SyriCalm™ CLR (PC)

To soothe, protect
and strengthen

confidential information © CLR - Chemisches Laboratorium Dr. Kurt Richter GmbH

SyriCalm™ CLR (PC)

Based on Phragmites and Poria

cocos, SyriCalm™ CLR (PC) soothes
after and protects against external
stresses.

Protects and re-establishes the skin’s

barrier function by
• reducing inflammation
• protecting against loss in cell
viability and energy state
• acting against photosensitivity
SyriCalm™ CLR (PC)

In vitro test results

Effect on UV-induced inflammation
Reduction of UV-induced proinflammatory
mediator production

TNFα (%)
Control 1.0% SyriCalm™ 1.5% SyriCalm™
Keratinocytes were irradiated
100
with UV light (2 J/cm² UVA; 0.2
J/cm² UVB) after pretreatment 90
with different dosages of 80
SyriCalm™. 70

The release of TNFα by 60

irradiated cells without 50
pretreatment with SyriCalm™ is 40
set at 100%.
30
Method: Luminescence ELISA
20
10
0

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

In vitro test results

Cellular resilience against
UV irradiation
Protection against UV-induced
ATP depletion
ATP (RLU)
250 Control 0.25% SyriCalm™ 1.0% SyriCalm™ 1.5% SyriCalm™
Keratinocytes were 240
irradiated with different
dosages of UV light after 230
pretreatment with 220
different dosages of
SyriCalm™. 210

200
Control cells were
irradiated without 190
pretreatment with 180
SyriCalm™.
170
Method: 160
Luciferase/Luceferin assay
150

1.0 J/cm2 UVA 2.0 J/cm2 UVA

0.1 J/cm2 UVB 0.2 J/cm2 UVB

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

In vitro test results

Effect on photosensitivity
Photosensitivity

Hypericin

UV Reactive oxygen species

• Decrease in cell viability

• Inflammation

SyriCalm™ CLR (PC)

Hypericin phototoxicity: Protection against
decrease in cell viability
Cell viability (%)
110 Control A Control B 0.25% SyriCalm™ 0.50% SyriCalm™
Keratinocytes were
treated with hypericin 105
(0.5 µM) and UV
irradiation (0.25 J/cm² 100
UVA; 0.025 J/cm² UVB)
95
with and without
pretreatment with 90
SyriCalm™ .
85
Related to control,
without presence of 80
hypericin and
75
SyriCalm™ (100%).
70
Method: MTT assay
(absorption: 570 nm) UV + + + +
Hypericin – + + +

SyriCalm™ CLR (PC)

Hypericin phototoxicity: Reduction of
IL-8 expression
Control 0.5% SyriCalm™ 1.0% SyriCalm™ 1.5% SyriCalm™
IL-8 (RLU)
7500
Keratinocytes were treated
with and without
SyriCalm™ ,then irradiated 6500
with 0.25 J/cm² UVA; 0.025
J/cm² UVB.
5500
RLU (relative luminescence
units)
4500

Method: Luminescence ELISA

3500

without hypericin with 0.5 µM hypericin

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

Effect on
epidermal integrity
Epidermal integrity

The barrier function of the skin

is not only dependent on the
quality of the Stratum Corneum.

External stress leads to

alterations in cell shape and
volume and intercellular
anchoring in the epidermis
• loss in barrier function
• inflammation

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

Effect on
epidermal integrity
In vitro test results
Determining epidermal integrity with ECIS

ECIS: Electric Cell-substrate Impedance Sensing

An automated non-invasive method to monitor cell behaviour and attachment
(barrier function)

Principle:
A confluent layer of keratinocytes will
have a certain resistance (impedance)
against conducting an electrical current.
Change in cell volume and shape and
loss of intercellular attachment lead to a
loss of impedance. Regaining the
impedance indicates recovery of
epidermal integrity.

SyriCalm™ CLR (PC)

Determining epidermal integrity with ECIS

External stress leads to a disturbance of epidermal integrity

• loss in cellular volume
• degradation of intra- and inter cellular structural proteins,
potentially leading to cell-detachment

External stresses

SyriCalm™ CLR (PC)

Protection against loss of epidermal integrity
after an electrical pulse
1.5% SyriCalm™ CLR Control

110
Impedance (%):

A confluent layer of 105

human keratinocytes
was grown on an 100
electrode array chip and
incubated with 95
SyriCalm™ for 24 hours.
Resistance was recorded 90
as a function of time
with ECIS. 85
The impedance before
the pulse is set at 80
100%.
75

70
-0,02 0 0,02
0.02 0,04
0.04 0,06
0.06 0,08
0.08 0,1
0.1 0,12
0.12 0,14
0.14
t (hours)

SyriCalm™ CLR (PC)

 Protection against loss of epidermal integrity
after UV irradiation
1.5% SyriCalm™ CLR Control

Impedance (%) 105

A confluent layer of
human keratinocytes
was grown on an 100

electrode array chip and

incubated with
SyriCalm™ for 24
hours. 95

Resistance was recorded

as a function of time
with ECIS.
The impedance before 90
the irradiation was set
at 100%.
85
-0,5 00 0.5
0,5 11 1.5
1,5 22 2.5
2,5

t (hours)

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

Effect on
epidermal integrity
UV-induced degradation of E-Cadherin
E-Cadherin in the epidermis

Trans-membrane protein, Actin

essential in the formation α-Catenin
and maintenance of cell p120ctn
junctions in the epidermis β-Catenin
E-Cadherin

• plays a crucial role in the

formation of tight
junctions and
desmosomes
• part of adherens
junctions

SyriCalm™ CLR (PC)

Reduction of UV-induced E-Cadherin degradation

Soluble E-Cadherin Ground state Control SyriCalm™ 0.5%

(pg/ml) 3200

Keratinocytes were
3000
irradiated with UV light (1
J/cm² UVA; 0.1 J/cm² UVB)
2800
after pretreatment with
0.5% of SyriCalm™.
2600
Method: ELISA
2400

2200

2000
UV - + +

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

Effect on
epidermal integrity
Cellular Taurine efflux during
hyperosmotic stress
Hyperosmotic stress - consequences

Keratinocytes in the Stratum Granulosum (SG) are the real

Stratum Corneum (SC) factory; they

• produce precursors of the NMF and Ceramides

• are in the process of cornification
• are in the process of formation of the cornified envelope

Healthy keratinocytes in the SG are vital to skin function.

SyriCalm™ CLR (PC)

Hyperosmotic stress in the epidermis

Water gradient in normal skin

Water gradient in dry and diseased skin

The higher the gradient in the SG, the more

hyperosmotic stress

SC SG SS SB

SyriCalm™ CLR (PC)

Hyperosmotic stress - Strategies

Isotonic Hypertonic
Electrolytes

fast

fast
Taurine
slow

Electrolytes

Taurine

SyriCalm™ CLR (PC)

Hyperosmotic stress - Strategies

Isotonic Hypertonic
Electrolytes

Taurine

Taurine

Electrolytes

SyriCalm™ CLR (PC)

Experiment – Protection against Taurine efflux

T T T T T T
T
Taurine influx

Washing of medium

T T T T T T
T

Taurine efflux

T T T T T T T T T T

T T T

SyriCalm™ CLR (PC)

Experiment – Protection against Taurine efflux

Change of medium Measurement

[cell model washed] A B

0h 24 h 48 h 48+2 h 48+24 h 48+48 h

Hyperosmotic stress

Taurine efflux
1st application
2nd
3rd

Method: evaluation of counts per minute with TopCount NXT™ Microplate Scintillation
counter (Perkin Elmer) in MicroScint 40 Scintilation cocktail (Perkin Elmer)

SyriCalm™ CLR (PC)

Protection against Taurine efflux

Taurine efflux (%)

non-treated treated (1st+2nd) treated (3rd) treated (1st+2nd+3rd)
3D epidermal skin models
100
were cultivated in a CELLnTEC
PCT medium, exposed to a 90
hyperosmotic medium (400
mOsm) and treated with 3% 80
SyriCalm™. 70

The osmolarity of the medium 60

was kept at 400 mOsm 50
during the whole experiment.
40
Taurine efflux is expressed as
30
percentage radioactivity of
the cell medium related to 20
total radioactivity (cell lysate +
consecutive samples of the 10
medium) 0
48+24 h 48+48 h
Measurement A B

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

Effect on
epidermal integrity
Hyperosmotic stress induced inflammation
Protection against IL-8 expression
during hyperosmotic stress

IL-8 (%) untreated Control 0.25% SyriCalm™ 1.5% SyriCalm™

1400

Keratinocytes were
1200
incubated in a
hyperosmolary medium
1000
(400 mOsm) in the
presence of different 800
concentrations of
SyriCalm™. 600

Method: Luminescence 400

ELISA
200

30 min 12 hours

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

In vivo test results

Effect on UV-induced increase
in TEWL
Effect on UV-induced increase in TEWL

TEWL (g/hm2)

On designated skin areas Positive control (Bepanthen®, 5% Panthenol and Lanolin)

on the inner side of the 3% SyriCalm™
16
thigh of 10 volunteers (33 -
Application of products
79 years old) the TEWL was 14
measured. Then these
areas were exposed to 12
2.0 MED of UVB irradiation 10
to induce loss of barrier
function. 24 hours after 8
that the TEWL was
6
measured again. Directly
after that the test products 4
were applied for 4 days,
2
twice daily. Every day the
TEWL was measured 0
(method: Tewameter TM ground max. day 1 day 2 day 3 day 4
210, Courage & Khazaka, status erythema
Germany)

SyriCalm™ CLR (PC)

Effect on UV-induced increase in TEWL - Trends

Positive control (Bepanthen®, 5% Panthenol and Lanolin)

TEWL (%)
100 3% SyriCalm™
Trends in reduction of
TEWL as a function of 95
time. 90

Comparison between 3% 85
SyriCalm™ and positive 80
control.
75
The TEWL measured at 70
maximum erythema was 65
set at 100%.
60
55
50
TEWL day 1 day 2 day 3 day 4
max
erythema

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

In vivo test results

Effect on UV-induced erythema
Effect on UV-induced erythema

Skin redness (%)

Positive control (Bepanthen®, 5% Panthenol and Lanolin)
On designated skin areas on
3% SyriCalm™
the inner side of the thigh of
100
10 volunteers (33 - 79 years
old) the skin redness was
measured. Then these areas 90
were exposed to 2.0 MED of
UVB irradiation to induce 80
erythema. 24 hours after that
the redness of the skin was 70
measured again. Directly after
that the test products were 60
applied for 4 days, twice daily.
Every day the skin redness was 50
measured. The maximum
erythema is set at 100%.
40
(Method: Chromameter CR day 1 day 2 day 3 day 4
200, Minolta, Japan)

SyriCalm™ CLR (PC)

SyriCalm™ CLR (PC)

In vivo test results

Effect on histamine-induced itching
Effect on histamine-induced itching

Itching (Scoring system) Positive control (Fenistil®, 0.1% Dimethindene Maleate)

2
Histamine was applied 3% SyriCalm™
intracutaneously on a
designated area on the inner 1,5
forearm of 10 volunteers
(33–79 years old) via a lancet
stitch. The application of the 1
test products took place
immediately after the insult
0,5
with histamine. The itching
was assessed by a scoring
system 15, 30 and 60 min
0
after application of the test 15 min 30 min 60 min
products.

SyriCalm™ CLR (PC)

Summary
In vitro:
• Protection of barrier function
o Increase in cell viability
o Increase of cell energy level INCI: Water, Phragmites Communis Extract,
o Maintenance of cellular integrity Poria Cocos Extract

o Fights against the effects of Dosage: 3%

hyperosmotic stress Recommended pH: 3.0 – 8.0

• Reduction of photosensitivity Preserved with: sodium benzoate

• Anti-inflammatory action Also available as SyriCalm ™ CLR

In vivo: INCI: Water, Phragmites Kharka Extract,

Poria Cocos Extract
• Protection against histamine & UV-induced stress
o Accelerated recovery of skin barrier function (TEWL)
o Accelerated reduction of erythema
o Accelerated reduction in itching
• Conforms to the ECOCERT Natural and Organic Cosmetics standard
• Certification for NATRUE

SyriCalm™ CLR (PC)