Early state detection of skin inflammation by AFM · Early state detection of skin inflammation by AFM Introd Peter Schön 1, Ercüment Unsurorlu , Cas Damen2, Christoph Riethmüller3,

Early state detection of skin inflammation by AFM

Introd

Peter Schön1, Ercüment Unsurorlu1, Cas Damen2, Christoph Riethmüller3, Martin Bennink1 1) NanoBioInterface and 2) NanoPhysics Group, Saxion University of Applied Sciences, Enschede 3) Serend-ip GmbH, CenTech Münster, Germany

2

Atomic Force Microsopy (AFM)

tactile method

marker-free

ultra-resolution (1 nm! in height)

Diagnosis

by

AFM ?

Atopic Dermatitis (AD)

Skin inflammations (in general):

230 million diagnoses in 2010

(3,5% world population)

Detection:

– Visible marks, itching, weeping

and crusting

AD Diagnosis: patch test

series of allergens to the skin surface, evaluation 1-3 days

Stratum corneum

Integrity of the skin barrier is a prerequisite for health !

Corneocytes = ‘messengers’

Dynamic equilibrium= proliferating epidermis balanced by permanent abrasion of cornified epithelial cells

Corneocytes = messengers

minute changes of their morphology might hint at out-of-balance processes underneath

Atopic Dermatitis

• Atopic dermatitis (AD) :

no clear knowledge about the exact cause

and mechanism

disturbed organization of the cytoskeleton on

desmosome disruption

immature and fragile cornified envelopes (CEs), and attachment sites of desmosomes

Materials and methods

No labelling or sample processing

Pain free method

Materials and methods

Nanoscale Topography

Atomic Force Microscopy

with network analysis based software

‘Biomarker’: Villus-like projections (VP)

Franz

et al.

394

topography recording computer vision number value

Dermal

Topographical

Index

DTI

On skin samples, the assay is termed

x/y = 40 µm, z = 500 nm

The extraction method is patented (US, Western Europe)

DERMATACT

VERUM INDEX SUI ET FALSI

Healthy vs inflamed skin

DERMATACT Index (DMI)

Risk factor: +100 corneocytes/400 µm²

Franz

et al.

Skin depth

– 1 patient, 1 position, 6 different layers

– Decrease in VPs in depth skin

Age and Pigmentation do not matter

Outliers reveal a hereditary background,

(elevated count hints at individual risk)

Age (years)

outliers are healthy, but have a

hereditary background in skin

disease

Skin Type

Study on healthy individuals (2013/14)

Franz

et al.

Clinical tests

Second batch:

– 9 different patients, same layer

– P1, P2, P6, P10 are risk factors

AFM vs clinics

Patient Risk factor Disease Match

P1 Yes Skin inflamation Yes

P2 Yes Skin inflamation Yes

P3 No Contact with skin inflamation Yes

P4 No P3 without contact (healthy) Yes

P5 No Healthy person Yes

P6 Yes Scar No

P7 No Healthy person Yes

P9 No Healthy person Yes

P10 Yes Skin inflamation in the past Yes

Scar vs healthy tissue

20 µm2

Healthy Scar

z= 1 µm

FLG mutation and DMI

Genetics:

Filaggrin LOF mutations

Immunogold labelling

targeting corneodesmosin

Dr. Marek Haftek, Uni Lyon

FLG

+/+

F

LG

-/-

F

LG

-/+

(6 weeks with topical glucocorticoids) THERAPY Monitoring

Conclusion

• DTI indicates (inflammatory) barrier damage

• Objective, accurate, painless method

• works at non-lesional skin areas

➤ predict an individual risk for skin diseases (subclinically)

➤ abbreviate clinical studies (reduce study population)

➤ enable effective therapy monitoring (pro-actively)

23

Atomic Force Microsopy (AFM)

tactile method

marker-free

ultra-resolution (1 nm! in height)

Diagnosis

by

AFM !

Questions

m.l.bennink@saxion.nl

p.m.schon@saxion.nl

Backup Slides

Detergent causes drastic DTI increase

20 µm2 z= 1 µm

Sodium Lauryl sulfate

Genetics:

Filaggrin LOF mutations

Biochemistry:

Natural Moisturizing Factor (NMF)

Physiology:

Barrier function as TransEpithelial Water Loss (TEWL)

DERMATACT Index (DTI) of non-lesional sites

correlates to severity of skin lesions (SCORAD)

Immunogold labelling

targeting corneodesmosin

Dr. Marek Haftek, Uni Lyon

FLG

+/+

F

LG

-/-

F

LG

-/+

Method Comparison

Intensity Height

SEM AFM

SE

M

20 µm

AF

M

80 µm 5 µm

Circular protrusions on corneocytes

kn

ow

n s

ince 1

97

9