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PubMed1991

The bonding behavior of calcite to bone.

Plates of calcite (CaCO3) were implanted in rabbit tibiae, and their biocompatibility and bonding ability to bone were studied. The plates were also implanted subfascially in rabbit muscle for 8 weeks, and changes on their surfaces in the body were examined. Contact microradiography and Giemsa surface stain demonstrated direct bonding between calcite and bone without interpositions. The average failure load of the interface between calcite and bone was 4.11 kg, indicating an adequate strength of bonding. However, a Ca-P-rich layer, which formed on the surfaces of other bioactive ceramics in vivo, was not detected by a scanning electron microscope-electron probe x-ray microanalyzer. Scanning electron micrographs of the surface of calcite implanted subfascially for 8 weeks showed marked degradation and a rough surface. However, the surface apatite layer was not detected by thin-film x-ray diffraction analysis and Fourier transform infrared reflection spectroscopy. Calcite is a biodegradable material that bonds to bone without a surface apatite layer. The mechanical bonding provided by the anchoring effect of the newly formed bone into the surface roughness of calcite is considered to be a major factor in calcite-bone bonding.

Animals↗
PubMed1991

Bone bonding mechanism of beta-tricalcium phosphate.

It has been proposed that the formation of a surface apatite layer in vivo on surface active ceramics is an essential condition for chemical bonding between ceramics and bone tissue. To clarify the difference in bone-bonding mechanisms between surface active ceramics and bioresorbable ceramics, two experiments were performed using plates of dense beta-tricalcium phosphate (beta-TCP). First, plates of beta-TCP were implanted subcutaneously in rats for 8 weeks. Surface change due to bioresorption was observed with scanning electron microscopy. Formation of the apatite layer on the surface was investigated using thin-film x-ray diffraction and Fourier transform infrared reflection spectroscopy. Second, plates of beta-TCP were implanted in tibiae of rabbits for 8 and 25 weeks and subjected to the detaching test to measure bone-bonding strength. beta-TCP bonded strongly to bone. Undecalcified sections of the interface of bone and beta-TCP were examined with SEM-EPMA. However, by physicochemical methods, no formation of surface apatite layer was observed. These results suggest that beta-TCP bonds to bone through microanchoring between bone and rough surface of resorbed beta-TCP.

Animals↗
PubMed1991

Apatite formation on the surface of Ceravital-type glass-ceramic in the body.

Previous studies on surface structural changes in vitro as well as in vivo of bioactive A-W-type glass-ceramics and Bio-glass-type glasses showed that the essential condition for glasses and glass-ceramics to bond to living bone is formation of a bonelike apatite layer on their surfaces in the body. Gross et al., however, had explained the bone-bonding mechanism of Ceravital-type apatite-containing glass-ceramic without mentioning formation of the surface apatite layer. In the present study, apatite formation on the surface of one of Ceravital-type glass-ceramics was investigated in vitro as well as in vivo. An apatite-containing glass-ceramic of the composition Na2O 5, CaO 33, SiO2 46, Ca(PO3)2 16 wt%, which was named KGS by Gross et al., was soaked in an acellular simulated body fluid which had ion concentrations almost equal to those of the human blood plasma. The same kind of glass-ceramic was implanted into a rabbit tibia. Thin-film x-ray diffraction, Fourier transform infrared reflection spectroscopy, and scanning electron microscopic observation of the surfaces of the specimens soaked in the simulated body fluid showed that Ceravital-type glass-ceramic also forms a layer of carbonate-containing hydroxyapatite of small crystallites and/or a defective structure on its surface in the fluid. Electron probe x-ray microanalysis of the interface between the glass-ceramic and the surrounding bone showed that a thin layer rich in Ca and P is present at the interface. These findings indicated that Ceravital-type glass-ceramics also form the bonelike apatite layer on its surface in the body and bond to living bone through the apatite layer.

Animals↗
PubMed1990

Time-resolved cryotransmission electron microscopy.

We describe a new technique, time-resolved cryotransmission electron microscopy (TRC-TEM), that can be used to study changes in microstructure occurring during dynamic processes such as phase transitions and chemical reactions. The sample is prepared on an electron microscope grid maintained at a fixed temperature in a controlled atmosphere. The dynamic process is induced on the grid by a change in pH, salt, or reactant concentration by rapid mixing with appropriate solutions. Alternatively, induction is by rapid change of specimen temperature, or by controlled evaporation of a volatile component. We call such procedures on-the-grid processing. The dynamic process is permitted to run for a defined time and then the thin-film specimen is thermally fixed by plunging into liquid ethane at its freezing point, producing a cryotransmission electron microscopy specimen. By repeating this procedure with varying delays between induction and sample fixation, we can observe transient microstructures. We demonstrate the use of TRC-TEM to study the intermediate structures that form during the transitions between L alpha, III, and HII liquid crystalline phases in phospholipid systems. We also identify several other possible applications of the technique.

Cryopreservation↗
PubMed1989

Lubrication of selected salivary molecules and artificial salivas.

The lubrication regime displayed by human salivas (parotid and submandibular-sublingual), purified salivary molecules (the mucins MG1 and MG2 and alpha-amylases), and selected artificial salivas (Oracare D, Saliva Substitute, and Orthana) was assessed in vitro using a friction-testing device. Thin-film (boundary) lubrication was observed for all of the salivary samples and two of the artificial salivas examined. Oracare D, a glycerol-based artificial saliva, was the exception since it lubricated by a thick-film (hydrodynamic) regime. On a molar basis, the best lubricants of the purified salivary molecules were MG1 greater than MG2 approximately nonglycosylated alpha-amylases approximately glycosylated alpha-amylases.

Amylases↗
PubMed1991

Lipid polymorphism as observed by cryo-electron microscopy.

Lipid polymorphism was studied with the aim to gain more insight in bilayer to non-bilayer phase transitions, with particular emphasis on the development of cubic structures on one hand and inverted hexagonal structures on the other hand. Thin vitrified films prepared from aqueous lipid suspensions were used in this study. The entire hydrated contents of these films can be visualized in their two-dimensional projection by cryo-electron microscopy. As the starting material, unilamellar vesicles were prepared from mixtures of dioleoylphosphatidylethanolamine, dioleoylphosphatidylcholine and cholesterol. By heating of the suspension, vesicle fusion (Frederik et al. (1989) Biochim. Biophys. Acta 979, 275-278) and lipid polymorphism was induced. From these suspensions thin films were prepared at various temperatures, and vitrified for low temperature observation. In a parallel series of experiments samples were fast-frozen for freeze-fracture analysis. In vitrified thin films bilayer structures were often observed in coexistence with an inverted hexagonal structure. The bilayer areas were frequently of a complex structure because multiple contacts between stacked membranes were found. These contact points were variable in size and shape and usually had the form of a diabolo (when viewed side-on) giving the impression of a bilayer contact with an aqueous channel. This structure is compatible with the interlamellar attachment site (ILA) proposed by Siegel ((1986) Biophys. J. 49, 1155-1170). In some specimens ILA's seemed to merge into arrays. After thermal cycling of the suspension, arrays of packed globules were observed, which are likely the result of close packing of ILA's. The arrays probably represent a cubic structure. A comparison of freeze-fracture replicas and vitrified thin films indicated that both techniques may provide valuable structural information on lipid polymorphism. Most of the lipidic particles observed by freeze-fracturing probably correspond to the ILA's (fractured around their waist region) as observed in vitrified thin films. The results obtained with vitrified thin films were interpreted in relation to the principles of thin-film formation. Finally, we speculate that lipid structures occurring close to each other in space may represent a developmental series of structures occurring successively in time.

Cholesterol↗
PubMed1990

Hippocampal cell death following ischemia: effects of brain temperature and anesthesia.

The effect of brain temperature and anesthesia on ischemic neuronal damage was studied in the hippocampal formation using the four vessel occlusion model in awake and anesthetized rats. Neuronal damage was assessed by immunocytochemistry and silver impregnation of tissue sections. The degree of ischemia was monitored by recording spontaneous and evoked electrical activity from the hippocampus and dentate gyrus in all animals. In addition, the hippocampal temperature and oxygen tension were also recorded using a chamber-type thin-film microelectrode in the anesthetized animals. Fifteen minutes ischemia in the awake animals caused greater neuronal damage and mortality of animals than 30 min ischemia in anesthetized rats. The temperature of the brain was found to drop by 4-6 degrees C during complete forebrain ischemia in the latter group. Neuronal damage was observed infrequently in the hippocampus of these animals. When the brain temperature was kept constant at the preischemic level during 30 min occlusion, all animals died within a day, while after 15 min occlusion the majority showed an almost complete degeneration of CA1 pyramidal cells and hilar somatostatin immunoreactive neurons. Following 15 min ischemia, the awake animals showed a similar cell loss in the CA1 region and the hilus. It is concluded that, in the anesthetized animals prepared for acute recording, the decreased temperature of the brain during ischemia is a major factor in protecting neurons from damage, but that Equithesin anesthesia also has a significant protective effect. Consistent ischemic degeneration occurs in awake animals by four vessel occlusion, if the brain temperature is controlled and the completeness of ischemia is monitored by recording spontaneous and evoked electrical activity with chronic electrodes.

Anesthetics↗
PubMed1990

Heterogeneous electron transfer of cytochrome c facilitated by polypyrrole and methylene blue polypyrrole film modified electrodes.

Polypyrrole and methylene blue incorporated polypyrrole thin-film modified electrodes were prepared by the electrochemical polymerization method. These modified electrodes may facilitate heterogeneous electron transfer of cytochrome c with high electrocatalytic activity and good stability. Optical thin-layer spectroelectrochemical techniques were used to determine the characteristics of these electrochemical processes such as formal redox potential (E0), electron transfer number (n), and the apparent rate constant (ks.h0).

Cytochrome c Group↗
PubMed1991

Shared-stimulus driving and connectivity in groups of neurons in the dorsal cochlear nucleus.

Extracellular spike discharges were recorded from ensembles of up to five neurons simultaneously in the DCN of guinea pig using solid-state, thin-film, multichannel electrodes having up to five recording sites spanning up to 600 microns. Responses from 73 unit pairs were collected of which 54 had both units responding to pseudorandom wideband noise stimulation. Shared-stimulus driving was present in 78% (42/54) of the unit pairs and could be attributed to an overlap in their spectral sensitivities. Effective connectivity was indicated for 87% (47/54) of the unit pairs. Wideband noise proved more useful than tonebursts for investigating shared-stimulus driving and connectivity because it evoked widespread, but not overly synchronous, responses in the ensembles.

Acoustic Stimulation↗
PubMed1990

Thermoelectric enzyme sensor for measuring blood glucose.

A new calorimetric sensor has been developed which employs a thin-film thermopile in association with an immobilized enzyme. The thermopile detects the minute temperature rise that occurs when a specific chemical substrate is catalyzed by the enzyme. A prototype sensor is described which generates an equivalent proportional voltage response to glucose concentrations present in either buffer solution or blood. These sensors have remained useful for up to 18 days when operated intermittently for measuring glucose in buffer solutions, or for up to 4 days when operated continuously. When implanted inside cardiovascular shunts on anesthetized dogs, the sensors responded appropriately to changes in the blood glucose concentration.

Animals↗
PubMed1979

Generation of aspartate aminotransferase multiple forms by deamidation.

The development of aspartate aminotransferase subforms in vitro was followed by densitometry after thin-film isoelectric focusing. At the same time ammonia production was measured. Each reaction can be expressed in terms of a first-order process in which 2 mol of glutamine or asparagine/mol of dimer are deamidated with a half time of 22 days. The more negatively charged subforms developed in vitro were almost fully active. Another process occurred leading to inactivation by coenzyme modification, and this was independent of deamidation. Although the enzyme formed absorbed maximally at 340nm, it was different from the naturally occurring inactive enzyme that absorbs at this wavelength.

Ammonia↗
PubMed1991

The UT 19-channel DC SQUID based neuromagnetometer.

A 19-channel DC SQUID based neuromagnetometer is under construction at the University of Twente (UT). Except for the cryostat all elements of the system are developed at the UT. It comprises 19 wire-wound first-order gradiometers in a hexagonal configuration. The gradiometers are connected to planar DC SQUIDs fabricated with a Nb/Al, AlO kappa/Nb technology. For this connection we developed a method to bond a Nb wire to a Nb thin-film. The SQUIDs are placed in compartmentalised Nb modules. Further, external feedback is incorporated in order to eliminate cross talk between the gradiometers. The electronics basically consist of a phase-locked loop operating with a modulation frequency of 100 kHz. Between SQUID and preamplifier a small transformer is used to limit the noise contribution of the preamplifier. In the paper the overall system is described, and special attention is paid to the SQUID module (bonding, compartments, external-feedback setup, output transformer).

Equipment Design↗
PubMed1977

Tear-film dynamics and oxygen tension under a circular contact lens.

The cornea requires a minimum level of oxygenation; this may be prevented by a contact lens, and corneal swelling and other undesirable effects may result. By employing thin-film lubrication theory, this study derives analytical solutions for tear-film velocity components and pressure under a circular contact lens. Oxygen-tension contours are then obtained for various values of squeeze action and parallel motion of the lens.

Contact Lenses↗
PubMed1992

Regeneration microelectrode array for peripheral nerve recording and stimulation.

A microelectrode array capable of recording from and stimulating peripheral nerves at prolonged intervals after surgical implantation has been demonstrated. The microelectrode array, fabricated on a silicon substrate perforated by multiple holes (referred to as via holes), is implanted between the ends of a surgically severed nerve. Regenerating tissue fixes the device in place to provide a stable mapping between the microelectrodes and the axons in the nerve. Processes were developed for the fabrication of thin-film iridium microelectrodes, micromachined via holes, and silicon nitride passivation layers. All fabrication methods were designed to be compatible with standard CMOS/BiCMOS processes to allow for on-chip signal processing circuits in future designs. Such arrays, implanted in the peroneal nerves of rats, were used to record from and stimulate the nerves at up to 13 months postoperatively.

Animals↗
PubMed1991

Perspective and limitations of cryo-electron microscopy. From model systems to biological specimens.

We investigated the possibility of vitrifying temperature-sensitive lipid phases as well as (small) biological specimens. From a suspension of unilamellar vesicles, prepared from dipalmitoyl-phosphatidylcholine (DPPC), thin aqueous films were formed at various temperatures. With cryo-electron microscopy vesicles were found to be smooth, rippled and faceted or faceted only, depending on the temperature of thin-film formation (318, 312 and 296 K respectively). The morphology and the electron diffraction patterns indicate that membranes can by physically fixed by vitrification in their high-temperature configuration and studied at low temperature by cryo-electron microscopy. This finding suggests that it may also be possible to preserve, in their original state, the more complex membrane systems found in living organisms by initiating rapid-cooling at a physiological temperature. This was explored by vitrification of thin films formed on specimen grids with (human) blood platelets adhering to collagen fibres. Low-temperature observation with an acceleration voltage of 120 kV revealed subcellular details, More details were observed when using higher accelerating voltages (200 and 300 kV) of the electron beam. The results presented in this paper illustrate the great potential of cryo-electron microscopy in the study of membrane dynamics, both in relatively simple model membrane systems and in more complex biological membrane systems.

Blood Platelets↗
PubMedDate not supplied

Demonstration of megavoltage and diagnostic x-ray imaging with hydrogenated amorphous silicon arrays.

Flat-panel imagers consisting of the first large area, self-scanning, pixelated, solid-state arrays made with hydrogenated amorphous silicon (a-Si:H) are under development by the authors for applications in diagnostic x-ray and megavoltage radiotherapy imaging. The arrays, designated by the acronym MASDA for multi-element amorphous silicon detector array, consist of a two-dimensional array of a-Si:H photodiodes and thin-film transistors and are used in conjunction with scintillating materials. Imagers utilizing MASDA arrays offer a variety of advantages over existing technologies. This article presents initial megavoltage and diagnostic-quality x-ray images taken with several such arrays including the first examples of anatomical-phantom images. The external readout electronics and imaging techniques required to obtain such images are outlined, the construction, operation, and advantages of the arrays briefly reviewed, and the future potential of this new technology discussed.

Diagnostic Imaging↗
PubMed1990

Evaluation of a silicon-substrate modiolar eighth nerve implant in a guinea pig.

The availability of thin-film multichannel electrodes provides new possibilities for implantation and direct stimulation of the modiolar portion of the auditory nerve. Electrodes in direct contact with the auditory nerve should be functional at lower thresholds and might require fewer remaining neurons for stimulation compared to electrodes in the scala tympani. This strategy would also provide close contact with neural elements subserving a greater frequency range. Implantation of the eighth nerve may also be advantageous in profoundly deaf subjects who lack an implantable scala tympani. In this study we evaluated the effect of surgical implantation and chronic placement of a silicon substrate implant in the modiolar portion of the auditory nerve of the guinea pig. Of six chronically implanted ears, five showed changes limited to the loss of spiral ganglion cells in the canal of Rosenthal, immediately adjacent to the implant. The sixth ear showed more extensive cochlear alteration in a pattern suggestive of vascular injury. In separate acute experiments, implants were placed in the modiolar portion of the auditory nerve and electrophysiologic analysis was performed. Middle latency responses with good morphology were obtained at thresholds below those found with scala tympani implants. Input-output functions exhibited a plateau in response amplitude at stimulus levels below thresholds for seventh or vestibular portion of the eighth nerve. Further modifications of the modiolar portion of the auditory nerve electrode design will include development of an electrode interconnect that will allow chronic implantation with stimulation.

Animals↗