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• W2949501144 abstract "It was the aim of this PhD-project to establish suitable models for investigating pharmacokinetic and pharmacodynamic properties of substances administered to the equine joint. Firstly, in order to assess the distribution of drugs either after intra-articular or systemic application, the ex vivo model of the isolated perfused equine distal limb was established. Distal forelimbs of horses obtained from a slaughterhouse were exarticulated in the carpal joint. Within 1 h after slaughter, the median artery was canulated and extremities were perfused with oxygenated tyrode solution containing sodium carboxymethyl cellulose at a flow rate of approx. 65 mL/min for up to 8 h. Tissue viability during the perfusion period was verified by determination of glucose consumption, lactate production and LDH liberation in the venous perfusate sampled from the radial vein. Furthermore, skin surface temperature and weight increase were monitored as indicators of peripheral blood supply and edema formation. The integrity of the joint capsule was assessed by histologic examination. Minimum requirements for parameters were formulated to ensure sufficient viability of the distal limb. The articular efflux rate of BM in the venous perfusate of the radial vein after intra-articular injection of BM disodium phosphate (4 mg BM) into the fetlock joint was measured with HPLC and UV detection (n=6). The average BM efflux rate per minute was calculated to be 5.1 µg/min with values ranging from 9 µg/min to 2.9 µg/min. 7.5 h after i.a.-application, 2.3 mg BM had left the joint. Also, the distribution of ASA and its metabolite SA into the fetlock joint after systemic administration of ASA was determined in the isolated perfused equine distal limb (n=8). Plasma concentrations of ASA and SA were adapted from an in vivo study and added to the perfusate (BROOME et al. 2003).  ASA and SA concentrations in the synovial fluid were determined using calibrated microdialysis probes and HPLC-UV. Two distal limbs were perfused with diluted autologous blood in order to give some orientation concerning the influence of plasma protein binding on the synovial distribution. For the tyrode-perfused limbs (n=6), ASA concentrations in the synovial fluid reached a maximum of 2.1x10-5 mol/L, maximum SA concentrations were 1.27x10-4 mol/L. Synovial salicylate concentrations in the hemoperfused limbs were about 50 % lower, presumably owing to the salicylate plasma protein binding of 50 %. The decrease of ASA and SA in synovial fluid did not parallel the decrease in plasma concentrations, indicating an accumulation in the joint. Secondly, in order to assess the anti-inflammatory potency of concentrations measured in the isolated perfused equine distal limb, a cell culture model of primary equine synoviocytes was established. Cells isolated from macroscopically healthy fetlock joints (n=6) were stimulated with LPS and treated with decreasing doses of BM, ASA and SA. The inhibition of prostaglandin E2 production in comparison with an untreated control was determined as an indicator of an anti-inflammatory effect. The lowest concentrations causing a significant inhibition of PGE2 production were 2.55x10-9 mol/L and 7.77x10-6 mol/L for BM and ASA, respectively. SA concentrations employed did not cause a significant PGE2 inhibition. These results may be a hint for a prolonged anti-inflammatory effect in the joint in comparison with plasma concentrations. In summary, the isolated perfused equine distal limb and primary equine synoviocyte cultures were successfully established as useful tools for the investigation of pharmacokinetic and pharmacodynamic properties of substances administered to the equine joint without the necessity to use living horses." @default.
• W2949501144 created "2019-06-27" @default.
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• W2949501144 date "2013-01-01" @default.
• W2949501144 modified "2023-09-27" @default.
• W2949501144 title "The isolated perfused equine distal limb and synoviocyte cultures as models for pharmacological studies" @default.
• W2949501144 hasPublicationYear "2013" @default.
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