AC MOISTURE-PLEX ADVANCED PF. Hyaluronic Acid Alternative + Potent Moisturizer + Improves Barrier Integrity

AC MOISTURE-PLEX ADVANCED PF Hyaluronic Acid Alternative + Potent Moisturizer + Improves Barrier Integrity

AC MOISTURE-PLEX ADVANCED PF Technical Information: Product Code: 16503PF INCI Name: Glycerin & Water & Sodium PCA & Urea & Trehalose & Polyquaternium-51 & Sodium Hyaluronate INCI Status: Conforms Suggested Use Levels: 1.0 10.0% Suggested Applications: Moisturizer, Barrier Formation, Hyaluronic Acid Alternative Solubility: Water Soluble

EPIDERMAL DEFENSE + HYDRATION Optimally Functioning Skin The epidermis is part of our innate immune system and serves as the body s primary defence mechanism against the environment Skin needs to protect us from UV damage, free radicals and hazardous microorganisms Must provide barrier protection from chemicals and liquids Skin s ability to function against environmental stress diminishes if it is not properly hydrated

DRY SKIN + DIMINISHED FUNCTION Ways skin becomes irritated, dry & damaged? Cold temperatures, humidity changes, wind in addition to cosmetic and personal care products can dry out the skin Soap, facial cleanser and toners are skin dehydrating culprits Many moisturizers contain alcohol which ironically dries the skin!

EPIDERMIS LAYERS + ROLE IN SKIN FUNCTION Epidermis Consists of several layers with the outermost layers being keratinized cells help together by epidermal lipids Sebum is secreted on the surface layer to provide moisturizing benefits Dry skin is caused by the stripping away of the surface sebum layer Integrity of the epidermal lipids becomes less substantive Keratinized cells separate Result is Red + Itchy + Inflamed + Cracked Skin

AC MOISTURE-PLEX ADVANCED PF Protecting the Skin AC Moisture-Plex Advanced PF was formulated to address skin issues by: Improving the skin s barrier integrity Reducing surface evaporation Improving moisture levels on the skin Regulating moisture and lipid balance Increasing cellular renewal

SKIN PROTECTION + INGREDIENTS AC Moisture-Plex Advanced PF Contains a complex of glycerine, sodium PCA, urea, plyquaternium-51 and sodium hyaluronate Glycerin: Hydroscopic + Humectant Helps hydrate skin and prevent lipid damage Sodium PCA: Hydroscopic Helps skin stay moist More hygroscopic than glycerine or sorbitol Urea: Moisturizing (NMF) + Exfoliant Capable of binding water in the strum corneum to prevent water loss Component of the body s natural moisturizing factor (NMF) Functions to smooth and soften skin

SKIN PROTECTION + INGREDIENTS AC Moisture-Plex Advanced PF Polyquaternium-51: Hydroscopic + Moisture Retainer Stabilized epidermal phospholipids while significantly improving moisture retention Sodium Hyaluronate: Cellular Repair + Cellular Renewal Minimizes cracking and damage Hyaluronic acid is known to minimize formation of scar tissue Trehalose: Osmo-Regulation + Protectant Glucose disaccharide Topical application may improve barrier function Possibly prevent heat shock

AC MOISTURE-PLEX In-vivo Moisturization Assay 120 110 Percent (%) Increase 100 80 60 40 65 48 95 Assay was conducted to determine the effectiveness of AC Moisture-Plex Advanced PF at increasing barrier function and improving moisture levels on the skin 7 panelists applied the control lotion (Cetaphil Moisturizing Lotion) and 3.0% AC Moisture- Plex Advanced PF in the base lotion to the respective skin patches on their volar forearms 20 Measurements were taken after 2 and 4 hours 0 Control 2 Hours 4 Hours 3% AC Moisture-Plex Advanced PF Figure 1. Improvements in moisturization following application of 3% AC Moisture- Plex Advanced PF after a 4 hour period.

AC MOISTURE-PLEX In-vivo 72 Hour Moisture Variation Assay Percent (%) Variation 90 70 50 30 10-10 80 75 66 44 25 27 23 19 12 12-5 -4-8 -2 1 2 4 6 24 48 72 Time (Hours) Assay was conducted to determine the effectiveness of AC Moisture- Plex Advanced PF at increasing barrier function and improving moisture levels on the skin over time 11 panelists applied the control lotion (Cetaphil Moisturizing Lotion) and 5.0% AC Moisture-Plex Advanced PF in the base lotion to the respective skin patches on their volar forearms Measurements were taken after 1, 2, 4, 6, 24, 48, and 72 hours Control 5% AC Moisture-Plex Advanced PF Figure 2. Improvements in moisturization following application of 3% AC Moisture- Plex Advanced PF during a 72 hour period.

AC MOISTURE-PLEX Percent (%) Variation 70 60 50 40 30 20 10 0 In-vivo Moisturization Comparison 30 Min 2 Hours 4 Hours 6 Hours 8 Hours 5 % AC Moisture-Plex Advanced PF 0.2% Hyaluronic Acid 0.1% Hyaluronic Acid Vehicle Figure 3. Variation in moisturization using AC Moisture-Plex Advanced PF against Hyaluronic Acid. Assay was conducted to determine the effectiveness of AC Moisture- Plex Advanced PF at increasing barrier function and improving moisture levels on the skin in comparison to Hyaluronic Acid 9 panelists applied the control lotion (Cetaphil Moisturizing Lotion), 5.0% AC Moisture-Plex Advanced PF in the base lotion, 0.2% Hyaluronic Acid and 0.1% Hyaluronic Acid to the respective skin patches on their volar forearms Measurements were taken at 30 minutes and again at 2, 4, 6, and 8 hours respectively

TEWL Assay AC MOISTURE-PLEX Open Chamber Water Levels 4.00 3.50 3.00 2.50 2.00 1.50 1.00 Experimental (2.0% AC Moisture-Plex Advanced PF in Base Lotion) Base Lotion Untreated Equipment: DermaLab Combo Principle of measurement: Open Chamber, vapor diffusion gradient Subjects: 10 (m/f) Test area: Volar forearms 0.50 0.00 T = 0 T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks Figure 4. TEWL measurements taken at individual test sites. Concentration of active used: 2.0% Frequency of application: Twice Daily

TEWL Assay AC MOISTURE-PLEX 0.00% -10.00% Base Lotion vs. Untreated Experimental (2.0% AC Moisture-Plex Advanced PF in Base Lotion) vs. Untreated Experimental (2.0% AC Moisture-Plex Advanced PF in Base Lotion) vs. Base Lotion Equipment: DermaLab Combo Percent (%) Difference -20.00% -30.00% -40.00% -50.00% -60.00% T = 0 T = 4 Weeks Principle of measurement: Open Chamber, vapor diffusion gradient Subjects: 10 (m/f) Test area: Volar forearms Concentration of active used: 2.0% -70.00% Figure 5. Comparison of percent reduction in water loss over time between two test sites.

AC MOISTURE-PLEX High Resolution Ultrasound Skin Imaging 35.00% 30.00% Equipment: DermaLab Skin Combo 25.00% Base Lotion vs. Untreated 10 volunteers M/F between the ages of 23 and 45 Percent (%) Difference 20.00% 15.00% 10.00% 5.00% 0.00% -5.00% T = 0 T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks Experimental (2.0% AC Moisture-Plex Advanced PF in Base Lotion) vs. Untreated Determine the skin density of the subject s volar forearms Apply 2 mg of each test material on their volar forearms Concentration: 2% Skin density was improved by 19.53% after one week and by 27.21% after 4 weeks when compared to the untreated control -10.00% Figure 9. Ultrasound Results Comparing Test Sites to Untreated Site.

AC MOISTURE-PLEX High Resolution Ultrasound Skin Imaging Percent (%) Difference 15.00% 10.00% 5.00% 0.00% -5.00% -10.00% -15.00% T = 0 T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks Figure 10. Ultrasound Results Comparing the Difference between the Test Site and the Control Site. Equipment: DermaLab Skin Combo 10 volunteers M/F between the ages of 23 and 45 Determine the skin density of the subject s volar forearms Apply 2 mg of each test material on their volar forearms Concentration: 2% AC Moisture-Plex Advanced PF improved skin density during each week of the trial, working 11.56% better than the base lotion after two weeks and 10.80% better than the base lotion after four weeks

AC MOISTURE-PLEX Change in Conductance After 24 Hours 30 Percent (%) Increase 25 20 15 10 5 Environment was kept at a constant temperature of 21 C and 19% Relative Humidity (RH) Measured on the skin of 6 volunteers before and after 24 hours The most effective level of AC Moisture-Plex Advanced PF after 24 hours is the 5.0% use level 0 1.0% AC Moisture-Plex Advanced PF 5.0% AC Moisture-Plex Advanced PF 10.0% AC Moisture-Plex Advanced PF Figure 11. Change in conductance due to increase in moisture from AC Moisture-Plex Advanced PF and the effects after 24 hours.

AC MOISTURE-PLEX Scratch Assay Analysis Scratch Area (mm 2 ) 2 1.8 1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 100ng/mL EGF Complete Media Serum-Free Media 16503PF AC Moisture-Plex Advanced PF Human dermal fibroblasts were allowed to grow to confluency in complete DMEM 1.0% AC Moisture-Plex Advanced PF was added to the serum free DMEM & incubated with fibroblasts The confluent cells were scratched 0 T=0 T=24 Hours T=48 Hours T=72 Hours Time Post Scratch Cells were stained for enhanced microscopy Figure 12. Area of scratch.

AC MOISTURE-PLEX Scratch Assay Analysis 100 90 80 Human dermal fibroblasts were allowed to grow to confluency in complete DMEM 70 Percent (%) Closure 60 50 40 100ng/mL EGF Complete Media Serum-Free Media 1.0% AC Moisture-Plex Advanced PF was added to the serum free DMEM & incubated with fibroblasts 30 20 16503PF AC Moisture- Plex Advanced PF The confluent cells were scratched 10 0 T=24 Hours T=48 Hours T=72 Hours Time Post Scratch Cells were stained for enhanced microscopy Figure 13. Percent scratch closure.

AC MOISTURE-PLEX Scratch Assay Analysis 80 70 60 100ng/mL EGF Human dermal fibroblasts were allowed to grow to confluency in complete DMEM Migration (µm/24 hours) 50 40 30 20 Complete Media Serum-Free Media 16503PF AC Moisture-Plex Advanced PF 1.0% AC Moisture-Plex Advanced PF was added to the serum free DMEM & incubated with fibroblasts The confluent cells were scratched 10 0 T=24 Hours T=48 Hours T=72 Hours Time Post Scratch Cells were stained for enhanced microscopy Figure 14. Cell migration rate.

AC MOISTURE-PLEX Scratch Assay Analysis Human dermal fibroblasts were allowed to grow to confluency in complete DMEM 1.0% AC Moisture-Plex Advanced PF was added to the serum free DMEM & incubated with fibroblasts The confluent cells were scratched Figure 15. Images at t=0 hours (A, D, G) and t=72 hours (B, E, H) for AC Moisture-Plex Advanced PF, positive control (EGF-1), and negative control (SFM). At experiment completion (t=72 hours), cells were fixed in paraformaldehyde and stained with crystal violet (C, F, I). Cells were stained for enhanced microscopy

ORAC Assay AC MOISTURE-PLEX 250 200 197 Trolox is used as the positive control Antioxidant Capacity (µmte) 150 100 153 Solutions were prepared at two concentrations for a reference Fluorescent measurements were taken every 2 minutes for 2 hours 50 0 12 200µM Trolox 12.5µM Trolox 0.5% 16503 AC Moisture-Plex Advanced 2 0.1% 16503 AC Moisture-Plex Advanced AC Moisture-Plex Advanced PF exhibited antioxidant activity comparable to 200µM Trolox Figure 6. Antioxidant capacities.

Cellular Viability AC MOISTURE-PLEX 100000 80000 MFU % Change 30% 20% Human dermal fibroblasts were seeded into 96-well tissue culture plates 10% Concentrations: 0.1%, 0.01% 60000 MFU 40000 0% -10% Ten microliters of viability reagent was added to 90µL of cell culture media in culture wells and a fluorometric measurement was taken at 560nm for excitation and 590nm for emission 20000 0 Complete Media 0.1% 16503PF AC Moisture-Plex Advanced 0.01% 16503PF AC Moisture-Plex Advanced -20% -30% AC Moisture-Plex Advanced PF had positive effects on cell metabolism at lower concentrations and slightly negative effects on metabolism at higher concentrations Figure 16. Cellular Metabolism of AC Moisture-Plex Advanced PF-treated fibroblasts expressed in terms of percent of control.

AC MOISTURE-PLEX Comparison to Hyaluronic Acid Percent (%) Increase 70 60 50 40 30 20 10 0 30 Minutes 2 Hours 4 Hours 6 Hours 8 Hours Measured the moisture levels of 9 volunteers before and after A solution of 5.0% AC Moisture- Plex Advanced PF, 0.2% Hyaluronic Acid, 0.1% Hyaluronic Acid, and a control vehicle on the forearm Measurements were taken at 30 minutes, 2 hours, 4 hours, 6 hours, and 8 hours 5.0% AC Moisture-Plex Advanced PF 0.2% Hyaluronic Acid 0.1% Hyaluronic Acid Unloaded Vehicle Figure 17. Comparative increase in skin moisturization over time when using AC Moisture-Plex Advanced PF vs. Hyaluronic Acid.

IL-6 ELISA Analysis AC MOISTURE-PLEX 3000 2500 2279.25 2500.21 2549.76 2599.40 Human dermal fibroblasts were seeded into 12-well tissue culture plates IL-6 Concentration (pg/ml) 2000 1500 1000 500 0 1% 16503PF AC MoisturePlex Advanced 0.1% 16503PF AC Moisture-Plex Advanced PF 0.01% 16503PF AC Moisture- Plex Advanced PF 1423.38 Positive Control Negative Control Concentrations: 1%, 0.2%, 0.01% AC Moisture-Plex Advanced PF were added to complete DMEM containing 1µg/mL LPS and incubated with fibroblasts for 24 hours AC Moisture-Plex Advanced PF, at all concentrations was able to decrease IL-6 production compared to our positive control Figure 18. AC Moisture-Plex Advanced PF-treated fibroblasts IL-6 concentrations.

IL-6 ELISA Analysis AC MOISTURE-PLEX % Decrease 14.00 12.00 10.00 8.00 6.00 4.00 2.00 0.00 1% 16503PF AC Moisture-Plex Advanced PF 0.1% 16503PF AC Moisture-Plex Advanced PF 0.01% 16503PF AC Moisture-Plex Advanced PF Series1 12.32 3.82 1.91 Figure 19. Percent decrease in IL-6 production compared to positive control. Human dermal fibroblasts were seeded into 12-well tissue culture plates Concentrations: 1%, 0.2%, 0.01% AC Moisture-Plex Advanced PF were added to complete DMEM containing 1µg/mL LPS and incubated with fibroblasts for 24 hours This decrease in IL-6 production indicates a reduced inflammatory environment which could decrease the signs of aging and reduce the formation of fine lines and wrinkles

Additional Data: AC MOISTURE-PLEX ADVANCED PF SAFETY DATA Dermal and Ocular Irritation Test: Non-Irritating

ACTIVE CONCEPTS LLC THANK YOU For more information Visit our website! www.activeconceptsllc.com