Molecular-Pharmacological Analysis of Periodontal Disease : Interaction between LPS and Kinins on Microcirculation of Oral Region

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The purpose of this study is twofold : 1) to design and develop an optical biosensor that can be used without renewing reagents in the probe for detection of biologically important analytes, and 2) to apply the hypoxanthine affinity optical biosensor to measuring the purine metabolism in an ischemia-reperfusion models. We developed the chemiluminometric bienzyme sensors on the basis of the horseradish peroxidase catalyzed H_2O_2/luminol reaction ; horseradish peroxidase was coimmobilized with xanthine oxidase. Optical biosensor was developed for the batch analysis system of hypoxanthine in the concentration range between 10^<-8> and 10^<-4> M.Optical biosensor probe with photomultiplier tube (PMP) with an integrated preamplifier was investigated to detect the chemiluminescence (which is enhanced by horseradish peroxidase). Horseradish peroxidase and xanthine oxidase were immobilized entrapping sodium alginate gel. The probe based superoxide anion radicals or H_2O_2 sensor achieve detection limits 10^<-8> M.The concentration of hypoxanthine in rabbit serum was also determined in peripheral blood samples (obtained from auricular vein). Increase in hypoxanthine levels was observed in serum obtained immediately after 90 min of auricular artery occlusion. Although there is conflicting evidence as to whether oxygen-derived free radicals play a major role in reperfusion injury, the relatively sustained nature of enhanced oxygen-derived free radical generation during reperfusion has potential clinical implications. With this optical biosensor, it should be possible to measure oxygen-derived free radical generation in a variety of conditions and with multiple interventions. Such studies may further characterize the role of oxygen free radicals in ischemiaミreperfusion injury.