Phytofuse Renew®Anti-Pollution + Film Forming + Moisturizing

Phytofuse Renew®

Technical Information

Product Code: 16586

INCI Name: Selaginella Lepidophylla Extract

INCI Status: Approved

Suggested Use Level: 1.0 - 10.0%

Suggested Applications: Moisturizing, Soothing, Antioxidant, Emolliency

• Unlike animals, plants are immobile

• As plants cannot escape from danger by taking motion, they need a special mechanism to withstand environmental stress

• They develop secondary metabolites to empower them to withstand harsh external influence and preserve life

• The desert provides a unique set of challenges to plant life - Very low levels of humidity- Difficulty for plant survival

• As a result, desert plants have had to develop more complex strategies for moisture retention

Phytofuse Renew®

Desert Plant: The Resurrection Plant

• Selaginella Lepidophylla

• These plants can endure almost total desiccation (loss of cellular water down to <5%) for long periods of time losing all appearance of life

• Under dry conditions SelaginellaLepidophylla is curled and wilted appearing almost as a fist

• Will fully recover when exposed to elevated moisture levels

Phytofuse Renew®

Phytofuse Renew®

The Resurrection Plant: Overcoming Stress

• Abscisic acid: Plant hormone that suspends primary & secondary metabolism during stress

• Stress Proteins: Provide anti-aggregation activity due to water stress

• Enzymes: Boost carbohydrate metabolism to repair damage caused by desiccation during rehydration

• Polysaccharides: Serves as energy storage while boosting metabolic activity, improving water holding capacity & enhancing membrane protection and stability

• Polyphenols: Work as antioxidants & membrane protectants

Phytofuse Renew®

The Resurrection Plant: Synergistic Effect

Traditional Cosmetic Approach• Focused on isolation of natural components

Novel Cosmetic Approach• Focused on isolation of intact systems to achieve a synergistic effect

All phyto-compounds in resurrection plants work synergistically to:

• Reduce cellular metabolism during desiccation

• Reduce damage during desiccation

• Resume cellular metabolism during re-hydration

• Repair damage during re-hydration

Phytofuse Renew®

The Resurrection Plant: Complex Iridoid Systems

• Reduces protein aggregation Improves integrity of structural proteins Promotes repair and improved barrier function

• Intense water holding and hydrating properties Potent moisturizing benefits Reduced Trans Epidermal Water Loss (TEWL)

• Antioxidant Protection

• Complex carbohydrates enhance epidermal Slip Emolliency Enhanced formulation aesthetics Natural Silicone Replacement

• Selaginella Lepidophylla Extract

• Unique manufacturing process- Allows for the extraction of intact systems as opposed to isolated compounds

• Provides a synergistic effect

• Globally Acceptable

• Sustainable

Phytofuse Renew®

Efficacy Data

In-vivo Efficacy Studies• Assessment of Hair Characteristics• Moisturization Comparison• Skin Density• TEWL Comparison• Pollution Protection Assay

In-vitro Efficacy Studies• High Resolution Ultrasound Skin Imaging• IL-6 ELISA• Oxygen Radical Absorbance Capacity Assay• Cellular Viability• Scratch Assay

Phytofuse Renew®

Protocol

• Baseline-A sensory evaluation was conducted

• Test Area: Hair

• Concentration of active used: 2.0% Phytofuse Renew® in a Serum

• Participant assessed hair characteristics

• Assessments were made using a rubicscale from 1-10

Figure 1. Rating of hair characteristics following sensory assessment.

Assessment of Hair Characteristics

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BaselineAssessment ofUntreated Hair

Half HeadAssessment ofControl Serum

Half HeadAssessment ofPhytofuseRenew® Serum

Phytofuse Renew®

Figure 2. Full head Baseline, Untreated Hair.

Protocol

• Subject washed & dried hair as normal

• Baseline-Sensory evaluation was conducted

• Test Area: Hair

• Concentration of active used: 2.0% Phytofuse Renew® in a Serum

Assessment of Hair Characteristics

Phytofuse Renew®

Protocol

• Subject washed & dried hair as normal

• Baseline-Sensory evaluation was conducted

• Test Area: Hair

• Concentration of active used: 2.0% Phytofuse Renew® in a Serum

Figure 3. Half-Head Zoomed-Out Photo of Hair Treated with Test Serum vs. Control Serum.

Half-Head Treated with PhytofuseRenew® Serum

Half-Head Control Serum

Assessment of Hair Characteristics

Phytofuse Renew®

Results

• Phytofuse Renew® Serum vs. Control Serum:

• Shine-14%

• Smoothness-28%

• Hydration-50%

• Manageability-43%

• Phytofuse Renew® Serum vs. Untreated Improved:

• Shine-60%

• Smoothness-29%

• Hydration-50%

• Manageability-25%

-30%

-20%

-10%

0%

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70%

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ExperimentalSerum vsUntreatedControl

ExperimentalSerum vs ControlSerum

Figure 4. Hair Assessment results for sensory characteristics.

Difference of Hair Sensory Assessment

Phytofuse Renew®

Protocol

• Equipment: DermaLab Combo

• Principle of measurement: Conductance, single frequency

• Subjects: 10 (m/f)

• Test area: Volar forearms

• Concentration of active used: 2.0%

• Frequency of application: Twice Daily

71%

110% 108%

95%

-3%

26% 27% 26%

-20%

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120%

t = 24 1 week 2 week 3 weeks

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Experimental vs. Untreated Control Experimental vs. Base Lotion

Figure 5: Improvements in moisturization following application of the test materials after a period of 3 weeks.

Moisturization Comparison

Phytofuse Renew®

Protocol

• Equipment: DermaLab Combo

• Principle of measurement: Ultrasound Probe

• Subjects: 10 (m/f)

• Test area: Volar forearms

• Concentration of active used: 2.0%

• Frequency of application: Twice Daily

Figure 6: Improvements in skin density following application of the test materials after a period of 3 weeks.

5%

8%

13%

15%

2%

6%

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14%

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t = 24 hours 1 week 2 week 3 weeks

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Experimental vs. Untreated Control Experimental vs. Base Lotion

Skin Density Comparison

Phytofuse Renew®

Protocol

• Equipment: DermaLab Combo

• Principle of measurement: Open Chamber, vapor diffusion gradient

• Subjects: 10 (m/f)

• Test area: Volar forearms

• Concentration of active used: 2.0%

• Frequency of application: Twice Daily

9%

22%23%

24%

3%

12%

14%

19%

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t = 24 1 week 2 week 3 weeks

Per

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ecre

ase

Experimental vs. Untreated Control Experimental vs. Base Lotion

Figure 7. Improvements in barrier function following application of the test materials after a period of 3 weeks.

TEWL Comparison

Phytofuse Renew®

Figure 8. Percent difference in skin density recordings between test materials.

High Resolution Ultrasound Skin Imaging

Protocol

• 10 M/F

• Equipment: DermaLab Skin Combo

• Apply 2 mg of each test material on volar forearms

• Measurements were taken 24 hours after application of test materials and then weekly for 4 weeks

• Concentration: 2.0% Phytofuse Renew®

0.00%

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t=24 1 week 2 week 3 week 4 week

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Experimental vs.Untreated

Base Lotion vs.Untreated Control

Experimental vs. BaseLotion

Phytofuse Renew®

Hair Pollution Protection Assay

Method

• 16 virgin hair swatches

• 4 testing groups – untreated, shampoo/conditioner, saturation with rinse, saturation without rinse

• Experimental – 2.0% Phytofuse Renew in Shampoo and Conditioner, 2.0% Phytofuse Renew aqueous solution

• Hair was exposed to cigarette smoke in a custom 22”x15”x6” smoke chamber for 30 minutes

• Lipid peroxidation assay was performed to quantitatively assess oxidative damage

• Malondialdehyde is useful for quantitatively measuring the end product of lipid peroxidation and determining oxidative stress.

• An increase in MDA indicates an increase in lipid peroxidation and oxidative stress.

Phytofuse Renew®

Phytofuse Renew®

Figure 9. Average values of MDA.

MDA Lipid Peroxidation

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Untreated Control Shampoo/ConditionerControl

2.0% PhytofuseRenew® in Shampoo

and Conditioner

2.0% PhytofuseRenew® Saturation

with Rinse

2.0% PhytofuseRenew® Saturation

without Rinse

nm

ol/

µL

Average MDA Concentration

Results

• 2.0% Phytofuse Renew® demonstrated significantly lower levels of MDA than the control standards

• 2.0% Phytofuse Renew® in Shampoo and Conditioner showed 41.8% less MDA than shampoo and conditioner alone.

• 2.0% Phytofuse Renew® with Rinse showed 69.0% less MDA than the untreated hair sample

• 2.0% Phytofuse Renew® without Rinse showed 41.9% less MDA than the untreated hair sample

Phytofuse Renew®

Figure 10. Average values of MDA.

MDA Lipid Peroxidation Results

• 2.0% Phytofuse Renew® demonstrated significantly lower levels of MDA than the control standards

• 2.0% Phytofuse Renew® in Shampoo and Conditioner showed 41.8% less MDA than shampoo and conditioner alone.

• 2.0% Phytofuse Renew® with Rinse showed 69.0% less MDA than the untreated hair sample

• 2.0% Phytofuse Renew® without Rinse showed 41.9% less MDA than the untreated hair sample

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2.0% Phytofuse Renew® inShampoo and Conditioner vs.Shampoo/Conditioner Control

2.0% Phytofuse Renew® Saturationwith Rinse vs. Untreated Control

2.0% Phytofuse Renew® Saturationwithout Rinse vs. Untreated

Control

Pe

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Dif

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Comparative Difference in Protection

Skin Pollution Protection Assay

Method

• Volunteers, male and female, between the ages of 23 and 45

• Apply 2 mg of each test material, experimental, control, and untreated on volar forearms

• Lotions were allowed to dry completely before the addition of 5 mg of micronized charcoal

• The micronized charcoal used has a particle size of 2.5 microns (PM 2.5) or less that mimics the small particulates found in polluted air

• Each treatment area was washed five times using deionized water

• Images were taken pre- and post-wash using a dissecting microscope

• The test material consisted of 2% Phytofuse Renew® in a Cetaphil Moisturizing for All Skin Types

Phytofuse Renew®

Pre-Wash Post-Wash

Test Sites (Left to Right): Untreated, Phytofuse® Renew, Control

Phytofuse Renew®

Pre-Wash Post-Wash

Phytofuse® Renew

Lotion Control

Untreated Control

Figure 11. Phytofuse Renew®-treated fibroblasts IL-6 concentrations.

IL-6 ELISA Analysis Protocol

• Human dermal fibroblasts were seeded into 12-well tissue culture plates

• Phytofuse Renew® were added to complete DMEM containing 1µg/mL LPS and incubated with fibroblasts for 24 hours

• Concentrations: 1%, 0.1%, 0.01%

• Phytofuse Renew®-exhibited anti-inflammatory effects on LPS-treated fibroblasts

• The changes in IL-6 production using PhytofuseRenew® appear to be dose dependent

1267.1

1432.9 1458.9

1917.9

450.1

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1% 16586 Phytofuse Renew™

0.1% 16586 Phytofuse Renew™

0.01% 16586 Phytofuse Renew™

Positive Control Negative Control

IL-6

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Phytofuse Renew®

Figure 12. Antioxidant capacity of Phytofuse Renew.

ORAC AssayProtocol

• Trolox was used as the positive control

• Solutions were prepared at two concentrations, as a reference

• Florescent measurements were taken every 2 minutes for 2 hours

• Phytofuse Renew® showed antioxidant activity

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200 µM Trolox 12.5 µM Trolox 10% Phytofuse Renew™

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Phytofuse Renew®

Figure 13. Cellular Metabolism of Phytofuse Renew®-treated fibroblasts expressed in terms of percent of control.

Cellular Viability AssayProtocol

• Human dermal fibroblasts were seeded into 96-well tissue culture plates

• Ten microliters of viability reagent was added to 90µL of cell culture media in culture wells

• Concentrations: 1%, .10%, 0.01%

• Phytofuse Renew® at all concentrations is able to increase cellular metabolism compared to the control

• The increase in fluorescent signal indicates an increase in cellular metabolism and viability post PhytofuseRenew® treatment

123.86%

100.95%

114.11%

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1% 0.10% 0.01%

Via

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ty (

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ntr

ol)

Concentration of Sample

Phytofuse Renew®

Protocol

• Human dermal fibroblasts were allowed to grow to confluency in complete DMEM

• 0.5% Phytofuse Renew® was added to the serum free DMEM & incubated with fibroblasts

• The confluent cells were scratched

• Cells were stained for enhanced microscopy

Figure 14. Images right after scratch, after 5 days and after 5 days but stained for enhanced microscopy.

Scratch Assay

A B

IHG

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Phytofuse Renew®

Phytofuse Renew®

Technical Information

Product Code: 16586

INCI Name: Selaginella Lepidophylla Extract

INCI Status: Approved

Suggested Use Level: 1.0 - 10.0%

Suggested Applications: Moisturizing, Soothing, Antioxidant, Emolliency

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Phytofuse Renew®