Hyalo-Oligo

Penetration type HA for Anti-aging

Penetrate into the skin and stay there with moisture

How Hyalo‐Oligo works in the skin

Effect on the gene expression and localization of Tight Junction related proteins like JAM

Strengthen Tight Junction

Moisturize, Anti-wrinkle, etc.

Hyalo-Oligo®

Common Hyaluronic acid

Size difference

Features of <Hyalo-Oligo®> （The differences from common hyaluronic acid）

Common Hyaluronic acid Hyalo-Oligo®

Granular layer

Horny layer

Published by Mandam in the 130th Nihon Yakugakkai

Penetration of Hyalo-oligo into the damaged skin model

Epidermis

Dermis

Hyalo-Oligo can partially penetrate into epidermis layer of the damaged skin

Feature． Skin moisture increase

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12

0 1 2 3

Hyalo-Oligo

CommonHyaluronic acid

before gauze day after days after days after

detach gauze detach gauze detach gauze detach

The gauze soaked in 1% solution of Hyalo-Oligo® was attached to the human

skin for 24 hours. （Control：1% solution of common hyaluronic acid）.After the

gauze was detached, the skin electric conductivity indicating the amount of

skin moisture was measured at three points ,namely 1 day after detaching

gauze(above 1), 2 days after(above 2), 3 days after (above 3).

(n =12)

Hyalo-Oligo® increases twice as much skin

moisture as common hyaluronic acid.

（Presented at Annual Meeting of the Pharmaceutical Society of Japan, March 2006 by Takushi

Yoshida Q.P.Corporation）

Ｒ

®

Increased

Am

ou

nt o

f Mo

isture(μ

S)

The functions of Tight Junction in the skin

１

２

Protect from an external stimulus

Barrier Function

Help the normal metabolism of the epidermis

Maintain the horny layer barrier

Horny Layer

Granular Layer

Tight

Junction Tight Junction controls

transfer of material

between the cells

First Barrier

Second Barrier

The main functions of Tight Junction in the skin

Human Epidermal Keratinocytes Add 1) Blank 2) Hyalo-Oligo® 0.001 μg/mL 3) Hyalo-Oligo® 0.01 μg/mL

6 hours later Extract RNA

Measure Skin related-gene expression 1. Aquaporin 2. Tight Junction

-Aquaporin-3 -Occludin -Claudin-1 -JAM-1

Aquaporin Localized to the cell membrane; controls the movement of water in and out of the cell.

Tight Junction Circles the cell in continuous band; critical for cell-cell connections; controls intercellular transit of liquid. Comprised of proteins such as Occludin, Claudin, and JAM.

Hyalo-Oligo® increases the expression level of

skin related genes

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1.4

1.6

対照 0.001μg/mL 0.01μg/mL

Aquaporin-3

Blank Blank 0

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1.2

1.4

1.6

Occludin

0.01 μg/mL 0.001 μg/mL

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0.2

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0.6

0.8

1

1.2

1.4

1.6

対照 0.001μg/mL 0.01μg/mL

Claudin-1

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6JAM-1

Blank 0.01 μg/mL 0.001 μg/mL Blank 0.01 μg/mL 0.001 μg/mL

Blank 0.01 μg/mL 0.001 μg/mL

＊＊ ＊＊

＊＊ ＊＊

＊

Significance: *, p < 0.1; **, p < 0.05

Hyalo-Oligo® increases the expression level of

skin related genes

Transfer cells to Hank medium to remove CaCl2 and culture in HuMedia KB2 medium containing 0.1 μg/mL Hyalo-Oligo® for 24 h.

Culture normal human epidermal keratinocytes in

HuMedia KB2 medium containing 1.6 mM CaCl2 for

24 h.

JAM-1 immunocytochemistry using a JAM antibody and

cell evaluation through confocal laser scanning

microscopy.

Wash the cells using phosphate buffered saline.

＜Test method＞

Hyalo-Oligo® increases JAM-1 protein expression and

promotes JAM-1 localization in epidermal cells

Control 0.1μg/mL Hyalo-Oligo®

Hyalo-Oligo® increases JAM-1 protein expression and

promotes its the localization in the epidermal cells

The damage of Tight Junction by UV

UV

Aging

Normal

horny layer

Disordered

horny layer

Wrinkle

Tight Junction is wasted away by

UV and aging....

Metabolism of the horny layer is disordered

→Water easily is evaporated from the skin

Tight Junction barrier is damaged

→Foreign materials easily enter the granular layer

Transfer cells to Hank medium to remove CaCl2 and then expose cells to UV-B light.

Culture normal human epidermal keratinocytes in

HuMedia KB2 medium containing 1.6 mM CaCl2 for

24 h.

JAM-1 immunocytochemistry with a JAM antibody

and cell evaluation using confocal laser scanning

microscopy.

＜Test method＞

Hyalo-Oligo® improves

the damage to JAM-1 caused by UV

Culture the cells in HuMedia-KB2

medium containing 1 μg/mL Hyalo-

Oligo® for 24 h.

Wash the cells with PBS.

Hyalo-Oligo® improves

the damage to JAM-1 caused by UV

Control 0.1μg/mL Hyalo-Oligo®

After exposure to UV-B

Before exposure to UV-B

<Sample

solutions>

1) Water

2) Hyalo-OligoⓇ

1% solution

Test Overview

• Inclusion Criterion: Wrinkle Grade：2.5 (Visual Evaluation/One side)

• Subjects: Females (N = 8), >45 years old

• Test Period: 8 weeks

• Procedure: Apply each sample solution (1 and 2) at morning and night

• Test measurements:

• 1) Analysis of replica (mean depth of a wrinkle)

• 2) Water content in stratum corneum (cheek)

• Equipment used: Corneometer

（２） 4cm

1 cm 4cm

※Consigned testing laboratory: Inforward Inc. Ebisu Skin Research Center

（Replica of Assessment Area)

＜Test method＞

Hyalo-Oligo® can reduce wrinkles

Water Hyalo-Oligo® 1% Solution

45-year-old female

Hyalo-Oligo® can reduce wrinkles 0

We

eks

8

We

eks

Water Hyalo-Oligo®

1% solution

＊：p<0.05

＊

Water Hyalo-Oligo®

1% solution

Rat

e o

f C

han

ge in

Incr

eas

ed

Mo

istu

re (

%)

Rat

e o

f C

han

ge in

Wri

nkl

e D

ep

th (

%)

Mean wrinkle depth Water content in

stratum corneum

(cheek)

Hyalo-Oligo® can reduce wrinkles

Hyalo-Oligo® penetrates

into the skin

Hyalo-Oligo® increases TJ-related proteins (JAM)

and promotes the localization of TJ proteins

Strengthens Tight Junctions and

improves the damage of the skin

Hyalo-Oligo® = Anti Photo-aging

How Hyalo‐Oligo® works in the skin

Wrinkle

UV

Aging

Water is easily evaporated

from the skin

Foreign materials easily enter

the granular layer

Tight Junctions decrease and

loosen due to aging and UV.

Before

After

Penetrate into the skin and stay there with moisture

How Hyalo‐Oligo works in the skin

Effect on the gene expression and localization of Tight Junction related proteins like JAM

Strengthen Tight Junction

Moisturize, Anti-wrinkle, etc.