biocareer malaysia(dr.fred zuli)

© Mibelle Biochemistry, Switzerland 2010

Plant stem cells to protect skin stem cellsFred Zülli - Mibelle Biochemistry Switzerland


Agenda

1. Introduction “aging”

2. Stem cell cosmetics

3. Activity of apple stem cells

4. New PhytoCellTec products based on grapes and alpine rose


1. Anti-Aging Cosmetics

Skin aging is based on

• Intrinsic aging

(chronological ageing, ROS,

reduced cell renewal)

• Extrinsic aging

(environmental influences:

UV, toxines, life style)


Theory of Evolution and Aging

General Considerations Natural selection (Darwin) favours surviving and reproduction in younger years versus vitality and beauty in older ages.

Many children vs. beauty at older age

Jeanne Calment, 20 /122 y


Theory of Evolution and Aging

Bad news:

Nature / evolution does not support today’s beauty trends

Good news:

Prevention and treatments with cosmetic products can counteract aging due to:

– High technology / science

– Consumer spend money for the products

Mimi Weddell, 93


Aging and Longevity in Different Countries

Genetic Control ?Environment?Life Style?


Why Can‘t we Live Forever?

• Mouse 4 y (90% dye<1y)

• Dog 29 y

• Chimpanzee 59 y

• Elefant 86 y

• Men 122 y

• Turtle 190 y

• Whale 211 y

• Bristlecone Pine 4838 y


Aging Postulate

• Limited cell divisions (stem cells)

telomer shortening

• Oxidative stress (caloric restriction)

fuel + oxygen energy to move a car

food + oxygen energy to move our body

oxygen ROS cell death


What are Stem Cells?

• Stem cells have two properties:

Self-renewal - the ability to go through numerous cycles of cell divisions in an undifferentiated state

Unlimited potency - the capability to differentiate into other cell types

• Toti-, pluri-, multi-, unipotent stem cells


Human: Embryonic Stem Cells (Pluripotent)

• Cells derived from the inner cell mass of an early stage embryo

• Blastocyst, 4-5 days post fertilization, 50-150 cells

• Pluripotent: Differentiate into one of the more than 220 different cell types of an adult body

• The use of embryonic stem cells in medicin is discussed contradictorily


Human: Adult Stem Cells (Multipotent)

• Undifferentiated cells among differentiated cells in a tissue or organ

• Generate all the cell types of the organ from which they originate — possibility to regenerate the entire organ from a few cells

• Function: Replenish dying cells and regenerate damaged tissues


2. Stem Cell Cosmetics


Epidermal Stem Cells

Stem cells are located in the basal layer where 2-7% of the cells are stem cells

Differentiating progeny of one epidermal stem cell

Epidermal stem cell

Transient amplifying cells

Basement membrane

Dermis

Basal layer

Spinous layer

Granular layer

Stratum corneum


Stem Cells and Aging

• Stem cells have a limited life expectation

• Environmental and intrinsic stress factors effect stem cells

• In aged skin the number and activity of stem cells is reduced

• Skin stem cells have to be protected and supported to delay preliminary aging


Epigenetic Factors Composition of Plant Stem Cells

• Plant stem cells contain similar epigenetic factors as human stem cells

• Plant stem cell extracts provide important nutrients to human stem cells

Plant stem cell nutrients improve the viability and healthiness of human stem cells


PhytoCellTecTM Malus Domestica 3 Stories

1. PhytoCellTec Technologie

2. Swiss Apple Story

3. Stem Cell Cosmetics


PhytoCellTecTM Malus Domestica Innovation Price

• BSB Innovation Price for most innovative natural active

• Received during incosmetics 2008

• Patent pending in CH, EU, USA and Korea


1. Plant Cell Cultures

• New cell culture technology makes it now possible to cultivate cells of endangered and rare species

• Selected plant material (leave, fruit) is wounded to induce the formation of a callus tissue

• The callus of a plant consists of dedifferentiated, totipotent cells (plant stem cells)


The 3 Steps of PhytoCellTec

1. Preparing a sterile plant preparation

2. Wounding the plant to induce callus tissue

3. Growing callus cells in liquid cultures


Plant Cell Culture Technology


Growth of Plant Cell Culture in a Reactor

0

2000

4000

6000

8000

10000

12000

0 5 10 15 20 25

days [d]

tota

l bio

mas

s [g

FS

]

0

5

10

15

20

25

30

35

40

tota

l su

gar

[g

/L],

co

nd

uct

ivit

y [m

S/c

m]

and

pH

-va

lue

Biomass sugar conductivity pH


PhytoCellTec

A Difficult and Tedious Process

• Preparation of sterile plants 2 months

• Callus induction 12 months

• Isolation of a stable stem cell line from callus 6 months

• Transfer into liquid media 3 months

• Transfer into bioreactors 3 months


2. Swiss Apple Story


Uttwiler Spätlauber

• An old cultivar is still alive in Switzerland but no longer cultivated

• The Spätlauber variety derives from the middle of the 18th century

• It was famous for its tremendous storability without shrinking

• Today this is a very rare and endangered apple variety with only a few trees left in the world


Uttwiler Spätlauber

The importance of the Uttwiler Spätlauber apple was honored by the release of a stamp.


Young and Old


Activity Assay for Epidermal Stem Cells

Stimulation of growth or over-expression of markers are not good parameters for the activity of stem cells

• Colony forming efficiency (CFE)

Cell cultures are seeded at low density. The number of colonies formed is a value for the concentration and activity of progenitor/stem cells

+ apple stem cell extract

Incubation

Determination of CFE


0

20

40

60

80

100

0.01% 0.04%

Concentration of Malus Domestica stem cell extract

Incr

ease

of t

he C

FE c

om

par

ed to

cont

rol i

n %

+ 92%

ResultNumber of active and healthy stem cells is increased compared to untreated culturesS-234

Increase of CFE by PhytoCellTecTM Malus Domestica


Capacity to Form 3D Epidermis

Study designComparison of 3D epidermis formed by early passage (young) and by late passage (old)epidermal progenitor cells.

Test substance• without test substance (control) • 0.01% PhytoCellTecTM Malus Domestica

S-244


Old cells (P14)(control)

Old cells (P14)+ PhytoCellTec MD

No cell growth No cell growth

Day 14 Day 21 Day 28

Young cells (P5)(control)

S-244


In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica

• Analysis of wrinkle depth in the crow‘s feet area• 20 volunteers • 28 days• Application twice daily • Against inital conditions


In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica

Day 0Day 282% PCT Malus Domestica

S-186


PhytoCellTecTM Malus Domestica Claims

• Delays senescence of essential cells

• Protects longevity of skin stem cells

• Protection of skin stem cells

• Rejuvenation of skin

• Maintains the power of regeneration

• Decrease in wrinkles

• Combats chronological aging

• With plant stem cells to maintain the power of regeneration

• With plant stem cells to delay senescence of essential skin cells


PhytoCellTecTM Solar VitisProtects epidermal stem cells against UV damage


PhytoCellTecTM - A new Technology to Produce Stem Cell Extracts

Stem cell culture from grapes A part of the flesh is selected

and cut


PhytoCellTecTM : Cell Culture Technology with Plant Stem Cells

Preparation of plant stem cells

• A part of a grape is selected and sterilized

• Induction of callus formation by cutting the plant

• Isolation of a stable stem cell line from the callus

• Incubation and proliferation on agar plates


UV Radiation and Skin Stem Cells

Epid

erm

isD

erm

is

Rad

iation

RO

S +

toxi

ns

Sun cream with UV-filters

Epidermal stem cell

Basal layer

Basement membrane

Reflection

Absorption

Keratinocyte PhytoCellTecTM Solar Vitis


Protection of Epidermal Stem Cells against UV Radiation

• Stimulation of CFE by 86% with PCT Solar Vitis

• 58% decrease of CFE after exposure to UV

• With PCT Solar Vitis stem cells were fully protected against negative effects of UV

S-245/©Mibelle Biochemistry 2009


PhytoCellTec™ Solar Vitis Protects against UV-Stress

Study design

• Determination of the individual MED in 20 volunteers (19 to 63 y).

• Application: cream with SPF 10 with and without 0.4% PhytoCellTec™ Solar Vitis.

• Irradiation with UV doses whose intensities were in the range of the MED multiplied by the SPF.

• 24 hours later, the redness of the skin was evaluated again.

PhytoCellTec™ Solar Vitis significantly increased the MED (MED+25% UV ~ SPF increase from 10 to 13).

S-268


PhytoCellTec™ Solar VitisClaim Ideas

• Protects skin stem cells against UV-stress

• Increases the skin‘s UV tolerance

• Fights photo-aging

• For a vital and healthy-looking skin


PhytoCellTecTM Alp RoseCharges skin stem cell resistance


The Swiss Alp RoseRhododendron ferrugineum L.

• In the Swiss Alps, the alpine rose is one of the most typical and prominent flowers

• Rhododendron (rhodon = rose, dendron = tree, ferrugo = rust)

• Ericaceae• 1500 – 2800 m • 1 m high, winter-green shrub, highly

branched• Pink, bell-shaped flowers• Egg-shaped, lanceolate leaves,

underneath with rust-colored scales


Production of PhytoCellTecTM Alp Rose

• Leaf of alpine rose is wounded to induce callus formation

• Callus cells are transferred to agar plates

• The developed callus with its stem cells is harvested and further cultivated on agar plates until complete dedifferentiation of the cells and generation of a homogenous culture


Viscous Circle Impairs Skin Barrier


Protection/Improvement of Skin Stem Cell Vitality with PhytoCellTecTM Alp Rose

Cell line: Epidermal stem cellsProduct: 0.15% Alp rose stem cell extractTreatment: Radiation with UVA+UVB at 450 kJ/m2 Analysis: Determination of colony forming efficacy (CFE)

Alp rose stem cell extract increases the vitality of epidermal stem cells and protects them against UV-induced stress.

S-245


Test products• Sun cream SPF 30

+ 0.4% PhytoCellTecTM Alp Rose• Placebo (sun cream SPF 30 without active)

Volunteers: 22 (20 – 52 y)

Application: 3 times daily (morning, noon, evening)

Test area: Face, half side test (blind, randomized)

PhytoCellTecTM Alp Rose Ski Holiday Study


Ski Holiday Study: Profile Photos (Visioface Quick®) Anti-Wrinkle Effect

D0 D17

S-2

86


Scoring:Visual scoring by dermatologists on day 0 and day 17

The skin side treated with PhytoCellTec™ Alp Rose showed an improvement of all 3 characteristics.

Ski Holiday Study: Visible Improvement of the Skin Quality

S-286


PhytoCellTecTM Alp Rose Claim Ideas

• Increases skin stem cell vitality

• Boosts epidermal regeneration

• Improves the skin barrier function

• Helps the skin to cope with climate changes


3 Different PhytoCellTecTM Products:Positioning


1. Use of plant stem cells to protect skin stem cellsThe components of plant stem cells such as from the apple, the grape and the alpine rose are rich in specific factors relevant to protect and maintain the function of skin stem cells

epigenetic factors

2. Use of plant specific factors for target activity “Uttwiler Spätlauber” apple > longevity “Gamay Teinturier Fréaux” grape > UV protection Swiss alpine roses > resistance

metabolites

PhytoCellTecTM ProductsProvide two Different Modes of Activity


3 Different PhytoCellTecTM ProductsKey Positioning

LongevityRegeneration

ResistanceBarrier function

UV-ProtectionPhoto-Aging

biocareer malaysia(dr.fred zuli)

Documents

y mibelle biochemistry

aging bad news

aging general considerations

plant stem cells

theory of evolution

nature evolution

cosmetic products

todays beauty trends