P Phyto Tell Pec Calus Tomestica (New Version)
Transcript of P Phyto Tell Pec Calus Tomestica (New Version)
© Mibelle Biochemistry, Switzerland 2009
PhytoCellTecTM Malus DomesticaPlant stem cells for skin stem cell protection
© Mibelle Biochemistry, Switzerland 2009
PhytoCellTecTM Malus Domestica3 Stories
1. PCT Technologie
2. Swiss Apple Story
3. Stem Cell Cosmetics
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PhytoCellTecTM Malus DomesticaInnovation Price
• BSB Innovation Price formost innovative natural active
• Received duringincosmetics 2008
• Patent pending in CH, EU, USA and Korea
© Mibelle Biochemistry, Switzerland 2009
1. Plant Cell Cultures
• New cell culture technology makes it now possible to cultivatecells of endangered and rare species
• Selected plant material (leave, fruit) is wounded to induce theformation of a callus tissue
• The callus of a plant consists of dedifferentiated, totipotentcells (plant stem cells)
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The 3 Steps of PhytoCellTec
1. Preparing a sterile plant preparation2. Wounding the plant to induce callus tissue3. Growing callus cells in liquid cultures
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Plant Cellculture Technology (PhytoCellTecTM =PCT)
• Suspension cultures from plant stem cells are grown in bioreactors
• Medium consists of sucrose, trace elements and plant hormones (cytokinins and auxins)
• Cultures grow with oxygen and without light (chlorophyll free)• Batch is harvested after all sugar has been metabolized• The cells are washed and homogenized to release the secondary
metabolites• Oil- and water-soluble ingredients are extracted by liposomal
structures
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2. Swiss Apple Story
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Storage Ability of Apples and Skin Aging
• Apples with very good storageabilities stay fresh over months
• This was an important factor forcultivar selection
• Modern apple cultivars are selectedfor intensive cultivation and for a sweet flavour since storage is no longer a problem
• Therefore cultivars with excellentstorage abilities disappeared fromthe market
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Uttwiler Spätlauber
• An old cultivar is still alive in Switzerland but no longercultivated
• The Spätlauber variety derivesfrom 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 withonly a few trees left in the world
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Uttwiler Spätlauber
The importance of the UttwilerSpätlauber apple was honoredby the release of a stamp.
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Young and Old
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The Secret of Storage Ability
• High acid and tannin concentrations• Longevity of cells• Undiscovered factors?
This apple can only be used by PCT
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PhytoCellTecTM Malus Domestica
CompositionMalus Domestica Stem Cells 9.0 %Phospholipids 0.14%Glycerin 0.4 %Phenoxyethanol 1.4 %Keltrol T 1.0 %Deionized water ad.100 %
CTFA/INCIMalus Domestica Fruit Cell Culture Extract (and) Xanthan Gum (and) Glycerin (and) Lecithin (and) Phenoxyethanol (and) Aqua/ Water
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3. Stem Cell Cosmetics
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What are Skin Stem Cells?
Properties of skin stem cells:
• Self-renewal - the ability to gothrough numerous cycles and celldivisions while maintaining theundifferentiated state
• Differentiation - the capacity to differentiate into other cell types of the same tissue
• Primary roles - replenish dying cellsand regenerate damaged tissues
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Adult Stem Cells in Skin
In skin, different types of stem cells have been found:
Type Place
Epithelial skin stem cells Basal layer of the epidermis
Hair bulge stem cells Hair follicle
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Epithelial 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
Epidermalstem cell
Transientamplifyingcells
BasementmembraneDermis
Basal layer
Spinous layer
Granular layer
Stratum corneum
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Hair follicle matrix
Epidermal layer
Sebaceous gland
Hair shaft
Inner root sheath
Outer root sheath
Basal layer
Dermal papilla
Hair bulgestem cells
Infundibulum
Lineages derived from bulge stem cells
Differentiation pathways supplied by bulge stem cells
Hair Bulge Stem Cells
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Stem Cells and Aging
• Stem cells have a limited life expection
• Environmental and intrinsicstress factors effect stem cells
• In aged skin the number and activity of stem cells is reduced
• Skin stem cells have to beprotected and supported to delay preliminary aging
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Epigenetic FactorsComposition 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
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PhytoCellTecTM Malus DomesticaProvides two Different Modes of Activity
1. Use of plant stem cells to protect skin stem cellsThe components of plant stem cells such as Malus domestica are rich in specific factors relevant to protect and maintain the function of skin stem cells
epigenetic factors
2. Use of longevity cell extract to delay skin agingThe apple Uttwiler Spätlauber is known to contain ingredients which insure longevity of cells
metabolites
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Protection of Stem Cells againstUV Damage – Study Design
Cell typeUmbilical cord blood stem cells
Study designIncubation for 24h with PhytoCellTec Malus Domestica Extract, UV radiation of cell cultures
Determination of living cellsMTS assay 48h after UV radiation
Control
+ 0.1% PhytoCellTecTM Extract
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Protection of Stem Cells againstUV Damage – Results
S-142
PhytoCellTec Malus Domestica improves thevitality of stem cells
Increased protectionagainst UV stress
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Novel Progenitor Cell Targeting Technology
• Cells: human epidermal stem cells (primary human keratinocytes) from a biopsie ca. 6% of cells are stem cells (optically not different from normal basal cells)
• Medium: specifically designed, fully defined medium that mimics the physiological environment of the in vivo stem cell niche
enrichment of so called keratinocyte progenitor cells (stem cells)
3x
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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
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0
20
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80
100
0.01% 0.04%
Concentration of Malus Domestica stem cell extract
Incr
ease
of t
he C
FE c
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%
+ 92%
ResultNumber of active and healthy stem cells is increased compared to untreated culturesS-234
Increase of CFE byPhytoCellTecTM Malus Domestica
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Result
PhytoCellTec apple stem cell extract enhanced CFE up to 92% compared to the control.
Maintenance of the stem cell vitality of epidermal stem cells was improved.
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Growth of Hair Follicle
Hair follicles From human skin fragments (from micro dissection)
Test substance PhytoCellTecTM Malus Domestica 0.2%
Design Length of hair follicle increases until day 14 → after day 14 induction of senescence→ length of hair follicle decreases
Day 0 Day 4 Day 7
S-157
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Length of Hair Follicle
-70
-60
-50
-40
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-20
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Day 16 Day 18 Day 20
Cha
nge
in le
ngth
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ay 1
4 in
%
Control 0.2% Malus Domestica stem cell extract
Senescence of hair (stem cells) can be delayed
S-157
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PhytoCellTecTM Malus DomesticaEffect on Gene Expression in SenescentDermal Fibroblasts
Cell line Normal Human Dermal Fibroblasts
Design • H2O2 induced senescence (2h)followed by a treatment with2% PhytoCellTecTM Malus Domestica or not (144 h)
Parameters • Gene profile (microarray)
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PhytoCellTecTM Malus DomesticaGene Profile (Microarray)
↑
↓ Gen, that is down-regulated by senescence induction (see control)
Gen, that is up-regulated or normalized in senescent cells by PhytoCellTec Malus Domestica
Gen After H2O2Control
After H2O2+ 2% Malus Domestica
stem cell extract
cyclin B1: induces proliferation 73 ↓ 130 ↑cyclin E1: cell cycle regulator 78 ↓ 135 ↑p53: tumor suppressor gene 63 ↓ 137 ↑insulin-like growth factor II: cell proliferation enhancer 71 ↓ 117 ↑
heme oxigenase 1: antioxidans enzyme 89 ↓ 211 ↑ S-194
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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
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In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica
0
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Day 14 Day 28
Dec
reas
e o
f wrin
kle
dep
th in
%
2% PhytoCellTec Malus Domestica
S-186
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In-Vivo Study: Anti-Wrinkle Effect of PhytoCellTecTM Malus Domestica
Day 0Day 282% PCT Malus Domestica
S-186
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PhytoCellTecTM Malus DomesticaApplications
• Stem cell cosmetics• Rejuvenation products for face and
body• Fresh cell treatments• Anti-Aging formulations• Rejuvenation of hair follicles• Delay of hair aging
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PhytoCellTecTM Malus DomesticaClaims
• 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
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PhytoCellTecTM Malus DomesticaPlant stem cells for skin stem cell protection
Patent pending