bios 7901 - 12

Energy-based Treatment of Tissue and Assessment VI

Modelling and characterization of photothermal

effects assisted with gold nanorods in ex-vivo

samples and in a murine model

AUTHORS:

Félix Rodríguez Jara1, Horacio Lamela Rivera2 and

Vincent Cunningham3

[email protected]@ing.uc3m.es

3vcunning.uc3m.es

OPTOELECTRONICS AND LASER TECHNOLOGY GROUP

ELECTRONIC TECHNOLOGY DEPARTMENT

San Francisco (CA), January 23th, 2011

mailto:[email protected]



Modelling and characterization of photothermal effects assisted with

gold nanorods in ex-vivo samples and in a murine model1

General Index:

1. Introduction

2. Opto-thermal modelling for Photo-Thermal Therapy

3. Experimental Results

4. Conclusions and Future Work

0. General Index.

1. Introduction

1.1 The Motivation

1.2 The hyperthermia

technique.

1.3 The Photo-thermal

therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.

3.1 Experimental Set-

up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

2

General Index:

1. Introduction


gold nanorods in ex-vivo samples and in a murine model

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

3

1.1 The Motivation.

CANCER, one of the main causes of mortality all around the world.

- 7.4 Millions out of the total deaths per year ≈ 13 % (WHO)

Need of investigation in new therapy techniques

Mortality and side effects

Number of patients that can be trated

Laser hyperthermia technique



Collaboration with a specialized Company in

Animal models

- Good Laboratory Practices (GLP)

- Qualified Staff

- Ethical Committee

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

ESTUDIO EXPERIMENTAL DE TÉCNICAS LÁSER PARA TERAPIA

CÁNCER EN RATONES UTILIZANDO NANOPARTÍCULAS DE ORO4

1.2 The hyperthermia technique: the GOAL

Hyperthermia temperature

(from 42ºC to 65 ºC)

held during various minutes

(3-10’)

Tumoral Cell death

GOAL: 80% Tumour

tissue ablation

Hyperthermia temperature selected:

55 ºC

Biological Tissue

Tumour

37 ºC

5-10 mm

42 ºC – 65 ºC

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

5

1.2 The Hyperthermia Technique: State-of-

The-Art

- Low selectivity

- Use high amounts of energy

- Very expensive devices with big dimensions

Tejido biológico

Tumor

Radiofrecuencia

Ultrasonidos (HIFU)

Láseres alta potencia

estado sólido (Nd:YAG, OPO)

Fibra óptica + Difusor

Registro Tª



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

6

1.3 The Photo-thermal Therapy System

Temperature

Register

HIGH

POWER

LASER

DIODE

Nanoparticle

infusion:

Intratumoural or

Intravenous ?

Interna

Superficial

λ= 808

nm

Biological

Tissue



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

7

1.4 Nanotechnology in Photo-Thermal Therapy

Gold Nanoparticles

GOLD NANORODS

Tuned optical

absorbance

EFFICIENCY POWER

400 500 600 700 800 900 1000 11000

0.5

1

1.5

Longitud de onda [nm]

Ab

so

rció

n [

cm

-1]

Fig. adapted of “Cancer Research”, 69(9):1-9, (2009)

- Passive

- Harmless



A Longitudinal

Surface Plasmon

Resonance Peak

B Axial

Surface Plasmon

Resonance Peak

A

B

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work



1.5 The Design: Experimental studies

- Number of animales for group -> Statistics

- System design in terms of efficiency:

(Power, Irradiance, Nanoparticle Concentration)

- Therapy parameters

(Exposure time, way of application)

- Ethical Committee (international directives of

animal handling) -> HEATING UP, NOT BURNING

OR CHARRRING

- Tumour size?

- Stops Tumor growing?

- Ablation of the tumoral tissue?

Efficiency Study

OPTIMIZE THE SET-UP

Efficacy Study

CT-26 COLON CANCER

XENOGRAFT STUDIES

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work



General Index:

2. Opto-thermal Modelling for

Photo-Thermal Therapy

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

11

2.1 Approach

Fuentes térmicas

Optical power,

Irradiance

P [W], I [W/cm2]

Optical properties

of the tissue

µa [cm-1], µs [cm-1]

Energy

Deposition Transferencia

térmica

Physical and thermal properties

of the tissue

ρ[kg/m3]C [JKg

-1K

-1, k [Wm

-1K

-1],

Temperature

dV

Thermal Energy

TransferredTemp

dV

Optical Power

Thermal Source

Tissue absorption + Nanoparticles

µtotal = µtissue+µnanoparticles



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

14

2.2 Finite Element Modelling (FEM)

Initial

Temperature

37 ºC

Energetic

Contribution.

Laser energy

absorption

Thermal

Energy

transference

Temperatures

update

Stop time

reached?

Final

Temperature

YES

NO



- Less computational resources needed

- It can be applied to complex geometries

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

15

General Index:

3. Experimental Results

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

16

3.1 Experimental Set-up (I)

Optoelectronic General Schema

Cabezal láser

(CNI-MDL-H-808-5000, CNI)

Fibra óptica

(CNI-SMA-Fibre-600, CNI)

d1

Termómetro infrarrojo

(OS-530LE, Omega)

Tejido irradiado

Lentes de

acoplo

l1 = l2

f1= f2 = 2.54 cm

D1 = D2= 2.54 cm

Diámetro del haz

(FWHM)

Sonda termopar

hipodérmica

(HYP-1, Omega)

Thermocouple

thermometer

d0

Driver

(PSU-H-LED, CNI)

Soporte

l1 l2 d2



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

17

3.1 Experimental Set-up (II)

Experimental Set-up for therapy application in ex-vivo tissue samples



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

18

3.1 Experimental Set-up (and III)

Experimental set-up for therapy application in mice (in-vivo)



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

19

3.1 Experimental Set-up:

Development Stages

Biological Model

ex-vivo(tissue samples)

Concept, Design

and Implementation

of the system

Clinical

Application

(Human Beings)

Develpment Stages of the Photo-Thermal Therapy System

Experimental

Results

Optimization

of the

System

Biological Model

in-vivo(ratones)

Optimization

of the

System

Experimental

Results

Requirements

and Goals



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

Optimization loop

20

3.2 Experimental results - ex-vivo (I)

Sample preparation:

- Fresh chicken muscle tissue.

- Previous marking for infrared thermometer alineation.

- Hypodermical infusion of nanoparticles (Ntracker 30-PM-850, NANOPARTz,

saline solution PH = 7.4, 0.1 ml, OD = 25).



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

22

3.2 Experimental results - ex-vivo (II)

Looking for the optimal irradiance, experimental results.

0 50 100 150 200 250 3000

5

10

15

20

25

30

35

40

Tiempo de exposición [sg]

T

[ºC

]

Phaz

= 1.25 W, IFWHM

= 2.38 [W/cm2], Láser+NRds

Phaz

= 0.5 W, IFWHM

= 0.95 [W/cm2], Solo Láser

Phaz

= 0.75 W, IFWHM


Phaz

= 1.00 W, IFWHM


Phaz

= 1.25 W, IFWHM


50 100 150 200 250 300

20

25

30

35

40

45

50

55


T

[ºC

]

Phaz

= 0.5 W, IFWHM


Phaz

= 0.75 W, IFWHM


Phaz

= 1.00 W, IFWHM


Phaz

= 1.25 W, IFWHM


Phaz

= 0.5 W, IFWHM


Phaz

= 0.75 W, IFWHM


Phaz

= 1.00 W, IFWHM


Phaz

= 1.25 W, IFWHM


Pbeam = 1.25 W

IFWHM = 2.38 W/cm2

ΔT = 31 ºC

ΔT = 3 ºC

Exposure Time [s]



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

27

3.3 Experimental Results - in-vivo (I)

Animal model

- Female mice, albines, BALB/c (BALB/cAnNHsd) , specific patogens free.

- Animals were handled by qualified staff in a company with the Good Laboratory Practice Certificate

(GLP)

Animal preparation

- Random distribution by weigth.

- Identification of each animal.

- Hair removing from the exposed area.

- Light anaesthesya Ketamine/Xilacine (10μl/g).

- Hypodermic infusion of nanoparticles

(Gold Nanorods, Ntracker).

- Laser irradiation exposure.

Identification

marks Shaving of

irradiated area

Positioning

platform

Laser beam

direction



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

28

3.3 Experimental Results - in-vivo (II)

Study of the thermal increment induced as a function of laser irradiance.

0 50 100 150 200 2400

5

10

15

20

25

30

35


T

[ºC

]

HYP: P = 0.5 W

SUP: P = 0.5 W

HYP: P = 1 W

SUP: P = 1 W

HYP: P = 1.25 W

SUP: P = 1.25 W

Pbeam = 1.25 W

IFWHM = 2.38 W/cm2

ΔT = 29.9 ºC

12 ºC

Exposure Time [s]



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

0 100 200 300 400 500 60025

30

35

40

45

50

55

60


Tem

pera

tura

[ºC

]

HYP: Gold Nanorods + Láser

SUP: Gold Nanorods + Láser

HYP: PBS + Láser

SUP: PBS + Láser

33

3.3 Experimental Results - in-vivo (III)

Proof of concept,

experimental results.

Phaz = 1.25 W

IFWHM = 2.38 W/cm2

T = 57.8 ºC

T = 38.1 ºC



Te

mp

era

ture

[ºC

]

Exposure time [sg]

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

34

3.3 Experimental Results - in-vivo (IV)

Aspect of the skin after irradiation.

Superficie de piel expuesta

a la radiación láser



Skin exposed to the laser

beam

0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

35

3.3 Experimental Results - in-vivo (V)

Proof of concept, FEM

Modelling.

0 100 200 300 400 500 6000

2

4

6

8

10

12

14

16


T

[ºC

]

Superficial. Laser + NRds. EXP

Superficial. Laser + NRds. FEM

The superficial thermal gradient registered

experimentally was of 14.75 ºC, while the

modelled one was of 14.86. This is an absolute

error less than 0.11 ºC (0.75 %)

Exposure time [sg]



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

37

General Index:

4. Conclusions and future

work



0. General Index.

1. Introduction

1.1 The Motivation


technique.


therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 The Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

38

4.1 Conclusions:

The laser hyperthermia therapy system using gold nanoparticles

has been demonstrated to be VIABLE in a mice model.

Only the tissue injected with nanoparticles reaches hyperthermia

temperatures.

The tissue exposed to the laser beam, but NO injected with

nanoparticles, remains unaltered (it does not reach hyperthermia

temperature)

The stablished irradiance does not induce tissue charring or skin

burning.

The computational model implemented allows to make accurate

estimations of the final temperature of the irradiated tissue with

nanoparticles.



0. General Index.

1. Introduction

1.1 Motivation


technique

1.3 Photo-thermal

therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Energy Balance

2.3 Implemention of the

solution. FEM.

2.4 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work

39

4.2 Future work

Modelling and implementation

Study and design of new techniques of optical energy irradiation:

contact applicators, more than one laser source, pulsed light.

Design and test of an integrated control and monitorization

system:

Experimental study

Tumor model in mice to test the efficacy of the system-Determine if the tumour growing is stopped and finally, the tumour is

ablated.

- 15 animals (BALB/c)

- Cell line CT-26 mice colon cancer (CT26.WT)

Tissue

temperature

Temp.

register

Laser power



0. General Index.

1. Introduction

1.1 Motivation


technique

1.3 Photo-thermal

therapy system.

1.4 Nanotechnology

in Photo-thermal

therapy.

1.5 Design.

2. Opto-thermal

modelling for

photo-thermal

therapy.

2.1 Approach

2.2 Energy Balance

2.3 Implemention of the

solution. FEM.

2.4 Finite Element

Modelling.

3. Experimental

Results.


up.

3.2 Experimental

Results – ex-vivo

3.3 Experimental

Results – in-vivo

4. Conclusions and

Future Work



Thank you for your attention!

Have a nice day.

bios 7901 - 12

Technology

photothermal therapy

photothermal therapy

experimental setup

experimentalresults

experimental results3

future workresults exvivo3

finite elementmodelling

introductiongeneral