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Biomedical subjects

J D Gaylor

Publications and source records attributed to J D Gaylor.

At least 19 recordsLinked to original sources

In vitro investigation of the blood response to medical grade PVC and the effect of heparin on the blood response.

This paper reports the results of an investigation into the blood response of polymers in vitro, using non-anticoagulated and heparinised blood and plasma. The materials studied were regenerated cellulose, (Cuprophan), an acrylonitrile-allyl sulphonate copolymer (AN69S), and medical grade polyvinyl chloride plasticised with di-2-ethyl-hexyl-phthalate (PVC/DEHP). Blood-material or plasma-material contact was achieved using a parallel plate flow cell, and C3a generation and FXII-like activity measured. The results of the study with non-anticoagulated human blood show that PVC/DEHP is a high complement activator. C3a concentration in the blood was higher after contact with PVC/DEHP than after contact with regenerated cellulose. The introduction of heparin in the blood induced complex alterations in the blood response. C3a generation could be elevated, decreased, or remain the same, depending on the material. The FXII-like activity on the surface of the PVC/DEHP after contact with plasma was also higher than the other two polymers. The introduction of heparin could increase or decrease FXII-like activity, depending on material. The patterns of response obtained with non-anticoagulated blood in vitro for AN69S and Cuprophan bore a strong resemblance with patterns of response obtained in the clinic, whereas those obtained with heparinised blood in vitro did not.

Acrylic Resins↗

Oxygen transfer in a diffusion-limited hollow fiber bioartificial liver.

A mathematical model was developed to predict oxygen transport in a hollow fiber bioartificial liver device. Model parameters were taken from the Hepatix ELAD configuration; a blood perfused hollow fiber cartridge with hepatocytes seeded in the extracapillary space. Cellular oxygen uptake is based on Michaelis-Menten kinetics, and nonlinear oxygen transport in the blood is considered. The effect of modulating three important parameters is investigated, namely, the Michaelis-Menten constants Vm (volumetric oxygen consumption of the hepatocytes) and Km (half-saturation constant), and hollow fiber oxygen permeability. A computer implementation of the model is used to assess whether a given cell mass could be maintained within such a device. The results suggest that liver cell lines possessing low rates of oxygen consumption could be maintained if membranes of sufficiently high oxygen permeability are used. For primary hepatocytes, which have much higher oxygen demands, radial transport of oxygen is rate limiting, and the axial-flow hollow fiber cartridge is thus an inappropriate design for use as a bioartificial liver with primary hepatocytes.

Algorithms↗

Cytotoxicity of bile in human Hep G2 cells and in primary cultures of rat hepatocytes.

There has been increasing interest in the development of a hepatocyte bioreactor for the treatment of acute hepatic failure; however, little is known about the effect of hepatocyte byproducts on the viability of the cells in the bioreactor environment. We investigated the effects of increasing concentrations of bile on the growth and viability of the human hepatoma cell line Hep G2 and on the cytochrome P-450 content and dependent mixed function oxidase (MFO) activities, reduced glutathione (GSH) content, and glutathione S-transferase (GST) activity of primary cultures of rat hepatocytes. Our purpose was to determine whether or not it would be necessary to pretreat the plasma from patients with acute liver failure to remove elevated bile concentrations which might be toxic to the hepatocytes in an artificial liver device. Bile was found to inhibit Hep G2 cell growth at concentrations as low as 0.1% and to decrease viability at concentrations above 0.5%. The cytochrome P-450 and GSH contents and the activities of the MFO system and of GST were decreased in the primary cultures of hepatocytes following 24 h treatment with concentrations of bile at and above 0.5%. The MFO activities associated with different cytochrome P-450 isoenzymes decreased to different extents in the presence of bile with the O-dealkylation of pentoxyresorufin being more labile than that of ethoxyresorufin. Our data indicate that elevated bile concentrations are cytotoxic to liver cells, and it may be necessary to pretreat patient plasma to decrease its bile content to protect the cells during the clinical operation of a hepatocyte bioreactor device.

Animals↗

The effects of serum from patients with acute liver failure on the growth and metabolism of Hep G2 cells.

In many bioartificial liver systems currently being designed and evaluated for use in fulminant hepatic failure, direct contact is required between the patient's blood and the liver cells in the device. The efficacy of such devices will be influenced by the interaction of fulminant hepatic failure (FHF) patient serum with the cells. We have found that FHF serum inhibits the growth rate and the synthesis of DNA, RNA, and protein; disturbs glutathione homeostasis; and induces morphological changes in cultured human Hep G2 cells. These interactions should influence the design of bioartificial liver devices based on proliferating cell lines and indicate the requirement to pretreat FHF patient plasma to reduce the toxin load.

Animals↗

Water vapour transmission rates in burns and chronic leg ulcers: influence of wound dressings and comparison with in vitro evaluation.

One of the main functions of wound dressings is to control water vapour transmission rate (WVTR) from wounded skin. In this paper, the influence of hydrocolloid, knitted viscose and gauze dressings was evaluated through in vivo measurement of WVTR in burns and chronic leg ulcers utilizing an evaporimeter. The results suggest that the evaporative water vapour loss from exposed skin wounds depends mainly on the wound depth, and that chronic leg ulcers have the same level of the WVTR as full thickness burns. Compared with the knitted viscose and gauze dressings, hydrocolloid dressing has a greater effect on reducing evaporative water loss, with WVTR being 20-30% of that of exposed wounds under the conditions used in this study. This result is in agreement with that obtained in an in vitro evaluation.

Adult↗

In vitro uptake and elimination of isoflurane by different membrane oxygenators.

OBJECTIVE: This study was designed to investigate the effect of membrane oxygenator design and composition on the uptake and elimination of isoflurane. DESIGN: Prospective, in vitro laboratory study. SETTING: Bioengineering laboratory. PARTICIPANTS: Three types of membrane oxygenator were tested: the SM-35 (polydimethylsiloxane in sheet form), the CML (polypropylene in sheet form), and the SAFE II (polypropylene in hollow-fiber form). The oxygenators were incorporated into a standard cardiopulmonary bypass circuit. INTERVENTIONS: Isoflurane was added to the oxygenator input gas and measured in exhaust gas and in (bovine) blood leaving the oxygenator at 1, 2, 3, 5, 7, 10, 15, and 20 minutes. The isoflurane vaporizer was then turned off, and samples were obtained at the same time intervals. The experiment was performed at 28 degrees C and 37 degrees C. MEASUREMENTS AND MAIN RESULTS: Uptake and elimination of isoflurane were slower via the SM-35 compared with the CML and the SAFE II (p < 0.01). CONCLUSIONS: If isoflurane is administered during cardiopulmonary bypass, knowledge of the influence of oxygenator membrane composition on its pharmacokinetics is essential if patient awareness and unexpected cardiovascular depression are to be avoided.

Anesthesia, Inhalation↗

Techniques for measurement of oxygen consumption rates of hepatocytes during attachment and post-attachment.

Three techniques for measuring oxygen consumption rate (OCR) of cultured cells relevant to the development of bioartificial liver devices are reported. In an oxystat apparatus, HepG2 cells immobilised on Cytodex 3 microcarriers at a concentration of 10(6) cells ml-1 had a mean OCR of 0.7 nmol s-1/10(6) cells. The OCR decreased with increasing cell density, a characteristic previously reported for other cell lines. Rat hepatocytes immobilised on single collagen layers in a flow cell and challenged with ammonia had a mean OCR of 0.59 nmol s-1/10(6) cells. A novel two-compartment oxystat system was used to determine the OCR of rat hepatocytes during the attachment phase. OCR declined from 1.0 nmol s-1/10(6) immediately after seeding to 0.7 nmol s-1/10(6) cells at nine hours. The low OCR for HepG2 reflects loss of certain oxygen dependent metabolic pathways. The OCR measured for rat hepatocytes during and post-attachment are significantly higher than those reported elsewhere and have major implications for the development of bioartificial liver devices.

Animals↗

In vitro blood compatibility evaluation of hollow fibre membrane using a controlled flow system: a comparative study.

A procedure has been established for the in vitro assessment of hollow fibre haemodialysis membranes. A 30 ml syringe containing 20 ml of fresh non-anticoagulated blood was mounted onto a non-pulsatile syringe pump and blood was perfused through minimodules constructed from 80 fibres retrieved from Cuprophan (Baxter ST15), cellulose acetate (M57-12, JMS Co Ltd, Hiroshima, Japan), and AN69HF (Filtral 20, Hospal, France) dialysers. Samples were collected before perfusion, 3, 6, 9 and 12 minutes. The modules were clamped vertically to minimise the effect of red cell pooling and the dialysate compartment was filled with 0.9% saline to minimise ultrafiltration. After sample processing, complement C3a, thrombin-antithrombin III complexes, prothrombin F1 + 2, and factor XII-like activity were evaluated. The results indicated that the system could discriminate between the membranes evaluated and therefore was a relevant procedure for the assessment of hollow fibre haemodialysis membranes.

Antithrombin III↗

In vitro contact phase activation with haemodialysis membranes: role of pharmaceutical agents.

Contact phase activation was investigated in vitro using flat sheet type of haemodialysis membranes, Cuprophan (Akzo, Faser, Germany) and AN69S (Hospal, France), and a negatively charged polyamide Ultipor NR 14225 membrane as a control. The investigation focussed on the determination of factor XII-like activity (FXIIA) as an indicator of contact phase activation in the supernatant phase and at the membrane surface after plasma-membrane contact using an incubation test cell. The findings were compared with the observations from a plasma-free system utilizing purified unactivated factor XII. The plasma FXIIA bound to the membrane surface was significantly different between the membranes, while the supernatant phase FXIIA exhibited no significant differences. In contrast, the plasma-free system exhibited significant differences in the supernatant FXIIA and membrane-bound FXIIA for all the materials used and the magnitude of the activity was significantly greater for negatively charged materials. This finding demonstrated the strong influence of the interaction of other plasma constituents on the membrane surface and as such the binding and subsequent activation of factor XII may be altered possibly due to competitive binding and steric hindrance. On the addition of anticoagulants such as heparin, low-molecular-weight heparin, citrate and hirudin, no significant differences were observed in plasma supernatant phase FXIIA. However, each anticoagulant appears to have a distinct influence on the magnitude of plasma membrane-bound FXIIA. On the addition of aprotinin (a kallikrein inhibitor), no significant differences were observed in the plasma supernatant FXIIA. In contrast, aprotinin appears to significantly reduce membrane-bound FXIIA on Cuprophan and polyamide NR, but significantly increase the magnitude of the membrane-bound FXIIA on AN69S.

Anticoagulants↗

In vitro assessment of water vapour transmission of synthetic wound dressings.

The water vapour transmission rate (WVTR) of 14 commercially available wound dressings (11 hydrocolloids, 2 hydrogels and 1 polyurethane film) was evaluated over 24 and 48 h periods using a modified ASTM standard method (ASTM E96-90). For the 48 h studies a novel microcomputer-controlled apparatus was employed. The dressings exhibited a wide range of WVTRs (76-9360 g m-2 d-1 at 24 h under forced air convection of 0.4 m s-1). The influence of air velocity of 0.4 m s-1 was not significant if the WVTR of the dressing was less than 880 g m-2 d-1 when measured under static air conditions. The influence of outer barrier layers and additional bandages on WVTR was also examined.

Bandages↗

Blood-contacting biomaterials: bioengineering viewpoints.

The investigation of blood-contacting biomaterials is an important challenge and is relevant for an improvement in the clinical application of biomaterials. With the purpose of improved clinical treatment, bioengineering viewpoints of blood-contacting biomaterials cover the material options and selection, the utilization of materials, the development of materials with better properties, and processing characteristics, and the design of relevant evaluation procedures. The bioengineering objective remains that of achieving an enhanced understanding of the relationship between a biomaterial and the biological response.

Biocompatible Materials↗

Endemic fluorosis: an analysis of needs and possibilities based on case studies in Kenya.

The decline in prevalence of dental caries in the western world is largely ascribed to the protective role of fluoride in water. However, in several Third World regions, its presence in excessive amounts has been detrimental to the health of resident communities due to the resulting endemic dental and skeletal fluorosis. As a prelude to introduction of preventive intervention among the affected communities, there is need to assess knowledge and perception, and affordable and effective possibilities. The results of such an assessment which was based on response of mothers from two affected communities showed that objectionable dental fluorosis was not viewed as a common health problem in the context of other more common diseases. A reawakening of interest and concern was evident when issues pertaining to oral health were addressed. Dental fluorosis was viewed as an important problem because of its unfavourable effects on an individual's personality by between 60.4 and 84.3% of the respondents. While 60% and over of the respondents attributed the problem to water, knowledge on perceived methods of prevention of fluorosis were significantly lower. Only 12% of respondents from a relatively higher income group were instituting relevant preventive strategies. Although defluoridation of water had been instituted in one area, problems relating to wrong choice of water for defluoridation to inadequate distribution of the defluoridated water were evident. Education of the communities on methods of reducing fluoride ingestion and the significance of the defluoridated water, rationalizing the distribution of defluoridated water, facilitating collection of rain water, protection of available low fluoride surface water from contamination with agro-chemicals and household defluoridation were recommended.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Culture of a differentiated liver cell line, Hep G2, in serum with application to a bioartificial liver: effect of supplementation of serum with amino acids.

Much effort has been directed toward the development of serum-free, hormonally defined culture conditions for the maintenance of differentiated functions in many cell types, including hepatocytes. However, in the development of a hepatocyte bioreactor for artificial liver support, many designs propose the maintenance of cells in plasma as opposed to defined culture medium. There is very little reported literature on the growth and function of cells cultured in plasma or serum; therefore, the effect of increasing serum concentrations was investigated using the human hepatoma, Hep G2, as a model cell line. It was found that Hep G2 can survive and grow in 100% serum if the serum is supplemented with L-glutamic acid, glycine, and L-cysteine.

Amino Acids↗

Biomaterials in cardiopulmonary bypass.

The improved utilization of biomaterials in cardiopulmonary bypass is dependent on polymer science and technology, procedures for blood compatibility assessment, optimization of biomaterial/antithrombotic agent combinations and the interpretation of clinical data.

Biocompatible Materials↗

Polymeric biomaterials: influence of phosphorylcholine polar groups on protein adsorption and complement activation.

The introduction to polymeric biomaterials of phosphorylcholine polar groups represents an approach towards the development of materials with improved blood compatibility. In this respect, two biomaterials, one a copolymer of butyl methacrylate and 2-methacryloyloxyethylphosphorylcholine (MPC), (poly(BMA-co-MPC) and the other, MPC-grafted Cuprophan, were examined with respect to their influence on protein adsorption and complement activation. Protein adsorption was studied by measurement of the adsorption of radiolabelled single proteins (albumin and fibrinogen), while complement activation was measured using radioimmunoassay for C3a des Arg. The investigation demonstrated that the polymers containing phosphorylcholine polar groups can achieve a marked reduction in protein adsorption and complement activation and supports the utilization of phosphorylcholine polar groups as a means of improving the compatibility of biomaterials for blood-contacting applications.

Adsorption↗