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1 Department of Anesthesiology, University of Munich, Munich, Germany
2 Institute for Surgical Research, University of Munich, Munich, Germany
3 Department of Anatomy II, University of Munich, Munich, Germany
* To whom correspondence should be addressed. E-mail: alwin.goetz{at}med.uni-muenchen.de.
Permeability of the endothelial barrier to large molecules plays a pivotal role in the manifestation of early acute lung injury. We here present a novel and sensitive technique that brings microanatomical visualization and quantification of microvascular permeability in line. White New Zealand rabbits were anesthetized and ventilated mechanically. Rabbit serum albumin (RSA) was labeled with colloidal gold particles. Macromolecular leakage of gold labeled RSA and thickening of the gas exchange distance were quantified by electron microscopy taking into account morphology of microvessels. The control group receiving a saline solution represented a normal gas exchange barrier without extravasation of gold labeled albumin. Infusion of lipopolysacharide (LPS) resulted in a significant displacement of gold labeled albumin into pulmonary cells, the lung interstitium and even the alveolar space. Correspondingly intravital fluorescence microscopy and digital image analysis indicated thickening of width of alveolar septa. The findings were accompanied by a deterioration of alveolo arterial oxygen difference (AaDO2), whereas wet/dry ratio and albumin concentration in the bronchoalveolar lavage fluid missed to detect that early stage of pulmonary edema. Inhibition of the nuclear enzyme poly (ADP-ribose) synthetase by 3-aminobenzamide prevented LPS induced microvascular injury. Summarizing, colloidal gold particles visualized by standard electron microscopy are a new and very sensitive in vivo marker of microvascular permeability in early acute lung injury. This technique enabling detailed microanatomical and quantitative pathophysiologic characterisation of edema formation can form the basis for evaluating novel treatment strategies against acute lung injury.
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