Poster ID

Logo

Emerging Trend In Use Of Skin Care And Sun Care, A Boost To Upgrade In Vitro Strategies For Local Tolerance

Evaluation Of Ingredients

Roso Alicia1, Puginier Mickaël1, Bergal Mathilde1, De Servi Barbara2, Ceriotti Laura2, Meloni Marisa2

1- Seppic, La Garenne Colombes, France; 2- VitroScreen, 20149 Milan, Italy

Introduction:

Materials & Methods:

Results & Discussion:

Conclusions:

References:

The logo or logos of your institution(s)

Formulation trends or new uses are also an opportunity to improve our technical knowledge and upgrade our practices. In recent years, the popularity of aerosol sprays has increased worldwide. Out of traditional use in fragrances, deodorant/antiperspirant, hairspray and spring waters, aerosol format expanded into new categories, especially skin care and sun protection, in parallel with packaging powered by air instead of the typical propellants. This trend raises a new challenge for local tolerance assessment as some ingredients, not present in traditional aerosol compositions, can be occasionally or frequently in contact with the upper airways. The objective of the study was therefore to investigate the tolerance of some key ingredients on a reconstructed upper airway epithelium model. The ultimate goal was to facilitate, at early development stages, the selection and appropriate dosages of ingredients widely used in skin care and sun care applications.

Add the more important results in tables and figures

Show the methodology used to perform the work, it is suggested to use a descriptive diagram

Summarize the finding

1. Maione AG, Jackson GR, Vinall JL, Simpson H, Storey EL, Debatis M, Klausner M, Roper C, Hayden PJ (2018) Comparative inhalation toxicity testing using in vitro organotypic rat and human airway epithelial models. 20th ESTIV Congress, poster 135, Berlin, Germany.

2. Roso A, Puginier M, Bergal M, Nunzi F, Alonso A (2021) In vitro Approach to Assess Local Tolerance of Ingredients Dedicated to Specific Topical Care Applications. J Dermatol & Skin Sci. 3(1):30-48.

3. Denis A, Coudert C, Despax S, Ben Arous J, Bulcourt C, Roso A (2019) Innovative Rheology Modifiers for Mucosal Formulations. Skin and Formulation 5th Symposium Reims, France.

Poster ID

Logo

Emerging Trend In Use Of Skin Care And Sun Care, A Boost To Upgrade In Vitro Strategies For Local Tolerance

Evaluation Of Ingredients

Roso Alicia1, Puginier Mickaël1, Bergal Mathilde1, De Servi Barbara2, Ceriotti Laura2, Meloni Marisa2

1- Seppic, La Garenne Colombes, France; 2- VitroScreen, 20149 Milan, Italy

Introduction:

Materials & Methods:

Results & Discussion:

Conclusions:

References:

The logo or logos of your institution(s)

Formulation trends or new uses are also an opportunity to improve our technical knowledge and upgrade our practices. In recent years, the popularity of aerosol sprays has increased worldwide. Out of traditional use in fragrances, deodorant/antiperspirant, hairspray and spring waters, aerosol format expanded into new categories, especially skin care and sun protection, in parallel with packaging powered by air instead of the typical propellants. This trend raises a new challenge for local tolerance assessment as some ingredients, not present in traditional aerosol compositions, can be occasionally or frequently in contact with the upper airways. The objective of the study was therefore to investigate the tolerance of some key ingredients on a reconstructed upper airway epithelium model. The ultimate goal was to facilitate, at early development stages, the selection and appropriate dosages of ingredients widely used in skin care and sun care applications.

Add the more important results in tables and figures

Show the methodology used to perform the work, it is suggested to use a descriptive diagram

Summarize the finding

1. Maione AG, Jackson GR, Vinall JL, Simpson H, Storey EL, Debatis M, Klausner M, Roper C, Hayden PJ (2018) Comparative inhalation toxicity testing using in vitro organotypic rat and human airway epithelial models. 20th ESTIV Congress, poster 135, Berlin, Germany.

2. Roso A, Puginier M, Bergal M, Nunzi F, Alonso A (2021) In vitro Approach to Assess Local Tolerance of Ingredients Dedicated to Specific Topical Care Applications. J Dermatol & Skin Sci. 3(1):30-48.

3. Denis A, Coudert C, Despax S, Ben Arous J, Bulcourt C, Roso A (2019) Innovative Rheology Modifiers for Mucosal Formulations. Skin and Formulation 5th Symposium Reims, France.

SC_152

Esterification

Leucine involved in skin pigmentation

Leucine—

Biosourced Aminovector™

Cutaneous bio-affinity

Isosorbide—

Derived from starch

Stability & efficacy

ELAIsosorbityl Octanoyl Leucine

—Biosourced Esters of

Capryloyl/Capryloyl Leucine Isosorbide Untreated control

IBMX(a) 0.02%

ELA 0.005%

EtOH 0.5%

µg m

elan

in /

µg p

rote

in

******

+47%

▌Evaluation on normal human Caucasian melanocytes

***p<0.001(a) 3-Isobutyl-1-methylxanthine

ELA significantly

boosts melanin synthesis

in both models

▌Melanosomes transfer by immunofluorescence

▌Melanosomes transfer by flow cytometry

Untreated condition α-MSH(b) 0.0002% ELA 0.005%

Quantity of keratinocytes with integrated melanosomes

+33.9% +33.7%Q1 gate: keratinocytes (cytokeratin staining) Q2 gate: keratinocytes with integrated melanosome (cytokeratin and PMEL17 staining)Q3 gate: non-staining cells Q4 gate: melanocytes (PMEL17 staining)

ELAsignificantly stimulates melanosome transfer

quality & quantity from melanocytes to

keratinocytes

Untreated Untreated

Tan boosting (Products + UV)

Long lasting effect(No product, no UV)

ELA Acetyl Tyrosine Untreated

ITA

varia

tion

vs D

0 (%

)

+15%*+6%*

* p<0.05 vs untreated* p<0.05, ** p<0.01 vs acetyl tyrosine* p<0.05 vs D0

ELA significantly

decreases ITA & boosts the tan-up vs untreated area & acetyl tyrosine from

day 4 to day 11

ELAsignificantly provides a

long-lasting tan (up to 15 days

after use)

Key points• Natural bio-inspired

AMINOVECTOR™• Boosts melanin

synthesis & transfer• Tan booster and extender

member of

zoom

ELAsignificantly boosts the expression of genes involved in

melanogenesis [3-14]

**

KERATINOCYTES

MELANOCYTES

STAGE I

STAGE II

STAGE III

STAGE IV

TYR TYROSINASETYRP1TYRP2PMEL17

ACTIN

Dendrite & Lamelipodes formation

MYO5A

RAC1

melanosome integration

by keratinoyctes

KNT1

microtubule movement

microtubule movement

MAPKUSF1 MITFPh Ph

KEYS MELANOGENIC ENZYMES

STX6

AIM1

AP1

PMEL17

C-KIT FGFR2

SCF

bFGFELA 0.005%F2RL1

MELANOSOME MATURATIONAND MELANIN SYNTHESIS

SYNTHESIS AND TRANSPORTOF MELANOGENIC ENZYMES

MELANOSOMES TRANSFER

(b) α-Melanocyte Stimulating Hormone

(a) 3-Isobutyl-1-methylxanthine

Going Further in Skin Tanning, a New Solution with Normal Daily Exposure

Chanut, Pauline1* ; Hernandez, Elsa1 ; Kern, Catherine1 ; Garcia, Christine1 1Seppic Research & Innovation, 50 boulevard National, 92257 La Garenne Colombes Cedex, France

*pauline.chanut@airliquide.com +33(0)563728634

Introduction

 BIOLOGICAL EVALUATION

Human skin pigmentation is a complex biological process responsible for the color and tanning of the skin. It is triggered by a signaling cascade leading to melanin synthesis by melanosomes in melanocytes, followed up by their transfer to surrounding keratinocytes [1, 2].

Our aim was to investigate the effect of a newly-developed esterified lipoaminoacid ELA on skin tanning.

Materials & Methods

Results & Discussions

ConclusionReferences: 1. Plonka et al.. (2009) What are melanocytes really doing all day long…?. Exp. Dermatol ., 18(9): 799-819.2. Cichorek et al.. (2013) Skin melanocytes: biology and development. Advances in Dermatology and Allergology/Postȩpy Dermatologii I Alergologii, 30(1): 30.3. Park et al. (2009) Cellular mechanisms regulating human melanogenesis. Cell Mol Life Sci 66(9): 1493-1506.4. Berson et al.. Pmel17 initiates premelanosome morphogenesis within multivesicular bodies. Mol Biol Cell. 2001, 12(11): 3451-645. Sommer L. Generation of melanocytes from neural crest cells (2011) Pigment Cell Melanoma Res. 24(3): 411-21.6. Theos et al.. Functions of adaptor protein (2005) (AP)-3 and AP-1 in tyrosinase sorting from endosomes to melanosomes. Mol Biol Cell. 16(11): 5356-727. D’Mello et al. (2016) Signaling pathways in melanogenesis. Int J Mol Sci. 17(7): e1144.8. Jung et al.. (2011) Depigmenting action of platycodin D depends on the cAMP/Rho‐dependent signalling pathway. Exp Dermatol. 20(12): 986-91.9. Kim et al.. (2010) MKK6 increases the melanocyte dendricity through the regulation of Rho family GTPases. J Dermatol Sci. 60(2): 114-9.10. Scott GA, Cassidy L. (1998) Rac1 mediates dendrite formation in response to melanocyte stimulating hormone and ultraviolet light in a murine melanoma model.

J Invest Dermatol. 111(2): 243-50. 11. Strom et al.. (2002) A family of Rab27-binding proteins. Melanophilin links Rab27a and myosin Va function in melanosome transport. J Biol Chem. 277(28):

25423-30.12. Westbroek et al.. (2003) Interactions of human Myosin Va isoforms, endogenously expressed in human melanocytes, are tightly regulated by the tail domain. J

Invest Dermatol. 120(3): 465-75. 13. Toyoshima et al.. (1992) Kinectin, a major kinesin-binding protein on ER. J Cell Biol. 118(5): 1121-31. 14. Seiberg et al.. (2000) The protease-activated receptor 2 regulates pigmentation via keratinocyte-melanocyte interactions. Exp Cell Res. 254(1): 25-32.

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 CLINICAL EVALUATION

• 20 Caucasian women (20-37 years old) – phototype III • Formula with 1% ELA, 1% acetyl tyrosine (Benchmark) or untreated • Application on thighs, twice a day during 11 days + 6 UV-A exposures (mean 16.6 J/cm²) during the study + 14 days without product nor UV • Measures: Pictures + measure of L* and b* values to calculate ITA (Individual Typology Angle: a pigmenting effect results in the decrease in ITA)

 BIOLOGICAL EVALUATION

 CLINICAL EVALUATION

D0 D2 D4 D7 D9 D11 D12 D18 D25

BIOLOGICAL ACTION

MELANOCYTE

KERATINOCYTE

MELANOSOME

ELA

1. Stimulation of Melanin synthesis

2. Stimulation of Melanosome transfer

CLINICAL BENEFITS

The bio-inspired esterified lipoaminoacid ELA can be considered as a key active ingredient acting biologically to boost and prolongate skin tanning while limiting sun exposure.

ELA 0.005% (w/w)or positive reference

Human melanocytes

Melanin extraction Measurement at 405 nm

8 daysTopical applicationELA 1% (w/w) or placebo

RNA extraction +TaqMan array

30 h

Melanized RHEphototype III-IV

ELA 0.005% (w/w)or positive reference

Human keratinocytes & melanocytes co-cultivation

Flow cytometry amount of keratinocytes integrating melanosomes

Immunofluorescenceamount of transferred melanosomes from melanocytes to keratinocytes

2 days PMEL17& Cytokeratin co-staining

Formula with 1% ELA+ 1% acetyl tyrosine + untreated

6 UV exposureMinimum Pigmentation Dose of UVA

> to standardize UV exposure

PHENOTYPIC ANALYSIS

GENE ANALYSIS

PHENOTYPIC ANALYSIS GENE ANALYSIS

+90%** of transferred melanosomes with ELA 0.005%

**p<0.01