Determination of regional bone blood flow by means of fluorescent microspheres using an automated sample-processing procedure
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vor 21 Jahren
The determination of regional blood flow utilizing fluorescent
microspheres (FMs) is an established method for numerous organs.
Recent progress, in particular the automation of sample processing,
has further improved this method. However, the FM method (reference
sample technique), which allows repetitive measurement of regional
organ blood flow, has so far not been used for the determination of
blood flow in bone. The aim of the present study was to establish
FM for the quantification of regional bone blood flow (RBBF).
Female, anesthetized New Zealand rabbits (n = 6) received left
ventricular injections of different amounts of FM at six subsequent
time points. In order to examine the precision of RBBF
determination, two different FM species were injected
simultaneously at the sixth injection. At the end of the
experiments the femoral and tibial condyles of each hind limb were
removed and the fluorescence intensity in the tissue samples was
measured by an automated procedure. In an in vitro study we have
shown that acid digestion of the crystalline matrix has no effect
on the fluorescence characteristics of FM. The determination of the
number of spheres per tissue sample revealed that depending on the
tissue sample size up to 3 x 10(6) spheres/injection were necessary
to obtain about 400 microspheres in the individual bone samples.
RBBF values of the tibial and femoral condyles did not differ at
various injection intervals. The tibial blood flow values varied
between 6.6 +/- 1.1 and 8.5 +/- 1.4 ml/min/100 g and were
significantly higher than those of the femur (4.3 +/- 1.1 to 6.0
+/- 1.8 ml/min/100 g). The bone blood flow values obtained by
simultaneous injection of two FM species correlated significantly
(r = 0.96, slope = 1.06, intercept = 0.05), the mean difference was
0.39 +/- 1.11 ml/min/100 g. Our data demonstrate that the
measurement of RBBF by means of FM allows a valid determination of
RBBF. Copyright (C) 2003 S. Karger AG, Basel.
microspheres (FMs) is an established method for numerous organs.
Recent progress, in particular the automation of sample processing,
has further improved this method. However, the FM method (reference
sample technique), which allows repetitive measurement of regional
organ blood flow, has so far not been used for the determination of
blood flow in bone. The aim of the present study was to establish
FM for the quantification of regional bone blood flow (RBBF).
Female, anesthetized New Zealand rabbits (n = 6) received left
ventricular injections of different amounts of FM at six subsequent
time points. In order to examine the precision of RBBF
determination, two different FM species were injected
simultaneously at the sixth injection. At the end of the
experiments the femoral and tibial condyles of each hind limb were
removed and the fluorescence intensity in the tissue samples was
measured by an automated procedure. In an in vitro study we have
shown that acid digestion of the crystalline matrix has no effect
on the fluorescence characteristics of FM. The determination of the
number of spheres per tissue sample revealed that depending on the
tissue sample size up to 3 x 10(6) spheres/injection were necessary
to obtain about 400 microspheres in the individual bone samples.
RBBF values of the tibial and femoral condyles did not differ at
various injection intervals. The tibial blood flow values varied
between 6.6 +/- 1.1 and 8.5 +/- 1.4 ml/min/100 g and were
significantly higher than those of the femur (4.3 +/- 1.1 to 6.0
+/- 1.8 ml/min/100 g). The bone blood flow values obtained by
simultaneous injection of two FM species correlated significantly
(r = 0.96, slope = 1.06, intercept = 0.05), the mean difference was
0.39 +/- 1.11 ml/min/100 g. Our data demonstrate that the
measurement of RBBF by means of FM allows a valid determination of
RBBF. Copyright (C) 2003 S. Karger AG, Basel.
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