SymHair™ Force 1631 PDF

SymHairTM Force 1631

MICROALGAE

NATURAL STRENGTH

GREATER HAIR

The information set forth herein does include proprietary and

confidential information of Symrise and must be treated in confidence

and not be used by the recipient hereof unless otherwise agreed to in

writing by Symrise.

PAGE 3

AGENDA

Identifying key consumer needs

Hair growth disorders

Hair loss causes & reasons

PRESENTATION APPENDIX

HAIR LOSS - CONSUMERS & MARKETS

SYMRISE‘S SOLUTIONS

Screening strategy & mechanism of action

In vivo objectivation

FOCUS ON SCIENCE

Human hair figures & structure

Hair growth cycle

Hair loss main reasons & female pattern

Symrise green technology

Microalgae biotechnology

FROM CONSUMERS NEEDS to MICROALGAE

PAGE 4

HAIR LOSSCONSUMERS & MARKETS

PAGE 5

CONSUMER INSIGHTSHAIR LOSS - ALOPECIA

Hair:

Socio-cultural phenomenon of great significance in every civilization

Important beauty criterion or sacred character

Hair loss or Alopecia:

Loss of self-esteem and self-confidence usually associated with healthy looking

Psychosocial impact influencing thepsychological well-being

PAGE 6

THE IMPORTANCE OF BEAUTYHEALTHY HAIR, NATURAL LOOK

Consumers agree that your hair is a defining characteristic of who you are

Two consumers out of three around the world are proud of their hair

What consumers want the most is healthy hair that look naturally beautiful

Healthy is the most popular quality they want

their hair to reflect (47% globally)*

Natural comes second (36% globally)*

They want to sustain their hair health and vitality

*Source: Symrise LE CMI global database 2012

PAGE 7

BEAUTY HAS ITS SEASONSPREPARING HAIR IS CAREFUL

Consumers care about their hair

31% use hair treatments & balms at least once

a week globally*

Many consumers know seasons and climate can affect the life of their hair

Temperature change can impact hair and scalp

Humidity as well

The wind can also generate or increase

problems, as well as carry pollution

The idea of preparing in advance for climate and seasonal changes is well-accepted

Many consumers believe that good products

need time to work

PAGE 8

HAIR STRENGTH AND VITALITYCONSUMERS ARE READY TO PAY

Thinning, lack of volume and/or hair loss are key problems consumers want solutions for

36% of consumers are ready to pay the

most they can afford for solutions that

work against hair loss*

This can be for different reasons

Beauty, health and vitality, strength

This can be at all ages

From young people who want to look

attractive and at their best to older ones

who are more concerned with aging

*Source: Symrise LE CMI global database 2012

PAGE 9

Affects males and females at any age

Higher prevalence for men

Androgenic alopecia :95% of all hair loss

Hair loss can be permanent or temporary

?

HAIR GROWTH DISORDERSAT THE HEART OF CONSUMER’S LIFE

PAGE 10

HAIR GROWTH DISORDERS HAIR LOSS CAUSES & REASONS

Causes:

Imbalance of the hair growth cycle

Genetic disposition

Natural aging process and / or disease

Anagen(2-6 years)

Catagen(4-6 weeks)

Telogen (2-3 months)

Exogen

Hair growth cycle of a human

scalp hair follicle

Reasons:

Hormonal (DHT)

Structural deficiency of hair roots

Inflammation and/or deficient scalp irrigation

Human scalp hair growth:

Anagen: active hair growth (2-6 years)

Catagen: regression (4-6 weeks)

Telogen: resting (2-3 months)

Exogen: release of dead hair at the end of telogen

PAGE 11

FROM CONSUMERS NEEDSTO MICROALGAE

PAGE 12

FROM CONSUMER NEEDS E.E TO MICROALGAE

Strong demand for innovative and natural solutions based on clearly identified mechanism of action

New approach to address disorders of hair growth


Natural active ingredient from micro algaeto prevent hair loss

Natural strength and greater hair

PAGE 13

ALGAE GENERAL

Algaeprokaryoteseukaryotes

pluricellular unicellular

CyanobacteriaMacroalgae Microalgae

PAGE 14

SYMRISE GREEN-TECHNOLOGYMICROALGAE BIOTECHNOLOGY

Screening and development of a new natural biological hair loss prevention from microalgae

Cutech Srl

Expertise in ex-vivo skin and ex-vivo hair follicle screening

Contact to experts in microalgae cultivation and production

Why microalgae?

Sustainable manufacturing process from renewable sources

From 25-40 thousand registered microalgae species only few are used commercially so far

Microalgal biotechnology is an already used technology to produce e.g. vitamins, carotenoids, polyunsaturated fatty acids, proteins, cosmetics and health foods

PAGE 15

Cyclic peptidese.g. Hassallidin A

MICROALGAE COMPOSITION

Huge biodiversity: Interesting source for common and rare compounds potentially active on human tissue

O

OH

O

OH

Carotenoidse.g. astaxanthin

NH

ON

CO2H

NHN

H

O

HO2C

Other pigmentse.g. phycocyanin

CO2H

Polyunsaturated fatty acidse.g. docosahexaenoic acid (DHA)

Proteins

PolysaccharidesMinerals

Amino acids

O

Alkenones e.g. heptatriaconta-8E,15E,22E-trien-2-one

PAGE 16

In the course of our extensive screening, we identified Isochrysis sp. var. Tahitian (T-ISO) Marine water microalgae

For decades successfully exploited in aquaculture for supporting the larvae rearing of delicate marine fish

Live food particularly rich in highly unsaturated fatty acids and

other micronutrients : T-Iso is rich in vit. PP and vit. B61

Indication of low toxicity

Known cultivability in aquaculture2

Tahitian Isochrysis is a strain of Isochrysis collectable at Mataiva (Tahiti). This strain has been isolated by K. Haines in 1977 at Mataiva, Tahiti.

Ideal candidate for producing high value active ingredients

1 De Roeck-Holtzhauer Y. et al., 1991, Journal of Applied Phycology, 3: 259-264

2 Reitan K.I. et al., 1997, Aquaculture 155: 207-221

MICROALGAE SELECTION

PAGE 17

MICROALGAE CULTIVATION OPTIONS

Depending on the specific strain, microalgae can be cultivated in either open ponds or closed photobioreactors

-Allows to produce large

quantities of microalgae at

low costs

-High risk of

contamination by other

species or protozoa

-Instable, uncontrolled

environmental conditions

-Only few algal species

are suitable to be cultured

using this technology

-Example: Spirulina

platensis (production of

3000 t/a)1

OP

EN

PO

ND

S

-Requires higher

investment and operating

costs

-Low risk of contamination

by other species

-Allows controlling of

environmental conditions

and stable production

-Possibility to cultivate a

large variety of microalgae

strains

-Example: Chlorella

vulgaris / beta-Glucan

(production of 1.5 t/a)1

CL

OS

ED

PH

OT

OB

IOR

EA

CT

OR

S

1 C. Griehl and S. Bieler, Nachr. Chem., 2011, 59, 942-947

Image source:

http://www.cyanotech.com/company.html

PAGE 18

MICROALGAE BIOMASS PRODUCTION

- F&M: Fotosintetica & Microbiologica, Professor M. Tredici, GWP (Green Wall Panel)

- Archimede Ricerche, Dr Silvio Mangini

Microalage supplied by Archimede Ricerche Plant

PAGE 19

Microalgae

inoculumSun CO2

Artificial sea

water

Microelements

Vitamins

Mineral

nutrients

Microalgae biomass Drying Extraction

MICROALGAE BIOMASS PRODUCTION

PAGE 20

A NEW, HIGHLY POTENT

HAIR LOSS PREVENTION ACTIVE

FROM MICROALGAE

SYMRISE‘S SOLUTIONS


PAGE 21

SYMRISE’S SCREENING MODELSHUMAN HAIR FOLLICLES

Screening strategy for hair loss prevention active:

Hair shaft elongation

ex vivo organ culture of human hair follicle

ex vivo human scalp culture

Mechansim of action

ex vivo organ culture of human hair follicle

Objectivation in vivo

PAGE 22

SCREENINGEX VIVO HUMAN HAIR FOLLICLES

Hair shaft elongation at day 9

Test protocol

Human micro-dissected scalp hair follicles obtained from plastic surgery were transferred into 24 well culture plates

After18 h of pre-incubation, hair follicles suitable for testing were selected(good vital stage and a growth of not less than 0.2 mm)

12-18 hair follicles per concentration and control were used

Test compounds were applied on day 1 and reapplied every second day*

On day 9 (after 8 days of treatment), hair follicles were photographed and the elongation of each hair follicles was measured

* Preparation of supplemented media by addition of DMSO dissolved test compounds into the standard culture medium

Micro-dissected

human hair follicles

PAGE 23


Test protocol schematically

Day 0 1 2 3 4 5 6 7 8 9

Pre-

incu-

bation

Treatment (8 days)

Test compound and medium renewal using the prepared supplemented media

(= medium with or without test compounds)

Selection of suitable

hair follicles

Photography and analysis

of growth performance

PAGE 24

Positive controls / benchmarks in the hair shaft elongation assay

1. Philpott et al, 1994, J Invest Dermatol

2. Taylor et al, 1993, J Invest Dermatol

3. Foitzik et al., 2007, Exp Dermatol

Micro-dissected

human hair follicles

CompoundEffect in the hair shaft

elongation assay (hHFs)Literature

Insulin Stimulation Stimulation1

Cyclosporine A Stimulation Stimulation2

Minoxidil no effect Contradictory results

Finasteride no effect Contradictory results

Carnitine tartrate*** Stimulation Stimulation3

*

* Blood cirulation enhancer

** 5-alpha reductase inhibitor

*** patented (Henkel)

**


PAGE 25

BIO1631 identified as highly potent candidate

Average hair growth stimulation

(based on responding donors)

at 0.004 ppm: +13%

Cyclosporin A (0.2 ppm): +10%

Insulin (20 ppm): +9%

Carnitine tartrate (0.2 ppm): +11%

Effective concentration in vitro:

0.004 - 0.4 ppm

Responsiveness: 73%

Number of tested donors: 15

(11 responders)

Hair shaft elongation in vitro of BIO1631 after 8 days of treatment

concentration [ppm]

+7%

+13%

+8% +10%+11%

0

4

8

12

16

0.004 0.04 0.4

BIO1631

0.2

Carnitine

tartrate

Ave

rag

e h

air

gro

wth

stim

ula

tion

[%

]

ba

se

d o

n re

sp

on

din

g d

on

ors

+9%

0.2

Cyclosporin

A

20

Insulin

*

*

*

* p < 0.01


PAGE 26

PROOF OF CONCEPTEX VIVO HUMAN SCALP MODEL

Hair shaft elongation at day 6

Test protocol

Human scalp skin samples obtained from plastic surgery were transferred into culture plates

Pictures of the skin samples were taken on day 0, 3 and 6 and each hair shaft was labeled and measured at day 0, 3 and 6

BIO1631 formulated in a tonic or placebo was once daily applied topically on the skin samples (3 samples per treatment)

The elongation of each growing hair follicle was obtained by calculating the difference in hair shaft length between day 6 and day 0

Hair follicles that did grow less than 50 µM in 3 days were considered to be dead and were not considered Human scalp

culture

PAGE 27

PROOF OF CONCEPTEX VIVO HUMAN SCALP MODEL

Results

BIO1631 confirmed as excellent hair growth stimulator on ex vivo human scalp skin

Formulation: aqueous alcoholic

tonic

Number of experiments /

donors: 2

Mean stimulation versus

placebo from both experiments:

+46% at 40 ppm BIO1631

+12% at 4 ppm BIO1631 Data from

1 experiment

with

representative

pictures

Control 4 ppm 40 ppm BIO1631

PAGE 28

MECHANISTIC INSIGHTS BIO1631

PAGE 29

ANTI-HAIR LOSSPROBLEM, CAUSE, SOLUTION

Anagen

Catagen

Telogen

Exogen

Anagen

Catagen

Telogen

Exogen

Hair

cycle

Aging Spontaneous hair loss Loss of switch telogen / anagen

Solution:

Prolongation of the anagen phase

Blocking of transition into catagen

Stress Stress-induced hair loss Premature entry into catagen

PAGE 30

MECHANISM OF ACTIONHAIR CYCLE STAGING

Test protocol


After 1 day of pre-incubation, hair follicles suitable for testing were selected

12 hair follicles per concentration and control were used

Test compounds were applied on day 1 and 3*

After 5 days of organ culture, hair cycle staging was performed by morphological analysis of the bulb region stained with haematoxylin-eosin

Extrapolation of the hair cycle stage of the examined hair follicle on the basis of the accepted morphologic criteria after 9 days of culture (Müller-Röver et al., 2001, J. Invest. Dermatol.)


PAGE 31

MECHANISM OF ACTIONHAIR CYCLE STAGING

Results

At 0.04 ppm BIO1631 increases the percentage of hair follicles in the anagenphase versus control by 27%

Hair cycle

staging by

morphologic

analysis at

day 5 of hair

follicle organ

culture

55%

45%

70%

30%

Control 0.04 ppm BIO1631

+27%

-33%

PAGE 32

MECHANISM OF ACTIONPROLIFERATION / APOPTOSIS

Test protocol


After 1 day of pre-incubation, hair follicles suitable for testing were selected

12 hair follicles per concentration and control were used

Test compounds were applied on day 1*

After 3 days of organ culture, TUNEL / Ki67 / DAPI triple immunostaining was performed

Quantitative investigation of the effect of the tested compounds on the induction of apoptosis and/or proliferation of the cell populations localized on the bulb region

TUNEL and Ki67 label apoptotic and proliferating cell nuclei, respectively

DAPI, staining all the cell nuclei, is used as a control


PAGE 33


Results:

BIO1631 significantly decreases the number of apoptotic cells by 59% and 63% versus control at 0.04 and 0.4 ppm, respectively

Ratio of

proliferation /

apoptosis at

day 3 of hair

follicle organ

culture

Control 0.04 ppm BIO1631 0.4 ppm BIO1631

p < 0.05

PAGE 34


Results II

Visable decrease in apoptotic cells (green) and increase in proliferating cells (red) versus control

DAPI (used as control): all nuclei in blue

Representative

pictures

Proliferation /

apoptosis at

day 3 of hair

follicle organ

culture

Control 0.04 ppm BIO1631 0.4 ppm BIO1631

PAGE 35

OBJECTIVATION IN VIVO HAIR LOSS PREVENTION

PAGE 36

HAIR LOSS PREVENTIONCLINICAL STUDY

Protocol:

30 male volunteers (age: 18 to 60 years) suffering from alopecia in stages II-IV of the Norwood-Hamilton scale divided into 2 groups of 15 subjects each

1 group applied the active tonic while the other group applied the placebo

Active tonic: 4% SymHairTM Force 1631 (= 160 ppm BIO1631) in water 30% and ethanol 66%, placebo: water 34% and ethanol 66%

Application mode: once daily approx. 2 ml on the whole scalp for 3 months

Reading: Start (T0) and after 3months

Assessment criterion:

Instrumental phototrichogram analysis: number of anagen and telogen hairs

Comparison of a photograph of a defined area of the scalp directly after hair clipping with a photograph taken after a certain period of time long enough to allow hair growth of the clipped segment (48 hours).

Grown hairs = in anagen phase , non-grown hairs = in telogen phase

Subjective analysis: Subject’s evaluation of the efficacy of the treatment by filling in a specific questionnaire

Clinical analysis: dermatologist judgement of the tolerability of the treatment and of the degree of dandruff, seborrhoea, erythema, itching and burning

PAGE 37

baseline Day1 Month1 Month2 Month3

Selection

of

subjects

2 x 15

Treatment (3 months)

Daily application of Hair Tonic with 4% SymHairTM Force 1631 group 1

versus placebo group 2

First application

Day 1

IN-VIVO CLINICAL STUDYPROTOCOL SCHEMATICALLY

Last application

Day 84- Phototrichrogram analysis

- Subjective evaluation

- Clinical analysis

PAGE 38

Instrumental results Phototrichogram analysis

After 3 months of daily treatment with an aqueous ethanolic tonic containing 4% SymHairTM Force 1631

+10.5% !! increase of anagen hairs versus placebo

4.6% enhancement of the number of anagen hair

versus start (D0)

Placebo: 5.9% decrease in anagen hair

versus start (D0)

11.7% !! decrease of telogen hairs versus placebo

15.1% increase in telogen hair versus start (D0)

for the placebo compared to only 3.4% increase

for the tonic containing the active

-8

-6

-4

-2

0

2

4

6

Placebo Active

Vari

ati

on

T3m

on

ths -

T0 [

%]

Anagen hair

+10.5% vs

placebo

-5.9%

+4.6%

0

2

4

6

8

10

12

14

16

Placebo Active

Vari

ati

on

T3m

on

ths -

T0 [

%]

Telogen hair

-11.7% vs

placebo

+15.1%

+3.4%


Subject 3

T0 T12wks

PAGE 39

Subject’s evaluation by questionnaire analysis

After 3 months treatment

80% of the subjects who applied the

active stated that they perceived

Anti hair loss efficacy

Stronger and more vital hair

Thicker/more voluminous hair


Dermatologist’s evaluation:

Very good tolerance after 3 months of

treatment

0

10

20

30

40

50

60

70

80

90

Anti-hair lossefficacy

Stronger andmore vital hair

Thicker/morevoluminous

hair

Pe

rcen

tag

e o

f s

ati

sfa

cti

on

[%

]

80% 80% 80%

67% 67% 66%

Placebo

Active

PAGE 40

SUMMARYEFFICACY

Discovery of BIO1631, a new, natural, highly potent anti-hair loss active

13% stimulation of hair growth versus untreated control at 0.004 ppm

after 8 days of treatment.

Successful proof of concept on ex vivo scalp skin

(12 and 42% hair growth stimulation at 4 and 40 ppm, respectively).

Decrease of the number of apoptotic cells in the hair bulb

and by that increase of the number of hair follicles in the anagen phase

were identified as mode of action

Proven in vivo efficacy with +10.5% increase anagen hairs !!

and -11.7% telogen hairs vs placebo !!

�� In 3 months, hair appear stronger, more vital, more dense !!

PAGE 41

SymHairTM FORCE 1631THE DIFFERENCE & USP

Natural active ingredient from

microalgae to prevent hair loss

Natural extract from a sustainable and

renewable source

Cost effective

Dual action benefits:

Strengthens and thickens hair

Prevents thinning and hair loss

Clinically shown to increase hair density

(*vs. placebo)

+10.5% increase of anagen hair

11.7% decrease of telogen hair

Prevents hair loss

Thicker and more luscious hair

Improves volume

Stronger, healthier more vital hair

MA

RK

ET

ING

Natural hair loss prevention

Naturally

Increases of anagen hair

Decreases of telogen hair

Mode of action identified :

Decreases significantly the

number of apoptotic cells in

the hair bulb

Increases the number of hair

follicles in the anagen phase

Easy to formulate (soluble in water,

alcohol and glycols)

Patent pending

Preservative freeT

EC

HN

ICA

L

PAGE 42

SymHair® Force 1631 SUMMARY & CONCLUSION

■ Natural hair loss prevention from microalgae

■ Sustainable manufacturing process from renewable sources

■ Blend of BIO1631 in pentylene glycol

■ INCI : Pentylene Glycol, Isochrysis Galbana Extract

■ Recommended dosage: 4%

■ Clear to opal black green liquid

■ Water soluble

■ Easy to formulate but has to be protected from light

■ Light sensitive and colored product due to contained chlorophylls and carotenoids

■ PN #510316

PAGE 43

HAIR SCIENCEBIOLOGY OF HAIR GROWTH

PAGE 44

HUMAN HAIR FIGURES

Some facts about hair

The full head of hair consists of 120 000 to 150 000 hairs

The hair density per cm2 of the scalp is 250 in average

The speed at which hair grows is 1 cm per month which corresponds to

0.3 to 0.5 mm per day

12 cm pear year

The number of hairs that we naturally lose each day is between 50 and 100

Keratin is the main constituent of hair

Hair is composed of 50% C, 20% O, 17% N, 6% H and 5% S

PAGE 45

HUMAN HAIR STRUCTURE

Hair has two separate structures - the follicle in the skin and the shaft that we see

Together with the sebaceous gland and thearrector pili muscle, the hair follicle is part of thepilosebaceous unit

The hair folicle is a complex mini-organ of itsown

Hair growth results from the proliferative activity of matrix keratinocytes which give rise to the hair shaft and the inner root sheath

Nutrition of the papilla and the overlying matrix cells is provided via the capillary loop located within the dermal papilla

PAGE 46

HUMAN HAIRGROWTH CYCLE

Human hair growth

Scalp hair grows in cycles

Each cycle consists of 3 phases

Anagen: active hair growth (2-6 years)

Catagen: regression (4-6 weeks)

Telogen: resting (2-3 months)

Exogen: release of dead hair at the

end of telogen

Typically, 90-85% of scalp hair are in anagen and approx. 10-15% in telogen

Normally each hair follicle cycles independently,

Thus, the total number of scalp hairs

remains stable Anagen

Catagen

Telogen

Exogen

Hair growth cycle of a human

scalp hair follicle

PAGE 47

Hair Growth and Disorders


PAGE 48


Each hair follicle undergoes 10 to 30 hair growth cycles per lifetime

The cyclic transformations are controlled by finely tuned changes in the local signaling milieu, based on change in the expression of

cytokines

hormones

neurotransmitters

their receptors

transcriptional factors

enzymes

which act via endocrine, paracrine and autocrine routes

Imbalances lead to hair growth disorders

PAGE 49

A DYSFUNCTION: HAIR LOSS3 MAIN REASONS

Structural deficiency of hair roots: weak implementation � effect of loss of extracellular matrix (ECM)

Inflammation & deficient scalp irrigation

Androgenic alopecia

Hormonal:Testosterone

5 α reductase

Dihydrotestosterone (DHT)

Miniaturization of follicles & regression of hair roots

Hair thinning / shorter, finer hairs / baldness

PAGE 50

A DYSFUNCTION : HAIR LOSSFEMALE PATTERN

Female pattern hair loss (FPHL) is also an androgenic alopecia it is the most common type of alopecia : 20% of women in their 50’s have alopecia.

The early onset is similar to male pattern hair loss while the late onset appears with menopause.

Causes :

Androgens :

Dehydroepiandrosterone sulfate (DHEA-S) is converted

into DHT

This mechanism requires 3 enzymes (17β-hydrosteroid

dehydrogenase, 5α-reductase and 3β-hydroxysteroid

deshydrogenase.

Most of the FPHL are androgen-dependent, patients

treated with anti-androgens or 5α-reductase inhibitors

show increased hair growth.

Hair Growth and Disorders

PAGE 51

Estrogens :

Hair follicles have estrogen receptors

(ERα and ERβ)

The precursor Andrestenedione requires 3

enzymes : the 17-hydroxysteroid

deshydrogenase, Aromase and the 5α-

reductase, to be converted in DHT.

How estrogens influence FPHL has not been

determined yet.

Genetics :

This is a polygenic disorder but the genes

involved have not been found.

Others :

Blood circulation

Stress, chemotherapy, malnutritionV

Endocrine reviews.

A DYSFUNCTION : HAIR LOSSFEMALE PATTERN

PAGE 52

SymHairTM FORCE 1631NATURAL STRENGTH for GREATER HAIR

SUMMARY

Thinning hair and hair growth disorders are a wide spread global issue

that can affect both sexes at any age

Anti hair loss actives are proven agents for preventing, slowing or treating

hair loss.

Discovery of BIO1631, a new, natural, highly potent anti-hair loss active

13% stimulation of hair growth versus untreated control at 0.004 ppm

after 8 days of treatment.

Successful proof of concept on ex vivo scalp skin

(12 and 42% hair growth stimulation at 4 and 40 ppm, respectively).

Decrease of the number of apoptotic cells in the hair bulb

and by that increase of the number of hair follicles in the anagen phase

were identified as mode of action

Proven in vivo efficacy with +10.5% increase anagen hairs !!

and -11.7% telogen hairs vs placebo !!

�� In 3 months, hair appear stronger, more vital, more dense !!

PAGE 53

SYMRISE,

ALWAYS

INSPIRING

MOREE

DISCLAIMERSymrise makes no warranties, either express or implied, as to the accuracy or completeness of the information set forth

herein. Symrise expressly disclaims any implied warranty of merchantability and fitness for a particular purpose. Prospective

users are requested to determine for themselves the suitability of Symrise materials and suggestions for any use prior to their

utilization. Any necessary approvals from regulatory authorities for finished products must be obtained by the prospective

user. Suggestions for applications involving our products or the reference to, or incorporation of, descriptive materials from

patents and the citation of specific patents in this document may not be understood as recommendation for the use of

Symrise products in violation of any patent or as a permission or licence to use any patent of Symrise or a third party.

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SymHair™ Force 1631 PDF