Segmental differences of microvascular permeability for FITC-dextrans measured in the hamster cheek pouch

Abstract

An intravital microscopic method for quantitative measurement of interstitial concentrations of fluorescent tracers has been applied to the investigation of microvascular permeability in the hamster cheek pouch. Some nanoliters of FITC-dextran mean mol wt ( M w ) 20,000, 3000, or sodium fluorescein ( M w 376) were injected into an arteriole of the exposed cheek pouch via micropipet. The extravasation of fluorochromes was measured by a photodensitometric method including two sets of calibration procedures ( in vitro and in vivo). Postcapillary and collecting venules exhibited the highest absolute increase of fluorochrome concentration in the tissue for all tracer molecules tested when compared to arterioles or capillaries. The permeability of the vascular wall was quantified, assuming that diffusion processes play the main role for the transport of the investigated molecules under the experimental conditions of a high concentration gradient across the membrane. Permeability coefficients P (cm/sec) and apparent diffusion coefficients D′ (cm 2/sec) of the microvascular wall were calculated using a mathematical model for one-dimensional diffusion in composite media. The analysis is based on measured data of interstitial diffusion coefficients of the tracers used. For all tracer molecules tested, the wall of the capillaries and postcapillary venules was significantly more permeable than the arteriolar wall. For the largest test molecule (FITC-dextran M w 20,000), the permeability coefficient of the vessel wall showed a maximum in the postcapillary venules. These findings support the concept of a “gradient of permeability” with a nonuniform distribution of exchange capacity only for the precapillary microvessels. A marked preponderance of venular over capillary permeability could, if at all, only be detected for FITC-dextran M w 20,000. The present study characterizes the vessel wall by apparent diffusion coefficients which are, for FITC-dextran M w 3000, and free fluorescein, roughly three orders of magnitude lower than the apparent diffusion coefficients in connective tissue.