Truth + Fiction + Path to Sustainability

Tomorrow’s Vision… Today!®

Truth + Fiction + Path to SustainabilityMeristemic Cells + Brand Differentiation + Phyto-Biotics

CONTENTS

1. Our Environment + Our Footprint + Our Responsibility

2. Sustainability in the Cosmetic + Personal Care Markets

4. A New Approach: Brand Differentiation + Sustainability

3. Meet the Stem Cell

5. Phyto-Biotics Product Line

OUR ENVIRONMENT + OUR FOOTPRINT

On average, American women use 12 personal care products a day, and men average six products daily. That means an adult is likely to be exposed daily to

126 unique chemical ingredients in personal care products alone.1

Environmental, social and economic impact of the personal care industry

Research shows the most significant footprints in the cosmetic industry lie with packaging and ingredients

The cumulative effects of wasteful and irresponsible habits associated with the personal care industry can significantly contribute to global warming

Reference: Good Sustainability Practice (GSP) for the Cosmetics Industry. Colipa – European Trade Association. Colipa – The European Cosmetics Association. Brussels. (2010) 1-35.

OUR NEGATIVE IMPACT

"Our global footprint now exceeds the world's capacity to regenerate by about 30 percent.“2

FACTS:

Extinction rates are rising by a factor of up to 1,000 above natural rates. Every hour, three species disappear. Every day, up to 150 species are lost. Every year, between 18,000 and 55,000 species become extinct. The cause: human activities. 3 ~ Ahmed Djoghlaf

Every day, an estimated 100 plant and animal species are lost to deforestation. 4

Many river systems approach the fate of those in China, where chiefly because of pollution 80 percent of the 50,000 kilometers of major channels can no longer support fish of any kind. 5

OUR RESPONSIBILITY

“The social, economic and environmental pillars of sustainability all have a place at the heart of the life cycle of cosmetic and personal care products. The

integration of these elements is not a marketing tool, it represents a genuine commitment to sustainability … at many levels of the business model.”

~ Bertil Heerink - Director-General of Cosmetics Europe

What this means?

Personal Care and Cosmetic companies need to think and act sustainably

Includes sourcing, producing, manufacturing, packaging, and distributing

Attention to health and well-being of consumers


WHAT IS SUSTAINABILITY?

Development that meets the needs of the present without compromising the ability of future generations to meet their own needs.

Three Pillars of Sustainability

Environmental

Economic

Social

Sustainability takes an honest look at how a company’s business practiceseffect each of these pillars on an individual scale.


ACHIEVING SUSTAINABILITY

Creating and maintaining a company that generates a profit

Use available resources wisely(employees, materials, money)

Reduce business’s effects on the environment

Integrate and remain a successful part of the community and how it operates

Manage, train and develop aconscientious staff

Social

Environment Economic

Bareable Equitable

Viable

Sustainable

Reference: Ten Steps to Sustainability; All you need to know and do for a successful start. Colipa – European Trade Association. Colipa – The European Cosmetics Association. Brussels. (2011) 1-15.

PERSONAL CARE MARKET + SUSTAINABILITY

The personal care and cosmetic industries directly affect all three pillars of sustainability

Consume natural resources at an unsustainable rate Diminishing biodiversity

Initiatives, specifically by the European Trade and Cosmetic Association have been taken to reduce negative impacts and increase positive impacts

Evaluate operations to identify the social, economic and environmental impacts at every stage in the business value chain:

Raw materials Product design + development Production Packaging Transport + retailing Use End of life

Reference: Ten Steps to Sustainability; All you need to know and do for a successful start. Colipa – European Trade Association. Colipa – The European Cosmetics Association. Brussels. (2011) 1-15.

BENEFITS OF SUSTAINABILITY

Reduce costs + sheds light on areas of wasted resourced

Inspires product development

Maintain/ increase sales

Fosters competitive position in the market + remain competitive in the future

Employee Motivation + Employee Satisfaction =Increased Productivity + Improved Company Climate

Boosts company reputation and community perception

Promotes + nurtures supplier and retailer relationships

Viewed as fiscally responsible leading to enhanced investments


A FACE-LIFT FOR THE STEM CELL

“The new age of anti-aging is how Cosmetic Design is describing plant stem cell technology, saying plant stem cell extracts are efficacy in a jar”

~ Eric Perrier/LVMH

The demand for stem cell technologies that promise potent anti-aging benefits has grown exponentially in the last decade

Cosmetic market has seen many incarnations of stem technology Plant stem cells Ingredients to activate endogenous adult stem cells

Plant stem cell technology facilitates the development of eco-friendly and sustainable cosmetic products. Technological advances allow cosmetic scientists to select, extract and improve natural molecules.

A FACE-LIFT FOR THE STEM CELL

WHAT ARE STEM CELLS?

Non-specialized cells capable of self-renewal…

What this means…

Each new cell has the potential to either remain a stem cell or become another type of cell with a more specialized function

Specialized cells define cell differentiation

Cellular Plasticity:

A specific characteristic of stem cells

The cell’s ability to move from an undifferentiated state toa specific cell type

Types of Plasticity:

Pluripotent-cells that can transform from a generic plant or animal cell into many different cell types

Totipotent-cells that can transform into any cell type

STEM CELL DIFFERENTIATION

Meristem Cells

• Pluripotent cells found in plants

• Have the ability to replicate beyond Hayflick’s Limit

• No ethical issues surround the use of plant stem cells (in contrast to embryonic stem cells)

• Currently two approaches to stem cells: Stimulation of adult stem cell proliferation The use of plant stem cells

STEM CELLS + CURRENT TECHONOLOGY

Meristemic Cell Technology in Cosmetics

Most popular form of stem cell technology

Said to slow aging by defending against extrinsic stress

This technology uses non-differentiated cells from simple cell extracts

Real problem for “anti-aging” cosmetic products… No specific activity No cosmetic benefits

STEM CELLS + COSMETIC MARKET TRENDS

The Green Approach: Plant Cell Cultures

Molecules of interest are highly concentrated in the meristem cells (buds)

In culture, all cells are dedifferentiated cells with meristematic character

Only requires a small amount of plant tissue to find active substance

Completely eco-friendly, protects biodiversity, no harvesting wild plants or crops

Full access to natural substances plus a higher presence of key plantmolecules

Numerous plants can be grown in culture in the presence of bacteria and yeast to enhance the production of secondary metabolites

PLANT CELL CULTURES + MARKET TRENDS

A NEW APPROACH:THE TRUE PATH TO SUSTAINABILITY

A sustainable and eco-friendly product line

• The use of biotic stress induces secondary metabolites: Allows for the sustainable production of extracts precisely

standardized for specific actives

• The use of secondary metabolites: Essential for the plant to interact with its environment Allows for adaptation, defense & ultimately survival in less than ideal

conditions

Biotic Stress

Damage caused by living organisms prompts plant to excrete stress responses

Examples: Bacteria, Fungi, Viruses, & Insects

Abiotic Stress

A negative impact on plants due to extreme environmentalconditions and/or pathogenic stress

Examples: Extreme Heat/ Cold, Intense UV Radiation, Flood/ Drought, Salicylate Production

A NEW APPROACH:BRAND DIFFERENTIATION + SUSTAINABILITY

Efficacious + Eco-Sustainable Extracts

Continued destruction of plants poses a major extinction threat

Secondary metabolites have no fundamental role in the maintenance of a plant’s life process

Isolating these organic compounds through solvent extraction for specific benefits

Numerous plants can be grown in culture in the presence of bacteria and yeast to enhance the production of secondary metabolites

Allows for the differentiation of plant stem cell trough pathogenic stress

A NEW APPROACH:BRAND DIFFERENTIATION + SUSTAINABILITY

PHYTO-BIOTICS PRODUCT LINE

Use biotic stress embodied by sustainable practice of co-culturing plant cells with Leuconostoc or Pseudomonas sp.

This approach induces cellular differentiation, or finely customized materials, standardized for specific activity and cosmetic skin benefits

Phyto-Biotics Product Line promotes brand differentiation & a means ofincluding patented, potent and efficacious actives Cosmetic benefit claims link to the product not the active Avoid intellectual property infringement

Phyto-Biotics Acai®

• Code - 16587

• INCI Name – Euterpe Oleracea Fruit Extract

• INCI Status – Approved

• Suggested Use Levels – 1.0-10.0%

• Suggested Applications – Anti-aging, Anti-wrinkle, Soothing, Antioxidant, ATP Synthesis, Increases Cellular Metabolism, Moisturizing, Decrease in Moisture Regression


Addresses:

Intrinsic + Extrinsic Aging

Dehydrated Skin

How:

Antioxidant protection

Increased ATP Synthesis

Enhanced Cellular Metabolism

Potent moisturizing benefits that continue to moisturize the skin even after application has ceased

PHYTO-BIOTICS ACAI®

Why is Acai of Interest?

This tree is known for its ability to survive in an extremely hot environment

Tree evolved by producing secondary metabolites to survive

What Secondary Metabolite Helps Acai Trees Survive?

Ferulic Acid (C10H10O4):

Found in the plants cell wall components

Readily forms a resonance stabilized phenoxy radical

Acts as a potent antioxidant & ROS scavenger

PHYTO-BIOTICS ACAI®

EFFICACY TESTING + PHYTO-BIOTICS ACAI®

In-vivo Moisturization AssayProtocol

• DermaLab Corneometer was used to measure moisture levels on the volar forearms

• Corneometer is an instrument that measures the amount of water within the skin

• The presence of moisture in the skin improves conductance & results in higher readings than dry skin

• Baseline readings were taken on day one of the study (T=0)

Graph 1. Average moisturization levels measured at each testsite.

Protocol

• DermaLab Corneometer was used to measure moisture levels on the volar forearms.



• Baseline readings were taken on day one of the study

20

40

60

80

100

120

Pe

rce

nt

(%)

Dif

fere

nce

Experimentalvs. UntreatedControl

Experimental vs. Base Lotion


In-vivo Moisturization Assay

Comparative Moisture Results Between Test Sites

0

t = 24 T= 1 week T= 2 weeks T= 3 weeks T= 4 weeks

Graph 2. Comparative moisture analysis between testsites

Protocol

• Genetically uniform, shoot-based clonal lines of Acai were isolated & co-cultured

• Inoculated with Leuonostoc sp. for 30 days

• Ferulic acid was extracted from both inoculated & un-inoculated

• Absorbance was measured at 333nm

• Improvements of Ferulic Acid content in the co-cultured Acai palms compared to the control

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

0-1 0-4 0-5 0-24 OM-1 OM-3 OM-8

Tota

lFe

rulic

Aci

dC

on

ten

t(m

g/g

FW)

Control Inoculated

Reference: Fu-Hsiang, L, et al. Ferulic Acid Stabilizes a Solution of Vitamin C & E & doubles its Photoprotection of Skin. Journal of Investigative Dermatology (2005)125;826-832.

In-vitro Ferulic Acid Assay

Improvements in Ferulic Acid Content in Acai Co-Cultured with Leuconostoc sp.


Graph 3. Compared to the control, Ferulic Acid content increases when Acai is co-cultured with Leuconostocsp.

Protocol

• Purpose of study was to evaluate the effect of dosage on skin protection using known antioxidant solutions

• Skin is pretreated with 75, 150 &250mL of 15.0% Vitamin C, 1.0%Vitamin E and 0.5% Ferulic Acid

• Skin is irradiated with solar-stimulated radiation at a MED

• Sunburn protection is dosedependent

0

5

10

15

20

25

30

1 2

SBC

/mm

skin

3

xMedControl

15.0% Vit C, 1.0% Vit E, 0.5% FA, 150mL

4 5

15.0% Vit C, 1.0% Vit C, 0.5% FA, 250mL

15.0% Vit C, 1.0% Vit E, 0.5% FA, 75mL




Effect of Dosage on Skin Photo-Protection

Graph 4. Effects of dosage on photo-protection when using vitamins C, E and Ferulic Acid.

Protocol

• Purpose of this study was to compare photo-protection by topical antioxidant formulations

• Skin was pretreated with vehicle, 0.5% Ferulic Acid, 15.0% Vitamin C + 1.0% Vitamin E, , 15.0% Vitamin C + 1.0% Vitamin E + 0.5% Ferulic Acid

• Skin is irradiated with solar-stimulated radiation 2x – 10x minimal erythema dose (MED) at 2 x MED intervals

• Evaluations 1 day later



0

5

10

15

20

25

30

35

2 4 8 10

Co

lori

met

er(%

)In

cre

ase

6

Minimal Erythema Dose


Colorimeter Measurements of Photoprotection by Antioxidant Solutions

Control Vehicle Ferulic Acid Vitamin C + Vitamin E Ferulic Acid + Vitamin C + Vitamin E

Graph 5. Colorimeter measurements of photoprotection by antioxidant solutions

Protocol

• Purpose of this study was to compare visual and antioxidant photo-protection by topical antioxidant formulations

• Skin was pretreated with vehicle, 0.5% Ferulic Acid, 15.0% Vitamin C + 1.0% Vitamin E, , 15.0% Vitamin C + 1.0% Vitamin E + 0.5% Ferulic Acid

• Skin is irradiated with solar-stimulated radiation 2x – 10x minimal erythema dose (MED) at 2 x MED intervals

• Evaluations 1 day later



0

5

10

15

20

25

30

35

40

2 4 8 10

SBC

/m

msk

in

6

Minimal ErythemaDose


Visual Erythema + Antioxidant Photo Protection by Antioxidant Solutions

Control Vehicle Ferulic Acid Vitamin C + Vitamin E Ferulic Acid + Vitamin C + Vitamin E

Graph 6. Visual erythema + antioxidant photoprotection by antioxidant solutions

Protocol

• Trolox used as a positive control

• Solution were prepared at three concentration as a reference

• Fluorescent measurements were takenevery two minutes for two hours

• Phyto-Biotics Acai® showed antioxidant activity at 0.025% concentrations

Graph 5. The results indicated that Ferulic Acid provides comparable antioxidant activity toTrolox.


In-vitro ORAC Assay

Protocol

• Human fibroblasts are allowed to grow to confluency in complete DMEM.

• 1.0%, 0.1% and 0.01% of Phyto-Biotics Acai® was used

• Incubated with fibroblasts for 24 hours

• Decrease in IL-6 production, indicates a reduce inflammatory environment

• Decrease signs of aging & reduceformation of lines & wrinkles

Graph 6. Decrease in IL-6 production when using Phyto-Biotics Acai® indicates a reduced inflammatoryenvironment.


In-vitro IL-6 ELISA Assay

Protocol

• Measures cell mediated cytotoxicity,cell proliferation & metabolic activity

• Human fibroblasts were seeded into a 96 well tissue culture with complete DMEM

• Concentrations tested were 1%, 0.1% & 0.01%

• The fibroblasts were incubated for 24 hours

• .01% Phyto-Biotics Acai® increased cell metabolism

Graph 7. Cellular metabolism of Phyto-Biotics Acai® treated in fibroblasts expressed in terms of percentcontrol


In-vitro Cellular Viability Assay

Phyto-Biotics Acai®

• Code - 16587

• INCI Name – Euterpe Oleracea Fruit Extract



• Suggested Applications – Anti-aging, Anti-wrinkle, Soothing, Antioxidant, ATP Synthesis, Increases Cellular Metabolism, Moisturizing, Decrease in Moisture Regression



Phyto-Biotics Perilla

• Code – 40600

• INCI Name – Perilla Frutescens Extract



• Suggested Applications – Anti-aging, Anti-wrinkle, Soothing, Antioxidant, ATP Synthesis, Increases Cellular Metabolism, Increases Moisturization

Addresses:


Dehydrated Skin

How:

Antioxidant protection

Increased ATP Synthesis

Reduces Appearance of Wrinkles

Enhanced Cellular Metabolism

Potent moisturizing benefit

PHYTO-BIOTICS PERILLA

Brand Distinction Using Perilla

Of particular interest – Perilla frutescens

This Chinese basil is a popular medicinal treatment

Can flourish in the most desolate conditions

Its secondary metabolites provide:

Potent antioxidant properties

Soothing properties

Protection from inflammation caused by free radicals


What Secondary Metabolite Protects Perilla?

Rosmarinic Acid (C18H16O6):

Ester of caffeic acid & 3,4 dihydroxyphenyllactic acid

Its four phenolic hydrogens contribute to its ability to control free radical oxidation

Contains two catechol (1,2 dihydroxybenzene) rings, giving it a quality of polarity

Thus can form intermolecular hydrogen bonds between the free hydrogen of its hydroxyl & its phenoxyl radical

Significantly improving its radical stability


0.0

50.0

100.0

150.0

200.0

250.0

T = 24 Hours T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks

Mo

istu

riza

tio

n(µ

Sie

men

s)

2.0% Phyto-Biotics Perilla in Base Lotion

Base Lotion

Untreated

Protocol

• DermaLab Corneometer was usedto measure moisture levels on thevolar forearms


• The presence of moisture in theskin improves conductance & results in higher readings than dry skin


Graph 1. Average moisturization measured at eachsite


Average Increase in Moisture Levels

EFFICACY + PHYTO-BIOTICS PERILLA

Protocol

• DermaLab Corneometer was used to measure moisture levels on the volar forearms




0.0

20.0

40.0

60.0

80.0

100.0

120.0

140.0

160.0



Pe

rce

nt

(%)

Dif

fere

nce

2.0% Phyto-Biotics Perilla + Base Lotion vs. Untreated

2.0% Phyto-Biotics Perilla in Base Lotion vs. Base Lotion




Protocol

• Genetically uniform, shoot-based clonal lines of Perilla frutescens were isolated & co-cultured

• Inoculated with Pseudomonas, sp for 30 days

• Rosmarinic acid was extracted from both inoculated & un-inoculated


• Improvements of Rosmarinic Acid content in the co-cultured Perilla frutescens compared to the control

0

1

2

3

4

5

6

7

0-1 0-4 0-5 0-24 OM-1 OM-3 OM-8

Tota

lPh

en

olic

Co

nte

nt

(mg

/gFW

)

ClonalLines


In-vitro Rosmarinic Assay

Improvements in Rosmarinic Acid in Perilla frutescens Co-Cultured with Pseudomonas sp.

Control Inoculated

Graph 3. Improvements in Rosmarinic Acid in Perilla frutescensco-cultured with Pseudomonassp.

Reference: Mi Yoo, Sun and Ran Kang, Jeong et al. Antioxidant Effects of Rosmarinic Acid on Human Skin Melanoma Cells Treated with Hydrogen Peroxide.Journal of Korean Society of Applied Biological Chemistry (2009) 52(3), 247-251

Protocol

• To evaluate antioxidant effect of Rosmarinic Acid (RA) on cultured human skin melanoma cells injured by ROS

• Cytotoxicity & antioxidant effects were analyzed by an XTT assay after cells were treated with H2O2

• Rosmarinic Acid increased cell adhesion activity & DPPH-radical scavenging activity in cells treated with H2O2

• Rosmarinic Acid showed antioxidant effects on ROS, such as H2O2

0

20

40

60

80

100

120

LDH

Act

ivit

y(%

of

Co

ntr

ol)

Protective Effects of Rosmarinic Acid on H2O2


In-vitro Rosmarinic Assay

Control 90nMH2O2 100 Mg/ mL 130 Mg/mL

Graph 4. The protective effects of Rosmarinic Acid againstROS.

Reference: Mi Yoo, Sun and Ran Kang, Jeong et al. Antioxidant Effects of Rosmarinic Acid on Human Skin Melanoma Cells Treated with Hydrogen Peroxide.Journal of Korean Society of Applied Biological Chemistry (2009) 52(3), 247-251

Protocol



• Fluorescent measurements were taken every two minutes for two hours

• Phyto-Biotics Perilla showed antioxidant activity at 0.1% concentrations

10

20

30

40

50

60

70

An

tiox

idan

tC

apac

ity

(nM

TE)

0

200nM Trolox 12.5nM Trolox Phyto-Biotics Perilla 0.1%

Graph 5. The results indicated that Phyto-Biotics Perilla providescomparable antioxidant activity to Trolox.


In-vitro ORAC Assay

Protocol

• Human fibroblasts are allowed to grow to confluency in complete DMEM

• 1.0%, 0.1% and 0.01% of Phyto-biotics Perilla was used


• A decrease in IL-6 production, indicates a reduce inflammatory environment

• Decrease signs of aging & reduce formation of lines & wrinkles

-80

-70

-60

-50

-40

-30

-20

-10

0

Phyto-Biotics Perilla1.0%



Perc

ent

(%)

Dec

reas

e

Graph 6. Decrease in IL-6 production when using Phyto-BioticsPerillaindicates a reduced inflammatoryenvironment.


In-vitro IL-6 Elisa Assay

Protocol

• Measures cell mediated cytotoxicity, cell proliferation & metabolic activity


• Fibroblasts were incubated for 24 hours

• 0.10% & 0.01% concentrations of Phyto-Biotics Perilla increased cell metabolism

104

106

108

110

112

114

116

118

Via

bili

ty(%

of

Co

ntr

ol)

Phyto-Biotics Perilla 0.10% Phyto-Biotics Perilla 0.01%

Graph 7. Cellular metabolism of Phyto-Biotics Perilla treated in fibroblasts expressed in terms of percentcontrol..




Phyto-Biotics Perilla

• Code – 40600

• INCI Name – Perilla Frutescens Extract



• Suggested Applications – Anti-aging, Anti-wrinkle, Soothing, Antioxidant, ATP Synthesis, Increases Cellular Metabolism, Increases Moisturization


Phyto-Biotics Quercus

• Code – 16588

• INCI Name – Quercus Alba Bark Extract



• Suggested Applications – Anti-aging, Anti-wrinkle, Soothing, Antioxidant, ATP Synthesis, Increases Cellular Metabolism, Increases Moisturization, Improves Skin Density

Addresses:


Dehydrated Skin

Sagging Skin + Wrinkles

How:

Antioxidant Protection

Enhances ATP Synthesis + Cellular Metabolism

Improves Skin Density

Potent Moisturizing Benefit

PHYTO-BIOTICS QUERCUS

Quercus – The Herbal Soother

Quercus alba, or White Oak bark, was traditionally used for its astringent & anti-inflammatory effects

Has high constituents of phenolic compounds, tannins & quercin

Potent astringent properties:

Helps absorb toxins

Sooth irritated/swollen skin

Can improve appearance of problem skin


What Secondary Metabolite Protects Quercus?

Tannic Acid (C72H52O46): Ester of caffeic acid & 3,4 dihydroxyphenyllactic acid

Its four phenolic hydrogens contribute to its ability to control free radical oxidation

Contains two catechol (1,2 dihydroxybenzene) rings,giving it a quality of polarity

Thus can form intermolecular hydrogen bongs between the free hydrogen of its hydroxyl & its phenoxyl radical

Significantly improving its radical stability


Protocol

• DermaLab Corneometer was used tomeasure moisture levels on the volarforearms



• Baseline readings were taken on dayone of the study

Graph 1. Average moisturization levels measured at each testsite.

0.00

50.00

100.00

150.00

200.00

250.00

300.00


2.0% Phyto-Biotics Quercus + Base Lotion

Base Lotion

Untreated

EFFICACY + PHYTO-BIOTICS QUERCUS


Average Increase in Moisture Levels

0.00

20.00

40.00

60.00

80.00

100.00

120.00

140.00

160.00

Pe

rce

nt

(%)

Dif

fere

nce

2.0% Phyto-Biotics Quercus+ Base Lotion vs. Untreated

2.0% Phyto-Biotics Quercus+ Base Lotion vs. Base Lotion

Protocol

• DermaLab Corneometer was used tomeasure moisture levels on the volarforearms









60.00

65.00

70.00

75.00

80.00

85.00

T = 0 T = 1 Week T = 2Weeks

T = 3Weeks

T = 4Weeks

Am

ou

nt

of

Co

llag

en

Experimental (2.0% Phyto-Biotics Quercus+ Base Lotion)

Base Lotion

Untreated

Protocol

• 10 M/F subjects, ages 23-45. High Resolution Ultrasound Skin imaging was used on the volar forearms

• The intensity of the signals arereceived refer to a color scale

• Dark colors represent areas of skin with low reflection-no or little change in skin density

• Bright colors represent areas with strong reflections or substantial change in skin density


In-vivo Skin Density Assay

Average Results Per Test Site

Graph 3. Skin density measured via a high resolution ultra sound which reports collagen level at each test site.

Protocol

• 10 M/F subjects, ages 23-45. High Resolution Ultrasound Skin imaging was used on the volar forearms

• The intensity of the signals arereceived refer to a color scale

• Dark colors represent areas of skin with low reflection-no or little change in skin density

• Bright colors represent areas with strong reflections or substantial change in skin density

-0.10

Graph 4. Percent Difference between test site collagen levels over 4 weeks.

-0.05

0.00

0.05

0.10

0.15

0.20

T = 0 T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks

Perc

en

t(%

) D

iffe

ren

ce


In-vivo Skin Density AssayCollagen Ultrasound

Experimental vs. Base Lotion Treatment

Protocol

• Genetically uniform, shoot-basedclonal lines of Quercus alba wereisolated & co-cultured

• Inoculated with Leuconostoc, sp for 30 days

• Tannic acid was extracted from both inoculated & un-inoculated


• Improvements of Tannic Acid content in the co-cultured Quercus alba compared to the control

0

1

2

3

4

5

6

Tota

lTan

nic

Aci

dC

on

ten

t (m

g/gF

W)


In-vitro Tannic Acid Assay

Improvements in Tannic Acid Content in White Oak Bark Co-Cultured with Leuconostoc sp.

AC81912 AC61712 AC51112 AC12513 AC22413 AC30813 AC42813

Control Inoculated

Graph 5. Improvements in Tannic Acid in Quercus alba co-cultured with Leuconostocsp.

Protocol



• Fluorescent measurements weretaken every two minutes for twohours

• Phyto-Biotics Quercus showed antioxidant activity at 0.5% concentrations

0

50

100

150

200

250

200nM Trolox 12.5nM Trolox 0.5% Phyto-Biotics™ Quercus

An

tiox

idan

tC

apac

ity

(nM

TE)


In-vitro ORAC Assay

Graph 6. The results indicated that Tannic Acid providescomparable antioxidant activity to Trolox.

Protocol

• Human fibroblasts are allowed to grow to confluency in complete DMEM

• 0.1% and 0.01% of Phyto-BioticsQuercus was used


• Decrease in IL-6 production, indicatesa reduce inflammatory environment

• Decrease signs of aging & reduce formation of lines & wrinkles

-90

-70

-50

-30

-10

Phyto-Biotics Quercus 0.10% Phyto-Biotics Quercus 0.01%

Perc

ent

(%)

Dec

reas

e


In-vitro IL-6 Elisa Assay

Graph 7. Decrease in IL-6 production when using Phyto-BioticsQuercusindicates a reduced inflammatoryenvironment.

Protocol

• Measures cell mediated cytotoxicity, cell proliferation & metabolic activity


• Concentrations tested were 0.01%

• The fibroblasts were incubated for24 hours

• Phyto-Biotics Quercus at a concentration of 0.01% increased cell metabolism

80

90

100

110

120

Control Phyto-Biotics Quercus 0.01%

Graph 8. Cellular metabolism of Phyto-Biotics Quercus treated in fibroblasts expressed in terms of percentcontrol..

Via

bili

ty(%

of

Co

ntr

ol)




Phyto-Biotics Quercus

• Code – 16588

• INCI Name – Quercus Alba Bark Extract



• Suggested Applications – Anti-aging, Anti-wrinkle, Soothing, Antioxidant, ATP Synthesis, Increases Cellular Metabolism, Increases Moisturization, Improves Skin Density

REFERENCES

1. Living Planet Report. World Wide Fund for Nature. Ropress. Gland, Switzerland. October 2008. http://www.yoursharemakesadifference.com/ysmad_statistics.html (2.14.2014)

2. Dojoghlaf, Ahmed. U.N. Convention on Biological Diversity. Message to the Board. (2007) http://www.yoursharemakesadifference.com/ysmad_statistics.html (2.02.14)

3. Problem: Extinction of Plant and Animal Species. The National Wildlife Federation. 2010. http://www.yoursharemakesadifference.com/ysmad_statistics.html (2.02.14)

4. Wilson, E.O. The Creation – An Appeal to Save Life on Earth. W.W. Norton & Company, Inc. New York, New York. 2006.

5. Good Sustainability Practice (GSP) for the Cosmetics Industry. Colipa – European Trade Association. Colipa – The European Cosmetics Association. Brussels. (2010) 1-35.

6. Ten Steps to Sustainability; All you need to know and do for a successful start. Colipa – European TradeAssociation. Colipa – The European Cosmetics Association. Brussels. (2011) 1-15.

7. Fu-Hsiang, L, et al. Ferulic Acid Stabilizes a Solution of Vitamin C & E & doubles its Photoprotection of Skin. Journal of Investigative Dermatology (2005)125;826-832.

8. Mi Yoo, Sun and Ran Kang, Jeong et al. Antioxidant Effects of Rosmarinic Acid on Human Skin Melanoma Cells Treated with Hydrogen Peroxide. Journal of Korean Society of Applied Biological Chemistry (2009) 52(3), 247-251

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