption on PEAUm films was analysed after its elution from heparinized samples, quantified by coagulation tests (aPTT), and related to the presence of the PAAs chains grafted on to the surface. Results indicate that PAA-grafted PEUAm elastomeric biomaterials, display enhanced heparin adsorption abilities.

Adsorption↗

Immobilization of human thrombomodulin onto poly(ether urethane urea) for developing antithrombogenic blood-contacting materials.

Thrombomodulin (TM) is a newly described endothelial cell-associated protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. In this study, focussing on the application of TM for biomedical materials, recombinant human TM (hTM) was immobilized onto the polymers for medical use, and the evaluation of their antithrombogenicity and the interaction with platelets were investigated. As the base polymer for immobilization reaction, poly(ether urethane urea) (PEUU), which was reported to have good blood compatibility, was used. hTM-immobilized PEUU showed superior antithrombogenic activity, such as the prolongation of plasma recalcification time and the inhibition of thrombin-induced platelet aggregation, though the amount of immobilized hTM was very small (i.e. less than 1 microgram/cm2). Platelet adhesions onto hTM-immobilized PEUU were not observed. These results show that the immobilization of hTM does not change the native good blood compatibility of PEUU, but provides excellent anticoagulant activity.

Antithrombins↗

Protein adsorption of poly(ether urethane) surfaces modified by amphiphilic and hydrophilic polymers.

A commercial biomedical poly(ether urethane), Pellethane 2363-80AE, was surface modified through the use of amphiphilic polymeric additives, and through surface grafting with poly(ethylene glycol), PEG. Two different amphiphilic polymers, Polymer C and Pluronic PE9400, were used as additives. Polymer C, a segmented polyurethane, was prepared from PEG1500, 4,4'-diphenylmethane diisocyanate and a C16-C18 monoglyceride chain extender. Pluronic PE9400 is a propylene oxide-ethylene oxide tri-block co-polymer obtained from BASF. Adsorption of human albumin and fibrinogen to the modified surfaces was studied by means of radiolabelled proteins. By contact angle measurements and X-ray photoelectron spectra the amphiphilic polymers were shown to accumulate at the polyurethane surfaces. Adsorption of fibrinogen, in particular, was significantly reduced by the amphiphilic additives to levels similar to those obtained for Pellethane surfaces grafted with PEG 20,000. In vitro clotting times for citrate-buffered blood in contact with the amphiphilic surfaces increased as compared with the unmodified ones.

Adsorption↗

Derivatization of a new poly(ether urethane amide) containing chemically active sites.

A novel poly(ether urethane amide) was synthesized by chain-extending the prepolymer with fumaric acid, resulting in a polymer exhibiting superior mechanical properties and containing chemically active derivatization sites, stemming from the fumaramide double bond. The derivatization of the active sites was performed via a Michael-like addition of amine-terminated poly(ethylene oxide) chains, which when performed in bulk, greatly affected the mechanical properties of the polymer, except in the case in which a relatively high molecular weight was grafted. Surface grafting of the polymer led to a significantly lower effect on its mechanical properties, which was minimal when a high molecular weight molecule was used.

Absorption↗

Poly(ether urethane) wound covering with high water vapour permeability compared with conventional tulle gras on split-skin donor sites.

The experimental poly(ether urethane) (PEU) wound covering with a high water vapour permeance was compared with tulle gras treatment on adjacent areas of the same 20 split-skin donor sites. All patients experienced little or no pain from the PEU-covered areas, while 70 per cent of the patients complained of more pain from the tulle gras-covered areas. The PEU covering did not absorb the wound exudate underneath, neither did it retain wound fluid, but turned the wound exudate into a jelly-like clot layer by allowing a high evaporative water loss from the wound. Tulle gras treatment also prevented wound desiccation, but the exudate was absorbed into the overlaying cotton pads, where it became dry at the outer surface. Microscopy revealed that re-epithelialization occurred at a similar rate under the PEU covering as under tulle gras. In conclusion, the high water vapour permeable PEU wound covering prevents fluid retention, induces clotting of the wound exudate and reduces pain in split-skin donor sites. Tulle gras dressed with gauzes and crêpe bandage prevents wound desiccation, but causes more pain.

Adolescent↗

Tonic desynchronisation of cortical electroencephalogram by electrical and chemical stimulation of superior colliculus and surrounding structures in urethane-anaesthetised rats.

Damage to the superior colliculus in rats impairs desynchronisation of the cortical electroencephalogram in response to light flashes. However, it is unclear which elements within the superior colliculus, and which efferent collicular pathways, might be involved in alerting cerebral cortex to visual stimuli. To investigate this problem, the superior colliculus and surrounding structures were stimulated either electrically (3 s trains of 0.2 ms 100 Hz cathodal pulses), or chemically (200 nl of 5 mM sodium L-glutamate), in rats anaesthetised with urethane. The cortical electroencephalogram was recorded bilaterally from frontal cortex. At each site tested with electrical stimulation the threshold current (up to 60 microA) required to produce tonic desynchronisation (outlasting stimulation-offset by at least 10 s) was determined. Comparison of the effects of electrical and chemical stimulation suggested the following: (1) stimulation of cells in the deep layers of the superior colliculus can desynchronise the cortical electroencephalogram. There may also be an additional effective area in the rostral part of the superficial layers, but this needs to be confirmed in unanaesthetised animals. (2) Stimulation of fibres in the deep white layers of caudal superior colliculus, and of cells in a wide area of caudal midbrain reticular formation, are also effective at desynchronising the cortical electroencephalogram. It is therefore possible that the ipsilateral descending pathway, that runs from the superior colliculus to terminate in the parabigeminal and cuneiform nuclei and surrounding reticular formation, is involved in mediating cortical desynchronisation initiated by the superior colliculus. Evidence from other studies indicates that some sites in this pathway may be part of a "defence arousal system". (3) Sites on the ascending pathways from the superior colliculus, to structures including dorsal thalamus, pretectum, zona incerta and rostral midbrain reticular formation, were relatively ineffective at tonically desynchronising the cortex. However, some of these pathways might mediate phasic, movement-related arousal of collicular origin.

Animals↗

Tetanus toxin blocks inhibition of granule cells in the dentate gyrus of the urethane-anaesthetized rat.

Field potentials of dentate granule cells in response to stimulation of the perforant path have been studied before and after injecting tetanus toxin (200 mouse LD50; or phosphate-buffered saline in controls) into the hilus of the dentate gyrus of rats under urethane anaesthesia. Within 1 h of toxin injection, the population spike, but not the slope of the excitatory postsynaptic potential, had increased markedly in amplitude and double or treble population spikes appeared in response to perforant path stimulation. Both paired pulse inhibition (15-ms interval between conditioning and test stimuli) and commissural inhibition (10-ms interval) were substantially reduced by the toxin. Neither multiple spikes nor the reduction in inhibition were seen in controls. Apparent inhibition of the excitatory postsynaptic potential, seen with paired stimuli to the perforant path, was not affected by the toxin. At later times after the injections, a progressive increase in the size of the spikes was seen in the controls while in the toxin animals there was often a secondary decrease in size. It is concluded that tetanus toxin can block both feed-back and feed-forward inhibitory components acting on dentate granule cells. The results are discussed with respect to the role of inhibitory processes in the control of epileptogenesis.

Action Potentials↗

Analysis and evaluation of a biomedical polycarbonate urethane tested in an in vitro study and an ovine arthroplasty model. Part II: in vivo investigation.

The polyurethane (PU) elastomer Corethane 80A (Corvita) is being considered as the acetabular bearing material in a novel total replacement hip joint. Its biostability was investigated in vitro (Analysis and evaluation of a biomedical polycarbonate urethane tested in an in vitro study and an ovine arthroplasty model. Part I: material selection and evaluation, Biomaterials, in press) together with three other commercially available biomedical PUs: Pellethane 2363-80A (DOW Chemical), a polyhexamethylene oxide based PU, PHMO-PU (CSIRO, not supplied as a commercial product) and ChronoFlex AL-80A (CardioTech). From the in vitro studies, Corethane 80A displayed the best overall resistance to hydrolysis, ESC, MIO and calcification, followed by ChronoFlex 80A and PHMO-PU, with Pellethane 80A being the least stable. Building on the in vitro investigation, the follow-up in vivo study (reported here) assessed Corethane 80A as the bearing layer in a prototype compliant layer acetabular cup, in a fully functioning ovine total hip arthoplasty (THA) model. PU degradation in the retrieved cups was analysed using a range of analytical and physical-testing methods including mechanical testing, differential scanning calorimetry, Fourier transform infrared spectroscopy and environmental scanning electron microscopy. The Corethane 80A functioned well in the THA model, with the bearing surfaces of the retrieved hip cups showing no significant evidence of biodegradation or wear damage after 3 years in vivo. The findings in this study provide compelling evidence for the biostability and effectiveness of acetabular cups incorporating a Corethane 80A compliant bearing layer.

Absorptiometry, Photon↗

Interactions between endothelial cells and a poly(carbonate-silsesquioxane-bridge-urea)urethane.

We have recently developed a polymer which contains silsesquioxane in the form of nano-bridges poly(carbonate-silsesquioxane-bridge-urea)urethane (PCBSU) for cardiovascular device applications. The polymer has been characterised and the durability has been confirmed with long-term in vivo tests. The aim of this study was to test the cytocompatibility of the new polymer and to investigate any potential cytotoxic effects. To assess the effect of direct contact with PCBSU sections of polymer material were cut and placed into a 24-well plate. Six discs were seeded with 2 x 10(5) human umbilical vein cells (HUVEC). As a positive control, six wells were seeded with the same number of HUVEC. In a further experiment to assess indirect contact with PCBSU a sample of the polymer was powdered using a Micro-Dismembrator. Cell culture medium was exposed to powdered polymer (1-100 mg/ml) for a period of 7 days. HUVEC seeded as above were then exposed to the treated cell culture medium for 24 and 96 h. Finally, cell proliferation was studied over 16 days by seeding 2 x 10(5) HUVEC on films of PCBSU cast in glass Petri dishes. Cell viability and growth were assessed using Alamar blue, lactate dehydrogenase and Pico green assays and morphology was studied by Toluidine blue staining and scanning electron microscopy. Viable cells were demonstrated to be present after 16 days seeded on PCBSU. Exposing cells to PCBSU-treated cell culture medium resulted in no apparent damage to the cells at concentrations of 1 or 10 mg/ml, and only a slight reduction at 100 mg/ml after 96 h exposure. This study demonstrates that PCBSU can support the growth of endothelial cells for a prolonged period and does not demonstrate any significant toxic effects to cells. Thus it has the potential to be used both as a medical device and as scaffolding in tissue engineering applications.

Apoptosis↗

PEO-PPO-PEO-based poly(ether ester urethane)s as degradable reverse thermo-responsive multiblock copolymers.

Aiming at developing biodegradable thermo-responsive polymers that display enhanced rheological properties, a family of PEO-PPO-PEO based poly(ether ester urethane)s, was developed. The materials were produced following a two-step synthetic pathway. The PEO-PPO-PEO triblocks were first end-capped with LA or CL oligo(ester)s whereby pentablocks were produced. Then, the different precursors were chain extended using hexamethylene diisocyanate to create the respective polymers. The length and type of the ester block influenced the behavior of the molecules in water, especially their viscosity versus temperature response. The gelation temperature increased from 23 degrees C for a 20wt% F127 solution to 26 and 31 degrees C for pentablocks with 4.4 and 7.5 lactoyl units, respectively. Materials containing longer LA units failed to show any reverse thermo-responsiveness. The presence of the oligo(ester) blocks also reduced the viscosity of the gel at 37 degrees C. While F127 displayed a viscosity of around 28,000Pas, pentablocks containing 4.4 and 7.5 LA units showed values of 15,400 and 12,600Pas. Also, the viscosity at 37 degrees C as well as the gelation temperature decreased as the molecular weight of the oligo(ester)s increased. Finally, the degradation process of the gels was studied by monitoring their viscosity at body temperature and determining the molecular weight of the polymers, over time. Polymers were tailored so to combine high initial viscosity values with diverse degradation rates, as a function of the length and type of the oligo(ester) present along the polymeric backbone.

Absorbable Implants↗

Biocompatibility and biostability of a series of poly(carbonate)urethanes.

In this work, we synthesized several MDI-based poly(carbonate)urethanes (PCU) by using four different soft segments, including two aliphatic macrodiols (poly(hexyl, ethyl)carbonate diols, MW 2017 and 865, respectively) and two aromatic macrodiols (MW approximately 2000 and 1000, respectively), in different molar ratios to MDI. We demonstrate that these polymers exhibited various degree of micro-phase separation that further influenced their surface protein adsorption, platelet activation as well as cellular attachment and growth. Polyurethanes based on poly(hexyl, ethyl)carbonate diol with MW 2017 in a molar ratio MDI/macrodiol/chain extender of either 3/2/1 or 4/3/1 resulted in greater micro-phase separation as well as superior biocompatibility and biostability.

Adsorption↗

Permeability of dye through poly(urea-urethane) microcapsule membrane prepared from mixtures of di- and tri-isocyanate.

Poly(urea-urethane) microcapsules were prepared by the interfacial polymerization with using mixtures of tri- and di-isocyanate monomers as wall forming materials, and dioctyl phthalate containing an oil-soluble dye as a core material. The time course of the dye release in dispersing tetrahydrofuran was measured as a function of the weight fraction of tri-isocyanate monomer in the total monomer w and the core/wall material-weight ratio g. The dye release curves were well represented by an exponential function C=Ceq(1-e-t/tau), where C is the concentration of the dye in the dispersing medium, Ceq that at equilibrium state, t the elution time and tau is a time constant. tau increased linearly against w at high g, suggesting controllability of the release rate of microcapsules by varying tri-isocyanate/di-isocyanate ratio.

Calorimetry, Differential Scanning↗

Cellular engineering of conduits for coronary and lower limb bypass surgery: role of cell attachment peptides and pre-conditioning in optimising smooth muscle cells (SMC) adherence to compliant poly(carbonate-urea)urethane (MyoLink) scaffolds.

OBJECTIVE: We are developing a hybrid arterial bypass graft of compliant poly(carbonate-urea)urethane (MyoLink), endothelial and smooth muscle cells (SMCs). To enhance adhesion of SMCs we assessed various attachment factors and the effect of pre-conditioning on cell retention. METHODS: MyoLink segments were coated with either RGD, superfibronectin, fibronectin, fibronectin-like engineered polymer protein (FEPP), FEPP plus or type 1 collagen overnight. (111)Indium-radiolabelled SMCs were placed onto MyoLink segments for 48 h before being aspirated, then lavaged off. All grafts, aspirates and lavages were counted in a gamma counter. SMC viability on the MyoLink segments was also assessed for viability using the Alamar blue redox assay. Separately, MyoLink grafts lined with radiolabelled SMCs were divided into a pre-conditioned group, exposed to subarterial pulsatile flow whilst another group were held in static culture. After 1-week, grafts were exposed to arterial pulsatile flow whilst radioactivity was assessed using a gamma camera. RESULTS: Only FEPP plus significantly enhanced SMC attachment: mean of 32+/-6% cell attachment compared to 21+/-5% for uncoated control. Cell viability was enhanced by all attachment factors except fibronectin. Pre-conditioning was shown to significantly enhance the retention of SMCs onto the MyoLink once exposed to pulsatile arterial flow: the final attachment was 57+/-7% for the static and 76+/-7% for the pre-conditioned group. CONCLUSIONS: FEPP plus enhances SMC attachment to MyoLink. We believe this is because of its repeating sequences of RGD and its positive charge. Pre-conditioning enhances the retention of SMCs to MyoLink once exposed to pulsatile arterial flow.

Blood Vessel Prosthesis↗

Fatigue resistance and stiffness of glass fiber-reinforced urethane dimethacrylate composite.

STATEMENT OF PROBLEM: Retentive properties of cast metal clasps decrease over time because of metal fatigue. Novel fiber-reinforced composite materials are purported to have increased fatigue resistance compared with metals and may offer a solution to the problem of metal fatigue. PURPOSE: The aim of this study was to investigate the fatigue resistance and stiffness of E-glass fiber-reinforced composite. MATERIAL AND METHODS: Twelve cylindrical fiber-reinforced composite test cylinders (2 mm in diameter and 60 mm in length) were made from light-polymerized urethane dimethacrylate monomer with unidirectional, single-stranded, polymer preimpregnated E-glass fiber reinforcement. Six cylinders were stored in dry conditions and 6 in distilled water for 30 days before testing. Fatigue resistance was measured by a constant-deflection fatigue test with 1 mm of deflection across a specimen span of 11 mm for a maximum of 150,000 loading cycles. The resistance of the cylinder against deflection was measured (N) and the mean values of the force were compared by 1-way analysis of variance (alpha = .05). The flexural modulus (GPa) was calculated for the dry and water-stored cylinders for the first loading cycle. Scanning electron microscopy was used to assess the distribution of the fibers, and the volume percent of fibers and polymer were assessed by combustion analysis. RESULTS: The test cylinders did not fracture due to fatigue following 150,000 loading cycles. Flexural modulus at the first loading cycle was 18.9 (+/- 2.9) GPa and 17.5 (+/- 1.7) GPa for the dry and water-stored cylinders, respectively. The mean force required to cause the first 1-mm deflection was 33.5 (+/- 5.2) N and 37.7 (+/- 3.6) N for the dry and water stored cylinders, respectively; however, the differences were not significant. After 150,000 cycles the mean force to cause 1-mm deflection was significantly reduced to 23.4 (+/- 8.5) N and 13.1 (+/- 3.5) N, respectively (P < .0001). Scanning electron microscopy highlighted fiber- and polymer-rich areas within the specimens and indicated that individual fibers were well impregnated with resin. The combustion analysis studies identified the fiber content to be 35.9 vol%. CONCLUSION: The results of this study suggest that the fatigue resistance of the fiber-reinforced material examined was increased; however, the reduction in flexural modulus of fiber-reinforced composites may restrict their use where high rigidity is required, such as in removable partial denture clasps.

Air↗

Humidity-dependent dynamic infrared linear dichroism study of a poly(ester urethane).

Fourier transform infrared techniques, infrared difference spectroscopy and dynamic infrared linear dichroism (DIRLD), have been utilized to explore the effects of humidity and water absorption on a poly(ester urethane). An environmental infrared microbalance cell was used to measure the infrared spectra as a function of humidity and accompanying weight change for the absorption-desorption processes. The infrared difference data indicate that exposure to humidity affects the hydrogen-bonding interactions in the polymer. Dynamic infrared linear dichroism studies in tensile deformation mode as a function of humidity demonstrate how changes in water content affect the orientational response of functional groups. Complex behavior as a function of humidity for functional groups involved in hydrogen bonding indicates that water absorbed by the polymer affects the micro-environments near these functional groups.

Humidity↗