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Facial Expression Wrinkles and Their Relaxation by a Synthetic Peptide

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Abstract

Expression wrinkles form over time due to repeated facial movements such as smiling and frowning. They have an imprint on facial skin in areas such as the corner of the eyes, where they take the form of crow’s feet, the forehead and the glabella, where they appear as frown lines, and around the mouth, as marionette lines. In the study presented here, we recruited two sets of volunteers. An older group of 57 volunteers aged 50 to 65 years, and a group of eight younger volunteers aged 21–35 who were the biological daughters of eight of the older volunteers. Using VISIA CR, we took images of the volunteers in relaxed, angry and smiling mode to assess similarities in expression wrinkle patterns. In addition, the older volunteers were split into a placebo group and an active group who applied a formulation of 4% of a cosmetic product containing the peptide diaminobutyroyl benzylamide diacetate (DABBA) for four weeks. Wrinkles were assessed by image analysis, expert grading and Primoslite measurements. Our study found striking similarities in the facial wrinkle patterns of mothers with relaxed faces and daughters with angry or smiling faces. We found a decrease in visible wrinkles in the group of older volunteers applying DABBA. We created a facial map for graded wrinkles showing these changes. Volunteers using the active formulation showed significantly less wrinkle area and length on the forehead when frowning compared to the placebo group (p < 0.05).
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International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
https://doi.org/10.1007/s10989-020-10146-z
Facial Expression Wrinkles andTheir Relaxation byaSynthetic Peptide
RemoCampiche1 · FrancescaPascucci1· LilyJiang2· ThibaultVergne3· MarieCherel3· SarahGougeon3·
ElodiePréstat‑Marquis3· GhislainFrançois3· GuillaumeLaurent1· MathiasGempeler1
Accepted: 19 November 2020 / Published online: 29 November 2020
© The Author(s) 2020
Abstract
Expression wrinkles form over time due to repeated facial movements such as smiling and frowning. They have an imprint
on facial skin in areas such as the corner of the eyes, where they take the form of crow’s feet, the forehead and the glabella,
where they appear as frown lines, and around the mouth, as marionette lines. In the study presented here, we recruited two
sets of volunteers. An older group of 57 volunteers aged 50 to 65years, and a group of eight younger volunteers aged 21–35
who were the biological daughters of eight of the older volunteers. Using VISIA CR, we took images of the volunteers in
relaxed, angry and smiling mode to assess similarities in expression wrinkle patterns. In addition, the older volunteers were
split into a placebo group and an active group who applied a formulation of 4% of a cosmetic product containing the peptide
diaminobutyroyl benzylamide diacetate (DABBA) for four weeks. Wrinkles were assessed by image analysis, expert grading
and Primoslite measurements. Our study found striking similarities in the facial wrinkle patterns of mothers with relaxed faces
and daughters with angry or smiling faces. We found a decrease in visible wrinkles in the group of older volunteers apply-
ing DABBA. We created a facial map for graded wrinkles showing these changes. Volunteers using the active formulation
showed significantly less wrinkle area and length on the forehead when frowning compared to the placebo group (p < 0.05).
Keywords Skin· Expression wrinkles· Relaxation· Facial color map· Peptide
Introduction
A hallmark of aging skin is the formation of wrinkles. Facial
wrinkles appear as fine lines, creases or deep wrinkles (Bazin
and Leveque 2011; Zouboulis etal. 2019). They show as
frown lines on the forehead and the glabella, crow’s feet on
the eye’s lateral edge and as nasolabial folds, upper lip lines or
marionette lines around the mouth (Bazin and Doublet 2007).
Wrinkles occur either through chronological aging, or photo-
aging and are particularly pronounced in photo-exposed areas
of the body such as the face (Nkengne and Bertin 2013), neck
(Hatzis 2004) and hands (Jakubietz etal. 2008). Moreover,
various additional factors to which skin is exposed contribute
to aging and wrinkling. These factors are collectively termed
the skin aging exposome (Krutmann etal. 2017). With respect
to wrinkles, the skin aging exposome mainly works on the
degradation of skin tissue structures such as the extracellular
matrix comprising elastic fibres, collagen and hyaluronic acid
(structural changes) (Langton etal. 2010; Shin etal. 2019;
Watson and Griffiths 2005). However, particular wrinkle pat-
terns, such as crow’s feet, smile lines, or frown lines imprint
in the skin due to repeated facial movements and expressions
(mechanical changes) (Miyamoto and Hillebrand 2007; Hille-
brand etal. 2010). Facial movements occur via contraction of
facial muscles. Here, nerve endings release neurotransmitters
like acetylcholine which binds to the nicotinic acetylcholine
receptor on the muscle surface. This leads to depolarization
and the opening of sodium ion channels. Consequently, an
action potential propagates along the cell causing calcium
channels to open which stimulates contraction via additional
calcium release from the sarcoplasmatic reticulum (Kuo and
Ehrlich 2015). Expression or mimic wrinkles were termed type
3 wrinkles by Piérard etal. (2003). Hillebrand etal. showed
how expression lines turn into persistent wrinkles on the
crow’s feet area in an 8-year longitudinal study (Hillebrand
etal. 2010). Various studies have also dealt with the in silico
simulation of expressive wrinkle formation (Kuwazuru etal.
* Remo Campiche
remo.campiche@dsm.com
1 DSM Nutritional Products, Personal Care & Aroma,
Kaiseraugst, Switzerland
2 SGS Stephens Inc., Richardson, TX, USA
3 Newtone Technologies, Lyon, France
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1010 International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
1 3
2008; Zhao etal. 2020; Chavoshnejad etal. 2020), but the
topic remains poorly investigated despite the consensus that
facial mimic wrinkles eventually turn into persistent wrinkles,
a concept already proposed back in the 1980s for the forma-
tion of frown lines (Pierard and Lapiere 1989). In addition,
preliminary evidence suggests that facial mimic wrinkles form
similarly in genetically related individuals such as mothers
and daughters (Hillebrand etal. 2010). Facial wrinkles, and
expression or mimic wrinkles in particular, can be of cosmetic
concern when they become persistent (Nkengne etal. 2008;
Blume-Peytavi etal. 2016). There are aesthetic concerns,
which can cause psychological distress in many individuals
because they show their age, or look older than they are, which
is generally considered less attractive (Samson etal. 2010), but
there is also the fear of misconception by others, i.e. giving an
impression of being constantly angry or annoyed (Willis and
Todorov 2006; Tian and Xiao 2016; Song 1998). Along these
lines, there is evidence that facial expressions not only con-
vey emotions to the outside world, but also modulate human
emotions (Alam etal. 2008; Ekman etal. 1983; Izard 1990).
Due to these many reasons, the cosmetic relaxation of facial
mimic wrinkles has become a sought-after treatment in aes-
thetic dermatology (Becker-Wegerich etal. 2001). Botulinum
toxin A has established itself as the number one treatment in
this area (Franca etal. 2017). It reversibly paralyzes facial
expression muscles by inhibiting the release of acetylcholine
on nerve endings, thus preventing muscle depolarization. The
aesthetic physician injects the toxin subcutaneously to induce
muscle relaxation at distinct facial areas (Small 2014). In addi-
tion, non-invasive treatments using cosmetic peptides such as
diaminobutyroyl benzylamide diacetate (DABBA) have been
developed and have shown skin smoothing activity invivo
(Campiche etal. 2017) by mimicking botox-like activity. Since
movement of facial muscles furthers the imprint of expression
wrinkles, it is likely that suppression of facial muscle move-
ment may delay the imprint of expression wrinkles over time.
Therefore, patients have started using botulinum toxin A at an
earlier age, usually at lower dosages (Kinonen 2017).
In the study presented here, we investigated facial mimic
wrinkle formation in biological mothers and daughters. We
provide new evidence for the peptide DABBA and its abil-
ity to relax facial expression wrinkles and also introduce a
novel facial color map for wrinkle grading to investigate and
display the efficacy of anti-aging treatments.
Materials andMethods
Test Formulations
The test formulations consisted of a placebo formulation
and an active formulation of 4% of a commercial product
containing 2500ppm of DABBA (Trade name SYN®-AKE,
DSM Nutritional Products, Switzerland). The composition
of both placebo and active formulation is outlined in Table1.
Study Outline andVolunteers
This human study was conducted at SGS Stephens Inc. (for-
merly Thomas J. Stephens & Associates Inc.), Richardson,
TX, United States. The study took place between 8th July
and 22nd September 2019. This was a single centre, pla-
cebo-controlled, randomized, and double-blind study. This
study adhered to the guidelines of the declaration of Hel-
sinki. Volunteers gave their informed consent to participate
in the study. The study was approved by the local ethics
committee, IntegReview Institutional Review Board (IRB)
on 25th June 2019. IntegReview IRB, located in Austin,
Texas, is a duly constituted IRB under Title 21 CFR Parts 50
and 56. Volunteers were female: 54 Caucasian, 2 Asian, and
1 mixed race, ages 50 to 65years. For the wrinkle pattern
analysis, we recruited eight additional volunteers (7 Cau-
casian, 1 Asian) who were biologically related daughters
to eight of the volunteers. Subjects were in good general
health (physical, mental, and social well-being, not merely
the absence of disease/infirmity), according to a self-report,
and had Fitzpatrick skin type I–IV. The 8 daughters were
21–35years old and had no to a mild score for wrinkles on
the overall face (score of 0–3 according to a modified Grif-
fiths scale, where 0 = none and 9 = severe). Volunteers in
the 50–65 age group had mild to moderate scores (score of
3–6 according to a modified Griffiths scale, where 0 = none
Table 1 Composition of formulations used in this study
INCI Product A
(placebo)
Product B
(containing
DABBA)
Aqua 66.72 66.72
Disodium EDTA, Aqua 0.05 0.05
Xanthan gum 0.20 0.20
Propanediol 10.00 10.00
Cetearyl olivate, sorbitan olivate 3.00 3.00
Polysorbate 20 0.50 0.50
Stearic acid, palmitic acid 1.00 1.00
Isopropyl myristate 4.00 4.00
Isostearyl isostearate 4.00 4.00
Dicaprylyl ether 4.00 4.00
Phenoxyethanol, ethylhexylglycerin 1.00 1.00
Dimethicone 1.00 1.00
Hydroxyethyl acrylate/sodium acry-
loyldimethyl taurate copolymer
0.50 0.50
Glycerin, aqua, dipeptide diaminobu-
tyroyl benzylamide diacetate
– 4.00
Aqua 4.00
Sodium hydroxide, Aqua 0.03 0.03
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1011International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
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and 9 = severe) for wrinkles in at least 3 of the following
locations: Forehead, Crow’s feet, Nasolabial (nose to cor-
ners of mouth), Marionette (corners of mouth down to chin).
Subjects were willing not to use any foundation during the
course of the 4-week study (lip and eye color cosmetics were
allowed), not had any facial treatments in the past 6months,
and were willing to withhold all facial treatments during the
course of the study.
Exclusion criteria: Standard exclusion criteria were
applied. In brief: having known allergies to facial skin care
products; to be nursing, pregnant, or planning to become
pregnant according to self-report; a history of skin cancer
within the past 5 years; to be taking or have taken anti-aging
or skin lightening products, or other products or topical or
systemic medication known to affect skin aging or dyschro-
mia, during the two weeks before the study.
Assignment andApplication ofTest Formulations
Randomization: Validated software (RITA, Randomization
in Treatment Arms) was used to assign the test products to
the subgroups in a way that minimized potential imbalance
regarding age, phototype and ethnicity. Product A received
28 subjects. Product B received 29 subjects. Application:
Twice per day (morning and evening), after facial cleans-
ing, a nickel-sized amount (2mgcm−2) of test material was
applied over the entire face and allowed to absorb.
Image Acquisition
Subjects had images taken at the baseline and week 4. Sub-
jects acclimatized to ambient conditions within the clinic
for at least 15min before any photographs were taken. Digi-
tal images of each subject’s face were taken using VISIA
CR photo station (Canfield Imaging Systems, Fairfield, NJ,
USA) with a Canon Mark II digital SLR camera (Canon
Incorporated, Tokyo, Japan). Subjects adopted a neutral,
unsmiling expression for images taken from left, center, and
right views. At the baseline and week 4, an additional center
view image was taken with an “angry” frowning expression,
and an additional left view image was taken with a wide
smiling expression. In order to standardize this procedure as
much as possible, subjects were asked to assume as angry an
expression as possible and to assume as wide a smile as pos-
sible at both timepoints. A 48 patches color chart (Newtone
Technologies, Princeton, NJ, USA) was added to the system
and present on each image.
Primoslite Imaging Procedures
Primoslite imaging was performed on crow’s feet area. The
Primoslite 45 × 30mm system (GFMesstechnik GmbH,
Teltow, Germany) is a hand-held 3D imaging device for
assessing the microtopography of skin. The field of view is
45 × 30 × 20mm with resolution of 61 × 61 × 6µm. Primoslite
images were analyzed using the Primoslite wrinkle analysis
software built-in function (Widgerow, Jiang, and Calame
2018).
Clinical Grading ofEfficacy Parameters
At the baseline and week 4, subjects were evaluated for
wrinkles on the forehead, crow’s feet, nasolabial folds, mari-
onette area, and upper lip according to a modified Griffiths
scale (Griffiths etal. 1992).
Wrinkle Analysis ofAngry andSmiling Faces
andVisual Enhancement
In order to study the evolution of wrinkles over time, the
wrinkles were segmented using specific image processing
algorithms. Then the wrinkles were characterized by com-
puting their conspicuous surface area, length, and volume
(Campiche etal. 2019). In addition to the analysis, two types
of illustration were created to visualize the product effect.
First, a visual enhancement was performed to highlight
wrinkles using an image processing pretreatment. Then,
delta values of expert grading scores between the baseline
and week 4 were mapped on a 3D face using a pink facial
colour map (Newtone Technologies, Lyon, France) (Voegeli
etal. 2015, 2019).
Results
Mimic Wrinkle Patterns inDaughters’ Expressive
Faces Resemble Patterns inMothers’ Relaxed Faces
We had eight biological mother-daughter pairs who resem-
bled each other visually as exemplified by the three pairs
shown here (Fig.1a). We compared daughters’ expressive
faces with their respective mothers in relaxed facial mode
and found a similar wrinkle pattern on the glabella when
the daughters were frowning (Fig.1b). In addition, when
daughters were smiling, a similar pattern could be seen on
their crow’s feet compared to their mothers in relaxed mode
(Fig.1c).
DABBA can Smoothen Persistent Facial Mimic
Wrinkles
We have shown previously that DABBA can smoothen facial
wrinkles on both forehead and crow’s feet (Campiche etal.
2017). We confirm these data here using a slightly differ-
ent measuring device (Primoslite) which delivers metric
values. Looking at the crow’s feet of subjects in the 50 to
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1012 International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
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65 age group, we measured a reduction in wrinkle length
of -3.48mm after 2weeks (p < 0.05 vs baseline), and of
3.38mm after 4weeks (p = 0.094 vs baseline) (Fig.2a).
In addition, we measured a decrease in wrinkle area of
0.66 mm2 after 2weeks and of − 2.1 mm2 after 4weeks
(p = 0.086 vs baseline) (Fig.2b). The visible effects can be
seen in the examples in Fig.2c (volunteer #8) and Fig.2d
(volunteer #52) which both show the crow’s feet area at the
baseline and week 4 with decreased wrinkles at week 4.
Expert Grading ofFacial Wrinkles in‘Mothers’ Shows
aWrinkle Decrease After 4weeks ofDABBA Use
In addition to Primoslite measurements, we employed
expert grading to assess the visual improvement of facial
wrinkles when using a formulation containing DABBA.
The results of the expert grading are listed in Table2. We
found a change vs the baseline for the group using DABBA
of − 0.26 units (p < 0.01) for forehead wrinkles, and − 0.21
units (p < 0.01) for crow’s feet, − 0.16 units (p < 0.01) for
the nasolabial fold, and − 0.12 units (p < 0.05) for mari-
onette lines. In order to visualize the effect of the DABBA
formulation, we created a facial wrinkle grading map
(Fig.3) based on our facial colour mapping technology
(Voegeli etal. 2015). We drew this colour map with the
delta values of the active formulation and assigned a pink
colour gradient to the map. The darker the pink colour, the
greater the delta to baseline was and thus the greater the
effect of the formulation. This shows that the main activ-
ity of the formulation was in the area of the forehead. In
Fig. 1 Resemblance of daughters and mothers. a Side view of bio-
logical mother–daughter pairs participating in the study. They clearly
look alike. b) Frown lines on the glabella of the daughters’ angry face
look similar to the ones on the mother’s relaxed face. Frown lines
1 and 2 form in the daughter when she pulls an angry face and are
extended at the same place on the mothers’ relaxed face. The lower
panel shows a wrinkle enhancement highlighting the wrinkles pre-
sent in the standard images of the upper panel. c Crows’ feet on the
edge of the eye of the daughters’ smiling face look similar to the ones
on the mother’s relaxed face. Crows’ feet 1, 2, 3, and 4 form in the
daughter when she smiles and are extended at the same place on the
mothers’ relaxed face. The lower panel shows a wrinkle enhancement
highlighting the wrinkles present in the standard images of the upper
panel
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1013International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
1 3
Fig. 2 Smoothing of crows’ feet wrinkles by a formulation contain-
ing DABBA as measured by Primoslite. a Crows’ feet length was sig-
nificantly reduced after 2weeks and stays about the same at 4weeks.
*p < 0.05 vs baseline. b Crows’ feet area was reduced time-depend-
ently after 2 and 4 weeks. c Volunteer #8 showing visibly reduced
crows’ feet (arrow) at week 4 compared to the baseline. d Volunteer
#52 showing visibly reduced crow’s feet (arrow) at week 4 compared
to the baseline. Error bars represent standard error of the mean. Stu-
dent’s t-test for paired samples was used
Table 2 Result of expert
grading for facial wrinkles
Wilcoxon-signed rank test was used
SEM standard error of the mean
Wrinkle area Formulation Delta to baseline at
week 4 [units]
SEM p-value to baseline
Forehead Placebo − 0.11 0.05 0.11
4% DABBA − 0.26 0.06 < 0.01
Crow’s feet Placebo − 0.21 0.05 < 0.01
4% DABBA − 0.21 0.05 < 0.01
Nasolabial fold Placebo − 0.11 0.04 < 0.05
4% DABBA − 0.16 0.05 < 0.01
Marionette lines Placebo − 0.09 0.05 0.13
4% DABBA − 0.12 0.04 < 0.05
General aging signs (global face)
Hyperpigmentation Placebo − 0.14 0.05 < 0.05
4% DABBA − 0.29 0.07 < 0.01
Perceived age Placebo − 0.82 0.35 < 0.05
4% DABBA − 1.03 0.42 < 0.05
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1014 International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
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addition to wrinkles, hyperpigmentation and perceived age
were reduced, too (Table2).
The Peptide DABBA isAble toSuppress Mimic
Wrinkle Formation ontheForehead
We analysed the images of the volunteers using a wrinkle
segmentation method similar to the one described previously
(Campiche etal. 2019). We found a significant reduction in
mimic wrinkle formation on the glabella after four weeks in
the group using the DABBA formulation compared to the
group using the placebo formulation (Fig.4a, b). Specifically,
the wrinkle area was reduced by 13.52% (p < 0.05 vs placebo),
wrinkle length was reduced by 14% (p < 0.05 vs placebo), and
wrinkle volume was reduced by 13.59% (p = 0.076 vs placebo)
(Fig.4c). This reduction in wrinkle formation was visible as
shown by volunteer #70 using the active formulation whereas
in volunteer #43 using the placebo formulation it was not
(Fig.4d). In contrast to the forehead, we did not find a reduc-
tion in expressive wrinkle formation on the marionette lines
(smiling lines) at the corner of the mouth (Fig.5). Here, the
wrinkle area decreased in the DABBA group compared to the
placebo group by 2.9%, wrinkle length decreased by 3.9%,
and wrinkle volume decreased by 3.6% (all non-significant)
(Fig.5a - c). This decrease was slightly visible only in some
volunteers, e.g. in #26 (Fig.5d).
Discussion
Expression wrinkles become permanent wrinkles over
time (Miyamoto and Hillebrand 2007). They form dis-
tinct patterns such as horizontal frown lines on the
forehead, vertical lines on the glabella, and crow’s feet
radiating from the edges of the eyes (Hillebrand etal.
2010). We have conducted a study including biological
mother–daughter pairs to investigate the formation, or
cutaneous imprinting, of such expressive wrinkle patterns
further. The mother–daughter link is not only genetically
evident, but also deemed strong across cultures (Bojczyk
etal. 2010; Owen-Smith 2003). Thus, daughters share
similar ways of behaving and expressing themselves to
their mothers. In line with this, we found an intriguing
similarity between expression wrinkle patterns on the gla-
bella (Fig.1b) and the crow’s feet (Fig.1c) of daughters
pulling an angry face or smiling and these same wrin-
kle patterns in their respective mothers when showing a
relaxed face. This, together with previously published data
(Hillebrand etal. 2010), provides further evidence that
expression wrinkles turn into permanent wrinkles over
time, and that they can form similar patterns in individu-
als sharing a similar genetic and emotional background.
In view of this, avoiding or suppressing the formation of
expression wrinkles has become a widely used method
for counteracting permanent wrinkles or smoothening the
skin. While in aesthetic dermatology botulinum toxin A is
the vehicle of choice for smoothing permanent expression
wrinkles (Becker-Wegerich etal. 2001; Small 2014; Song
1998), the cosmetic industry has focused on topical, non-
invasive treatments (Campiche etal. 2017; Blanes-Mira
etal. 2002). We developed the peptide DABBA which
we showed was able to smooth frown lines and crow’s
feet invivo (Campiche etal. 2017). Here, we show fur-
ther evidence for this action on crow’s feet wrinkles using
Primoslite (Fig.2). To emphasize the visible anti-wrinkle
activity of the peptide, we performed expert grading on
Fig. 3 Facial color map of
changes in visible wrinkles as
assessed by expert grading for
the group using the DABBA
formulation. Front and side
view are shown. The corre-
sponding color map is displayed
on the right. The more intense
the color, the greater the
improvement in wrinkle severity
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1015International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
1 3
various areas of the face. There was a visible decrease
in wrinkle severity in these facial areas (Table2). The
greatest decrease was seen on the forehead, which is con-
sistent with previous findings for this peptide (Campiche
etal. 2017). In addition, perceived age and visible hyper-
pigmentation decreased, too; indeed, hyperpigmentation
decreased by double in the DABBA group compared to the
placebo group. Interestingly, we had previously found that
the peptide DABBA was able to modulate skin tone in the
facial skin of Asian volunteers (Seroul etal. 2020). Often
in cosmetic clinical studies we see that the placebo formu-
lation shows a benefit to skin hydration which influences
the appearance of skin from fine lines, small wrinkles up
to radiance and skin tone, possibly. Here the placebo even
yielded significance for some facial areas when assessed
by expert grading (Table2). However, the peptide formu-
lation still showed a better performance for most param-
eters which together with the other results presented here
suggests the peptide to additionally contribute to the for-
mulation’s activity. In order to better visualize the changes
in wrinkle severity, we chose to create a wrinkle grading
map (Fig.3) based on our previously published facial color
map technology (Voegeli etal. 2015). This technology
makes it possible to readily appreciate even small changes
in facial parameters such as skin hydration or skin surface
pH (Voegeli etal. 2019). In line with this, the changes in
facial wrinkles recorded by the expert graders can eas-
ily be appreciated on the wrinkle grading map. Since the
peptide DABBA is able to reduce expression wrinkles,
we investigated whether the ability to express was indeed
reduced in volunteers using a formulation containing 4%
of the commercial product after four weeks. Interestingly,
we found a significant decrease in wrinkle parameters on
the glabella for the active group compared to the placebo
group (Fig.4). However, no clear reduction was found
for marionette lines (smile lines) (Fig.5). We hypothesise
Fig. 4 Effect of DABBA on expression wrinkles on the glabella
of an angry face. a Significant decrease of expressive wrinkle area
compared to placebo. *p < 0.05 vs placebo. b Significant decrease of
expressive wrinkle length compared to placebo. *p < 0.05 vs placebo.
c Decrease of expressive wrinkle volume compared to placebo. d
Images of volunteer #43 using placebo formulation and volunteer #70
using DABBA formulation. There was a visible decrease in wrinkles
(blue segmentation) in volunteer #70 (lowerimages) which was not
seen in volunteer #43 (upperimages). Error bars represent standard
error of the mean. Student’s t-test for unpaired samples was used for
a, b, and c
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1016 International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
1 3
that this may be due to the thicker skin around the mouth
compared to the forehead, and that the thicker skin masked
the effect of the peptide in this area. We propose this as
substantiation that the peptide is able to modulate facial
expression invivo.
In conclusion, by using a mother-daughter compari-
son of expressed and relaxed faces, we have shown how
expression wrinkles imprint facial skin. We provide further
evidence using different methodologies that the peptide
diaminobutyroyl benzylamide diacetate helps smoothen
facial expression wrinkles, and thus counteracts visible
signs of aging. In addition, we present a novel facial colour
map based on expert grading for augmented visualization
of anti-wrinkle treatments.
Acknowledgements We would like to thank the study volunteers for
their participation. We would also like to acknowledge our colleagues
for fruitful discussions and comments throughout the project. The
peptide diaminobutyroyl benzylamide diacetate (DABBA) is marketed
by DSM Nutritional Products under the trade name SYN®-AKE.
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copy of this licence, visit http://creat iveco mmons .org/licen ses/by/4.0/.
Fig. 5 Effect of DABBA on expression wrinkles on the marionette
lines of a smiling face. a through c Only a very small decrease in
wrinkles was seen in the active group compared to the placebo group.
d Images of volunteer #17 using placebo formulation and volunteer
#26 using DABBA formulation. There was a small visible decrease in
wrinkles (blue segmentation) in volunteer #26 (lower images) which
was not seen in volunteer #17 (upper images). Error bars represent
standard error of the mean. Mann–Whitney U test was used for a and
b, Student’s t-test for unpaired samples was used for c
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1017International Journal of Peptide Research and Therapeutics (2021) 27:1009–1017
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... Wrinkles are partially due to excessive stressor stimuli on the facial muscles; unsurprisingly, it is usual to refer to them as "expression lines" [392][393][394]. ...
... In detail, a variation in membrane potential enhances the entry of Ca 2+ ions into the neuron. Then, once these ions enter the presynaptic terminal, the release of acetylcholine from the vesicles occurs [301,393]. SNAP-25 (synaptosomalassociated protein 25)-a membrane receptor protein associated with vesicles, involving the SNARE (soluble N-ethylmaleimide-sensitive factor activating protein receptor) complex-modulates this process, regulating the binding and fusion of the vesicles [301,395,396]. In detail, to form the SNARE complex, Munc-18 (mammalian uncoordinate-18) proteins must bind to syntaxin, followed by SNAP-25 and VAMP (vesicle-associated membrane protein, necessary for the synaptic vesicle's docking and fusion to the presynaptic membrane for the acetylcholine release) [397,398]. ...
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