Karolina Bazela, PhD Dr Irena Eris Cosmetic Laboratories, Centre for Science and Research, Piaseczno, Poland

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1 Activation of mitok ATP in skin cells for vitiligo skin treatment Karolina Bazela, PhD Dr Irena Eris Cosmetic Laboratories, Centre for Science and Research, Piaseczno, Poland

2 POTASSIUM CHANNELS Potassium channels have been found in the inner mitochondrial membranes of various cells Potassium channels regulate mitochondrial membrane potential, respiration and synthesis of reactive oxygen species The activation of these channels is cytoprotective

3 IN VITRO STUDY The purpose of in vitro study: detection of mitochondrial ATPregulated potassium channels in human fibroblasts identification of a substance modulating the activity of these channels.

4 RESULTS IDENTYFICATION OF mitok ATP CHANNEL IN FIBROBLASTS BIOPHYSICAL PROPERTIES OF THE mitok ATP CHANNEL

5 POTASSIUM CHANNELS-PUBLICATIONS

6 FLAVONOIDS - NARINGENIN flavonoid abundant in genus Citrus anti-oxidant, free radical scavenger anti-inflammatory anti-allergic

7 RESULTS ACTIVATION OF THE mitok ATP CHANNEL BY NARINGENIN

8 RESULTS IN VITRO STUDIES - NARINGENIN Regulatory effect in mitochondria membrane potential after UVA Regulatory effect on Ca 2+ level (with and w/o UVA) Reduction of ROS (with and w/o UVA) Induction of skin antioxidant defence system (MnSOD with and w/o UVA) Reduction in oxidative skin protein damages after UVA Protection of mitochondrial proteins against UVA Regulatory effect on mitochondrial biogenesis after UVA

9 NARINGENIN INHBITOR OF IL-6 PRODUCTION (ELISA ANALYSIS)

10 NARINGENIN IN SILENCING THE VITILIGO-ACTIVATING GENES (BMAL1, PER1)

11 VITILIGO dysfunction or damages in skin cells mitochondria (e.g. by free radicals formation) could be one of the reason of vitiligo development mitochondria might be a unifying target of ROS generation, cytokine production, catecholamine release and/or alteration of Ca(2+) metabolism that leads to melanocyte loss The hypothesis was that free radicals are removed by the antioxidants and mitochondrial stimulating agents (presented in the cream), turning the melanocytes functional and starting the production of melanin in the area of the vitiligo.

12 CLINICAL STUDY The purpose of in vivo study was to evaluate the efficacy of skin care products containing e.g. naringenin dedicated for vitiligo skin.

13 CLINICAL STUDY MATERIALS AND METHODS Skin care products: day cream, night cream

14 CLINICAL STUDY MATERIALS AND METHODS 38 patients with vitiligo were enrolled for the trials; they use emulsions every day for a period of 3 months Each patient was used: NB-UVB (311 nm) therapy only or cream only or combination of NB-UVB + cream in three different lesions for 3 months Researchers, in three independent dermatological clinics, were estimating the localisation and area of vitiligo lesion in %

CLINICAL STUDY MATERIALS AND METHODS In vivo evaluation: VASI (Vitiligo Area Scoring Index) assessment Clinical evaluation of skin condition in 10-point analogues scale (skin elasticity and dryness)

15 CLINICAL STUDY MATERIALS AND METHODS In vivo evaluation: VASI (Vitiligo Area Scoring Index) assessment Clinical evaluation of skin condition in 10-point analogues scale (skin elasticity and dryness) Patient s self-evaluation VASI scale (Vitiligo Area Scoring Index), repigmentation degree from 0 to 100%: Full repigmentation (100%), Very large repigmenation (76%-99%), Large repigmentation level (51%-75%), Medium repigmentation (26%-50%), Minimal repigmentation (1%-25%), Lack of repigmentation (0%) (-) Without Assessment Depigmentation within each area was estimated to the nearest of 1 of the following percentages: 0, 10%, 25%, 50%, 75%, 90%, or 100%.

16 RESULTS CLINICAL STUDY NIGHT CREAM CLINICAL ASSESSMENT USING THE VITILIGO AREA SCORING INDEX (VASI) Patient assignment NB-UVB Night cream Night cream + NB-UVB Day 84 Summary 5 out of 18 patients using light therapy (28% of test subjects) 3 out of 28 patients using the cream (11% of test subjects) 15 out of 28 patients using the cream in combination with light therapy (53% of test subjects) Average degree of repigmentation 11% Average degree of repigmentation 12.5% Average degree of repigmentation 30%

17 RESULTS CLINICAL STUDY The most spectacular improvement was observed when using the cream-gel for the night in combination with NB-UVB therapy. Image 07_b_1 before Image 07_a_2 after 3 months of product use

18 RESULTS CLINICAL STUDY NIGHT CREAM EFFECTIVENESS CONFIRMED BY CLINICAL TEST ON VITILIGO SKIN 100% of test subjects - prevents the enlargement of patches 100% - it moisturises the skin 78% - it nourishes the skin and replenishes lipids 63% - it regenerates the skin and restores its elasticity

19 CONCLUSIONS For the first time the mitok ATP has been characterized in skin cells Naringenin acts as a modulator of mitok ATP in fibroblasts Cosmetic products containing naringenin can safely and effectively restore vitiligious skin condition and what is more enhance medical treatment with NB-UVB therapy ACKNOWLEDGMENT This study was supported by a grant MERIS PBS1/B8/1/2012 from National Centre of Research and Development.

20 THANK YOU FOR YOUR ATTENTION!

Katarzyna Równy. Combination of antioxidants, plant extract enhancing VDR synthesis and NB-UVB therapy for vitiligo skin care

Katarzyna Równy. Combination of antioxidants, plant extract enhancing VDR synthesis and NB-UVB therapy for vitiligo skin care Combination of antioxidants, plant extract enhancing VDR synthesis and NB-UVB therapy for vitiligo skin care Katarzyna Równy Scientific Specialist @ dr Irena Eris Centre for Science and Research Research