Biomedical subjects

T D Turner

Publications and source records attributed to T D Turner.

At least 19 recordsLinked to original sources

PubMed1998

Biocompatibility of potential wound management products: hydrogen peroxide generation by fungal chitin/chitosans and their effects on the proliferation of murine L929 fibroblasts in culture.

Agaricus bisporus, Fusarium graminearum, Phycomyces blakesleeanus, unbleached and bleached, Rhizomucor miehei, and Rhizopus oryzae were examined as sources of fungal chitin/chitosan. The nitrogen content of the alkalitreated mycelia/sporangiophores obtained after optimization of culture conditions, and of similarly treated A. bisporus stipes, was 2.87, 1.29, 6.27, 6.50, 4.80, and 4.95% w/w, respectively, which relates to an estimated chitin content of 42, 19, 91, 94, 70, and 72%, respectively. The hydrogen peroxide (H2O2)-generating ability of the treated fungal materials after 8 h at pH 7.4 and 37 degrees C decreased in the order R. oryzae > P. blakesleeanus unbleached approximately R. miehi > F. graminearum > A. bisporus > P. blakesleeanus bleached. This did not correlate with estimated chitin content. The effect of these fungal materials on the rate of proliferation of murine L929 fibroblasts in culture also was examined. Both pro- and antiproliferant effects were observed. Significant (P < .05) proproliferant effects were observed on day 6 with R. miehei, R. oryzae, and P. blakesleeanus (unbleached and bleached) at 0.01% w/v. The greatest antiproliferant effect was observed with R. oryzae at 0.05% w/v on day 6 (-63% relative to the control, P < .05; cell viability, 95%). In contrast, A. bisporus failed to affect cell yield significantly at either 0.01 or 0.05% w/v. Addition of catalase to cultures containing R. oryzae or R. miehei at 0.05% w/v failed to abolish the antiproliferant effect on day 3, instead producing a small but significant (P < .05) increase in the effect. Catalase also failed to affect significantly the antiproliferant effect of F. graminearum at 0.05% w/v, but did abolish the proproliferant effect of P. blakesleeanus (unbleached and bleached) on day 3. Overall, our results suggest that the H2O2 being generated by the fungal materials modulates cell proliferation but that this effect is superimposed upon a H2O2-independent antiproliferant effect manifesting itself at the higher concentrations of fungal material. The antiproliferant effect was not attributable to Ca2+, Mg2+, or Fe2+ depletion although chelation of Fe2+ did correlate with H2O2-generating ability. Only P. blakesleeanus appears to lack this antiproliferant activity while retaining H2O2-generating activity. These results may aid the selection of fungal chitin/chitosan for further evaluation as a potential wound management material.

Animals↗

PubMed1998

Penetration of dynorphin 1-13 across the blood-brain barrier.

Previous studies have demonstrated neuroprotective effects of the opioid peptide dynorphin (dyn) 1-13 in focal cerebral ischemia. The passage of dyn 1-13 across the blood-brain barrier (BBB) was studied by a modification of the Oldendorf technique in the normal rat and cat, as well as in a feline model of experimentally induced focal cerebral ischemia. In the rat, dyn 1-13 penetration of the BBB could not be detected by this technique, even in the presence of peptidase inhibitors. In contrast, dyn 1-13 did cross the BBB into the normal cat hippocampus, cortex and cerebellum. The passage of dyn 1-13 across the BBB was greater in cats with experimentally induced focal cerebral ischemia. Some of the tritium-labeled material which crossed the BBB was confirmed by high performance liquid chromatography to be dyn 1-13. These studies support the hypothesis that the therapeutic effects observed after the peripheral administration of dyn 1-13 to cats with focal cerebral ischemia can be produced by a central mechanism of action.

Analgesics, Opioid↗

PubMed1995

Management of veterinary wounds.

A comparison is made between the management of human and animal wounds. Wounds should be classified in order to aid the selection of the most appropriate form of treatment. The concept of the 'golden period' is now less accepted and effective early surgical treatment is advocated. The application of newer dressing materials is recommended together with the initial results of a pilot study based on the application of these materials to animal wounds. An enthusiasm was demonstrated in this study and the need for further research has been identified.

Animals↗

PubMed1994

Biocompatibility of potential wound management products: fungal mycelia as a source of chitin/chitosan and their effect on the proliferation of human F1000 fibroblasts in culture.

Aspergillus oryzae, Mucor mucedo, and Phycomyces blakesleeanus cultures were examined as sources of chitin/chitosan. The nitrogen content of the alkali-treated mycelia/sporangiophores of A. oryzae, M. mucedo, and P. blakesleeanus was 2.52, 3.61, and 6.27% w/w, which relates to an estimated chitin content of 37, 52, and 91%, respectively. The effect of these fungal materials on the rate of proliferation of human F1000 fibroblasts in culture was examined. At 0.01% w/v, all three materials exhibited significant (P < .05) proproliferant activity over a period of 13 days. However, at 0.05% w/v, P. blakesleeanus further enhanced cell proliferation, whereas A. oryzae and M. mucedo produced a significant (P < .05) antiproliferant effect. Higher concentrations of P. blakesleeanus (0.1 and 0.5%) caused marked inhibition of F1000 cell proliferation when measured on days 3 and 6. Only the proproliferant effect of these fungal materials appears to correlate to their chitin content. Furthermore, the cytomorphology of the fibroblasts indicated that P. blakesleeanus, and to a lesser extent M. mucedo, possessed cell attractant properties, again correlating with chitin content. If developed for use as wound management materials, the sporangiophores of P. blakesleeanus and the mycelium of M. mucedo could possibly promote the growth of fibroblasts and provide a matrix for their anchorage, thus contributing to the granulation phase of the healing cascade.

Aspergillus oryzae↗

PubMed1994

U50488 reduces the severity of tissue damage in a rabbit model of focal cerebral ischemia.

Many pharmacotherapies for stroke that have been successful in the laboratory have proven to be ineffective in the clinical setting, often because patients do not arrive for treatment until hours after the onset of the ischemic insult. Kappa opioid treatment of cerebral ischemia has been successful in the cat and mouse with treatment delays of up to 6 h. The purpose of the present study was to develop a model of delayed kappa opioid treatment of cerebral ischemia in the rabbit. Fourteen rabbits underwent permanent, unilateral occlusion of the internal carotid, middle cerebral, and anterior cerebral arteries via a transorbital, microsurgical approach. At 6 h postocclusion, animals received a blinded bolus injection and continuous infusion of either saline or the kappa agonist, U50488. Survival was not improved after U50488 treatment. U50488 treatment did, however, reduce areas of severe tissue damage and increase areas of modest tissue damage. This suggests U50488 arrested the progression of damage from noninfarcted to fully infarcted tissue. The present results show beneficial effects of delayed treatment with kappa agonists in a species similar in vasculature to humans, and much less costly than primates or cats.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

PubMed1994

Antibacterial activity of Actisorb Plus, Actisorb and silver nitrate.

Activated Charcoal Cloth with silver (Actisorb Plus) and solutions of silver nitrate, but not Actisorb (Activated Charcoal Cloth without silver), demonstrated antibacterial activity against Gram-negative and Gram-positive bacteria. This activity was unimpaired in the presence of plasma. Sodium thioglycollate was an effective neutralizer of Actisorb Plus and of silver nitrate, indicating that the release of silver from Actisorb Plus contributed to the antibacterial activity of the dressing.

Charcoal↗

PubMed1993

Toxicity of L-ascorbic acid to L929 fibroblast cultures: relevance to biocompatibility testing of materials for use in wound management.

Fibroblast cultures are often used to evaluate materials intended for medical use, cytotoxicity being taken as an indicator of bioincompatibility. Such an approach has previously been taken with ascorbic acid in determining its value in wound healing. We have now reexamined the toxicity of L-ascorbic acid to L929 fibroblast cells in culture. Concentrations of ascorbic acid between 0.5 mM and 11 mM were tested. At concentrations above 2 mM, ascorbic acid was found to inhibit cell proliferation, with cell viability decreasing as the concentration was increased. This effect could be prevented by the addition of either superoxide dismutase or catalase to the culture medium. Assays of glutathione and glutathione disulfide were carried out on 8 day old cultures exposed for 24 h to the same concentrations of ascorbic acid. A dose-related depletion of glutathione occurred whilst glutathione disulfide levels remained essentially constant. Lactate dehydrogenase and glucose-6-phosphate dehydrogenase activities were induced by ascorbic acid at all concentrations tested but the ratio of NADP to NADPH nevertheless increased as the concentration of ascorbic acid increased. Finally, ATP in cells from 8-day-old cultures became depleted in the presence of ascorbic acid at concentrations in excess of about 5 mM when assayed after 24 h incubation. These biochemical changes and the concomitant cytostatic/cytotoxic effects may be ascribed to the reactive oxygen species produced by the autoxidation of ascorbic acid in the culture medium. Ascorbic acid breakdown products appeared not to be directly involved. In addition, our results suggested that superoxide acted cooperatively with hydroxyl to elicit these effects on the fibroblasts. It is evident from this study that the microenvironment surrounding fibroblasts in culture may differ fundamentally from that surrounding fibroblasts in a healing wound, making it impossible to extrapolate directly to an in vivo situation and hence to make any recommendations from these results concerning the use of ascorbic acid in wound healing.

Adenosine Triphosphate↗

PubMed1993

A study of hydrogen peroxide generation by, and antioxidant activity of, Granuflex (DuoDERM) Hydrocolloid Granules and some other hydrogel/hydrocolloid wound management materials.

The effect of Granuflex Hydrocolloid Granules (0.01-0.50% w/v) on the rate of proliferation of murine (L929) fibroblasts was examined. The dose-response curve showed a significant (P < 0.02) pro-proliferant effect at 0.05%, and a significant (P < 0.02) antiproliferant effect at 0.50%, mirroring the dose-response curve produced by hydrogen peroxide in the concentration range 10(-9) - 10(-4) mol/l. The antiproliferant effect at 0.20% w/v was abolished by catalase, suggesting that the biological activity of Granuflex was mediated by the in situ generation of hydrogen peroxide. Formation of hydrogen peroxide by Granuflex was confirmed by performing the scopoletin-horseradish peroxidase assay in the presence and absence of catalase. The total concentration of hydrogen peroxide detected was about 8 x 10(-6) mol/l (using 0.5% w/v Granuflex) after 48 h at 37 degrees C. In contrast, when hydrogen peroxide itself was added to L929 cultures, a similar antiproliferant activity was observed at concentrations between 10(-4) and 10(-5) mol/l. These results suggested that Granuflex was undergoing autoxidation in the culture medium, and hence that it might possess antioxidant activity. In assays for antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH), Granuflex, and two other hydrocolloid dressings (Comfeel Powder and Bard Absorption Dressing) showed significant ability to reduce DPPH to DPPH2. These three dressings also displayed superoxide scavenging activity in a nitroblue tetrazolium reduction assay. We conclude that, in addition to providing a moist wound-healing environment, Granuflex and certain other hydrocolloids might contribute to the establishment and maintenance of the reducing environment necessary for energy production and hence cell division.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Quantification of hydrogen peroxide generation by Granuflex (DuoDERM) Hydrocolloid Granules and its constituents (gelatin, sodium carboxymethylcellulose, and pectin).

The hydrogen peroxide generating capacity of Granuflex Hydrocolloid Granules and its constituents (porcine gelatin, sodium carboxymethylcellulose and pectin) was examined using the scopoletin-horseradish peroxidase assay in the presence and absence of catalase. Oxygen purging reduced the formation of hydrogen peroxide by 77-96%. The total concentrations of hydrogen peroxide detected were 1.9 x 10(-6), 1.2 x 10(-6) and 2.3 x 10(-6) mol/l for Granuflex, pectin and gelatin (using 0.5% w/v), respectively, after 48 h incubation in a phosphate buffer, pH 7.4, at 37 degrees C. No hydrogen peroxide was formed by sodium carboxymethylcellulose. The results indicate that hydrogen peroxide generation by Granuflex may be ascribed to its gelatin and pectin components, but not to the sodium carboxymethylcellulose. The release of low levels of hydrogen peroxide into the wound environment could conceivably contribute both to the inflammatory phase and to fibroblast proliferation, and hence to the granulation phase of wound healing.

Animals↗

PubMed1993

Biocompatibility of wound management products: a study of the effects of various polysaccharides on murine L929 fibroblast proliferation and macrophage respiratory burst.

An in-vitro screening method to examine the biocompatibility of materials used in wound management has been evaluated. This involved the use of a macrophage respiratory-burst assay and a fibroblast proliferation assay to represent respectively the inflammatory and the granulation phases in wound healing. Standard polysaccharides (calcium and sodium alginates, l-carrageenan, chitin, chitosan lactate, chondroitin sulphate and pectic acid) were used as test compounds. None of the polysaccharide samples caused a significant increase in L929 fibroblast cell numbers relative to control after 6 days incubation. The overall effect of exposure of the fibroblast cultures to the alginates, carrageenan and chondroitin sulphate was an extension of lag phase followed by an enhanced rate of cell proliferation in the logarithmic phase. Only calcium and sodium alginates and chondroitin sulphate enhanced the respiratory burst activity of murine macrophages; l-carrageenan and chitosan lactate were markedly inhibitory. The results suggest that a macrophage activity assay should be included as part of an in-vitro screening program to evaluate the biocompatibility of wound management materials and to detect intrinsic biological activity.

Alginates↗

PubMed1989

Biocompatibility of wound management products: standardization of and determination of cell growth rate in L929 fibroblast cultures.

To facilitate the development of a bioassay procedure by which the biocompatibilities of materials used in wound management may be assessed and compared, those environmental factors affecting cell growth in mouse L929 fibroblast cultures have been identified. Standardization of the initial cell number and frequency of change of medium resulted in the virtual elimination of variation of growth curves of L929 cells cultured in flasks of specified surface area. In addition, three methods for assessing fibroblast growth rate in the presence of alginate products used in wound management were evaluated. These were the haemacytometer counting chamber method, the Coulter counting method, and a liquid scintillation counting method. The first two methods determine the number of cells in a given volume of a cell suspension, whereas the third method determines the rate of synthesis of deoxyribonucleic acid (DNA), and hence cell growth, by measuring the incorporation of [3H]thymidine. The haemacytometer method had significant advantages over the other two procedures in providing both qualitative and quantitative data on culture morphology and cell growth response.

Alginates↗

PubMed1989

Biocompatibility of wound management products: the effect of various monosaccharides on L929 and 2002 fibroblast cells in culture.

The effects of various monosaccharides on the growth of human 2002 and mouse L929 fibroblast cultures have been investigated. Eleven monosaccharides having acidic, neutral, and basic characteristics were evaluated in a bioassay procedure developed for the investigation of biocompatibility of wound management materials. Rate of growth in both cell lines was inhibited by D-galacturonic acid and by D-glucuronic acid. Although most neutral sugars produced no significant change in the growth rate or in the morphology of the cells, galactose produced a significant increase in the growth rate of both cell lines whilst L-fucose caused a significant decrease in growth of the L929 cells but did not significantly affect the growth of 2002 cells; xylose increased the growth rate of L929 but not 2002 cells. D-Glucosamine, a basic sugar, produced inhibition of growth which followed a different pattern from that produced by the acidic sugars; N-acetylglucosamine produced a species specific increase in cell growth of L929 cells. The results show that the effects produced by the monosaccharides on the cultured fibroblasts are related to their chemical structure and to cell line, and suggest that the use of galactose as a possible aid to wound healing should be investigated.

Animals↗

PubMed1988

An assay procedure to compare sorptive capacities of activated carbon dressings: the detection of impregnation with silver.

A semiquantitative assay procedure for the estimation of uptake of diethylamine by activated carbon dressings has been developed. The procedure involves the use of Nessler's reagent as a comparative indicator. The use of diethylamine allows the concomitant detection of silver with which the dressings may be impregnated. The implications of silver/amine complex formation during dressing use are discussed. Quantification of the silver content of one such fabric was carried out by atomic absorption spectrophotometry. A silver content of 2770 ppm was found. It is suggested that this method might be suitable for adoption as the basis for a pharmacopoeial assay procedure for activated carbon dressings.

Adsorption↗

PubMed1985