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In vivo sledování doxorubicinu ve svalové tkání
Ing. Iva Blažková; doc. RNDr. Libuše Trnková, CSc.
Reg.č.projektu: CZ.1.07/2.3.00/20.0148
Název projektu: Mezinárodní spolupráce v oblasti "in vivo" zobrazovacích technik
Název:
Školitel:
Datum: 18.1.2013
Anthracycline cytostatic
Cardiotoxicity
Fluorescence properties
Materials:
Doxorubicin hydrochloride infusion solution (Doxorubicin-Teva 0,2)
Chicken muscle tissue
Methods: ◦ fluorescence spectroscopy
◦ Carestream In vivo imaging system
B) Doxorubicin in water
A) Doxorubicin in methanol
2000 μg/ml
2000 μg/ml
1000 μg/ml
500 μg/ml
1000 μg/ml
500 μg/ml
250 μg/ml
250 μg/ml
125 μg/ml
125 μg/ml
63 μg/ml
63 μg/ml
31 μg/ml
31 μg/ml
16 μg/ml
16 μg/ml
8 μg/ml
8 μg/ml
4 μg/ml
4 μg/ml
0
0,5
1
1,5
2
2,5
3
3,5
4
4,5
0 0,5 1 1,5 2
Flu
ore
scence in
tensit
y x
10
3 [a
.u.]
cm [mg/ml]
The dependence of the intensity of the fluorescence
on the concentration of doxorubicin in water
The best fluorescence properties: ◦ Dox in methanol: 250 – 500 μg/ml
◦ Dox in water: 63 – 250 μg/ml
- Pure doxorubicin - In methanol: 2000 μg/ml
- In water: 1000 μg/ml
The application:
◦ Injection
◦ Infusion
◦ Tube with doxorubicin
Intensity of the fluorescence of the doxorubicin in the tissue depends on:
◦ the concentration and the solvent
◦ the time
◦ the volume (mass of dox)
◦ the depth
500 µg/ml dox (water)
500 µg/ml dox (methanol)
C
0
1
2
3
4
5
6
7
520 620 720 820In
tensit
y o
f th
e f
luore
scence x
10
4 [a.u
.]
wavelength [nm]
Intensity of the fluorescence
Meat + dox Meat
Meat + dox unmixing
Meat
0
5
10
15
20
50
100
150
200
250
300
350
400
450
500
Are
a x 1
04 [p
ixel]
Volume [µl]
Area
0
100
200
300
400
500
600
700
800
50
100
150
200
250
300
350
400
450
500
Max in
tensit
y [a.u
.]
Volume [µl]
Max intensity
50 µl
400 µl
100 µl 150 µl
350 µl
300 µl 250 µl 200 µl
500 µl
450 µl
Fig. The growing volume of the doxorubicin (500 µg/ml) in the tissue: 50 – 500 µl (25-225 µg).
The doxorubicin diluted in the water
is detectable in the depth of 3 mm
The doxorubicin diluted in the methanol is detectable in the depth of 5 mm
Fluorescence of the doxorubicin (500 µg/ml in methanol) in the tube in the depth 4-6 mm.
Fluorescence of the doxorubicin (63 µg/ml in water) in the tube in the depth 3mm.
4 mm 5 mm 6 mm
3 mm
0
500
1000
0 1 2 3 4 5 6
Mean in
tensit
y [a.u
.]
Depth [mm]
The dependence of the fluorescencence intensity on
the depth of the tube in the tissue
1 min 2 min
4 min
3 min
12 min 10 min
500
700
900
1100
1300
1500
1700
0 1 2 3 4 8 10 12 21Inte
nsit
y o
f t
he f
luore
scence
time [minutes]
Intensity of the fluorescence
• Intravenous application of the 100 µl doxorubicin (2000 µg/ml) • The highest intensity of the fluorescence was observed after 3 minutes
muscle
heart
liver
lungs
kidney
muscle
liver
lungs
spleen
kidney
heart
Organs – mouse without doxorubicin Organs - mouse with doxorubicin
spleen
Intraperitoneal injection of 100 µl of the doxorubicin (500 µg/ml in methanol)
Mouse with doxorubicin
Dox
The doxorubicin has got good fluorescence properties
Organic solvents ehance this fluorescence
The In-Vivo imager can detect the doxorubicin in the living organism, so it is posible to continue with the application of the doxorubicin into the living organisms and observe the distribution of the doxorubicin in the body
to my collegues from the Laboratory of Applied nanobiotechnology
Mgr. Markéta Ryvolová, Ph.D.
prof. Ing. René Kizek, Ph.D.
and other collegues
