Tracer kinetics analysis of the extracellular spaces in saline perfused hearts

Precise estimation of the cellular water content presupposes a correct definition of the water fraction in tissue extracellular space. Low molecular weight markers (LMM), such as sulphate ion and sucrose, are widely used to define extracellular space size despite indications that they penetrate the cell. In contrast, inulin, with molecular weight of about 5000, is commonly regarded as a cell impermeable extracellular marker.

To compare LMM with inulin as markers in determining extracellular space size.

The size of extracellular space in guinea pig hearts perfused with crystalloid solution (hydrated hearts) was determined morphometrically and by mathematical model analysis of washout kinetics of LMM ((35)SO(4), (14)C-sucrose) or (3)H-inulin.

Morphometrically, the sizes of vascular and interstitial spaces in the hydrated hearts were estimated to be 102+/-8 mL/kg wet mass (wm) and 452+/-17 mL/kg wm, respectively. Comparable data were obtained from model simulation of tracer washout: 67 mL/kg wm for vascular space and 439 to 462 mL/kg wm for interstitial space. Tracer penetration into cellular water, as shown by model analysis, was 28% for LMM and, reported here for the first time, 18% for inulin. The observed edema was probably due entirely to fluid accumulation in the interstitial space.

Intracellular penetration of LMM must be taken into account, especially in modern nuclear magnetic resonance spectroscopic methods of cellular water monitoring in isolated perfused hearts.