IFSCC_Magazine_0121_low.pdf

MAGAZINE
The Scientific Publication of the International Federation
of Societies of Cosmetic Chemists
Volume 24 | Number 1 | March 2021
New Horizon in Skincare Targeting the
Facial-Morphology-Retaining Dermal “Dynamic Belt”
A Perfusable Vascularized Full-Thickness Skin Model
for Topical and Systemic Applications
Do-It-Yourself Cosmetics – The Pleasure of Creating Your Own Emulsions
Evolution of Antiperspirants and Deodorants
supported by

2 IFSCC Magazine 1 | 2021
© 2021 International Federation of Societies of Cosmetic Chemists
SCIENTIFIC PAPERS
Editorial�� 3
Biographies�� 4
New Horizon in Skincare Targeting the Facial-Morphology-Retaining Dermal “Dynamic Belt”
Tomonobu Ezure, Satoshi Amano, Kyoichi Matsuzaki, Nobuhiko Ohno (Japan) ��5
A Perfusable Vascularized Full-Thickness Skin Model for Topical and Systemic Applications
Sacha Salameha, Nicolas Tissot, Kevin Cache, Joaquim Lima, Itaru Suzuki, Paulo André Marinho,
Maité Rielland, Jérémie Soeur, Shoji Takeuchi, Stéphane Germain, Lionel Breton (France, Japan) ��11
Do-It-Yourself Cosmetics – The Pleasure of Creating Your Own Emulsions
Megumi Kaji, Tomoyuki Iwanaga, Yuichiro Takeyama, Kazuki Matsuo, Toshihiro Arai,
Kenichi Sakai, Hideki Sakai (Japan) ��17
Evolution of Antiperspirants and Deodorants
Fujihiro Kanda (Japan) ��27
Abstracts of Papers published in the JSCCJ, Volume 55, No� 2, 2021 ��35
Author‘s Guidelines ��40
AD INDEX
IFSCC Conference 2021��10 IFSCC Congress 2022 ��16
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EDITORIAL
IFSCC Magazine 1 | 2021 3
Dear readers, dear friends,
One of the traditional values of the IFSCC is that not only does the Praesidium usually have a balanced composition but also the
expectations and needs of our members from the different zones are perceived. The introduction of a co-editing team is another
step in this direction. In this way, the IFSCC Praesidium thinks it can better address members and scientists from all zones and
promote international exchange.
We already mentioned in the last issue the handover of the pen to Dr. Kazutami Sakamoto, Ph.D., (Japan) and Dr. Giulio Pirotta (Italy).
Let’s welcome them heartily! Accordingly, please address in the future all contributions for Zone 1 (Europe, Africa, and Israel) and
Zone 3 (the Americas) to Dr. Giulio Pirotta and for Zone 2 (Asia, including Australia and New Zealand, and the Middle East) to
Dr. Kazutami Sakamoto.
Both gentlemen will have the opportunity to present themselves and their huge ideas and prospects in this small but beautiful issue.
You will already recognize their handwriting with the exciting review article by our highly esteemed Dr. Fuji Kanda (Japan).
And we are proud to finally present to you two award papers and complete the series of awardees 2020.
Once more we want to invite you as future authors to share your insights and knowledge with us.
Please take a look at our recently revised Authors’ Guidelines found at the end of this issue.
Let’s go to new shores!
Petra Huber
Chair of publication IFSCC
Our mission to move forward
It is our honor to serve the IFSCC Magazine as co-editors. Although it took over three months after the last publication to deliver the
first issue of 2021, we learned a lot about our mission and are now ready to move forward.
We appreciate our predecessors, especially Dr. Claudie Willemin, for their dedication to making the Magazine a fixed value among
the abundance of information on cosmetic science and technology. However, it was not easy to smoothly take over the role under
the current disruptive circumstances. We thank all the support we received from authors, reviewers and colleagues in the publishing
team: Petra as chair, Marcia as language manager and Bernhard as layout manager.
Our mission is to make the IFSCC Magazine be the first journal for members to look at to find the appropriate information they are
seeking, and tasks we found during the preparation period to achieve this goal are:
1) Define the precise scope of the Magazine.
2) Upgrade the editorial quality.
3) Update the manuscript management system.
4) Activate a Scientific Editorial Board.
5) Strengthen collaboration with other committees and member societies.
We hope that you, as a reader of the Magazine, will discover some of the preliminary trials for these tasks. One example is the review
by our IFSCC Fellow Dr. Fujihiro Kanda based on his webinar interview. The idea for this new type of review was inspired by the sug-
gestion given to us by one of the fellows: “There is a real need for a review in cosmetic science for current awareness and a balanced
opinion to fill a void at this time of change in the cosmetics industry.“ Another new entry is the abstracts of the J. Soc. Cosmet. Chem.
Jpn. provided by the Society of Cosmetic Chemists of Japan.
We have just started to move forward and welcome your feedback so we can make our Magazine better together.
Sincerely yours,
Dr. Giulio Pirotta
Dr. Kazutami Sakamoto
Co-Editors IFSCC

4IFSCC Magazine 1 | 2021
BIOGRAPHIES
Dr. Kazutami Sakamoto
Dr. Kazutami Sakamoto is currently Guest Professor of the Department of Pure and Applied Chemistry,
Faculty of Science and Technology, Institute for Colloid and Interface Science Research Center for Space
Colony at the Tokyo University of Science, Japan. He is a former professor of the Graduate School of
Pharmacy at Chiba Institute of Science for Cosmetic Science. Dr. Sakamoto received his Ph. D. from the
Faculty of Science at Tohoku University in 1980 in Japan. He has extensive R D experience in the indus-
try (from 1971 to 2008 with Ajinomoto, Shiseido, and Seiwa Kasei) and in academia since 2008. He has
elucidated pioneering research on the physicochemical properties of amino acid based chiral surfactants,
chiral mesoporous silica and cosmetic science for new material development and substantiations.
Dr. Sakamoto received the 39th Scientific Award from the Japan Oil Chemists’ Society in 2005. He was
Chairman of the Division of Colloid and Surface Chemistry at The Chemical Society of Japan for 2007 - 2008
and a fellow of The Chemical Society of Japan. Since 2018 he has served ISO as a chairman of ISO / TC91
Surface Active Agents.
For the IFSCC, he was an Awards Committee member for IFSCC Congresses, presented a plenary lecture at the 27th IFSCC Congress
(2012), was an EMCEP Philippines Speaker (2018), and is currently co-editor of the IFSCC Magazine (from 2021).
Dr. Giulio Pirotta
Born in northern Italy near the Alps, Dr. Giulio Pirotta still lives in the old family house. With a
PhD in Pharmaceutical Chemistry and Technology as well as a one in Pharmacy with specialization in
Cosmetic Science from the State University of Milan, he has been active as a professional consultant,
technical manager in cosmetic companies and regulatory expert for cosmetic and medical devices.
His specialties are regulatory cosmetics, cosmetics GMP, safety assessment, quality compliance, develop-
ment of new products, relationships with control authorities, import-export cosmetics, medical devices,
REACH and CLP.
Back in 1982, Dr. Pirotta started working in the cosmetic field with an early focus on natural products. He
has cooperated in projects with some very renowned international cosmetic companies, universities and
specialized manufacturers, gaining experience in the regulatory field, safety assessment and controlled
release systems. In 1999, he launched a company to produce cosmetic patches with controlled release
delivery. Among other things, he worked on the application of a new technology delivery from fabrics (medical and cosmetic). With
his office he provides support for medical devices and biocides.
Dr. Pirotta’s publications include duplicate university lecture notes, articles in various fields and different specialized and scientific
journals, specialized chapters in books (regulatory and delivery systems) and congress presentations. He has been a board member of
the Italian Society of Cosmetics Chemists since 1989 as the regulatory officer and is a member of the Steering Committee for SCCI-
accredited courses. He is also a member of the Society of Cosmetic Chemists (USA) as well as MENSA International. From 1981 -1982
he was an Officer of the Regiment of Horse Artillery and since 1982 has been a Reserve Officer UNUCI. Dr. Pirotta is a consultant for
reserve projects of the Italian Senate Defense Commission and serves as a scientific consultant for Italian law courts.
Dr. Pirotta was Chairman of the International IFSCC Conference in Florence in 2005 and on the Committee for the Milan Conference
in 2019.

SCIENTIFIC
PAPER
New Horizon in Skincare Targeting the Facial-
Morphology-Retaining Dermal “Dynamic Belt”
– Revolution in Skin Analysis: “4D-Digital Skin” Technology –
Tomonobu Ezure1, Satoshi Amano1, Kyoichi Matsuzaki2 and Nobuhiko Ohno3,4
1 Shiseido Co., Ltd. MIRAI Technology Institute, Yokohama, Japan
2 International University of Health and Welfare, Narita, Japan
3 Jichi Medical University, Shimotsuke, Japan
4 National Institute for Physiological Sciences, Okazaki, Japan
Keywords: 4D, aging, sagging, gravity, hair muscle, dynamics
This publication was the 2020 Basic Research Award winner at the 31st (virtual) IFSCC Congress in Yokohama,
Japan, October 21-30, 2020.
of their dynamics, namely their movement
during the whole deformation process,
because we would need to process a huge
spatiotemporal dataset. Thus, adding the
extra dimension of time (3D to 4D) is not
easy, and we need to overcome some
challenging technological limitations.
There are a variety of structures inside
the skin, including hair follicles and se-
baceous glands. Although their physi-
ological functions have been studied
INTRODUCTION
Human beings have evolved a range of
systems to fight gravity, such as the inner
muscles that have supported the back-
bone since we stood up and started to
walk. Age-dependent deterioration of
these internal systems results in loss of
upright posture and difficulty in walking.
The skin, which forms the outside of the
body, is also at the frontline of the battle
with gravity by helping to retain the inner
organs in the correct positions as well as
also maintaining the morphology of the
body.
Deterioration of this antigravity ability with
aging allows the skin to be deformed by
gravity, which results in sagging (ptosis).
We have established that this gravity-in-
duced sagging causes a variety of morpho-
logical changes, such as wrinkles, concave
cheeks, and loss of facial contours, name-
ly an aged appearance [1]. Thus, the skin’s
antigravity system is critical for aesthetics.
However, the nature of this system in the
skin has not been clarified.
Why is the skin’s antigravity system un-
clear? The major reason is that there has
been no way to investigate it. To under-
stand the system in detail, we need to
visualize all of the skin’s internal structures
in three dimensions (3D) at an ultrafine
resolution level, and moreover we need to
analyze the dynamics of these structures
comprehensively, i.e., we require 4D in-
formation. Technology to observe various
target structures inside the skin, such as
blood vessels [2], in 3D has become avail-
able, and we reported 3D visualization of
skin at the cellular level, but it remains
difficult to observe all of the structures.
Furthermore, even if we could visualize all
the internal structures in the skin, it would
still be extremely challenging to analyze all
Abstract
Gravity is a fundamental cause of fa-
cial aging, because it deforms skin
and leads to sagging (aged appear-
ance). However, it is unclear how skin
resists gravity, since current technol-
ogy cannot access its “antigravity sys-
tem”. Thus, there is no rational basis
of skincare for facial rejuvenation.
Here, we established a breakthrough
technology called “4D-digital skin” to
visualize whole-skin dynamics during
deformation at ultrahigh resolution.
To do this, we observed movement
of skin using micro-CT and then ana-
lyzed the skin dynamics. The 4D-
digital skin technology revealed that
young facial skin resists deformation
in the direction of gravity due to a re-
sistant area where hair muscles (arrec-
tor pili muscles) “lock” the skin in place
and resist deformation. These muscles
align with gravity but decrease with
aging, resulting in skin deformation
and an aged appearance. Therefore,
hair muscles act as the skin’s anti-
gravity system (designated the dermal
“dynamic belt”), and loss of this belt
is critical for facial aging. 4D-digital
skin technology represents a revolu-
tion in skin analysis from static 3D to
comprehensive spatiotemporal 4D
analysis, dramatically expanding the
targets, depth and speed of research.
The discovery of the dermal dynamic
belt is a breakthrough in skincare,
leading to novel antigravity skincare
for facial rejuvenation by targeting the
skin’s antigravity system itself.
This paper received the
Basic Research Award 2020
at the IFSCC Congress in Yokohama

extensively, little is known about their
contributions to skin dynamics. For ex-
ample, hair follicles and sweat glands
are composed of tightly bound epider-
mal cells and located from the skin sur-
face to the deepest part of the dermal
layer. Therefore, it seems likely that they
would contribute to the physical proper-
ties of the skin. Hair muscles (arrector
pili muscles) are also a component of
the dermal layer. They are composed
of smooth muscle, bind to the hair fol-
licles, and function to raise the hair from
the skin surface (“goose bumps”) in re-
sponse to cold or emotional stimulation
via noradrenergic nerve fibers [3]. How-
ever, besides the physical properties, the
nature of age-dependent changes in hair
muscles and the mechanism of their re-
generation are unclear.
In this context, we aimed to establish a
breakthrough in the rational design of cos-
metics to promote a youthful appearance
by taking the following steps:
1)
establishing a 4D analysis system to
monitor the dynamics of whole skin, in-
cluding all of its internal structures, dur-
ing skin deformation, and
2)
using it to uncover the system in the
skin that resists the effects of gravity and
its contribution to facial aging.
EXPERIMENTAL
Subjects and study protocol
Facial skin specimens (age: 0
-103) were
obtained from surplus skin excised during
plastic surgery. All studies were approved
by the relevant ethics committees.
X-ray micro-CT analysis
for 3D observation
X-ray micro-CT was conducted under the
following condition, (50 kV, 80 μA; Xra-
dia; Zeiss, Oberkochen, Germany) [4].
4D reconstruction of
whole skin structure and movement
Individual structures in each micro-CT
image of skin were auto-classified by
an artificial intelligence (AI)-based deep-
learning system, Dragonfly (Object Re-
search Systems, Montréal, Canada).
Classified images were reconstructed in
3D by Dragonfly. Then, landmark points
were placed at high density on the sur-
faces of all the skin structures, and their
movement during skin deformation was
analyzed.
Histological observation
Skin sections were prepared by the Amex
procedure [5]. Immunohistochemical ob-
servation was conducted with the EnVi-
sion system (Agilent, Santa Clara, CA,
USA). Antibodies were purchased from
Abcam (Cambridge, UK).
Statistical analysis
Differences between groups, expressed
as the mean ±
S.E.M., were evaluated by
means of Student’s t test, Dunnett’s test
or the Wilcoxon rank test. P

0.05 was
considered significant. Correlations were
evaluated using Pearson’s correlation co-
efficient or Spearman’s correlation coef-
ficient by rank test.
RESULTS
Establishment of
“4D-digital skin” technology
Skin specimens were deformed in various
ways, including pressing and stretching,
and changes in their internal structures
were measured by taking successive X-
ray micro-CT images (Figure 1). Then,
an AI deep learning system was applied
to identify the huge numbers of complex
internal structures on each CT image au-
tomatically. This process afforded for the
first time a series of 3D reconstructions of
whole skin during deformation. Next, we
analyzed the spatiotemporal movement of
each skin structure in the 3D reconstruc-
tions during deformation. We placed a
large number of landmark points on the
surface of all the structures and followed
their movement in the successive 3D im-
ages. This enabled us to reconstruct the
Figure 1 Reconstruction of whole skin dynamics on computer. Skin samples were observed
successively by X-ray micro CT during a variety of deformation processes. The 3D structure of
the skin was reconstructed from hundreds of images using an AI deep learning system for skin structure
auto-identification. Huge numbers of landmarks were set on the skin structure surfaces, and their
movement was traced during skin deformation.

SCIENTIFIC
PAPER
whole skin deformation process. This is
the first visualization of the spatiotempo-
ral (4D) dynamics of internal skin, and we
designated this technology as “4D-digital
skin” (Figure 2).
An anti-deformation system in skin
that functions vertically to resist gravity
Using4D-digitalskin,wefoundauniquefea-
ture of skin dynamics in young skin. The use
of skin specimens with directional informa-
tionwasthekeytothisfinding.Whenyoung
skinispressed(Figure 3),itdoesnotdeform
uniformly. Namely, young skin resists defor-
mation in the vertical direction. In contrast,
old skin deforms uniformly, showing that
the resistance to the vertical deformation
force was significantly decreased in aged
skin. Therefore, our results suggest that the
skin contains a vertical anti-deformation sys-
tem, which is lost with aging, leading to skin
deformation under the influence of gravity
and resulting in sagging.
Prevention of vertical deformation
of skin by hair muscles
How does young skin resist vertical de-
formation? The 4D-digital skin analysis of
the pressing process showed that a resis-
tant area exists in young skin (Figure 4).
4D-Digital anatomy showed that hair mus-
cles exist in the resistant area, suggesting
that hair muscles contribute to resist skin
deformation. Indeed, hair muscles them-
selves resist deformation, whereas other
structures are drastically deformed. Fur-
thermore, there was no specific direction
or deviation of collagen fibers (the main
component of the dermal layer) in this
area, and moreover, collagen fibers as a
hole were deformed in the same manner
as other structures, suggesting that they
make only a minor contribution to this re-
sistance area. Therefore, hair muscles ap-
pear to be the predominant contributors
to preventing vertical deformation of skin.
Hair muscles form a gravity-resisting
dermal “dynamic belt” that maintains
a facial youthful appearance
How does this anti-deformation system con-
tribute to facial appearance? 4D-digital skin
analysis revealed a high density of hair mus-
cles aligned in the vertical direction much
like a belt in young skin (Figure 5a), while
the hair muscles are drastically decreased in
aged skin (Figure 5b). We then estimated
the hair muscle direction (Figure 6a, b).
Only the forehead area showed a variety
of patterns of muscle direction, whereas
other areas showed similar patterns of
vertical muscle direction. Taken together,
these results suggest that a high density of
hair muscles in the face contributes to pre-
venting skin deformation and maintaining
Figure 2 4D-digital skin. a) 4D-digital skin
reconstructed on computer with spatiotemporal
information during a variety of skin movements
and b) at an ultrahigh resolution (magnified
single sweat gland as an example).
The displacement amount, i.e., 3D movement,
is indicated with a color map (for pressed skin
as an example). Movements of skin structures
are indicated by arrows.
Figure 3 Young skin contains a vertical anti-deformation system.
Top view of young skin a) before and b) after pressing. The skin is less deformed in the vertical direction.
Figure 4 Hair muscles prevent vertical deformation of skin: a) 4D-digital skin of deformed skin with
displacement information (color map) showing the presence of hair muscles (not colored) in the
resistant area (blue), and b) schematic illustration of a hair muscle preventing vertical skin deformation.

a youthful appearance by resisting vertical
deformation due to the force of gravity.
Thus, we call this antigravity system of the
skin the dermal “dynamic belt”. The loss of
hair muscles with aging, namely loss of the
dynamic belt, leaves the skin unable to resist
vertical deformation due to gravity and re-
sultsinsagging.Thisisanovelmechanismof
facial aging and suggests that the dynamic
belt can be a critical target of anti-aging
skincare for facial rejuvenation.
DISCUSSION
To understand how the skin resists the
effect of gravity, we first developed a
technology to comprehensively visualize
the dynamics of internal skin structures
during deformation. This 4D-digital skin
technology is based on our new, intact
skin micro-CT technology followed by AI
to process the enormous spatiotemporal
data set generated during the whole de-
formation process. This technology takes
skin analysis into the fourth dimension,
enabling full 4D spatiotemporal analysis.
Conventional skin dynamics analysis has
generally measured deformation of the
skin as a whole in terms of numerical data
(e.g., Young’s modulus). Thus, it does not
address the dynamics of individual skin
components and cannot identify the key
determinants of skin physical properties. In
contrast, our novel 4D technology can vi-
sualize the whole process of skin deforma-
tion at an ultrafine resolution level, reveal-
ing the dynamics of each skin component
both visually and numerically. Another ad-
vantage of this technology is that this skin
is reconstructed digitally (each structure
has its own 4D information) instead of just
providing a 4D image. Therefore, we can
analyze skin dynamics in unprecedented
ways, such as on-computer anatomy and
sorting, at any time point during deforma-
tion. This is critical to handling complex 4D
spatiotemporal datasets quite easily.
Furthermore, conventional skin dynamics
studies have mainly targeted just collagen,
and 3D visualization studies have been re-
stricted to one or two targets in a limited
area of skin. Our methodology enables us
to target both the dynamics and structure
of all components of whole skin. Therefore,
we can comprehensively analyze skin struc-
ture dynamics. This makes it possible to
identify key factors affecting skin dynamics
as well as causal relationships among struc-
tures associated with particular skin physi-
cal properties. For example, we identified
hair muscle as a critical factor for maintain-
ing facial skin morphology by comprehen-
sively comparing its physical properties with
those of other skin components.
Once we digitally reconstruct 4D skin, we
can archive the data with information on
gender, ethnicity, age and so on, to con-
struct a digital skin library, which would
be inaccessible to manual analysis. Indeed,
we used AI technology to process an astro-
nomic volume of complex data in order to
reconstruct skin dynamics (4D-digital skin)
on computer to show the importance of
utilizing big data and processing technol-
ogy for cosmetics RD. This approach is a
fundamental change in research style, dra-
Figure 5 A high density of hair muscles forming a gravity-resisting dermal “dynamic belt”:
a) Young skin contains a high density of vertically aligned hair muscles that b) decreases in aged skin.
Figure 6 Patterns of hair muscle direction: a) Similar patterns are seen in most facial areas (red),
except for the forehead (blue). b)Different directional patterns (arrows) and their ratio in the forehead
are shown.

SCIENTIFIC
PAPER
[3] Torkamani, N., Rufaut, N.W., Jones, L.,
and Sinclair, R.D., Beyond goosebumps:
does the arrector pili muscle have a role
in hair loss?, Int. J. Trichology, 6 (2014)
88-94.
[4] Kim, L.G., Park, S.A., Lee, S.H., Choi, J.S.,
Cho, H., Lee, S.J. Kwon, Y.W., and Kwon,
S.K., Transplantation of a 3D-printed tra-
cheal graft combined with iPS cell-derived
MSCs and chondrocytes, Sci. Rep., 10
(2020) 4326.
[5] Sato, Y., Mukai, K., Watanabe, S., Go-
to, M., and Shimosato, Y., The AMeX
method. A simplified technique of tissue
processing and paraffin embedding with
improved preservation of antigens for im-
munostaining, Am. J. Pathol., 125 (1986)
431-435.
[6] Ali AlHamdi, Facial Skin Lines, Iraqi. JMS
(2015) 103-107.
[7] Ruden, D.M., Bolnick, A., Awonuga, A.,
Abdulhasan, M., Perez, G., Puscheck,
E.E., and Rappolee, D.A., Effects of Grav-
ity, Microgravity or Microgravity Simula-
tion on Early Mammalian Development,
Stem Cells Dev., 27 (2018) 1230-1236.
[8] Vandenbrink, J.P., and Kiss, J.Z., Plant re-
sponses to gravity, Semin. Cell Dev. Biol.,
92 (2019) 122-125.
[9] Burke, S., and Hanani, M., The actions of
hyperthermia on the autonomic nervous
system: central and peripheral mecha-
nisms and clinical implications, Auton.
Neurosci., 168 (2012) 4-13.
[10] Xiao, L.J., and Tao, R., Physical Therapy,
Adv. Exp. Med. Biol., 1010 (2017) 247-
260.
Corresponding Author
Tomonobu Ezure
Shiseido Co., Ltd.
MIRAI Technology Institute
Yokohama
Japan
tomonobu.ezure@shiseido.com
G
matically reducing the time and resources
required for skin studies.
The 4D-digital skin technology described
here enabled us to discover the skin’s an-
tigravity system, the dermal dynamic belt,
and also to identify novel anti-aging targets
and solutions. Various physical lines in the
face, such as Langer’s line, have been pro-
posed to determine the direction of face
incisions in surgery, but they are different
from each other [6]. The most commonly
utilized line in facial plastic surgery is the
relaxed skin tension line (RSTL), which takes
account of subcutaneous physical proper-
ties, or the Kraissl line, which is determined
by subcutaneous facial muscle contraction
(mimetic muscle). On the other hand, the
dynamic belt represents the physical prop-
erties and direction of the skin itself, inde-
pendently of subcutaneous tissue. It main-
tains the facial morphology against gravity
and is highly correlated with facial appear-
ance. Thus, the dynamic belt appears to be
a critical system for maintaining a youthful
appearance, and it, rather than the contro-
versial surgical lines, is therefore important
for cosmetics.
The dynamic belt has a unique property in
that it prevents vertical deformation, but
it allows horizontal deformation. Indeed,
wrinkles form along the belt direction.
Thus, the dynamic belt enables the skin to
fulfil two functions, i.e., maintaining the
morphology of the body against gravity
while at the same time leaving the skin suf-
ficient flexibility to engage in subtle facial
expressions for communication.
Although gravity is ubiquitous, it has re-
ceived less attention than other environ-
mental factors, such as sunlight or oxygen
(oxidation), in relation to cosmetics. But
our results clearly show that considering
the influence of gravity can provide im-
portant new opportunities for skincare
(antigravity skincare). Since gravity can in-
fluence the movement and distribution of
components in the skin, such as water and
blood, a variety of physiological reactions
(not only edema) can be affected by it [7].
Indeed, gravity-controlled hormone distri-
bution and its effect on growth have been
well established in plants [8]. As a benefi-
cial side effect, changes in the direction
and strength of gravity while bathing and
sleeping can potentially positively affect
the skin condition and could be important
in developing new cosmetics.
Conventional targets of anti-aging skincare
are mainly collagen and fibroblasts (dermal
cells). However, the discovery of the dy-
namic belt provides a novel skincare target,
the hair muscle. This opens up possibilities
for many new skincare solutions, inglding
hair muscle stimulators (agonists or natu-
ral medicines), drug delivery through hair
follicles to hair muscle, beauty methods,
fragrances, and bath items. Furthermore,
as hair muscle is a smooth muscle, we can
apply a variety of solutions established
for smooth muscle in the field of physical
therapy, such as heat, electrical stimulation,
and ultrasound [9, 10]. As described above,
applying the concept of the direction of
skin can drastically update conventional
beauty treatments and devices.
CONCLUSION
We have established a new paradigm of
skin analysis, “4D-digital skin”. This moves
us from current 3D static analysis to com-
prehensive spatiotemporal 4D analysis of
whole skin dynamics, drastically expanding
the range of study targets and speeding up
research. This methodology allowed us to
discover a skin antigravity system consist-
ing of hair muscles, and we found that
this dermal “dynamic belt” is critical for
maintaining a youthful appearance. This
discovery can drastically change skincare by
establishing antigravity skincare as a novel
skincare field and providing new solutions
(drug targets and physical approaches).
References
[1] Ezure, T., Hosoi, J., Amano, S., and
Tsuchiya, T., Sagging of the cheek is relat-
ed to skin elasticity, fat mass and mimetic
muscle function, Skin Res. Technol., 15
(2009) 299-305.
[2] Shibata, T., Kajiya, K., Sato, K., Yoon, J.,
and Kang, H.Y., 3D microvascular analysis
reveals irregularly branching blood ves-
sels in the hyperpigmented skin of solar
lentigo, Pigment, Cell Melanoma Res., 31
(2018) 725-727.

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SCIENTIFIC
PAPER
A Perfusable Vascularized Full-Thickness Skin Model
for Topical and Systemic Applications
Sacha Salameh1,2, Nicolas Tissot1, Kevin Cache1, Joaquim Lima1, Itaru Suzuki1, Paulo André Marinho1,
Maité Rielland1, Jérémie Soeur1, Shoji Takeuchi4, Stéphane Germain3, Lionel Breton1
1 L’Oréal Research and Innovation, Aulnay-sous-Bois, France
2 Sorbonne Université, Collège Doctoral, F-75005 Paris, France
3 Center for Interdisciplinary Research in Biology (CIRB), College de France, CNRS, INSERM, PSL Research University, Paris, France.
4 Department of Mechano-Informatics, Graduate School of Information Science and Technology, The University of Tokyo,
7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan
Keywords: Reconstructed skin, tissue engineering, vascularization, vasculogenesis, angiogenesis, perfusion
This publication was the 2020 Applied Research Award winner at the 31st (virtual) IFSCC Congress in Yokohama,
epidermis containing all the layers of the
in vivo structure.
EXPERIMENTAL
Reconstruction of the skin equivalent
The culture device was as previously de-
scribed [6]. Briefly, three nylon wires
(0.55mm diameter) were strung across
the connectors before sterilization using
70% ethanol and UV light. Devices were
then treated with a plasma etcher (Har-
rick Plasma, Ithaca, NY, USA), filled with
neutralized solution of bovine type I col-
lagen and NHDFs prepared as previously
described [10,11] and then incubated at
37°C for 30/60mins until the collagen gel
formed a dermis-like layer. After 4 to 5
days, the nylon wires were removed and
the hollow channels were filled with 200
μl ECGM-2 containing HUVECs using a sy-
ringe pump (KD Scientific, Holliston, MA,
USA). Subsequently, keratinocytes were
seeded on the dermal layer. The central
channel was then perfused using a peri-
staltic pump (SJ-1211II-L, ATTO Corp., To-
kyo, Japan). After 3 days of immersion, the
skin equivalent was lifted to the air-liquid
interface to induce stratification of the
epidermal layer. Perfusion was maintained
until the end of culture (17 days in total).
The media was changed every other day.
To induce vasculogenesis, HUVECs trans-
duced with Turbo-RFP lentiviruses (Vec-
INTRODUCTION
Skin models are used in the evaluation of
efficacy and safety of active compounds
and drugs in cosmetic and pharmaceuti-
cal research, especially after the ban of
animal testing in cosmetics research in
2013 [1-3]. Many advances have been
made in skin engineering but challenges
still exist for constructing thick or complex
tissues with all in vivo functions. One of
the main challenges is vascularization. In
the human body, cells are fed with nutri-
ents and oxygen from capillaries located
not further than the 100-200μm range
[4,5]. Therefore, skin constructs thicker
than 200μm have a diffusion limitation.
The closer we get to the physiological vas-
culature, the closer we are to reaching rel-
evant in vitro tests. Nonetheless, the few
existing models of perfusable vascularized
full-thickness skin are limited. These mod-
els either rely on macrovascular channels
that lack cutaneous-like microvascular-
ization or contain a poorly differentiated
epidermis [6-9]. To address this issue, our
study focused on the development of a
perfusable vascularized full thickness skin
equivalent with more complex vasculature
than existing models and a differentiated
Abstract
Vascularization of reconstructed tissues
is one of the remaining hurdles to be
considered to improve both the func-
tionality and viability of skin grafts and
the relevance of in vitro applications.
Our study therefore sought to develop
a perfusable vascularized full-thickness
skin equivalent that comprises a more
complex blood vasculature then exist-
ing models. We here combined mold-
ing, auto-assembly and microfluidics
techniques in order to produce a skin
equivalent that recapitulates a prop-
erly differentiated epidermis as well
as complex vascular networks. Indeed,
three perfusable vascular channels that
sprouted via angiogenesis were created
and eventually connected to a capillary
plexus generated by endothelial cell
auto-assembly, i.e. vasculogenesis.
We then evaluated the skin permeabili-
ty for compounds with different chemi-
cal properties and systemic delivery of
the benzo[a]pyrene pollutant. Our re-
sults demonstrated that perfusion of
the vascular plexus resulted in a more
predictive and reliable model to assess
both topical and systemic applications.
This model is therefore aimed at fur-
thering drug discovery and improving
clinical translation in dermatology.
This paper received the
Basic Research Award 2020
at the IFSCC Congress in Yokohama

talys, Toulouse, France) (HUVECS-RFP)
were seeded as a monolayer between two
layers of collagen/fibroblasts. Then the
same protocol as described above was fol-
lowed, except that hollow channels were
seeded with HUVECs transduced with
EGFP lentiviruses (HUVECS-GFP).
Histological analyses
Morphologies of the skin equivalent and
vascular channels were analyzed using he-
matoxylin, eosin and saffron (HES) staining
or immunostaining. Immunofluorescent
stainings were performed on 7 µm frozen
sections fixed in cold acetone, blocked in
Bovine Albumin Serum (BSA)-10
% nor-
mal goat serum (SP-004-vx10, Diagomics,
Blagnac Cedex, France) and incubated for
1 h at room temperature (RT) or overnight
at 4
°C with diluted primary antibodies
in 1% BSA: Collagen IV (COLL IV) (Dako
M0785, 1:50, CiteAb, Bath, UK,), Per-
lecan (ab26265, 1:100, Abcam, Boston,
MA, USA,), CD31 (Dako M0823, 1:20),
Keratin 14 (K14) (10003, 1:20, Progen,
Heidelberg, Germany,), Keratin 10 (K10)
(Dako M7002, 1:100), Involucrin (INV)
(J64013-AB, 1:50, BTI, Penicuik, UK),
transglutaminase 1 (TGM1) (NBP2-34062,
1:500, Novus Biol., Littleton, CO, USA),
Filaggrin (FIL) (sc66192, 1:200, Santa-Cruz
Biotechnology, Santa Cruz, CA, USA).
Sections were washed with PBS- and in-
cubated with secondary antibody solu-
tion (A21422 or A21428) diluted 1:500
in 1
% BSA for 1 h at RT. Sections were
then mounted using Prolong with DAPI
(P36931, Life Technologies, Carlsbad, CA,
USA) and observed with a Nanozoomer
S60 (Hamamatsu, Japan) or Nikon Micros-
copy Eclipse 80i (Tokyo, Japan).
Measurement of skin permeability
To estimate skin barrier function, we mea-
sured skin permeation of a mix of 2 chemi-
cals with different physicochemical param-
eters: caffeine and minoxidil (purity 100 %)
obtained from L’Oréal (Aulnay-sous-Bois,
France). Around 25
mg of each chemical
were dissolved in 10
ml of PBS containing
0.25
% (w/w) Tween 80 (723-M-156-84,
Figure 1 Histological analysis and immunostaining of the skin equivalent with perfusable vascular channels and a secondary vascular plexus
A- Vasculogenesis of a secondary vascular plexus. (a) Two-photon fluorescence microscopy showing the HUVECS-RFP microvascular beds formed between
the three HUVECS-GFP perfusable vascular channels, (b) anastomosis of the HUVECS-GFP capillary network and HUVECS-GFP angiogenic sprouts from
the main channels, and (c) high magnification showing hollow space between the borders of the capillaries.
B- HES staining of the skin equivalent with 3 perfusable vessels and microvascular bed. Immunostaining of CD31 marker of the endothelial cells, Coll IV
and perlecan markers of the basal lamina.
C- HES staining showing epidermal differentiation layers. Immunostaining of K14 marker of the basal layer, K10 marker of the intermediate differentiation,
and involucrin, filaggrin and TGM1 markers of the late differentiation.

SCIENTIFIC
PAPER
Fisher, Hampton, NH, USA). An infinite
dose of 300 µL
/
cm² of the buffered solu-
tion was applied topically to the skin. Each
hour for the next 6 hours, 300 µL were
sampled from the receptor fluid under the
skin equivalent. Samples were then dilut-
ed (1/10) in a fresh culture medium and
quantified with standard curves prepared
in the same culture medium. All samples
were analyzed with an LC/MS-MS system
(Shimadzu Nexera LC system coupled with
a mass spectrometer API 3500 (Sciex, Con-
cord, Ontario, Canada).
Systemic exposure
to benzo[a]pyrene pollutant
To evaluate the systemic transport of
benzo[a]pyrene (BaP) to the dermis and
epidermis of skin substitutes, tissues ei-
ther in static or perfused conditions were
exposed to 10 µM BaP (B1760, Sigma, St.
Louis, MO, USA). BaP was added to the
emersion medium at Day 3 of the em-
ersion phase. After 48
h, the media was
changed to fresh media containing 10 µM
BaP. Samples were cultured for an addi-
tional 48
h before harvest. At the end of
the culture period, media of each device
was sampled to evaluate the remaining
BaP after skin culture. The dermis and epi-
dermis parts were separated and proteins
were precipitated using acetonitrile (ACN)
(271004, Sigma). The supernatant was
then extracted and the BaP concentration
was analyzed using liquid chromatogra-
phy with fluorescence detection HPLC-FLD
(Shimadzu Prominence- RF-20A-XS).
RESULTS
Development of a full-thickness skin
equivalent with three vascular channels
and a secondary vascular network
Taking advantage of the auto-assembly
properties of endothelial cells seeded into
3D collagen matrices, HUVECS were seed-
ed between two layers of fibroblasts em-
bedded in a collagen matrix at the same
level as the perfused vascular channels. To
identify the capillaries formed by angio-
genic sprouting from the fluidic channels
from those developed by vasculogenesis
in the lattice, we used HUVECS-GFP to
reconstruct the vascular channels and HU-
VECS-RFP for the microvascular network
between the channels. Under these con-
ditions a complex HUVECS-RFP microvas-
cular network was formed between the
three HUVECS-GFP perfusable vascular
channels (Figure 1A). This dense capillary
network displayed connected branches
(Figure 1Aa) that could reach and con-
nect to the angiogenic sprouts emanating
from the fluidic channels (Figure 1Ab).
The formation of a lumen could be identi-
fied by hollow spaces between vascular
borders (Figure 1Ac). The two layers of
collagen and fibroblasts were connected
with a visible line of cells expressing CD31,
perlecan and collagen IV at the location of
the microvascular bed (Figure 1B). The
three channels were preserved opened.
CD31, collagen IV and perlecan were
properly expressed (Figure 1B). More-
over, epidermal differentiation was well-
maintained and each layer was expressing
its specific markers: K14 for the basal lay-
er, K10 for the spinous layer, INV, TGM1
and FIL for the granular layer and stratum
corneum (Figure 1C).
Evaluation of skin permeability
We evaluated the effect of vasculariza-
tion and perfusion on epidermal barrier
function by studying skin permeation of
caffeine and minoxidil, two chemicals
with different structures and penetration
potentials. Six different skin models were
evaluated: control samples with no chan-
nels (Lat), channels with endothelial cells
but without perfusion (ECnoP), perfused
channels with endothelial cells (ECP), per-
fused channels without endothelial cells
(noECP), channels with endothelial cells
and secondary microvascularization with-
out perfusion (SandnoP), and perfused
channels without endothelial cells and
secondary microvascularization (SandP).
These models were also compared with
porcine ear skin (PES), which is tradition-
ally used as a surrogate for human skin in
the assessment of topical drug bioavail-
ability [12]. Thus, a model with a closer Kp
to porcine ear skin would be a more ap-
propriate alternative for the assessment of
skin permeation and penetration in vitro.
The results showed that for either caf-
feine or minoxidil, Kp is lower in perfused
samples (noECP, ECP, SandP) than in non-
perfused samples (Lat, ECnoP and Sand-
noP). While Kp obtained with porcine ear
skin remained statistically lower than all
skin equivalents, these results showed that
perfused reconstructed skin have a lower
Kp than non-perfused samples. However,
no differences were observed between
noECP, ECP and SandP (Figure 2). There-
fore, we showed for both chemicals that
Figure 2 Evaluation of skin permeability
Coefficient of penetration Kp (cm / h) of each of the 2 chemicals and each skin model.
Lat = control samples with no channels, ECnoP = channels with endothelial cells but without perfusion,
ECP = perfused channels with endothelial cells, noECP = perfused channels without endothelial cells,
SandnoP = channels with endothelial cells and secondary microvascularization without perfusion,
SandP = perfused channels without endothelial cells and secondary microvascularization.
PES = porcine ear skin * = P 0.1, ** = P 0.05, # = moderate effect size, ## = strong effect size,
### = very strong effect size. N = 3 for each condition.

the perfusion of skin equivalents improved
the epidermal barrier independently of the
presence of ECs.
Evaluation of systemic exposure
of a pollutant
Besides topical applications, having a
perfusable vascularized skin model holds
many interests for systemic exposure. We
therefore explored the advantage of hav-
ing, for the first time, perfusable vascular
channels, but also a complex microvas-
cular network coupled to them in a full
thickness skin equivalent, to estimate der-
mal and epidermal transport of systemic
compounds, specifically, pollutants.
To do this, 10 µM of benzo [a] pyrene BaP,
an extremely widespread environmental
and industrial pollutant, was added to the
culture media of each skin model. After 5
days of exposure, we measured the BaP
concentration in the epidermis, dermis and
culture media. In order to observe the im-
pact of a vascular plexus on BaP distribu-
tion, five different models were evaluated:
Control samples with no channels (Lat),
non-perfused channels with endothelial
cells (ECnoP) or with endothelial cells and
the secondary vascular plexus (SandnoP),
perfused channels with endothelial cells
(ECP) or with endothelial cells and the sec-
ondary vascular plexus (SandP) (Figure 3).
Under both static (Figure 3a) and fluidic
conditions (Figure 3b), the presence of a
secondary vascular plexus (SandnoP and
SandP) increased BaP delivery either in the
dermis or the epidermis compared with
samples without channels (Lat) or with
the three vascular channels only (ECnoP
and ECP). These results proved that the
presence of a complex vascular plexus im-
proves the efficacy of systemic diffusion
of the pollutant either in the dermis or
the epidermis.
DISCUSSION
Currently, 2D and 3D in vitro skin models
are used to replace animal testing both in
some pharmaceutical and in all cosmetic
studies. Many organizations (ECVAM in
Europe, ICCVAM in the US, JaCVAM in
Japan, KoCVAM in Korea, and braCVAM
in Brazil) were established worldwide to
validate the use of these alternative mod-
els for regulatory concerns. However,
these models still lack the complexity of
the in vivo skin in terms of appendages,
vasculature, cell variety and distribution.
To overcome this significant limitation
of existing skin models, we developed a
vascularized full thickness skin model that
comprises a differentiated epidermis, per-
fusable vessels and a secondary microvas-
cular network.
Compared to the published perfusable vas-
cularized skin models [6,
7,
9], we showed
for the first time a differentiated epidermis
expressing all the markers of the different
layers of differentiation. Vessel forma-
tion was also proven by CD31 staining,
a protein encoded by PECAM1 gene, in
order to determine the early formation of
tight junctions between endothelial cells.
The basal lamina of the microvessels was
marked with collagen type IV, perlecan and
laminin, which are principle components
of the basement membrane. These pro-
teins are important for development and
homeostasis of the vascular network [13].
As reconstructed skin models are used for
safety and efficacy evaluation of chemicals
applied systemically or topically to skin tis-
sues, the functionality of the vascularized
skin equivalent for in vitro assessments
was evaluated with topical and systemic
applications.
For topical application, it is important to
be able to estimate the permeation of new
compounds through the skin, resulting in
its absorption, distribution, metabolism,
and excretion (ADME). However, exist-
ing skin models have a higher permeation
than in vivo skin [14]. We therefore evalu-
ated the effect of vascularization and per-
fusion on epidermal barrier integrity by
evaluating the percutaneous absorption
of caffeine (high permeablity chemical)
and minoxidil (low permeability chemi-
cal). For both chemicals, we showed that
the perfusion of skin equivalents improved
the epidermal barrier independently of the
presence of endothelial cells.
Besides topical applications, having a
perfusable vascularized skin model holds
many interests for systemic applications.
Most of the existing skin models are cul-
tured in static conditions, which is not
representative of the in vivo reality. Ad-
ditionally, substances are usually added to
the static culture medium in contact with
all the dermis equivalents, whereas in vivo,
systemic substances are delivered via the
vascular network, meaning that they are
exposed to blood flow and have the ves-
sel wall as an additional diffusion barrier.
To better mimic the physiological reality,
we used our vascularized full thickness
skin models to estimate dermal and epi-
Figure 3 Evaluation of systemic delivery of pollutants
Graphical box plot presentation of BaP concentrations (ng / mL) in the dermis or epidermis either in static
models (a) or perfused models (b). Lat = control samples with no channels, ECnoP = channels with
endothelial cells but without perfusion, ECP = perfused channels with endothelial cells,
SandnoP = channels with endothelial cells and secondary microvascularization without perfusion,
SandP = perfused channels without endothelial cells and secondary microvascularization.
* = P 0.1, ** = P 0.05, # = moderate effect size. N = 6 for each condition.

SCIENTIFIC
PAPER
[11] Asselineau, D., Bernhard, B., Bailly, C.,
and Darmon, M., Epidermal morphogen-
esis and induction of the 67 kD keratin
polypeptide by culture of human kerati-
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mental cell research 159 (1985) 536-539.
[12] Flaten, G. E., Palac, Z., Engesland, A., Fil-
ipovic-Grcic, J., Vanic, Z., Škalko-Basnet,
N., In vitro skin models as a tool in opti-
mization of drug formulation. European
journal of pharmaceutical sciences: offi-
cial journal of the European Federation for
Pharmaceutical Sciences 75 (2015) 10-24.
[13] Marchand, M., Monnot, C., Muller, L.,
and Germain, S., Extracellular matrix
scaffolding in angiogenesis and capil-
lary homeostasis, Semin Cell Dev Biol, 89
(2019) 147-156.
[14] Schafer-Korting, M., Bock, U., Diembeck,
W., Düsing, H.J., Gamer, A., Haltner-Uko-
madu, E., Hoffmann, C., Kaca, M., Kamp,
H., Kersen, S., Kietzmann, M., Korting,
H.C., Krächter, H.U., Lehr, C.M., Liebsch,
M., Mehling, M., Müller-Goymann,
C., Netzlaff, F., Niedorf, F., Rübbelke,
M.K., Schäfer, U., Schmidt, E., Schreiber,
S., Spielmann, H., Vuia, A., and Weimer,
M., The use of reconstructed human epi-
dermis for skin absorption testing: Results
of the validation study, Alternatives to labo-
ratory animals: ATLA, 36 (2008) 161-187.
[15] Soeur, J., Belaïdi, J.P., Chollet, C., Denat,
L., Dimitrov, A.., Jones, C., Perez, P., Zani-
ni, M., Zobiri O., Mezzache, S., Erdmann,
D., Lereaux, G., Eilstein, J., and Marrot,
L., Photo-pollution stress in skin: Traces
of pollutants (PAH and particulate mat-
ter) impair redox homeostasis in keratino-
cytes exposed to UVA1, Journal of derma-
tological science 86 (2017) 162-169.
Corresponding Author
Sacha Salameh
L’Oréal Research and Innovation
Aulnay-sous-Bois
France
sacha.salameh@rd.loreal.com
G
dermal transport of systemic compounds,
specifically pollutants. In fact, pollutants
like benzo[a]pyrene (BaP) are phototoxic
and detected in the blood, serum, urine
and hair shafts of smokers and individu-
als living in polluted areas [15]. This sug-
gests that the skin could be exposed to
these pollutants from the surface but also
from the systemic circulation. Therefore,
having a complex vascular structure in a
dermal equivalent allows a better under-
standing of BaP transport and effects on
the dermis before reaching the epidermal
layer. Indeed, we found that both under
static and fluidic conditions, the presence
of the secondary vascular plexus increased
the transport of benzo[a]pyrene BaP into
the dermis and the epidermis compared
to samples without vessels or with the 3
vascular channels alone. This suggests that
having a finer vascular structure makes
the evaluation of systemic delivery of sub-
stances to or from the skin more efficient.
CONCLUSION
We developed for the first time a full-
thickness human skin model with a ma-
ture epidermis and three perfusable tu-
bular structures with angiogenic sprouts
that are associated with a complex micro-
vascular network. The integrity of each
compartment was confirmed by histo-
logical immunofluorescence analysis and
compared with that of normal human
skin. We proved that having a perfusable
vasculature closer to the in vivo vascular
plexus resulted in a more reliable model
for topical and systemic assessments. In
our study, systemic delivery of compounds
and transdermal absorption were evalu-
ated. However, a broad range of applica-
tions is possible either for skin knowledge
studies or efficacy and safety evaluations.
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SCIENTIFIC
PAPER
Do-It-Yourself Cosmetics –
The Pleasure of Creating Your Own Emulsions
Megumi Kaji1, Tomoyuki Iwanaga1, Yuichiro Takeyama1, Kazuki Matsuo1, Toshihiro Arai1,
Kenichi Sakai2 and Hideki Sakai2
1 POLA Chemical Industries, Inc., 560 Kashio-cho, Totsuka-ku, Yokohama, Kanagawa, Japan
2 Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, Japan
Keywords: Do-It-Yourself cosmetics, handmade cosmetics, at-home emulsification,
polymer emulsifier, core-shell particles
This publication was presented as a podium presentation at the 31st (virtual) IFSCC Congress in Yokohama,
However, they have not yet achieved
massive adoption, largely due to dissat-
isfaction with the difficulty of preparing
emulsions and customizing the mixtures;
this makes clear that preparing a technol-
ogy that empowers consumers to easily
prepare and customize emulsions is vital
to DIY cosmetic projects.
Cosmetic engineers use emulsifiers to pre-
pare emulsions - namely, a state in which
oil components are dispersed and remain
in water as oil particles without aggre-
gating [6]. Emulsifiers must be selected in
such a way that they adsorb to the oil and
stabilize the molecular structure. In addi-
tion, the optimal humectant must be se-
lected and managed at optimal tempera-
tures to successfully prepare the emulsion
[7-10]. With all aspects considered, it is
clear that a new emulsifier is required that
can be used to create emulsions at home
that is adaptable to a variety of oils and
humectants and requires no temperature
management.
Therefore, in this study we addressed
these issues, designing a new core-shell
type emulsifier that forms smaller particles
than the oil particles in order to promote
the adhesiveness by heterocoagulation
[11-13]. We called this molecule M-poly-
mer (Figure 1). Importantly, we tested
the stability of this new emulsifier over
a range of conditions and verified that it
is suitable for use in home DIY cosmetic
projects.
INTRODUCTION
There is much room for improvement
in terms of the enjoyment of cosmetics.
One possibility is with do-it-yourself (DIY),
where people can enjoy their own creativ-
ity. DIY is defined as activities in which
individuals engage raw and semi-raw
materials and component parts to pro-
duce, transform, or reconstruct material
possessions [1]. With a future projected
market of 143.3 billion US dollars [2], DIY
can not only create a unique product but
also offer the satisfaction of creation and
the joy of customization [1]. Additionally,
the Internet has made information regard-
ing DIY more accessible, and its popular-
ity is growing in many industries. Finally,
COVID-19 has changed lifestyles globally,
and DIY projects of everyday goods fulfill
time at home, and improve physical and
mental wellbeing [3-5].
In cosmetics, some emulsions, which are
the majority of cosmetic products, can
already be purchased for home projects.
Abstract
Do-it-yourself projects (DIY) are in-
creasingly popular, but DIY projects
involving cosmetics are still rare. To
enable consumers to create their
own cosmetics, the field still requires
a technology that allows easy prepa-
ration of emulsions as well as cus-
tomization of the composition and
feel of the mixtures. To address this
issue, we developed a new emulsifi-
er, a methoxy polyethylene glycol-23
methacrylate/glyceryl diisostearate
methacrylate copolymer we called
M-polymer, that is able to emulsify
various types of oil using convention-
al home cooking appliances without
temperature management. This is
because M-Polymer forms nanosized
particles in water and covers the oil /
water interface, as we inferred from
light scattering and electron micros-
copy data. Additionally, the oil droplet
size of the emulsions was stable for
3 months under temperatures from
5 °C to 50 °C. Finally, 21 test subjects
with no experience in cosmetic formu-
la development tested the effective-
ness of M-polymer in DIY cosmetics
and demonstrated their satisfaction
with the simplicity of the process and
the customization flexibility as well as
their willingness to repeat the experi-
ence. In summary, we successfully de-
veloped a new emulsifier that enables
consumers to create emulsions with
a customizable composition and feel.
We anticipate that this will contribute
to DIY cosmetics projects and inject
new life into the cosmetics industry.
Megumi Kaji has been nominated
for the Henry Maso Award
for the work on this paper.

EXPERIMENTAL
Design of a new emulsifier
with a new emulsion function
Synthesis of the new emulsifier
An emulsifier with the hydrophilic and
hydrophobic groups on the main chain
was designed. The designed emulsifier
(methoxy polyethylene glycol-23 meth-
acrylate/glyceryl diisostearate methacry-
late copolymer) was synthesized by NOF
in Japan. Used for the hydrophilic group
monomer was methoxy polyethylene
glycol monomethacrylate (NOF, Tokyo,
Japan) and for the hydrophobic group
monomer glyceryl diisostearate methac-
rylate (NOF, Tokyo, Japan). These compo-
nents were copolymerized in 2-propanol
with a radical polymerization initiator, and
the 2-propanol was removed, leaving a
copolymer with different weight ratios of
the hydrophilic groups and hydrophobic
groups. Gel permeation chromatography
(HLC-8220GPC, Tosoh, Tokyo, Japan) was
used for size exclusion chromatography
to measure the weight average molecu-
lar weight (Mw) and the number average
molecular weight (Mn).
Formulation of the new emulsifier
aqueous solution and evaluation
of the dispersion properties
Copolymers with weight ratios (m:n) of
the hydrophilic to hydrophobic groups
of 5:5, 6:4, 7:3, and 8:2 were used with
1,3-butanediol (1,3-BG, Daicel, Osaka,
Japan) and glycerin (Emery Oleochemi-
cals, Selangor, Malaysia) as humectants
together with purified water to formulate
the copolymer aqueous solution. The com-
position for each weight ratio of copoly-
mer was 1 wt% and the humectants were
adjusted to 0 or 29.7
wt%. The emulsi-
fier, humectants and purified water were
weighed and mixed in a glass test tube
and stored overnight at room temperature
to hydrate. Then the mixture was heated
(above 40
°C), mixed with a vortex mixer
(GENIE2, Scientific Industries, Bohemia,
NY, USA) for several seconds and finally
cooled to room temperature.
To measure the particle size of the copo-
lymers in the aqueous solution, on the
day after the solution was prepared it was
filtered at 0.45
µm (Millex LCR 13
mm,
Merck Millipore, Darmstadt, Germany)
and measured with a dynamic light
scattering (DLS) analyzer (ELSZ-2000ZS,
Otsuka Electronics, Osaka, Japan). The
temperature was set to 5, 25, 40, 50
and 60
°C, the light source was a high
output semiconductor laser (640 nm) and
the scattering angle was set to 165
°C
for this analysis. The particle size was
analyzed using CONTIN analysis, and the
median diameter with interquartile range
was calculated from a histogram of the
particle size distribution.
To evaluate the dispersion stability, the
copolymer morphology was examined in
the aqueous solution by transmission elec-
tron microscopy with the freeze-fracture
technique (FF-TEM). Using a copolymer
in which m:n was 7:3 (hereafter called
M-polymer) with the methods above,
aqueous solutions of 1, 10 and 90
wt%
were rapidly frozen by plunging into
liquid propane (less than -170
°C) a the
cryo preparation system (Leica EM CPC,
Leica Microsystems, Tokyo, Japan). Then
the freeze-replica preparing apparatus
(FR-7000
A, Hitachi High-Technologies,
Tokyo, Japan) was used to fracture the
frozen sample with a glass knife and a
replica film prepared by evaporating plati-
num and carbon on the fractured surface.
The replica film was cleaned with acetone
and water and then placed on a TEM grid
and dried for observation. The replica film
was observed using a transmission elec-
tron microscope (H-7650, Hitachi High-
Technologies, Tokyo, Japan) at an accel-
erating voltage of 100 kV.
Analysis of the internal structure
of the emulsifier particles
Using the same method as already de-
scribed, M-polymer aqueous solutions
of 1, 5, 10, 20, 30, 40, 50, 60, 70, 80
and 90 wt% were formulated. Structural
analysis of the M-polymer particles in the
aqueous solution was conducted using a
small-angle X-ray scattering system (SAX-
Sess, Anton-Paar, Graz, Austria) at 25°C.
The apparatus was operated at 40 kV
and 50
mA using Cu-Kα X-rays (wave-
length 0.154
nm). The scattering curve
was obtained by plotting the scattering
intensity against the scattering vector
(q). The background contributions from
a capillary cell and solvent were sub-
tracted. The absolute scattering intensity
was calibrated using water as a secondary
standard. Using the obtained scattering
curve, SasView (version 4.1.2) was used
for model fitting analysis. In addition, for
determination of liquid crystal phase, the
q ratio of a scattering curve was esti-
mated.
Investigation of the new
emulsification mechanism for DIY
Preparation of the emulsions
We used M-polymer and three oils of dif-
ferent structure: squalane (Nikko chemi-
cals, Tokyo, Japan), caprylic/capric triglyc-
eride (BASF, Ludwigshafen, Germany),
and dimethicone (Shin-Etsu Chemical,
Figure 1 Concept of the emulsifier design
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,ǇĚƌŽƉŚŽďŝĐ
ŐƌŽƵƉ

SCIENTIFIC
PAPER
Tokyo, Japan). To prepare an oil-in-water
(O/W) emulsion with a composition of M-
polymer 1
wt%, oil 65
wt% and purified
water 34
wt%, the oil was added to the
M-polymer aqueous solutions and mixed
for 5 minutes at 30,000 rpm with a hand
homogenizer (NS360D, Microtec, Chiba,
Japan). Immediately after preparation, the
emulsion was observed visually for separa-
tion of the oil and water phases to evalu-
ate if an emulsion had been successfully
prepared.
Emulsion properties of
the new emulsifier particles
Emulsions formulated using the same
method as previously described were
stored at 20 °C for 1 day and for 3 months
at 5, 20 and 50 °C. The oil droplets were
observed at a magnification of 400x using
an optical microscope (BX63, Olympus,
Tokyo, Japan) to evaluate the emulsion
stability. One hundred oil droplets were
randomly selected from each sample
stored for 1 day at 20 °C and for 3 months
at 5, 20, and 50
°C. Using imaging soft-
ware (cellSens, Olympus, Tokyo, Japan), a
histogram of the oil droplet size distribu-
tion was made and the median diameter
and interquartile range were calculated.
Additionally, M-polymer and oil (squalane
or dimethicone) were used with the same
method as already described to formu-
late emulsions consisting of M-polymer 1
wt%, oil 30 wt%, and water 69 wt%. To
evaluate the morphology of M-polymer
in emulsions as a function of oil type, the
oil-water interface of the emulsion was
observed by FF-TEM, as described above.
Comparison with conventional
technology - emulsification potential
Emulsions were formulated with M-poly-
mer, a mixed humectant of 1,3-BG and
glycerin in a 1:1 ratio and 11 types of oil
with different structures. The emulsifier
and humectant were mixed as an aqueous
solution at room temperature, and the oil
was added and mixed using a home cook-
ing mixing instrument (HB-1230, Had-
inEEon, Shenzhen, China) for 30 seconds
at high speed. To test the feasibility of
emulsion-making at home, a formulation
of M-polymer 1 wt%, oil 65 wt%, humec-
tant 12 wt% and water 22 wt%, which is
difficult to prepare as a DIY formulation,
was tested. For comparison, a polyoxyeth-
ylene ester-ether type nonionic surfactant
(POE surfactant) and acrylic acid meth-
acrylic acid copolymer (AAMA polymer)
were chosen as conventional emulsifiers,
and emulsions were formulated under the
same conditions. Immediately after pre-
paring the emulsion, separation of the oil
and water phases was assessed visually to
evaluate if an emulsion had been success-
fully prepared.
Comparison with the conventional
technology – texture and cosmetic
film structure
As a typical composition for actual use,
2
wt% M-polymer, 10
wt% 1,3-BG,
10
wt% glycerin and 20
wt% oil (squal-
ane, caprylic/capric triglyceride, or di-
methicone) were used to formulate an
emulsion according to the same methods
as described above. A thickener and a
neutralizer were used to adjust the fluid-
ity to the same level and pH 6.0 - 7.0. For
comparison, the POE surfactant and the
AAMA polymer were chosen as conven-
tional emulsifiers, and emulsions were for-
mulated under the same conditions. Nine
expert panelists evaluated the formulated
emulsions for factors such as stickiness
and smoothness when applied.
Additionally, several drops of the formu-
lated emulsion were dropped on a hy-
drophobic substrate (APS01, Matsunami
glass, Osaka, Japan) that had been washed
with a facial cleanser, and after spread-
ing evenly using a 76.2
µm thick doctor
blade (Yoshimitsu Seiki, Tokyo, Japan), the
applied surface was stored at an angle of
45 ° at 35 °C for 2 hours to prepare a cos-
metic membrane formed by the residue
remaining after evaporation of water and
volatile substances. In this experiment the
cosmetic membrane consisted of humec-
tants, oil, thickener and M-polymer. The
component distribution of the cosmetic
membrane was examined using a Raman
microscope (alpha300, WITec, Ulm, Ger-
many) and Raman scattering was examined
with a spectrometer (UHTS300, WITec,
Ulm, Germany). A spectro camera (New-
ton DU970N-BV, ANDOR, Belfast, UK) was
used as the detector, and imaging of the
cosmetic membrane cross-section was
generated using basis analysis in imaging
software (Project FIVE, WITec, Ulm, Ger-
many). The image analysis was based on
the fitting algorithm of the Raman spectra
of the components. The resolution in the
x- and z-directions was 500
nm
/
pixel and
750 nm / pixel, respectively.
Examination of effectiveness for DIY
To evaluate the DIY procedure, 21 adult
male and female subjects with no back-
ground in cosmetic formulation who were
interested in experiencing DIY cosmetics
were recruited. The subjects selected ei-
ther a cleansing cream, cleansing milk,
milky lotion or cream. Then they designed
their formula using M-polymer 1 wt%, six
kinds of humectants with the water phase
in the 0 - 34 wt% range, and 16 types of oil
in the 1 - 65 wt% range. To formulate their
emulsion, they used the same methods as
Figure 2 Chemical structure of a new polymer emulsifier [15]
We synthesized copolymers with a main chain containing hydrophilic and hydrophobic groups.
We selected methoxypolyethylene glycol monomethacrylate for the hydrophilic group monomer and
glyceryl diisostearate methacrylate for the hydrophobic group monomer.
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those used for comparison of the emul-
sification potential and a home kitchen
mixing appliance to mix the water and
oil phases. The satisfaction was scored in
five grades for difficulty of the steps, flex-
ibility of customization, experience of DIY
cosmetics, and willingness to repeat the
experience. Additionally, it was checked if
the emulsifications were successful as de-
scribed above. This survey was conducted
with the approval of the ethical review
board of POLA Chemical Industries, Inc.
(approval number: 2020-F-065).
RESULTS
Designing an emulsifier particle
for a new emulsion function
Synthesis of a new emulsifier
To synthesize the new copolymer emulsi-
fier, we chose isostearic acid for the hydro-
phobic group, polyethylene glycol (PEG)
for the hydrophilic group, and methacryl-
ic acid for the main chain and designed
an emulsifier with the hydrophobic and
hydrophilic groups on one main chain
(Figure 2) [15]. We synthesized emulsi-
fiers with weight ratios of the hydrophilic
to the hydrophobic group of 5:5, 6:4, 7:3,
and 8:2 (Table I ).
Formulation of the new emulsifier
aqueous solution and evaluation
of the dispersion properties
We formulated copolymer aqueous solu-
tions as described in the EXPERIMENTAL
section. The m:n 5:5 copolymer aqueous
solution was suspended and thus removed
from the analysis. When the m:n 6:4 or m:n
7:3 (M-polymer) copolymer aqueous solu-
tion was formulated at 25
°C, the median
diameter (interquartile range) of particles
in the solution was 11.7 nm (9.6 -15.1) and
12.4 nm (10.0 -16.2), respectively, and the
particle size distributions was monophasic
(Figure 3a-b). However, when the copo-
lymer aqueous solution was formulated at
m:n 8:2, no data could be obtained. Ad-
ditionally, the particle size distributions of
the copolymer aqueous solution with m:n
6:4 and the M-polymer (m:n 7:3) aqueous
solution were observed at 5, 40, 50 and
60
°C. All results showed monophasic dis-
tributions with a peak at 10 - 20 nm, except
for m:n 6:4 at 60 °C, where it was bimodal
with a peak at around 21 nm and another
at around 260 nm. Furthermore, when 1,3-
BG or glycerin was added as a humectant
Table I Weight Average Molecular Weight and
Polydispersity of the Newly Designed Emulsifiers
Figure 3 Particle size distribution of 1 wt% new emulsifier aqueous solutions at different temperatures
measured by dynamic light scattering. (a) When the hydrophilic and hydrophobic group weight ratio
(m:n) was 6:4 at 60 °C, the particle size distribution in the new emulsifier solution was bimodal (red
arrow head). (b) At m : n 7 : 3 (M-polymer), the median diameter (interquartile range) of particles in
the solution was 12.4 nm (10.0 -16.2) and the particle size distribution monophasic. The particle size
distribution of the M-polymer aqueous solution was monophasic at temperatures of 5, 25, 40, 50
and 60 °C and with the humectants 1,3-butanediol (1,3-BG) or glycerin (29.7 wt%).
Figure 4 Freeze fracture-transmission electron microscopy images of the new emulsifier at different
concentrations [15]
The M-polymer particles were observed in (a) 1 wt% and (b) 10 wt% M-polymer aqueous solutions
(white arrows). No particles were observed in (c) 90 wt% M-polymer aqueous solution and a mesh-
patterned aggregate structure was observed.
M-polymer: New emulsifier with a hydrophilic to hydrophobic group weight ratio of 7:3.
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0
5
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1 10 100 1000
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WĂƌƚŝĐůĞƐŝǌĞͬŶŵ
ŵ͗Ŷсϲ͗ϰ ;ďͿŵ͗Ŷсϳ͗ϯϭ
ZĞůĂƚŝǀĞĨƌĞƋƵĞŶĐǇŝŶƚĞŶƐŝƚǇ
0
5
10
1 10 100 1000
5℃
25℃
ŵ͗Ŷ͗tĞŝŐŚƚƌĂƚŝŽŽĨƚŚĞŚǇĚƌŽƉŚŝůŝĐĂŶĚŚǇĚƌŽƉŚŽďŝĐŐƌŽƵƉƐ
Dǁ͗tĞŝŐŚƚĂǀĞƌĂŐĞŵŽůĞĐƵůĂƌǁĞŝŐŚƚ
DŶ͗EƵŵďĞƌĂǀĞƌĂŐĞŵŽůĞĐƵůĂƌǁĞŝŐŚƚ
dĂďůĞI: tĞŝŐŚƚǀĞƌĂŐĞDŽůĞĐƵůĂƌtĞŝŐŚƚĂŶĚ
WŽůǇĚŝƐƉĞƌƐŝƚǇ ŽĨƚŚĞ EĞǁůǇ ĞƐŝŐŶĞĚŵƵůƐŝĨŝĞƌƐ
ŵ͗Ŷ Dǁ DǁͬDŶ
ϴ͗Ϯ ϱϵ͕ϳϬϬ Ϯ͘ϰϬ
ϳ͗ϯ ϲϯ͕ϱϬϬ Ϯ͘ϱϬ
ϲ͗ϰ ϲϰ͕ϬϬϬ Ϯ͘ϯϮ
ϱ͗ϱ ϱϵ͕ϬϬϬ Ϯ͘ϮϬ
10 100 1000
5℃
25℃
ƐŝǌĞĚŝƐƚƌŝďƵƚŝŽŶŽĨϭǁƚйŶĞǁĞŵƵůƐŝĨŝĞƌĂƋƵĞŽƵƐƐŽůƵƚŝŽŶƐĂƚĚŝĨĨĞƌĞŶƚƚĞŵƉĞƌĂƚƵƌĞƐŵĞĂƐƵƌĞĚďǇĚǇŶĂŵŝĐ
ŽƉŚŝůŝĐĂŶĚŚǇĚƌŽƉŚŽďŝĐŐƌŽƵƉǁĞŝŐŚƚƌĂƚŝŽ;ŵ͗ŶͿǁĂƐϲ͗ϰĂƚϲϬ℃͕ƚŚĞƉĂƌƚŝĐůĞƐŝǌĞĚŝƐƚƌŝďƵƚŝŽŶŝŶƚŚĞŶĞǁ
ǁĂƐďŝŵŽĚĂů;ƌĞĚĂƌƌŽǁŚĞĂĚͿ͘;ďͿƚŵ͗Ŷϳ͗ϯ;DͲƉŽůǇŵĞƌͿ͕ƚŚĞŵĞĚŝĂŶĚŝĂŵĞƚĞƌ;ŝŶƚĞƌƋƵĂƌƚŝůĞƌĂŶŐĞͿŽĨ
ƵƚŝŽŶǁĂƐϭϮ͘ϰŶŵ;ϭϬ͘ϬͲϭϲ͘ϮͿĂŶĚƚŚĞƉĂƌƚŝĐůĞƐŝǌĞĚŝƐƚƌŝďƵƚŝŽŶŵŽŶŽƉŚĂƐŝĐ͘dŚĞƉĂƌƚŝĐůĞƐŝǌĞĚŝƐƚƌŝďƵƚŝŽŶŽĨ
ƵĞŽƵƐƐŽůƵƚŝŽŶǁĂƐŵŽŶŽƉŚĂƐŝĐĂƚƚĞŵƉĞƌĂƚƵƌĞƐŽĨϱ͕Ϯϱ͕ϰϬ͕ϱϬĂŶĚϲϬ℃ ĂŶĚǁŝƚŚƚŚĞŚƵŵĞĐƚĂŶƚƐϭ͕ϯͲ
ͿŽƌŐůǇĐĞƌŝŶ;Ϯϵ͘ϳǁƚйͿ͘
;ďͿŵ͗Ŷсϳ͗ϯϭ
ZĞůĂƚŝǀĞĨƌĞƋƵĞŶĐǇŝŶƚĞŶƐŝƚǇ
0
5
10
1 10 100 1000
5℃
25℃
to the copolymer aqueous solutions, the
particle size distributions at 25
°C were
both monophasic with a peak at 8 - 12 nm
(Figure 3a-b).
When M-polymer aqueous solutions at
1
wt% and 10
wt% were measured using
FF-TEM, the size of the M-polymer particles
agreed with the dynamic light scattering
measurements (Figure 4a-b) [15]. Howev-
er, in the 90 wt% M-polymer aqueous solu-
tion instead of particles a mesh-patterned
structure was observed (Figure 4c).

SCIENTIFIC
PAPER
polymer and the following oils of different
structure: squalane, caprylic/capric triglyc-
eride and dimethicone.
(see the EXPERIMENTAL section)
Emulsification properties of the new
emulsifier particles
After the emulsion obtained with squal-
ane (see the EXPERIMENTAL section) had
been stored for 1 day at 20 °C and for 3
months at 5, 20 and 50 °C, the oil drop-
let median diameters (interquartile range)
were 9.9
µm (5.8-12.8), 9.8
µm (7.1-
13.7), 9.4
µm (5.6-14.9) and 10.5
µm
(7.5-13.2), respectively. The oil size
showed no significant change over time
at each temperature. Similarly, samples
obtained using caprylic/capric triglycer-
Figure 5 Scattering curves of the new emulsifier
aqueoussolutionsusingsmallangleX-rayscattering
(SAXS).AfterformulatingtheM-polymersolutions,
SasView (version 4.1.2) was used for model fitting
analysis. The scattering curves of 1 and 5 wt%
M-polymer solutions matched with a core shell
type model. Additionally, at over 50 wt% aqueous
solution, the obtained scattering curve had two
peaks (red arrows heads) and we found the scat-
teringvector(q)ratioofthepeakswas1 : 1.7to1.9.
M-Polymer: New emulsifier with hydrophilic and
hydrophobic group weight ratio 7:3
Intensity: Scattering Intensity
Analysis of the internal structure of
the emulsifier particles
Model fitting analysis of the scattering
curves obtained by SAXS measurements re-
vealed scattering curves of M-polymers in 1
and 5 wt% aqueous solution that matched
those of a core-shell type model that has a
core inside and a shell outside. The core size
of the M-polymer particles in 1
wt% and
5
wt% aqueous solutions was 4.4
nm and
3.4 nm, respectively, and the shell thickness
1.1
nm and 1.2
nm, respectively. At over
50
wt% aqueous solution, the obtained
scattering curve had two peaks and the q
ratio of the peaks was 1:1.7 to 1.9, which
is characteristic of a discontinuous micellar
cubic phase (Figure 5).
Investigation of the new
emulsification structure
Production of emulsions
O/W type emulsions were successfully
prepared with all formulations using M-
Figure 6 Optical microscope observation images and size distribution of the emulsion of droplets
An emulsion was prepared with 1 wt% M-polymer, 65 wt% oil (squalane,caprylic / capric triglyceride
or dimethicone) and 34 wt% water. To evaluate the stability of thec emulsion, the samples were stored
at 20 °C for 1 day and then for 3 months at 5, 20, and 50 °C and the change in median diameter of
the oil droplets was measured. The oil droplet diameters showed no change over time under each
temperature condition. The scale bar shows 20 µm.
M-polymer: New emulsifier with a 7 : 3 weight ration of hydrophilic and hydrophobic parts

ventional emulsifiers POE surfactant and
AAMA-polymer and 11 different oils (see
the EXPERIMENTAL section). The POE sur-
factant could not emulsify 8 of the 11 oils,
and the AAMA polymer emulsified none
the oils, as shown by separation of the oil
to the top layer (Table II, Figure 8a-b).
In contrast, M-polymer could emulsify all
compositions successfully without the oil
phase separating (Table II, Figure 8c).
The emulsion formulated with M-polymer
(the EXPERIMENTAL section) was com-
pared with emulsions formulated with the
conventional emulsifiers, and the feel was
evaluated. Eight out of nine expert pan-
elists evaluated the M-polymer emulsion
as less sticky and smoother. The results
of cosmetic membrane structure imag-
ing showed that the membrane structure
of the emulsions prepared with the con-
ventional emulsifiers had a mix of oil and
humectant, and the emulsifier was the
oil/humectant interface (Figure 9a-b). In
contrast, the membrane structure analysis
of the emulsion formulated with M-poly-
mer showed that the humectant was on
the base side and the oil on the surface,
forming a 2-layer structure (Figure 9c).
Examination of the effectiveness for DIY
All subjects who participated in the DIY
cosmetics study were successful in for-
mulating their designed emulsion. Addi-
tionally, all subjects said they were ‘very
satisfied’ with the freedom choosing in-
gredients. Moreover, 20 of 21 subjects
answered that they were ‘very satisfied’
or ‘satisfied’ with the difficulty mixing the
ingredients, their satisfaction with the DIY
cosmetic experience and their willingness
ide or dimethicone showed no significant
change over time at each temperature
(Figure 6). As shown in Figure 6, emul-
sification was possible regardless of the
oil type.
An emulsion was formulated (see the
EXPERIMENTALsection)withM-polymer
at 1 wt%, the oil (squalane or dimethi-
cone) at 30 wt%, and water at 69 wt%.
The oil-water interface of the emulsions
was observed using FF-TEM. In the squal-
ane emulsion, bumps of several nano-
meters were observed on the oil surface
(Figure 7a). In contrast, in the emul-
sion with dimethicone, granular parti-
cles of a few nanometers were observed
in the water phase near the oil-water
interface instead of on the oil surface
(Figure 7b).
Comparison with
conventional technology
To compare the effectiveness of the new
emulsifier and the conventional technol-
ogy, we prepared emulsions with the con-
Figure 7 Freeze fracture-transmission electron microscopy (FF-TEM) images of the state of M-polymer
particles at the oil-water interface
An emulsion was prepared with M-polymer at 1 wt%, oil at 30 wt%, and water at 69 wt% and replica
films of the freeze-fractured samples were observed using FF-TEM. (a) Emulsion with squalene: Bumps
of several nanometers were observed on the fractured surface. (b) Emulsion with dimethicone: The
fracturedsurfaceoftheoilwassmoothandparticlesofafewnanometerswereobservedinthewaterphase.
M-Polymer: New emulsifier with a hydrophilic/hydrophobic group weight ratio of 7:3.
Figure 8 Photographs of the exterior of emulsions made using conventional emulsifiers and the new
emulsifier. Emulsions with compositions of 12 wt% humectants (1,3-BG and glycerin in a 1:1 ratio)
and 65 wt% oil were prepared using (a) POE surfactant, (b) AAMA polymer and (c) M-polymer and
pictures taken immediately after emulsion preparation. The conventional emulsifiers could not emulsify
the oils, whereas M-polymer emulsified all the oils.
POE surfactant: Polyoxyethylene ester-ether type nonionic surfactant
AAMA polymer: Acrylic acid methacrylic acid copolymer
M-Polymer: New emulsifier with a hydrophilic:hydrophobic group weight ratio of 7:3
ŝŐƵƌĞϴ͗WŚŽƚŽŐƌĂƉŚƐŽĨƚŚĞĞǆƚĞƌŝŽƌŽĨĞŵƵůƐŝŽŶƐŵĂĚĞƵƐŝŶŐĐŽŶǀĞŶƚŝŽŶĂůĞŵƵůƐŝ
ƚŚĞŶĞǁĞŵƵůƐŝĨŝĞƌ͘
ŵƵůƐŝŽŶƐǁŝƚŚĐŽŵƉŽƐŝƚŝŽŶƐŽĨϭϮǁƚйŚƵŵĞĐƚĂŶƚƐ;ϭ͕ϯͲ'ĂŶĚŐůǇĐĞƌŝŶŝŶĂϭ͗ϭƌĂƚŝŽͿĂŶĚ
ǁĞƌĞƉƌĞƉĂƌĞĚƵƐŝŶŐ;ĂͿWKƐƵƌĨĂĐƚĂŶƚ͕;ďͿDƉŽůǇŵĞƌĂŶĚ;ĐͿDͲƉŽůǇŵĞƌĂŶĚƉŝĐƚƵƌĞƐ
ŝŵŵĞĚŝĂƚĞůǇĂĨƚĞƌĞŵƵůƐŝŽŶƉƌĞƉĂƌĂƚŝŽŶ͘dŚĞĐŽŶǀĞŶƚŝŽŶĂůĞŵƵůƐŝĨŝĞƌƐĐŽƵůĚŶŽƚĞŵƵůƐŝĨǇƚŚĞ
ǁŚĞƌĞĂƐDͲƉŽůǇŵĞƌĞŵƵůƐŝĨŝĞĚĂůůƚŚĞŽŝůƐ͘
WKƐƵƌĨĂĐƚĂŶƚ͗WŽůǇŽǆǇĞƚŚǇůĞŶĞ ĞƐƚĞƌͲĞƚŚĞƌƚǇƉĞŶŽŶŝŽŶŝĐƐƵƌĨĂĐƚĂŶƚ
DƉŽůǇŵĞƌ͗ĐƌǇůŝĐĂĐŝĚŵĞƚŚĂĐƌǇůŝĐ ĂĐŝĚĐŽƉŽůǇŵĞƌ
;ĂͿ ;ďͿ ;ĐͿ
KŝůƉŚĂƐĞ
ŵƵůƐŝŽŶƉŚĂƐĞ

SCIENTIFIC
PAPER
to repeat the experience. However, 16 of
21 participants felt it was difficult to de-
sign the composition (Table III).
DISCUSSION
In this study we developed a new emulsi-
fier that allows the easy preparation of
emulsions and customization of the com-
position and the feel of cosmetics in DIY
projects. The new emulsifier needed to
have the ability to form smaller particles
than the oil particles and to be uniformly
dispersed at 5
-
50
°C without aggrega-
tion. Thus, we designed a new emulsifier
with both hydrophobic groups, which as-
semble in water to spontaneously form
nanoparticles, and hydrophilic groups
[14]. For the hydrophobic group, isostearic
acid [16] was chosen because it hardly
changes in molecular fluidity, character-
istics, and viscosity in home environment
temperature ranges. Also, it is so hydro-
phobic that it easily form nanoparticles
in water. For the hydrophilic group, we
chose PEG [17-18] for its steric repulsion
characteristic that prevents the emulsifier
particles from aggregating.
The particle size distribution of 1
wt%
aqueous solutions of the new copolymer
with m:n ratios of 5:5, 6:4, 7:3 and 8:2
was evaluated. The suspension obtained
with the 5:5 copolymer suggests that it
formed aggregates. The fact that the 6:4
copolymer particle size distribution was
split into two peaks at 60 °C suggests that
some particles aggregated at this temper-
ature. No data could be obtained for the
8:2 copolymer particle size distribution,
suggesting that no particles were formed
because the increase in the ratio of the hy-
drophilic to hydrophobic group enhanced
dissolution in water. On the other hand,
we found that M-polymer (m : n = 7 : 3)
was monophasic at a temperature of
25
°C with a peak at 12.4
nm. We also
found that M-polymer kept its monopha-
sic peak at temperatures of 5, 40, 50 and
60 °C, as well as when 1,3-BG or glycerin
was included. Thus, we chose M-polymer
as our new emulsifier for DIY use.
The results of FF-TEM analysis showed
that M-polymer formed particles about
10 to 40 nm in size at 1 wt% or 10 wt%
aqueous solution. On the other hand, no
particles were observed at a concentra-
tion of 90 wt%. Furthermore, model fit-
ting using SAXS suggested that core-shell
type particles that have the hydrophobic
DͲWŽůǇŵĞƌ͗EĞǁĞŵƵůƐŝĨŝĞƌǁŝƚŚĂŚǇĚƌŽƉŚŝůŝĐ͗ŚǇĚƌŽƉŚŽďŝĐ ŐƌŽƵƉǁĞŝŐŚƚƌĂƚŝŽŽĨϳ͗ϯ
dĂďůĞ//͗ ǀĂůƵĂƚŝŽŶŽĨŵƵůƐŝĨŝĐĂƚŝŽŶĨĨŝĐĂĐǇŝŶĂ/zŽŵƉŽƐŝƚŝŽŶ
KŝůƚǇƉĞ WKƐƵƌĨĂĐƚĂŶƚ DƉŽůǇŵĞƌ DƉŽůǇŵĞƌ
EŽŶƉŽůĂƌ ŚǇĚƌŽĐĂƌďŽŶƐ
^ƋƵĂůĂŶĞ ʹ ʹ 〇
DŝŶĞƌĂůŽŝů 〇 ʹ 〇
WŽůĂƌŚǇĚƌŽĐĂƌďŽŶƐ
/ƐŽƐƚĞĂƌŝĐ ĂĐŝĚ ʹ ʹ 〇
KůĞŝĐĂĐŝĚ ʹ ʹ 〇
ŝŶŽůĞŝĐĂĐŝĚ ʹ ʹ 〇
ŝŶŽůĞŶŝĐĂĐŝĚ ʹ ʹ 〇
ĂƉƌǇůŝĐͬĐĂƉƌŝĐ ƚƌŝŐůǇĐĞƌŝĚĞ ʹ ʹ 〇
dƌŝĞƚŚǇůŚĞǆĂŶŽŝŶ 〇 ʹ 〇
KůĞĂĞƵƌŽƉĂĞĂ ;ŽůŝǀĞͿ ĨƌƵŝƚŽŝů 〇 ʹ 〇
^ŝůŝĐŽŶĞŽŝů
WŽůǇ;ĚŝŵĞƚŚǇůƐŝůŽǆĂŶĞͿ ʹ ʹ 〇
ǇĐůŽƉĞŶƚĂƐŝůŽǆĂŶĞ ʹ ʹ 〇
〇͗ŵƵůƐŝĨŝĐĂƚŝŽŶƐƵĐĐĞƐƐĨƵů͕×͗ŵƵůƐŝĨŝĐĂƚŝŽŶĨĂŝůĞĚ
DͲWŽůǇŵĞƌ͗EĞǁĞŵƵůƐŝĨŝĞƌǁŝƚŚĂŚǇĚƌŽƉŚŝůŝĐ͗ŚǇĚƌŽƉŚŽďŝĐŐƌŽƵƉǁĞŝŐŚƚƌĂƚŝŽŽĨϳ͗ϯ
dĂďůĞ//͗ ǀĂůƵĂƚŝŽŶŽĨŵƵůƐŝĨŝĐĂƚŝŽŶĨĨŝĐĂĐǇŝŶĂ/zŽŵƉŽƐŝƚŝŽŶ
KŝůƚǇƉĞ WKƐƵƌĨĂĐƚĂŶƚ DƉŽůǇŵĞƌ DƉŽůǇŵĞƌ
EŽŶƉŽůĂƌ ŚǇĚƌŽĐĂƌďŽŶƐ
^ƋƵĂůĂŶĞ ʹ ʹ 〇
DŝŶĞƌĂůŽŝů 〇 ʹ 〇
WŽůĂƌŚǇĚƌŽĐĂƌďŽŶƐ
/ƐŽƐƚĞĂƌŝĐ ĂĐŝĚ ʹ ʹ 〇
KůĞŝĐĂĐŝĚ ʹ ʹ 〇
ŝŶŽůĞŝĐĂĐŝĚ ʹ ʹ 〇
ŝŶŽůĞŶŝĐĂĐŝĚ ʹ ʹ 〇
ĂƉƌǇůŝĐͬĐĂƉƌŝĐ ƚƌŝŐůǇĐĞƌŝĚĞ ʹ ʹ 〇
dƌŝĞƚŚǇůŚĞǆĂŶŽŝŶ 〇 ʹ 〇
KůĞĂĞƵƌŽƉĂĞĂ ;ŽůŝǀĞͿ ĨƌƵŝƚŽŝů 〇 ʹ 〇
^ŝůŝĐŽŶĞŽŝů
WŽůǇ;ĚŝŵĞƚŚǇůƐŝůŽǆĂŶĞͿ ʹ ʹ 〇
ǇĐůŽƉĞŶƚĂƐŝůŽǆĂŶĞ ʹ ʹ 〇
〇͗ŵƵůƐŝĨŝĐĂƚŝŽŶƐƵĐĐĞƐƐĨƵů͕×͗ŵƵůƐŝĨŝĐĂƚŝŽŶĨĂŝůĞĚ
DͲWŽůǇŵĞƌ͗EĞǁĞŵƵůƐŝĨŝĞƌǁŝƚŚĂŚǇĚƌŽƉŚŝůŝĐ͗ŚǇĚƌŽƉŚŽďŝĐŐƌŽƵƉǁĞŝŐŚƚƌĂƚŝŽŽĨϳ͗ϯ
Table II Evaluation of emulsification efficacy in a DIY composition
ŝŐƵƌĞϵ͗ZĂŵĂŶƐƉĞĐƚƌŽƐĐŽƉǇŝŵĂŐĞƐŽĨƚŚĞĐƌŽƐƐͲƐĞĐƚŝŽŶŽĨƚŚĞĐŽƐŵĞƚŝĐŵĞŵďƌĂŶĞ͘
dŚĞĐŽƐŵĞƚŝĐŵĞŵďƌĂŶĞƐƚƌƵĐƚƵƌĞŽĨƚŚĞĞŵƵůƐŝŽŶƐǁŝƚŚĐŽŶǀĞŶƚŝŽŶĂůĞŵƵůƐŝĨŝĞƌƐƐŚŽǁŶŝŶ;ĂͿWKƐƵƌĨĂĐƚĂŶƚƐ
ĂŶĚ;ďͿDƉŽůǇŵĞƌĐŽŶƚĂŝŶƐĂŵŝǆŽĨŽŝůĂŶĚŚƵŵĞĐƚĂŶƚƐĂŶĚƚŚĞŝŶƚĞƌĨĂĐĞĂŶĚŚƵŵĞĐƚĂŶƚƐĐŽŶƚĂŝŶ
ĞŵƵůƐŝĨŝĞƌƐ͘/ŶƚŚĞĐŽƐŵĞƚŝĐŵĞŵďƌĂŶĞƐƚƌƵĐƚƵƌĞǁŝƚŚ;ĐͿDͲƉŽůǇŵĞƌ͕ƚŚĞŚƵŵĞĐƚĂŶƚƐĂƌĞŽŶƚŚĞďĂƐĞƐŝĚĞĂŶĚ
ƚŚĞŽŝůƉŚĂƐĞŝƐŽŶƚŚĞƐƵƌĨĂĐĞ͕ĐƌĞĂƚŝŶŐĂϮͲůĂǇĞƌƐƚƌƵĐƚƵƌĞ͘dŚĞƐĐĂůĞďĂƌƌĞƉƌĞƐĞŶƚƐϭϬђŵ͘
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Figure 9 Raman spectroscopy images of the cross-section of the cosmetic membrane
The cosmetic membrane structure of the emulsions with conventional emulsifiers shown in(a) POE
surfactants and (b) AAMA polymer contains a mix of oil and humectants and the interface and
humectants contain emulsifiers. In the cosmetic membrane structure with (c) M-polymer, the
humectants are on the base side and the oil phase is on the surface, creating a 2-layer structure.
The scale bar represents 10 µm.
POE surfactant: Polyoxyethylene ester-ether type nonionic surfactant
AAMA polymer: Acrylic acid methacrylic acid copolymer
M-Polymer: New emulsifier with a hydrophilic:hydrophobic group weight ratio of 7:3

IFSCC_Magazine_0121_low.pdf

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