The overall objective of the project is to correlate the biochemical and cellular events of ocular inflammation with the physiological and clinical manifestations. This will enable us to study not only the mechanism of action of anti-inflammatory compounds in current clinical use but also to evaluate the contribution of specific biochemical pathways in the inflammatory response. We will employ three models of ocular inflammation: intravitreal injection of bacterial endotoxin, intrastromal injection of clove oil into the cornea, and the alkali burned eye. Each model has unique characteristics. We will employ three models of ocular inflammation: intravitreal injection of bacterial endotoxin, intrastromal injection of clove oil into the cornea, and the alkali burned eye. Each model has unique characteristics. The time course of appearance of inflammatory mediators will be determined. Mediators in ocular tissues and fluids will be identified by bioassay, thin layer chromatography and high performance liquid chromatography. Additionally, we will further characterize and quantify mediators by gas liquid chromatography in conjunction with mass spectrometry. The time course of cellular infiltration into inflamed tissue will be determined by measuring the myeloperoxidase activity in ocular tissues (myeloperoxidase is an enzyme marker for polymorphonucleal leukocytes), performing direct cell counts in ocular fluids and parallel morphological studies. The vascular integrity of the eye will be evaluated by measuring extravasation of protein into the aqueous and by fluorescein angiography. The ocular response to intraocular administration of specific inflammatory mediators will be evaluated to demonstrate that different mediators can elicit discrete components of the physiological response. Pharmacological manipulation of the ocular inflammatory response will be studied using known steroidal and non-steroidal inflammatory agents and investigational drugs with potential anti-inflammatory properties. Studies in vitro will examine the effect of certain drugs upon arachidonic acid metabolism.
