Biomedical subjects

L C Kirkwood

Publications and source records attributed to L C Kirkwood.

6 recordsLinked to original sources

PubMed2003

Wettable phosphorylcholine-containing polymers useful in blood filtration.

The use of phosphorylcholine (PC)-containing polymers has long been acknowledged as a method to improve the haemocompatibility of blood contacting devices. Such polymers were investigated for coating leukocyte filters, as a means of preserving precious platelet numbers and function. It was demonstrated that by use of such coated filters, the platelet recovery could be significantly increased by some 30%. This was however, balanced by a decrease in the leukocyte removal efficiency of the filter. More problematic however, was the poor critical wetting surface tension (CWST) of the filter (45 mNm(-1)). This was due to the surface expression of the hydrophobic groups of the polymer in air. In order to obtain a filter that could be immediately wetted by the blood and hence remove the need for any detrimental priming of the filter, the PC polymer was modified in order to reduce its hydrophobic content. A series of new cross-linkable polymers was developed that contained other hydrophilic monomers, yet could film-form to produce a coating stable to fluid contact. These materials demonstrated the required wetting characteristics (a CWST of >78 mNm(-1)), whilst retaining the excellent blood-contacting properties.

Journal Article↗

PubMed2001

Crosslinkable coatings from phosphorylcholine-based polymers.

2-Methacryloyloxyethyl phosphorylcholine (MPC) was synthesised and then used in the preparation of crosslinked polymer membranes with lauryl methacrylate, hydroxypropyl methacrylate and trimethoxysilylpropyl methacrylate (crosslinker) comonomers. Some physical aspects of the membrane properties were evaluated in order to establish the basis for the synthesis of a series of post-crosslinkable polymers. These materials were made by copolymerisation of the constituent monomers via a free radical method, and characterised using NMR, FT-IR, viscometry and elemental analysis. The optimum crosslink density and conditions required for curing coatings of these polymers were investigated using atomic force microscopy (AFM) and showed the inclusion of 5 mol% silyl crosslinking agent to be ideal. A nanoindentation technique was employed to determine if the coating developed elasticity upon crosslinking. The biological properties of the coatings were evaluated using a variety of protein adsorption assays and blood contacting experiments, and an enzyme immunoassay was developed to detect E. coli in order to assess the level of bacterial adhesion to these biomaterials. Polymers of this type were shown to be very useful as coating materials for improving the biocompatibility of, or reducing the levels of adherent bacteria to medical devices.

Bacterial Adhesion↗

PubMed2000

Synthesis and characterisation of phosphorylcholine-based polymers useful for coating blood filtration devices.

Copolymers of 2-methacryloyloxyethylphosphorylcholine (MPC) and lauryl methacrylate (LMA) of molar ratios MPC: LMAX where x = 1, 2 or 4, have been synthesised by two different free-radical polymerisation techniques. The solubility characteristics of the resulting materials were investigated in a variety of water: alcohol solvent mixtures and found to be influenced not only by the molar ratio of MPC: LMA, but also the method of synthesis. A window of solubility was observed for certain copolymers and the alcohol used in the solvent mixture was also found to have a profound influence on the solubility profile of the polymers. These materials were soluble in a wider range of aqueous methanol mixtures compared to aqueous mixtures of higher aliphatic alcohols, such as ethanol or isopropyl alcohol, which was rationalised in terms of the affinity of the phosphorylcholine headgroup for the various alcohols relative to water. 1H nuclear magnetic resonance spectroscopy was used to further examine the solution properties of the copolymers in various solvents. The copolymer MPC: LMA2 was coated onto a variety of substrates from both alcohol-only and water: alcohol solvent systems and the surface properties of the films compared by static and dynamic contact angle, atomic force microscopy (AFM) and attenuated internal reflectance Fourier transform infrared spectroscopy (ATR-IR). The coating formed from the water: alcohol solvent was found to be hydrophilic in nature, possessing spontaneous wettability, whereas films formed from alcohol-only solvents were hydrophobic, and only on conditioning with water were more wettable surfaces attained. This phenomenon was applied in the coating of leukocyte filtration material, where the aqueous-based systems demonstrated lower critical wetting surface tension (CWST) and shorter wetting times relative to both uncoated filters and those coated from alcohol-only systems. The haemocompatibility of the coated filters was equivalent for both coating solvent systems. employed, and far superior when compared to the uncoated control.

Alcohols↗

PubMed1997

Determination of the dihydrocodeine metabolites, dihydromorphine and nordihydrocodeine, in hepatic microsomal incubations by high-performance liquid chromatography.

A high-performance liquid chromatographic assay for the oxidative metabolites of dihydrocodeine, nordihydrocodeine and dihydromorphine, formed in human liver microsomal incubations, is described. A simple solvent extraction followed by reversed-phase high-performance liquid chromatography with UV detection allows quantification of both metabolites in a single assay. Standard curve concentration ranges for dihydromorphine and nordihydrocodeine were 0.05-5 and 0.2-20 microM, respectively. Assay performance was assessed by intra- and inter-day accuracy and precision of quality control (QC) samples. The difference between the calculated and the actual concentration and the relative standard deviation were less than 15% at low QC concentrations and less than 10% at medium and high QC concentrations for both analytes. The method provides good precision, accuracy and sensitivity for use in kinetic studies of the oxidative metabolism of dihydrocodeine in human liver microsomes.

Analgesics, Opioid↗

PubMed1997

Characterization of the human cytochrome P450 enzymes involved in the metabolism of dihydrocodeine.

AIMS: Using human liver microsomes from donors of the CYP2D6 poor and extensive metabolizer genotypes, the role of individual cytochromes P-450 in the oxidative metabolism of dihydrocodeine was investigated. METHODS: The kinetics of formation of N- and O-demethylated metabolites, nordihydrocodeine and dihydromorphine, were determined using microsomes from six extensive and one poor metabolizer and the effects of chemical inhibitors selective for individual P-450 enzymes of the 1A, 2A, 2C, 2D, 2E and 3A families and of LKM1 (anti-CYP2D6) antibodies were studied. RESULTS: Nordihydrocodeine was the major metabolite in both poor and extensive metabolizers. Kinetic constants for N-demethylation derived from the single enzyme Michaelis-Menten model did not differ between the two groups. Troleandomycin and erythromycin selectively inhibited N-demethylation in both extensive and poor metabolizers. The CYP3A inducer, alpha-naphthoflavone, increased N-demethylation rates. The kinetics of formation of dihydromorphine in both groups were best described by a single enzyme Michaelis-Menten model although inhibition studies in extensive metabolizers suggested involvement of two enzymes with similar Km values. The kinetic constants for O-demethylation were significantly different in extensive and poor metabolizers. The extensive metabolizers had a mean intrinsic clearance to dihydromorphine more than ten times greater than the poor metabolizer. The CYP2D6 chemical inhibitors, quinidine and quinine, and LKM1 antibodies inhibited O-demethylation in extensive metabolizers; no effect was observed in microsomes from a poor metabolizer. CONCLUSIONS: CYP2D6 is the major enzyme mediating O-demethylation of dihydrocodeine to dihydromorphine. In contrast, nordihydrocodeine formation is predominantly catalysed by CYP3A.

Analgesics, Opioid↗

PubMedDate not supplied

Glucuronidation of dihydrocodeine by human liver microsomes and the effect of inhibitors.

1. Glucuronidation is the major route of metabolism of dihydrocodeine (DHC) and accounts for 25-30% of an oral dose in urine. The kinetics of DHC-6-glucuronide formation in liver microsomes from five human donors and the effect of a number of potential inhibitor drugs were examined using a newly developed and validated HPLC assay. 2. The formation of DHC-6-glucuronide exhibited atypical kinetics that conformed to the Hill equation. The mean intrinsic dissociation constant (Ks) and maximum velocity (Vmax) values were 1566 micromol/L and 0.043 micromol/min per g, respectively. The Ks and Vmax values varied 1.5- and 3.5-fold, respectively. 3. Seven drugs were tested for inhibitory effects on DHC glucuronidation at low (50 micromol/L) and high (500 micromol/L) concentrations. At 50 micromol/L, only diclofenac produced greater than 50% inhibition, while at concentrations of 500 micromol/L inhibition was greater than 35% for diclofenac, amitriptyline, oxazepam, naproxen, chloramphenicol and probenecid, but not paracetamol. 4. The present study found little interindividual variation in the activity of human liver microsomes for glucuronidation of DHC. Comparison of the results from the inhibition studies with those reported previously for codeine and morphine suggest that the UDP-glucuronosyltransferase isoform UGT2B7 is involved in the glucuronidation of DHC.

Adult↗