Transcript Слайд 1 - Eventry

Saratov State University
Department of Optics
& Biophotonics
PHYSICAL AND CHEMICAL METHODS OF
SKIN DRUG DELIVERY ENHANCEMENT:
COMPARATIVE STUDY OF HEALTHY SKIN
AND SKIN WITH DERMATITIS
Ekaterina A. Kolesnikova, Elina A. Genina, Georgy S. Terentyuk,
Natalya A. Tsyganova , Alexey N. Bashkatov, Daniil S. Chumakov,
Marina V. Basko, Valery V. Tuchin
Saratov State University, Saratov State Medical University,
Ulianovsk State University, Institute of Precise Mechanics and Control of RAS,
University of Oulu
e-mail: [email protected]
Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Motivation
The
main
advantages
of
transcutaneous
administration
of
preparations are:
1) minimal invasiveness or even noninvasiveness;
2) improved drug pharmacokinetics; and
3) targeted drug delivery
However, living epidermis and its upper layer stratum corneum (SC)
represent a major barrier making delivery of drugs deep into the skin a
rather difficult problem
Goal of the study is to investigate the effect of low-frequency
US, DMSO, TSO and their combination on transdermal permeation
of gold nanoparticles suspension through intact skin and skin with
the partly removed epidermis
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
 For this study laboratory rats with a healthy skin and experimental
allergic contact dermatitis were used
 As a potential drug carrier, a suspension of gold nanoshells (GNSs) in
immersion solution of glycerol (50%) and PEG-400 (50%) was used
 To enhance drug delivery into the skin dermis the suspension was
mixed with dimethyl sulfoxide (DMSO) or thiophane sulfoxide (TSO)
 For physical enhancement of the transdermal transport, the skin sites
were treated with low-frequency ultrasound (US)
 Monitoring of skin optical properties was implemented using an
optical coherence tomography (OCT)
 All measurements were performed at room temperature (about 20°C)
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Saratov Fall Meeting 2013
Materials and methods


Saratov State University
Department of Optics
& Biophotonics
Experimental setup
OCT system
Commercial spectral optical coherence tomography system OCP930SR
(Thorlabs, USA) (fig.1) working at the central wavelength 930 ± 5 nm
with 100 ± 5 nm full width at half maximum spectrum, an optical power of
2 mW, a maximum image depth of 1.6 mm, and a length of scanned
area 6 mm
Axial and lateral resolutions were 6.2 and 9.6 µm in air, respectively
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Saratov Fall Meeting 2013
Materials and methods


Saratov State University
Department of Optics
& Biophotonics
Experimental setup
OCT system
Fig.1. A general view of the spectral optical coherent tomography (left image)
and tomography probe with the sample stage (right image)
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Saratov Fall Meeting 2013
Materials and methods


Saratov State University
Department of Optics
& Biophotonics
Experimental setup
US system
Sonicator “Dynatron 125” (Dynatronics, USA) (fig.2) equipped with a 2.2cm diameter probe
The US frequency was 1 MHz, the power was 1.1 W in the continuous
mode, and the time of sonication was 4 min. During sonication, the US
probe was immersed into applied solutions.
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Fig.2. US system “Dynatron 125”
Saratov Fall Meeting 2013
Materials and methods

Saratov State University
Department of Optics
& Biophotonics
Immersion agents
As an OCA the mixture of dehydrated glycerol and polyethylene glycol with the MW
400 (PEG-400) in equal proportion was prepared. Refractive index of the prepared
mixture was evaluated the mean wavelength of the used OCT system as 1.421. The
mixture was divided into 3 parts. Dimethyl sulfoxide (DMSO, 99%, Sigma, USA) and
thiophane sulfoxide were added to the 1st and to the 2nd part, respectively, to obtain
9%-DMSO-OCA and 9%-TSO-OCA solutions.

Test objects
In vivo experiments were carried out with white outbred laboratory rats. The age of
the animals was nearly 12 months. Weight was 150-200 g. Before the experiment
they were subjected to general anesthesia by intramuscular injection of Zoletil 50
(Virbac, France). The dose was 0.18±0.02 mL. The hair on the chosen skin areas was
depilated previously using the depilatory cream Nair.
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Materials and methods

Saratov State University
Department of Optics
& Biophotonics
Test objects
In dependence on exposure type laboratory rats were divided into 8
II group - 20 minutes of mixture OCA - TSO exposure
III group - mixture OCA - DMSO application and single ultrasonic exposure
IV group - mixture OCA - TSO application and single ultrasonic exposure
V group - 20 minutes of mixture OCA - DMSO exposure
VI group - 20 minutes of mixture OCA - TSO exposure
VII group - mixture OCA - DMSO application and single ultrasonic exposure
Skin with
dermatitis
I group - 20 minutes of mixture OCA - DMSO exposure
Healthy skin
investigated groups:
VIII group - mixture OCA - TSO application and single ultrasonic exposure
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
SC
E
D
a
OCA
b
OCA
c
F
d
e
Fig.3. OCT images of healthy skin before the treatment (a), after 20 minutes of
mixture OCA - DMSO application (b), after 20 minutes of mixture OCA - TSO
application (c), after mixture OCA - DMSO application and single ultrasonic exposure
(d), and after mixture OCA - TSO application and single ultrasonic exposure (e). SCstratum corneum, E - epidermis, D - dermis, OCA – optical clearing agent, F – hair
follicle. The vertical label corresponds to 500 microns.
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Saratov State University
Department of Optics
& Biophotonics
Results
OCA
SC
D
a
b
OCA
c
d
e
Fig.4. OCT images of skin with dermatitis before the treatment (a), after 20 minutes of
mixture OCA - DMSO application (b), after 20 minutes of mixture OCA - TSO
application (c), after mixture OCA - DMSO application and single ultrasonic exposure
(d), and after mixture OCA - TSO application and single ultrasonic exposure (e). SCstratum corneum, E - epidermis, D - dermis, OCA – optical clearing agent.
The vertical label corresponds to 500 microns.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
SC
E
GNSs
GNSs
D
a
GNSs
b
GNSs
c
d
e
Fig. 5. OCT images of healthy skin before the treatment (a), after 20 minutes of
mixture GNSs - DMSO application (b), after 20 minutes of mixture GNSs - TSO
application (c), after mixture GNSs - DMSO application and single ultrasonic exposure
(d), and after mixture GNSs - TSO application and single ultrasonic exposure (e). SCstratum corneum, E - epidermis, D - dermis, GNSs - gold nanoshells.
The vertical label corresponds to 500 microns.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
GNSs
GNSsGNSs
SC
D
a
GNSs
b
GNSs
c
d
e
Fig.6. OCT images of skin with dermatitis before the treatment (a), after 20 minutes of
mixture GNSs - DMSO application (b), after 20 minutes of mixture GNSs - TSO
application (c), after mixture GNSs - DMSO application and single ultrasonic exposure
(d), and after mixture GNSs - TSO application and single ultrasonic exposure (e). SCstratum corneum, E - epidermis, D - dermis, GNSs - gold nanoshells.
The vertical label corresponds to 500 microns.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
GNSsGNSs
GNSs
a
b
Fig.7. Microphotographs of the rat skin with dermatitis after 20 minutes of mixture
GNSs - DMSO application. (a) – dyeing by hematoxylin-eosin, ×160;
(b) – dyeing by silver nitrate on Hacker G.W. method, × 1000, 1 - clusters of GNSs on
the skin surface, 2 - clusters of GNSs in the hair follicle.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
GNSsGNSs
GNSs
a
b
Fig.8. Microphotographs of the rat skin with dermatitis after 20 minutes of mixture
GNSs - TSO application. (a) – dyeing by hematoxylin-eosin, ×160;
(b) – dyeing by silver nitrate on Hacker G.W. method, × 600, 1 - clusters of GNSs on
the boundary between the epidermis and dermis and in the hair follicle area.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
GNSsGNSs
GNSs
a
b
Fig.9. Microphotographs of the rat skin with dermatitis after mixture GNSs - DMSO
application and single ultrasonic exposure, dyeing by hematoxylin-eosin, ×160 (a);
(b) –subcutaneous musculature, dyeing by silver nitrate on Hacker G.W. method with
toluidine blue addutional dyeing, × 600, 1 - clusters of GNSs in the myosymplasts
sarcoplasm.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
GNSsGNSs
GNSs
a
b
Fig.10. Microphotographs of the rat skin with dermatitis after mixture GNSs - TSO
application and single ultrasonic exposure. (a) – dyeing by hematoxylin-eosin, ×160;
(b) – dyeing by silver nitrate on Hacker G.W. method, × 600, 1 - clusters of GNSs in
the sebaceous glands.
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
Table 1. The characteristic optical probing depth lt of
the dermis at different methods of OCA suspension
delivery
lt, mkm
Exposure type
Intact skin
Skin with
dermatitis
Without exposure
119±5
123±4
20 min after
application
DMSO
127±3
154±4
TSO
132±3
153±3
Single US
exposure
DMSO
142±6
175±6
TSO
152±3
174±11
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Saratov Fall Meeting 2013
Saratov State University
Department of Optics
& Biophotonics
Results
Table 2. The characteristic optical probing depth lt of
the dermis at different methods of GNSs
suspension delivery
lt, mkm
Exposure type
Intact skin
Skin with
dermatitis
Without exposure
119±5
123±4
20 min after
application
DMSO
115±6
132±4
TSO
118±7
137±7
Single US
exposure
DMSO
112±5
150±6
TSO
108±6
162±7
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Summary





Saratov State University
Department of Optics
& Biophotonics
The analysis of the results has shown that OCA usage in both cases
of intact skin and skin with dermatitis led to the characteristic optical
probing depth (OPD) increase because of the matching of refractive
indices of collagen and elastin fibers and of interstitial fluid
The effectiveness of DMSO and TCO as amplifiers of diffusion
through the SC is about the same
OPD of the skin with dermatitis is higher than that of intact skin for
all exposure types. It can be explained by damage of SC - natural
skin barrier
GNSs delivery into the skin led to OPD decrease regardless of
delivery method because of the formation reflecting and scattering
screen on the skin surface that prevented deep light penetration into
the dermis.
Our results can be used for the development of new methods and
optimization of the existing ones of drug delivery into the skin
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Acknowledgements
This research was supported by:
Grant of President of RF NSH-1177.2012.2
FiDiPro, TEKES Program (40111/11), Finland
RF State contracts № 14.512.11.0022 and 14.B37.21.0728
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Thanks for your attention!