PPhotothermalhotothermal and photodynamic and photodynamic effects at laser heating with gold effects at laser heating with gold

nanoparticles and nanocomposits nanoparticles and nanocomposits Au-SiOAu-SiO22-Hp-Hp

Saratov Fall Meeting 2012

Georgy S. Terentyuk1,3, Alla B. Bucharskaya1, Andrey V. Ivanov2, Elina A. Genina3, Alexey N. Bashkatov3, Irina L. Maksimova3, Valery V. Tuchin3, Boris N. Khlebtsov4, Nikolay G. Khlebtsov4

1Saratov State Medical University n. a. V.I. Razumovsky, Saratov2Russian Cancer Research Center n. a. N.N. Blokhin , RAMS, Moscow

3Saratov State University n. a. N.G. Chernyshevsky, Saratov4Institute of Biochemistry, Physiology of Plants, Microorganisms RAS, Saratov

2

Determination of gold by atomic absorption Determination of gold by atomic absorption spectroscopyspectroscopy

Atomic -adsorbtion Spectrometr AAS-3 (С. Zeiss) with lamp LT-6М

Ashing с H2SO4

t=630оС

Resonance line 242,8 nm

Detection limit 0,02 μg/ml

Range of the linear region 0,2-20,0 μg/ml Terentyuk et al.

2009, No. 5, 292-302

gold accumulation in organs

24 hrs after intravenous injection

bra

in

kid

ne

ys

lun

gs

liv

er

sp

lee

n

blo

od

control15 nm50 nm160 nm

Au,

μg/

mL

3

Dynamics of gold Dynamics of gold concentration in blood concentration in blood after intravenous after intravenous injection of gold injection of gold nanoparticlesnanoparticles

Dynamics of gold Dynamics of gold concentrations in tumor concentrations in tumor tissue intertwined after tissue intertwined after intravenous injection of gold intravenous injection of gold nanoparticlesnanoparticles

○- control, ●- tumor, - skin and muscle tissue

Au,

μg/

mL

t, min t, hrs

The value of tumor vascularization for the passive delivery of The value of tumor vascularization for the passive delivery of nanoparticlesnanoparticles

5

brain

blood separation

Size-dependent changes in the organs 24 hours after Size-dependent changes in the organs 24 hours after intravenous injection of gold nanoparticlesintravenous injection of gold nanoparticles

kidneys 15 nm 50 nm 160 nm

plethora ++ ++ + degeneration of tubular epithelium

++ + +

violation of the vascular wall

+ - +

pigment in the lumen of blood vessels

+ + _

damage to the glomeruli

_ ++ +

liver 15 nm 50 nm 160 nm

violation of the structure

- - -

plethora of sinusoids + + + pigment in Kupfer cells

+ + +

pigment in parenchyma

+ _ +

blood separation - + - dystrophy of hepatocytes

+++ + -

spleen 15 nm 50 nm 160 nm

violation of the structure

- ++ _

plethora ++ +++ + pigment in folliculs + ++ + pigment in red pulpa +++ ++ + apoptosis - ++ - Violation of artery wall

- - +

6

Gold nanoparticles Gold nanoparticles synthesized in the synthesized in the laboratory of biosensors laboratory of biosensors and nanoscale structures and nanoscale structures of IBPPM RAS, Saratov, of IBPPM RAS, Saratov, RussiaRussia

Transparency window of Transparency window of biological tissuesbiological tissues

Setting resonancesSetting resonancesE

xtin

ctio

n

Wavelength, nm

Epidermis

Dermis

Hypodermis

UV Vis NIR

Comparative dynamics of heating of gold nanoparticles Comparative dynamics of heating of gold nanoparticles solutionssolutions

Comparative dynamics of heating of gold nanoparticles Comparative dynamics of heating of gold nanoparticles solutionssolutions

Te

mp

era

ture

, oC

Time, min

0.5 Wt/cm2

0.25 Wt/cm2

0.5 Wt/cm2

The concentration of gold per gram of tissue a The concentration of gold per gram of tissue a day after the intravenous administrationday after the intravenous administration

С – nanocages, R - nanorods

Au,

μg/

g

Au,

μg/

g control control

control controltumor muscle

liver spleen

Laser thermolysis of transplanted Ehrlich tumor in miceLaser thermolysis of transplanted Ehrlich tumor in miceT

, °C

T, °

C

Time of heating, min Time of heating, min

intratumoral intravenous

intravenous

control

720 μg/mL

90 μg/mL

Intravenous 400 μg/mL

Intravenous 50 μg/mL

control

The maximum temperature on the surface after laser heatingThe maximum temperature on the surface after laser heating

The kinetics of laser heating of the tumor (1) and health (2) tissues in its maximum one day after intravenous injection of gold nanorods at 2 (a) and 8 mg / kg (b). Curves 3 is the kinetics of maximum heating of healthy tissue without introducing nanoparticles (control)

a b

The temperature distribution on the surface of the The temperature distribution on the surface of the animal's skin with intravenous injection of nanoparticle animal's skin with intravenous injection of nanoparticle

- surface laser heating of the tumor- surface laser heating of the tumor

0 - before heating; 1-5 after 1-5 minutes after the start of heating, respectively; 6-8 after 1-3 minutes after the heating, respectively

T,

°C

Control - heating Control - heating without nanoparticles, without nanoparticles, in a dayin a day

Experiment - heating Experiment - heating with nanoparticles in with nanoparticles in a daya day

Transplanted Ehrlich tumor -Transplanted Ehrlich tumor -external irradiationexternal irradiation

experiment control

0 min 1 min

5 min

Interstitial hyperthermia with nanorods using a catheter with a Interstitial hyperthermia with nanorods using a catheter with a fiber diffuserfiber diffuser

Р=1.5 Vt t=15 min L=10 mm

15

Pathology of tumor tissues afterPathology of tumor tissues after controlled laser controlled laser thermolysis using plasmon-resonant gold nanoparticlesthermolysis using plasmon-resonant gold nanoparticles

Tumor tissue after laser exposureTumor tissue after laser exposure

Without nanoparticlesWithout nanoparticles With gold nanoparticlesWith gold nanoparticles

Tissue of transplanted rat kidney Tissue of transplanted rat kidney cancer before exposurecancer before exposure

hematoxylin-eosin stainhematoxylin-eosin stain

Increase Х150Increase Х150

16

thermogram of tumor interstitial introduction of nanoparticles to a depth of 7 mm

Growth dynamics of subcutaneously transplanted kidney Growth dynamics of subcutaneously transplanted kidney cancer in rats before and after laser hyperthermia, CW, 1 cancer in rats before and after laser hyperthermia, CW, 1 Watt, 30 minWatt, 30 min

○ ○ - control, ■ - laser heating without - control, ■ - laser heating without nanoparticlesnanoparticles

▲ ▲ - laser heating after intravenous injection - laser heating after intravenous injection of of nanoparticlesnanoparticles , 1 ml, 1 ml

- laser heating after interstitial injection of - laser heating after interstitial injection of nanoparticles, 0.2 mLnanoparticles, 0.2 mL

Tum

or s

ize,

cm

3

A day after tumor transplantation

Laseraction

thermogram of tumor without nanoparticles

Dependence of the mean tumor volume on the number of daysDependence of the mean tumor volume on the number of dayspassed after PPTTpassed after PPTT

experimental groups with doses of administered gold nanorods: 8 (1) and 2 mg / kg (2), control groups under laser irradiation without introducing nanoparticles (3) and without laser irradiation (4)

Tum

or v

olum

e, c

m3

Number of days

Nanocomposites: vitrified gold-silver nanocages Nanocomposites: vitrified gold-silver nanocages modified Yb-HPmodified Yb-HP

Au-NRAu-NR--SiOSiO22--HPHP

IR-laser ~2.3 Wt/cm2 He-Ne laser ~160 mWt/сm2

Tumor before exposure (V= 3 сm3)

Tumor (20 min of exposure)

Tumor after 72 hours(V= 0.16 сm3)

Three variants of the protocol for Au-NRThree variants of the protocol for Au-NR--SiO2-HPSiO2-HP PTDPTD + + LTTLTT

a) LTT accelerates chemical reactions

b) Synergistic effects on blood vessels (stasis, collapse)

c) Synchronicity subsequent processes in the tumor tissue

PTDPTD→→12 12 hourshours→→LTTLTT

a) absence of optical postthermal obstaclesb)exclusion of thermal effectsc) hemostasis (2-12 hours): it is benefit for thermotherapy as heat loss reducesd) decrease in the partial pressure of oxygen and pH increases thermal sensitivitye) LTT inhibits repair of necrotic cells

LTTLTT→→24 24 hourshours→→PDT PDT

a) direct necrosisb) hemostasis - partial

reduction of oxygen and pH

c) 24 hours restore of blood flow

d) 24 hours - pic HSP70e) resumption of protein

synthesis 12-26 hours

f) PDT inhibits repair of cells

1) the size of tumors < 3 cm3

2) Temperature 50-75 °C - to avoid the formation of coagulation optical barrier

Thank you!