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Clinical Interventions in Aging 2015:10 387–403
Clinical Interventions in Aging Dovepress
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ORIGINAL RESEARCH
open access to scientific and medical research
Open Access Full Text Article
http://dx.doi.org/10.2147/CIA.S69752
Mécano-Stimulation™ of the skin improves
sagging score and induces benecial functional
modication of the broblasts: clinical, biological,
and histological evaluations
Philippe Humbert1,2
Ferial Fanian1,2
Thomas Lihoreau1,2
Adeline Jeudy1,2
Ahmed Elkhyat1,2
Sophie Robin3
Carol Courderot-Masuyer3
Hélène Tauzin3
Christine Lafforgue1,2,4
Marek Haftek5
1Research and Studies Center on the
Integument (CERT), Department of
Dermatology, Clinical Investigation
Center (CIC 1431), Besançon
University Hospital; 2I NSE RM
UMR1098, FED4234 IBCT, University
of Franche-Comté, Besançon, France;
3SARL BIOEXIGENCE, Besançon,
France; 4Dermopharmacology and
Cosmetology Unit, University of Paris
Sud, France; 5University of Lyon 1,
EA4169, Experimental, clinical and
therapeutic aspects of the skin barrier
function, INSERM US7 – CNRS
UMS3453, Lyon, France
Background: Loss of mechanical tension appears to be the major factor underlying decreased
collagen synthesis in aged skin. Numerous in vitro studies have shown the impact of mechanical
forces on fibroblasts through mechanotransduction, which consists of the conversion of mechanical
signals to biochemical responses. Such responses are characterized by the modulation of gene
expression coding not only for extracellular matrix components (collagens, elastin, etc.) but also for
degradation enzymes (matrix metalloproteinases [MMPs]) and their inhibitors (tissue inhibitors of
metalloproteinases [TIMPs]). A new device providing a mechanical stimulation of the cutaneous
and subcutaneous tissue has been used in a simple, blinded, controlled, and randomized study.
Materials and methods: Thirty subjects (aged between 35 years and 50 years), with clinical
signs of skin sagging, were randomly assigned to have a treatment on hemiface. After a total of
24 sessions with Mécano-Stimulation™, biopsies were performed on the treated side and control
area for in vitro analysis (dosage of hyaluronic acid, elastin, type I collagen, MMP9; equivalent
dermis retraction; GlaSbox®; n=10) and electron microscopy (n=10). Furthermore, before and
after the treatment, clinical evaluations and self-assessment questionnaire were done.
Results: In vitro analysis showed increases in hyaluronic acid, elastin, type I collagen, and
MMP9 content along with an improvement of the migratory capacity of the fibroblasts on the
treated side. Electron microscopy evaluations showed a clear dermal remodeling in relation
with the activation of fibroblast activity. A significant improvement of different clinical signs
associated with skin aging and the satisfaction of the subjects were observed, correlated with
an improvement of the sagging cheek.
Conclusion: Mécano-Stimulation is a noninvasive and safe technique delivered by flaps
microbeats at various frequencies, which can significantly improve the skin trophicity. Results
observed with objective measurements, ie, in vitro assessments and electron microscopy, confirm
the firming and restructuring effect clinically observed.
Keywords: skin rejuvenation, skin sagging, mechanical stimulation, fibroblast synthesis
Introduction
Fibroblasts are the most abundant cells in the dermis. They play a major role in
synthesizing collagen and elastic fibers, glycoproteins, and proteoglycans in the
extracellular matrix. They also develop contractile forces required for matrix
remodeling and migration activities.
Skin aging is characterized by changes in the function and structure of the dermis.
With aging, the skin tends to become thinner, drier, and less elastic. Fibroblastic
dysfunction is one of the main markers of skin aging. This abnormal proliferation,
Correspondence: Philippe Humbert
Department of Dermatology, Research
and Studies Center on the Integument
(CERT), CHRU MINJOZ, 3 Boulevard
Fleming, Besançon, France
Email philippe.humbert@univ-fcomte.fr
Journal name: Clinical Interventions in Aging
Article Designation: Original Research
Year: 2015
Volume: 10
Running head verso: Humbert et al
Running head recto: Clinical, biological, and histological evaluations of Mécano-Stimulation™
DOI: http://dx.doi.org/10.2147/CIA.S69752
This article was published in the following Dove Press journal:
Clinical Interventions in Aging
2 February 2015
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differentiation, and synthesis of fibroblasts result in structural
and functional skin changes. Phenotypic studies about
wrinkle fibroblasts showed that the remarkable change in
mechanical properties of the skin with aging is indeed due
to reduced contractile capacity as well as reduced migration
activity of wrinkle fibroblasts.1
Early in 1911, Dr Lucien Jacquet observed that progressive
gradual mechanical excitation of the skin (gradual progression
in frequency and duration of treatment) is supposed to coun-
teract the age-related loss of the firmness of the face, neck,
and upper chest skin.2
More recently, it has been shown that fibroblasts contain
a molecular machinery to convert mechanical stimuli into
responses that influence tissue remodeling. In vitro studies
demonstrated that they are able to change their behavior and
induce complex cascades of biochemical events in extracel-
lular matrix.3–5 Thus, mechanical stimuli lead to a “synthetic”
fibroblast phenotype characterized by the induction of con-
nective tissue synthesis while simultaneously inhibiting
matrix degradation; in mechanically “stressed” lattices,
stress fiber and focal adhesion formation by fibroblasts have
been observed.6
Mechanical stimulation performed on the skin surface
sends a signal to the distant cells such as keratinocytes,
fibroblasts, and adipocytes and causes production of collagen,
elastin, and activation of lipolysis. Loss of mechanical tension
appears to be the major factor underlying decreased collagen
synthesis in aged skin; an aging-related loss of fibroblast
interaction with the surrounding fibrous matrix has also been
shown, leading to the vicious cycle of diminished procollagen
synthesis, disorganization of collagen bundles, and further
loss of mechanical stimulation of fibroblasts. Myofibroblasts
(active fibroblasts characterized by α-smooth muscle (SM)-
actin protein expression) play a central role in scarring and
cell migration and motility within the connective tissue.7
Fibroblast and collagen fibers presented alignment along
the stretching axis upon axial stimulation, compared to free-
floating lattices.8
A previous study using a technique based on mechanical
stimulation of the cutaneous and subcutaneous tissues has
shown that this technique is able to restart their natural
production of collagen and elastin by delivering microbeats
to the skin’s surface, which can stimulate cell rejuvenation
in-depth.9 However, the evaluations were performed on the
arm skin, and it was not clear if these results could be gen-
eralized to the face as well.
In the present study, a mechanical stimulation has been
applied on the facial skin for 24 sessions over 8 weeks.
The primary aim was to assess the clinical and macroscopic
effect of mechanical stimulation on the facial skin. The
secondary aim was to investigate the mechanism of action of
this technique at the cellular level by measuring contractile
activity of the fibroblasts and quantifying the skin changes
induced by fibroblastic activity.
Materials and methods
This is a monocentric, double-blinded, controlled, and
randomized interventional study on the effects of a technique
called Mécano-Stimulation™.
Volunteers
Thirty healthy subjects (20 females and 10 males) aged
35–50 years were enrolled in this study. The inclusion
criteria were Fitzpatrick skin phototype I–IV,10 with mild-
to-moderate facial skin sagging (cutaneous sagging grade
1–3, according to Ezure et al11) (Figure 1).
Exclusion criteria were sun or ultraviolet (UV) exposure
during the last 15 days, any facial dermatologic pathologies
such as rosacea, any chronic or acute systemic affections,
any topical or systemic treatment that could interfere with
the results of the study, and any contraindications to the
Mécano-Stimulation (eg, herpes, vitiligo, acne: inflamma-
tory and infective phase). All volunteers signed an informed
consent form in agreement with the Declaration of Helsinki
after having enough time to understand the modalities and
the aim of the study. The study protocol was approved by
the local ethical committee (CPP [Comités de Protection des
Personnes] Est II) and the National French Health Authori-
ties (ANSM [L’Agence nationale de sécurité du Médicament
et des produits de santé]).
Treatment
The treatment was applied on one side of the face (randomized
hemiface: chin, nasolabial fold, cheek, cheekbone, temple,
forehead) during 15 minutes, three times per week for 8 weeks
(24 sessions). In order to standardize the treatment, all treat-
ments were performed by the same practitioner. The treatment
was carried out via the Mécano-Stimulation system (LPG
Systems, Valence, France), which transforms manual mas-
sage movements through mechanical procedures by means
of different treatment heads. The treatment heads used in this
study are composed of two motorized flaps inside a treatment
chamber, which are dedicated to the treatment of the face or
small areas. The treatment forces are exerted without any
pinching or twisting of the skin but through combining both
horizontal and vertical stimulation (Figure 2). These heads
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
Figure 1 Ezure scoring scale.
Notes: Skin sagging is evaluated through the Ezure scoring scale:11 facial cutaneous sagging is graded from 0 to 5 on three areas of the hemiface (A), upper cheek (B), lower
cheek (C), and lateral cheek (D). Arrows and arrowheads point out specic areas of interest dening skin sagging stages. Volunteers selected in the study were of grade 1–3.
Vertical stimulation
Sequential suction Beating flaps
Horizontal stimulation
Figure 2 Double stimulation.
Notes: The Mécano-Stimulation™ technique consists of two motorized aps located inside a treatment chamber and dedicated to the treatment of the face or small areas.
The treatment forces combine sequential aspiration and beating aps creating, respectively, vertical and horizontal stimulation. Settings allow obtaining different intensity
and rhythmicity of the aspiration or beating frequency of the aps. Lift heads used in this study were “motorized”: heads contain a small engine allowing the aps to beat in
a regular and permanent way, synchronized specically with the sequential aspiration.
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Humbert et al
contain a small engine allowing the flaps to vibrate in a regular
repeated way, which is accompanied with aspiration.
Different parameters (intensity and rhythm of the
aspiration and beating frequency of the flaps) can be modu-
lated at any time according to the individual sensitivity and
tissue state (infiltrated, loosened, or wrinkled skin).
Lift heads used in this study were “motorized”: heads
contain a small engine allowing the flaps to beat in a regu-
lar and permanent way, synchronized specifically with the
sequential aspiration.
Measurement
Evaluations were performed before the treatment (T0), after
24 sessions (T1), and 1 month after the end of the treatment
(T2). Volunteers were evaluated in the standard skin situa-
tion (last face washing the night before measuring sessions,
without any cosmetic application until the measures [no
water, soap, cream, or makeup]). During the study, they
were also asked to not change their facial skin care habits,
hygiene, and cosmetic products, to avoid sun exposure and
UV treatment even occasionally, to avoid any medical and
invasive treatment on the face, and to report systemically
any medication consumption to the investigator. All evalu-
ations and measurements were performed under standard
conditions (constant temperature 20°C–23°C and hygrometry
40°C–60°C) after a 20-minute rest period.
The main evaluation criterion was clinical scoring for skin
sagging using a photographic scale. The secondary outcomes
were clinical scoring of skin relief and wrinkles, skin color and
radiance, and subject self-assessments. The biopsies were taken
from both sides of the face (treatment and control side) from
19 subjects, nine for histopathologic changes and ten for the in
vitro evaluation of the fibroblasts for their contractile capacity.
Primary outcome
The primary outcome, measured at T0 and T1, was the facial
skin sagging quotation performed by a blinded dermatolo-
gist, thanks to a clinical scoring based on the six-grade scale
of Ezure et al11 and carried out on three different locations:
upper part of the cheek/nasolabial folds (B zone), lower part
of the cheeks (C zone), and lateral part of the cheek (D zone),
as described in Figure 1.
Secondary outcomes
Clinical scoring
A blinded dermatologist evaluated the different signs of skin
aging, with each criterion being assessed on both treated and
nontreated side.
1. Skin radiance: A clinical scoring of skin radiance
was calculated through a visual analysis scale (CLBT™)
which includes seven descriptors. Actually, four different
colors (pink red, olive, beige, light pink) between 10% and
100% lead to the final analysis of three different parameters
(skin transparency, brightness, and luminosity) on a 0–10
scale.12
2. Skin relief: An analogical 0–9 scale was used to
evaluate the skin relief parameters. This allowed the calcula-
tion of a skin relief score defined as the sum of the following
scores: surface irregularity score + wrinkles/lines/blemishes
score + skin texture score + under-eye dark circles score +
papules/microcysts score. The lower the score indicates lower
skin relief, indicating better appearance.
3. Skin color: The same 0–9 scale was used to evaluate the
skin color through the following parameters: heterogeneity,
eye circle, redness/rosacea, skin spots, and papules/scars. The
sum of these parameters present the global color score.
Self-assessment
The volunteers were asked to fill a 9-point questionnaire at
T0 (before) and T1 (after 24 sessions of treatment) for the
following parameters (absence/presence and the intensity):
• wrinkles and fine lines;
• roughness;
• nasolabial folds;
• elasticity;
• firmness/softness;
• radiance;
• under-eye dark circles.
They were also asked to compare their skin with pretreat-
ment status for firmness, elasticity, smoothness, wrinkles,
sagging, and brightness. They were supposed to answer the
satisfaction questionnaire as well.
Photography
At each visit, the front, left, and right lateral photographs
were taken of the whole and lower face in normal daylight,
with eyes closed (Canon EOS Rebel XS with a 50-mm lens
and two flashes positioned on a Faraghan Medical System).
All photos were standardized for light (using Gretag MacBeth
Colour Checker Chart) and position (stereotaxic table; Eotech
SA, Marcoussis, France).
In vitro study
Two 2 mm punch biopsies were taken from nine subjects
(four males and five females) from both sides of the face
(treatment and control side) at T1. The samples were placed
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
in sterile gauze moistened with saline and immediately
treated: Each biopsy was placed in a 25 cm2 culture dish and
was allowed to adhere to the plastic surface for approximately
30 minutes at 37°C. Three milliliters of Dulbecco’s Modi-
fied Eagle’s Medium supplemented with 10% fetal calf
serum, 40 mg/mL of gentamicin, and 2 mg/mL of fungizone
(DMEMc) were added to each dish. The dishes were then
incubated at 37°C in 5% CO2. Subculture of fibroblasts was
established after 3–4 weeks of outgrowth by trypsination
(trypsin-EDTA (ethylene diamine tetra acetique) [1x]; PAA
laboratory, France) of explant cultures. The culture medium
was changed two times a week. Fibroblasts cultivated in
passages lower than ten were used.
Migratory capacity
Fibroblasts that were either subjected or not subjected to
Mécano-Stimulation were embedded in three-dimensional
hydrated collagen, using a modified version of the technique
developed by Bell et al13, to study their mechanical behavior
by measuring their capacity to contract free-floating lattices.
Collagen lattices were made by mixing six volumes of 1.76×
concentrated medium, three volumes of rat tail type I collagen
solution (2 mg/mL), and one volume of fibroblasts suspension
(8×105 cells/mL). This mixture was poured into plastic Petri
dishes (60 mm diameter). The gel polymerized in 30 minutes
at 37°C. Culture medium was then added and changed every
48 hours. To measure the retracted lattice diameter, the culture
dishes were placed on a transparent metric scale. The measure-
ment of the lattice diameter was performed for 10 days. The
evaluation of the migratory capacities of fibroblasts makes it
possible to show the possible reorganization of extracellular
matrix in the presence of Mécano-Stimulation.
Measurements of contractile forces developed by
broblasts in GlaSbox® device (Growing Lattice
Study box)
The cell chamber was composed of eight rectangular culture
cavities in which the lattices developed. Two silicon beams
were placed opposite to each other with strain gauges hanging
apart, down into each cavity at 27 mm distance. The lattice
was attached to this sensor through a grid directly etched
onto the lower part of the beams. The output signal from the
strain gauges was amplified, then converted, and collected
by a computer, which included an acquisition card and a
specific software giving the forces in real time. Fibroblasts
were prepared as defined in the previous paragraph; the lat-
tice mixture was poured into a rectangular culture cavity of
the GlaSbox® and polymerized in 30 minutes at 37°C. Two
milliliters of culture medium was added. The GlaSbox® was
then placed into a humidified incubator at 37°C, and force
measurements were started immediately and for 24 hours. The
results were expressed as contractile forces (arbitrary unit) of
the fibroblasts either subjected or not subjected to Mécano-
Stimulation according to the time. These measurements quan-
tify the contractile forces of fibroblasts and make it possible
to know whether a treatment applied for 15 minutes, three
times per week during 8 weeks (24 sessions) already shows
a “lifting” effect at the scale of fibroblasts.
Synthesis studies
Fibroblasts either subjected or not subjected to Mécano-
Stimulation were seeded in 12-cavity plates at 0.04×106 cells
per cavity. The plates were incubated at 37°C. The next day,
500 µL of DMEMc was poured into each cavity and the plates
were incubated at 37°C. After 48 hours, the supernatant part
was placed in a protease inhibitor cocktail (10% v/v, Merck
Millipore, Fontenay sous bois, France). The samples were
stored at −80°C until analysis.
Collagen I, hyaluronic acid, matrix
metalloproteinase-1, matrix
metalloproteinase-9, and tissue inhibitor
of metalloprotei nase-1 (TIMP-1) assay
The quantification was evaluated using an enzyme-linked
immunosorbent assay kit (USCN, Life Science and Ray-
Biotech®, Inc). Standards or samples were added into the
precoated cavities with a specific biotin-conjugated antibody.
After washing the samples, avidin (collagen I and hyaluronic
acid) or streptavidin (matrix metalloproteinase [MMP]-1,
MMP-9, and TIMP-1) conjugated to horseradish peroxidase
were added to each cavity and then incubated. After adding
the tetramethylbenzidine (TMB) substrate solution, only
those cavities that contained study substances showed a color
change. The enzyme–substrate reaction is terminated by the
addition of sulfuric acid solution and the color change is mea-
sured spectrophotometrically at a wavelength of 450 nm. The
concentration is then determined by comparing the optical
density of the samples to the standard curve.
Elastin assay
The total soluble elastin was assayed by a colorimetric method
(Fastin™ Elastin Assay; Biocolor). The samples were precipi-
tated with a solution of trichloroacetic acid and arginine, then the
reagent 5,10,15,20-tetraphenyl-21,23-porphine tetra-sulfonate
was added, which can particularly bind to elastin. The formed
complex was dissociated by a solution of guanidine-HCl and
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Humbert et al
propan-1-ol. The developed color was measured at 550 nm
using a spectrophotometer (Multiscan EX; Thermo).
Ultrastructural studies
Two-millimeter punch biopsies were taken under local
anesthesia (1% xylocain) from the jugular area of ten other
subjects (five males and five females) at T1. Biopsies from
the treated and nontreated sides were coded and examined
pair by pair in a blinded manner.
Each biopsy was cut into small fragments, some of which
were fixed in 3% paraformaldehyde in phosphate-buffered
saline (PBS), the others in 2% glutaraldehyde in sodium
cacodylate buffer at 4°C for 3 days. For standard morpho-
logical examination, the glutaraldehyde-fixed fragments
were washed and then postfixed in 1% osmium tetroxide
for 2 hours. After dehydration in graded ethanol series, the
fragments were impregnated and embedded in epoxy resins
(60°C for 3 days). Ultrafine sections were contrasted with
uranyl acetate and lead citrate before observation.
The paraformaldehyde-fixed fragments, used for immu-
nocytochemistry, were washed, dehydrated, and embedded in
LR White resin (UVA polymerization at 4°C; EMS, Hatfield,
PA, USA). Ultrafine sections on formvar-coated nickel grids
(EMS) were exposed to the α-SM-actin antibody (diluted
1/50, overnight at 4°C), washed in PBS, and incubated for
1 hour with goat anti-mouse antibodies coupled to 10 nm
colloidal gold (diluted 1/10; Amersham, Little Chalfont, UK).
After final washes in PBS and distilled water, the sections
were counterstained with uranyl acetate alone.
Transmission electron microscopy of all specimens was
performed, and digital images were obtained and analyzed.
Density and overall quality of the dermal fibrous structures,
ie, collagen bundles and elastic fibers, were evaluated semi-
quantitatively. Also, the morphological signs of fibroblast
metabolic and secretory activity as well as the intensity of
the α-SM-actin expression were noted.
Statistical analysis
Data are presented as mean ± standard deviation. The normal-
ity of the data distribution was tested with a Shapiro–Wilk test.
Differences between groups at baseline and at the end of the 24
sessions were tested with a Student’s t-test for paired values (or
a Wilcoxon test in case of nonnormally distributed data).
Variance analysis with repeated measurements (analy-
sis of variance) completed by a Student–Newman–Keuls
test (normal distribution of parameters) or a Friedman
test completed by a Dunn’s test (abnormal distribution of
parameters) was applied to analyze the evolution of the
studied parameters over the chosen time period.
Statistical significance was set at 5% and at 5%–10%
for trends.
For the in vitro study, one-way variance analysis followed
by a Fisher’s exact test (if needed) was performed for synthesis
studies. Two-way analysis of variance (groups and time) fol-
lowed by a Fisher’s exact test (if needed) was performed for the
measurements of migratory capacities and contractile forces.
Results
Thirty subjects (20 females and 10 males) with a mean age of
44.5±4.6 years finished the 24 sessions of treatments. Among
the 20 volunteers, 13 were under contraceptive treatments.
Depression (n=4), seasonal allergies (n=2), hiatal hernia
(n=1), hand eczema (n=1), familial hypercholesterolemia
(n=1), epilepsy (n=1), allergy to nickel (n=1), allergy to tra-
madol (n=1), emphysema (n=1), and chronic bronchitis (n=1)
were recorded as the past medical history of the volunteers.
Primary outcome: skin sagging
A clinical improvement of skin sagging of at least one
parameter of the facial skin sagging (Ezure scale) was observed
in 73% (n=22) of the subjects on the treated side versus in 53%
(n=16) of the subjects for the nontreated area (P=0.108). The
same calculation for the C and D zones revealed 70% (n=15)
improvement on the treated side versus 50% (n=15) improve-
ment for the control side (P=0.114) (Table 1).
Secondary outcomes
Clinical scoring
CLBT color scale
At T0, the treated and untreated hemiface were similar for
the clinical scores of skin complexion radiance evaluated
with the CLBT scale.
Table 1 Results of the skin sagging scores
Zones Number of volunteers
with improved score
(out of 30 subjects)
P
Treated
hemiface
Nontreated
hemiface
B10 (33%) 8 (27%) 0.573
C 7 (57%) 11 (37%) 0.120
D13 (43%) 8 (27%) 0.176
At least one among three 22 (73%) 16 (53%) 0.108
At least C or D 21 (70%) 15 (50%) 0.114
Notes: Number of volunteers with improved clinical sagging score (Ezure scale11)
is presented. A clinical improvement of skin sagging of at least one parameter of the
score was observed in 73% (n=22) of the subjects on the treated side. B zone is the
upper part of the cheek/nasolabial folds, C zone is the lower part of the cheeks, and
D zone is the lateral part of the cheek. Clinical improvement of the sagging of at least
C or D areas (where the massage was the more efcient) was observed in 70% of
treated hemiface of the subjects.
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
Table 2 Results for CLBT™ scoring before (T0) and after (T1) the 24 treatment sessions
Score (Musnier et al12) Treated hemiface Nontreated hemiface
T0 (mean ± SD) T1 (mean ± SD) T0 (mean ± SD) T1 (mean ± SD)
Color score (the lower the score the
more homogeneous the skin is)
12.60±3.10 10.90±2.98* 12.03±2.89 11.83±2.89§
Pink red 40.33±19.56 33.00±18.03* 40.00±17.42 38.33±16.42§
Olive 36.67±10.93 24.67±9.00* 36.00±10.37 32.67±9.07§
Beige 54.00±12.76 45.00±13.06* 53.33±13.73 50.67±13.67§
Light pink 69.67±12.17 59.00±17.09* 69.67±12.99 66.33±13.2§
Transparency 3.56±1.93 4.57±1.50* 3.55±1.84 3.59±1.45§
Brightness 2.43±1.88 3.04±1.64* 2.44±1.80 2.57±1.53§
Luminosity 2.08±1.12 4.11±1.69* 2.01±1.06 2.94±1.38*,§
Notes: The visual analysis scale (CLBT scoring12) follows seven descriptors that evaluate four different colors (scale between 10% and 100%) and three parameters of skin
transparency, brightness, and luminosity (0–10 scale). Compared to baseline, all scores were signicantly improved after treatment on the treated hemiface (P0.05); at T1, the
color scores as well as skin transparency, brightness, and luminosity were signicantly improved on the treated compared to the nontreated hemiface. All the parameters presented
a signicant improvement in terms of time/treatment evolution. *Signicantly different from T0 within each hemiface. §Signicantly different from treated hemiface at T1.
Abbreviations: CLBT, Color, Luminosity, Brightness and Transparency Scoring; SD, standard deviation; T, time.
A
Clinical scoring
(pink red)
C
Clinical scoring
(beige)
B
Clinical scoring
(olive)
D
Clinical scoring
(light pink)
50
45
40
35
30
25
20
40
45
50
55
60
40
35
30
25
20
50
55
60
65
70
75
80
T0 T0 T1
T0 T1
T1
T0 T1
*
**
*
Treated hemiface
Nontreated hemiface
Figure 3 CLBT™ scoring before (T0) and after (T1) the 24 treatment sessions: results for the color components.
Notes: The visual analysis scale (CLBT scoring12) follows seven descriptors that evaluate between 10% and 100% of four different colors: pink red (A), olive (B), beige (C),
and light pink (D). All these color parameters presented a signicant improvement (decrease) in terms of time/treatment evolution, in favor of the treated (blue) versus
nontreated (red) side. *P0.05.
Abbreviations: CLBT, Color, Luminosity, Brightness and Transparency Scoring; T, time.
Compared to the baseline, all scores were significantly
improved on the treated hemiface (P0.05). At T1, the
color scores (skin transparency, brightness and luminosity)
were significantly improved on the treated compared to the
nontreated hemiface (Table 2, Figures 3 and 4). Finally, the
seven evaluated parameters presented a significant improve-
ment in terms of time/treatment evolution.
From a descriptive point of view, we can notice an
improvement for the treated versus nontreated sides on all
parameters:
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Humbert et al
• “pink red”: 18.2% on the treated side versus 4.2% on the
nontreated side;
• “olive”: 32.7% versus 9.2%;
• “beige”: 16.7% versus 5%;
• “light pink”: 15.3% versus 4.8%;
• transparency: 28.4% versus 1.1%;
• brightness: 25.1% versus 5.3%;
• luminosity: 97.6% versus 46.3%.
Relief
At baseline (T0), no significant differences were observed
in surface irregularity, fine lines, wrinkles and blemishes,
skin texture, under-eye dark circles, buttons/microcysts,
and relief scores between the treated and the untreated
hemiface. Compared to the baseline, at T1 all scores
decreased on the treated hemiface (P0.05) (except for
buttons/microcysts) while no change was observed on the
nontreated hemiface. A significant improvement in terms of
time/treatment evolution was observed for all the evaluated
parameters (Table 3 and Figure 5). An improvement was
observed for treated versus nontreated side on the follow-
ing parameters:
• surface irregularity: 20.7% on the treated side versus
2.5% on the nontreated side;
• fine lines, wrinkles, and blemishes: 21.1% versus 0.6%;
• skin texture: 15% versus 6.37%;
• under-eye dark circles: 14.2% versus 4.2%;
• relief score: 19.2% versus 5.4%.
Color
At T0, no difference between the treated and the untreated
hemiface was observed for the clinical scores of skin color
(except for buttons/spots, P=0.086). At T1 compared to
baseline, heterogeneity, under-eye dark circles, papules/
scars, and color score were significantly improved on the
treated hemiface, while they were stable on the nontreated
hemiface. A significant improvement in terms of time/
treatment evolution was observed for all parameters (skin
complexion heterogeneity, under-eye dark circles, redness,
and color score) (Table 4 and Figure 6). An improvement
A
Clinical scoring
(transparency)
B
Clinical scoring
(brightness)
1T0
T0 T1
T1 T0
Treated hemiface
Nontreated hemiface
T1
2
3
**
*
4
1
2
3
4
5
6
C
Clinical scoring
(luminosity)
1
2
3
4
5
6
Figure 4 CLBT™ scoring before (T0) and after (T1) the 24 treatment sessions: results for transparency, brightness, and luminosity.
Notes: The visual analysis scale (CLBT scoring12) evaluates three parameters of skin transparency (A), brightness (B), and luminosity (C) on a 0–10 scale; a signicant
improvement (increase) in terms of time/treatment evolution is pictured in all cases, in favor of the treated (blue) versus nontreated (red) hemiface. *P0.05.
Abbreviations: CLBT, Color, Luminosity, Brightness and Transparency Scoring; T, time.
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
for treated versus nontreated side was observed on the fol-
lowing parameters:
• skin complexion heterogeneity: −13.8% on the treated
versus +1.2% on the nontreated side;
• eye circles: 15.1% versus 1.7%;
• redness/rosacea: −9.4% versus +2.5%;
• color score: 13.5% versus 1.7%.
Subjects self-assessment
Subjects’ self-assessment evaluation revealed an improve-
ment for 9 out of 10 parameters on the treated side while
the skin status on the nontreated side was considered to
show little improvement or even worsen compared to
baseline. Differences at T1 between treated and untreated
sides were significant for 7 of the 10 parameters and trend
for 1. Significant evolution of treated side was noticed for
roughness, firmness, radiance, and softness, compared to
untreated hemiface. A significant improvement in terms of
time/treatment evolution was observed for all parameters
(skin complexion heterogeneity, under-eye dark circles,
redness, and color score) (Table 5).
Table 6 shows the results of the self-assessment question-
naire allowing the volunteers to rate their skin state at T1 and
T2, compared to T0.
The results demonstrated a significant effect on the treated
side compared to the nontreated side for all parameters
(firmness, elasticity, smoothness, wrinkles, and sagging),
except brightness at T1.
Satisfaction questionnaire
At T1, 90% of the volunteers were satisfied by the mechano-
stimulation while the satisfaction rate was 100% at T2. Of all
the volunteers, 70% at T1 and 75% at T2 declared they wish
to continue the treatment, and 77% of the volunteers judged
the treatment as “very good” or “good” at T1. At T2, 87% of
the subjects still felt the positive effects of the treatment.
Photographs
Standardized photographs illustrated an improvement of the
oval face shape in 60% of the subjects on the treated area
(Figure 7).
In vitro study
Migratory capacity
The migration of fibroblasts was evaluated through
their capacity to retract free-floating collagen lattices.
The retraction of collagen lattices obtained with fibroblasts
subjected or not subjected to Mécano-Stimulation was
time dependent. The retraction of collagen lattices obtained
with fibroblasts subjected to Mécano-Stimulation was
significantly increased compared to nonstimulated fibroblasts
(P0.001; Figure 8).
Clinical scoring
(surface irregularity)
3.5
T0 T1
*
4.0
4.5
5.0
5.5
Treated hemiface
Nontreated hemiface
Figure 5 Clinical scoring of skin radiance: relief surface irregularity.
Notes: An analogical scale (from 0 to 9) was used to score the radiance through
relief parameters: among those, surface irregularity was evaluated from small (0) to
large (9) before (T0) and after (T1) the treatment. The gure represents a signicant
improvement (decrease) in terms of time/treatment evolution observed; surface
irregularity score was improved by 20.7% on treated side (blue) against 2.5% on
nontreated side (red). *P0.05.
Abbreviation: T, time.
Table 3 Clinical scoring of skin radiance: evaluation through relief before (T0) and after (T1) the 24 treatment sessions
Score Treated hemiface Nontreated hemiface
T0 (mean ± SD) T1 (mean ± SD) T0 (mean ± SD) T1 (mean ± SD)
Surface irregularity 5.13±1.20 4.07±1.05* 5.13±1.20 5.00±1.05§
Fine lines, wrinkles, and blemishes 4.73±1.26 3.73±1.05* 4.80±1.24 4.77±1.04§
Skin texture 3.73±1.84 3.17±1.88 * 3.77±1.87 3.53±2.03§
Under-eye dark circles 4.63±1.71 3.97±1.85* 4.77±1.72 4.57±1.74§
Buttons/microcysts 1.23±1.07 0.80±0.85* 1.17±1.02 0.70±0.79§
Relief score 19.47±3.85 15.73±4.02* 19.63±3.74 18.57±3.41§
Notes: An analogical scale (from 0 to 9) was used to score the radiance through relief parameters, before and after the treatment: surface irregularity, ne lines/wrinkles/
blemishes, skin pores, under-eye dark circles, and buttons/microcysts. A global relief score was dened as the sum of the previous scores (the lower the score, the better the
relief is). A signicant improvement in terms of time/treatment evolution was observed for all the evaluated parameters. *Signicantly different from T0 within each hemiface.
§Signicantly different from treated hemiface at T1.
Abbreviations: SD, standard deviation; T, time.
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Humbert et al
Measurements of contractile forces
developed by broblasts
No significant difference of the development of contractile
forces was observed between the Mécano-Stimulation group
and the control group (Figure 9).
Collagen I, elastin, and hyaluronic acid
synthesis
For each product, synthesis results are expressed as %
respective control.
At T1, a nonsignificant increase in the synthesis of collagen
I was observed between the treated and untreated hemiface
(107.8%±69.8% vs 100.0%±0.0%/control; Figure 10).
For elastin synthesis by the fibroblasts, a nonsignificant
difference was observed on the treated side comparing to the
untreated side (145.6%±32.1% vs 100.0%±0.0%/control for
the treated and untreated hemiface, respectively, P=0.175;
Figure 10). The same result was noticed for the synthesis of
hyaluronic acid (180.2%±69.8% vs 100.0%±0.0%/control for
the treated and untreated hemiface, respectively, P=0.268;
Figure 10).
MMP-9, MMP-1, and TIMP-1 synthesis
For each MMP, results are expressed as % respective
control.
A significant increase in the synthesis of MMP-9
(+115.4%, P0.05 vs control group, P=0.033; Figure 11)
was observed in fibroblasts from patients subjected to
Mécano-Stimulation compared to control fibroblasts. No
significant changes were observed in the synthesis of MMP-1
and TIMP-1 (Figure 11).
Ultrastructural results
After decoding of the samples, the treated side presented
ultrastructural signs of improvement in four of the ten
volunteers. These were characterized by:
• changes in the fibroblast morphology comprising
the enlargement of the intracellular network of ribo-
some-decorated rough endoplasmic reticulum, of the
electron-lucent mitochondria with well-developed
cristae, and of the presence of numerous small plasma
membrane-bound vesicles suggestive of an increased
secretory activity; none of these were observed in the
nonresponders;
• restructuring of the fragmented elastic fibers, ie, evidence
of the elastic fiber recycling by “activated” fibroblasts;
• increased α-SM-actin expression by the latter in 50% of
the cases (Figure 12).
Clinical scoring
(skin complexion heterogeneity)
2.5
3.0
3.5
*
T0 T1
Nontreated hemiface
Treated hemiface
Figure 6 Clinical scoring of skin radiance: color skin complexion heterogeneity.
Notes: The scoring of the radiance was also assessed through color parameters:
skin complexion heterogeneity score was evaluated with an analogical scale (from
0= homogeneous to 9=heterogeneous) and is presented before (T0) and after (T1)
the 24 sessions of treatment. At T1, the score was signicantly improved (decrease)
on the treated side (blue) compared to the nontreated hemiface (red); a signicant
improvement in terms of time/treatment evolution was observed. Skin complexion
heterogeneity score was improved by 13.8% on the treated side while a worsening
was noticed on the nontreated side (+1.2%). *P0.05.
Abbreviation: T, time.
Table 4 Clinical scoring of skin radiance: evaluation through skin color before (T0) and after (T1) the 24 treatment sessions
Score Treated hemiface Nontreated hemiface
T0 (mean ± SD) T1 (mean ± SD) T0 (mean ± SD) T1 (mean ± SD)
Skin complexion heterogeneity 3.40±0.86 2.93±0.98* 3.23±0.73 3.27±0.87§
Under-eye dark circles 3.57±1.52 3.03±1.50* 3.53±1.66 3.47±1.59§
Redness/rosacea 2.87±1.59 2.60±1.45 2.80±1.52 2.87±1.36
Skin spots (hyperpigmentation) 2.03±1.19 1.97±1.22 1.97±1.35 1.93±1.34
Buttons/scars 0.73±0.87 0.37±0.49* 0.50±0.73 0.30±0.53
Color score 12.60±3.10 10.90±2.98* 12.03±2.89 11.83±2.89
Notes: The scoring of the radiance was also assessed through color parameters with an analogical scale (0–9): skin complexion heterogeneity, under-eye dark circles,
redness/rosacea, skin spots, and buttons/scars. A global color score was dened as the sum of the previous scores (the lower the score, the better the radiance is). Results
are presented before (T0) and after (T1) the 24 sessions of treatment. A signicant improvement in terms of time/treatment evolution was observed for skin complexion
heterogeneity, under-eye dark circles, redness, and color score. *Signicantly different from T0 within each hemiface. §Signicantly different from treated hemiface at T1.
Abbreviation: T, time.
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
Table 5 Results of self-assessment evaluation
Improvement on treated side
at T1 compared to T0
(vs untreated side)
Signicant difference
at T1 between treated
and untreated
Signicant difference in
evolution between the
treated and untreated
Wrinkles 13.6% (4.8%) Yes (P=0.004)
Roughness 3.8% (−5.1%) Yes (P=0.028) Yes (P=0.016)
Fine lines −3.1% (−10.5%)
Nasolabial fold 21.3% (11.6%) Yes (P=0.089)
Elasticity 11.2% (0.5%) Yes (P=0.003)
Firmness 23.1% (2.5%) Yes (P=0.003) Yes (P=0.001)
Radiance 26.3% (14%) Yes (P=0.011) Yes (P=0.026)
Softness 2.1% (−7.4%) Yes (P=0.018) Yes (P=0.016)
Under-eye dark circles 18.8% (0%) Yes (P=0.001)
Notes: At T0 and T1, volunteers were asked to score some parameters which characterized their skin (described as presence/absence and the intensity of each parameter).
Self-assessment showed improvement on treated side for nine of the ten parameters evaluated; in all cases, nontreated side was felt as less improved or even to have
worsened. Difference at T1 between treated and untreated sides were signicant for seven of the ten parameters, and trend for one. Signicant evolution of treated side was
noticed for roughness, rmness, radiance, and softness, compared to untreated hemiface.
Abbreviation: T, time.
Facial skin of the control side showed changes related
to the chronologic and actinic aging, such as elastic fiber
fragmentation, diminished density and organization of the
fibrous extracellular matrix, and quiescence of the dermal
fibroblasts. These structural signs of tissue senescence were
particularly accentuated in seven of the ten subjects.
Discussion
Cutaneous aging is a complex process that depends on
intrinsic factors (chronologic aging, genetic basis) and
on detrimental influences from the environment (mainly
UV radiation).14 The resulting effects such as skin laxity,
sagging, and wrinkle formation are clinically obvious and
concern mainly the superficial dermis.15 Fibroblasts are the
most abundant cells in the dermis, playing a major role in
extracellular matrix components synthesis; matrix remodel-
ing, and migration activities; because several in vitro studies
have demonstrated that mechanical stimulation on fibroblasts
could alter their behavior and induce complex cascades of
biochemical events,3,4,6 the idea of performing mechanical
stimulation on the skin surface has emerged to positively
affect the dermis.
With aging, the skin tends to become thinner, less elas-
tic, and drier. Phenotypic studies about wrinkle fibroblasts
showed that the mechanical properties of the skin change
with aging: reduced contractile capacities as well as reduced
migration activities were observed in wrinkle fibroblasts.1
A major feature of aged skin is fragmentation of the der-
mal collagen matrix.16 Fragmentation results from actions of
specific enzymes (MMPs, collagenases, and elastases) and
impairs the structural integrity of the dermis. Fibroblasts that
produce and organize the collagen matrix cannot attach to
fragmented collagen. Loss of attachment prevents fibroblasts
from receiving mechanical information from their support,
and they collapse. Stretch is critical for normal balanced pro-
duction of collagen and collagen-degrading enzymes.17 Then,
in aged skin, collapsed fibroblasts produce low levels of col-
lagen and high levels of collagen-degrading enzymes.7
During the aging process, the proteoglycans such as
hyaluronic acid, which has a high affinity for water, are
less synthesized, and the contractile activity of fibroblasts
decreases. Fibroblasts are losing their ability to express
α-SM-actin and to differentiate into myofibroblasts. This
Table 6 Self-assessment of subjects skin at T1 (end of the
treatment) and T2 (1 month after the end of the treatment),
compared to T0
% of subjects with positive
answer
Treated Not treated
Firmer skin T1 70 23
T2 75 37
More elastic skin T1 73 20
T2 75 24
Smoother skin T1 67 23
T2 87 12
Less wrinkled skin T1 87 10
T2 75 12
Less relaxed skin T1 77 20
T2 75 25
Brighter skin T1 20 53
T2 62 24
Notes: The result of the self-assessment questionnaire allowing the volunteers to rate
their skin state at T1 and T2, compared to T0 is shown. The proportion of positive
answers for each item is presented; the results demonstrated a strong effect on the
treated side compared with the nontreated side, for all parameters, except brightness
at T1. After treatment, patients found their skin rmer, smoother, less wrinkled and
less sagged, with a remanent effect 1 month after the last treatment.
Abbreviation: T, time.
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Humbert et al
Figure 7 Skin sagging evolution during the study: examples on two volunteers treated on the left side of the face before and after 24 sessions of treatment.
Notes: Standardized photographs illustrated an improvement of the face oval: Two examples are pictured: before treatment (A, C) where skin sagging is indicated with
arrows on lateral and lower cheek; after 24 sessions of treatment, and (B, D) skin slackening is reduced.
induces lower forces of contraction on the extracellular
matrix and a decrease of mechanical tension of the skin,
producing a loss of elasticity and tonicity of the dermis.
Previous trials studied the effect of Mécano-Stimulation
and showed its ability to reinduce natural production of col-
lagen and elastin.9
In the present study, 30 subjects underwent 24 sessions
of treatment on the face with the Mécano-Stimulation in
order to understand its effects through clinical, biological,
and histological evaluations.
The results indicated that, while nonstatistically sig-
nificant, a trend for a clinical improvement of skin sagging
(the primary outcome) was observed, with an improvement
of at least one parameter of the facial skin sagging
clinical score11 in 73% of subjects. The location where the
mechanostimulation was less efficient was the upper part of
the cheek/nasolabial fold, while a clinical improvement of
Diameter of retracted collagen
lattices (%/D0)
Time (days)
100
Control
Mécano-Stimulation™
80
60
40
20
D0 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10
Figure 8 Effect of Mécano-Stimulation™ on broblast retraction according to
time.
Notes: Fibroblasts capacity to retract free-oating collagen lattices reects their
migration, and is presented to time. The retraction of collagen lattices obtained with
broblasts exposed to Mécano-Stimulation™ was signicantly increased (decreasing
of the diameter of retracted collagen lattices) compared to nonstimulated broblasts
(P0.001).
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
Forces (arbitrary unit/millions
of cells)
Time (hours)
1.4e+5
1.2e+5
1.0e+5
8.0e+4
6.0e+4
4.0e+4
2.0e+4
0.0 0510 15 20 25
Control
Mécano-Stimulation™
Figure 9 Quantication of contractile forces developed by broblasts after the
action of Mécano-Stimulation™ using GlaSbox® device.
Notes: The GlaSbox® device contains eight rectangular culture wells in which lattices
have been developed; two opposite silicon beams with strain gauges allow force
measurements to be obtained. The results are expressed as contractile forces (arbitrary
unit) developed by broblasts subjected or not subjected to Mécano-Stimulation™
according to the time. No signicant difference of the development of contractile
forces was observed between Mécano-Stimulation group and control group.
the sagging was observed in 70% of subjects either on the
lower part or the lateral part of the cheek. This result is in
accordance with the fact that structural skin changes, such as
sagging and wrinkling, clearly originate due to alterations in
the dermis, which is a tissue with slow turnover.
Indeed, the macroscopic clinical signs of wrinkles and
firmness did not show any particular change over a 6-month
period18 while in the present study only 24 sessions of treat-
ment over 8 weeks were performed. The wrinkle formation
and sagging of the facial skin are long-term cumulative
effects and would not be particularly affected by 8 weeks of
treatment, so it is recommended to perform long-term studies
in this field, as already reported in other studies.19,20 A longer
treatment period could also enhance the visible changes over
the treatment period.
In accordance with the latter observation, all the clinical
scores for skin complexion radiance evaluated by the CLBT
A
140
120
100
80
60
40
20
00
0
50
100
150
200
250
20
40
60
80
100
120
140
160
180
ControlMécano-Stimulation™ ControlMécano-Stimulation™
ControlMécano-Stimulation™
Collagen I (%/control)
B
Elastin (%/control)
C
Hyaluronic acid (%/control)
Figure 10 Collagen I, elastin, and hyaluronic acid synthesis of broblasts with and without the action of Mécano-Stimulation™.
Notes: The quantication of collagen I and hyaluronic acid was evaluated using an enzyme-linked immunosorbent assay kit. The concentration in the samples was determined
by comparing the optical density of the samples to the standard curve. The total soluble elastin was assayed by a colorimetric method (Fastin Elastin assay). Results are
expressed as % respective control. At T1, nonsignicant increase in the synthesis of collagen I (A) was observed between the treated and untreated hemiface (107.8%±69.8%
and 100.0%±0.0%/control). At T1, an increase in the synthesis of elastin (B) and hyaluronic acid (C) was observed between the treated and untreated hemiface (P=0.175 and
P=0.268, respectively).
Abbreviation: T, time.
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Humbert et al
scale12 were significantly upgraded after treatment. Indeed,
color transparency, brightness, and luminosity scores were
significantly improved at T1 on the treated compared to the
nontreated hemiface.
Such radiance improvement was observed after 6 months
with the use of a topical treatment in aged human skin,21
suggesting a very efficient impact of the Mécano-Stimulation
on radiance.
Accordingly, clinical scores for surface irregularity,
fine lines, wrinkles and blemishes, skin texture, under-eye
dark circles, papules/microcysts, and relief were signifi-
cantly improved compared to baseline on the treated side,
while no significant change was observed on the nontreated
hemiface.
The skin color evaluation focused on heterogeneity,
under-eye dark circles, and global color score showed
improvement on the treated side compared to the untreated
side after treatment.
The evolution of both skin relief and color scores is along
the same direction as other results. This could reinforce the
skin global improvement of the subjects from a clinical point
of view, considering that skin relief, color, radiance, and sag-
ging are strongly interrelated in the aging process also.
Self-assessment questionnaires expressed significant
results in efficiency of the treatment for parameters such as
wrinkles, roughness, nasolabial folds, elasticity, firmness,
skin radiance, softness, and under-eye dark circles. After
24 treatment sessions, the patients found their skin firmer,
smoother, less wrinkled and less sagged, with stable effect
1 month after last treatment session.
These results, together with the fact that between 90%
and 100% (T1 and T2) of the volunteers were satisfied by
C
TIMP-1 (%/control)
ControlMécano-Stimulation™
140
120
100
80
60
40
20
0
A
MMP-1 (%/control)
ControlMécano-Stimulation™
120
100
80
60
40
20
0
B
MMP-9 (%/control)
ControlMécano-Stimulation™
0
50
100
150
200
250
300
*
Figure 11 MMP-1, MMP-9, and TIMP-1 synthesis of broblasts with and without the action of Mécano-Stimulation™.
Notes: The quantication of MMP-1, MMP-9, and TIMP-1 was obtained using an enzyme-linked immunosorbent assay kit. Results are expressed as % respective control. No
signicant change was observed in the synthesis of MMP-1 (A) and TIMP-1 (C). At T1, a signicant increase in the synthesis of MMP-9 (B) was observed between the treated
and untreated hemiface (P=0.033). *P0.05 versus control.
Abbreviations: MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinases; T, time.
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Clinical, biological, and histological evaluations of Mécano-Stimulation™
this technique and that between 70% and 75% (T1 and T2) of
the volunteers declared their wish to continue the treatment
underlined both positive objective and subjective efficacy
of this method.
Beyond these aspects, the use of Mécano-Stimulation is
considered to increase the synthesis of some extracellular
matrix components, particularly MMP-9. The composition
and organization of extracellular matrix components are
governed by MMPs.22 MMPs play major roles in tissue regen-
eration and remodeling. MMP-9 protein expression has been
reported in the epidermal cells of acute wounds.23 MMP-9
catalyzes the cleavage of denatured collagens of all types
present in aged skin and the components of natural base-
ment membrane as well.24 Stimulation of MMP-9 synthesis
explained no significant change in collagen I production in
this study. No change in contractile forces developed by
fibroblasts subjected to the action of Mécano-Stimulation
was shown compared to control fibroblasts. These results
indicate that the 24 sessions of Mécano-Stimulation are not
enough in order to show a “lifting” effect at the scale of
fibroblasts. However, the cell migration was faster due to
the mechanical stimulations and could suggest a beneficial
effect of Mécano-Stimulation on migratory and remodeling
capacities. A significant increase in the synthesis of MMP-9
Figure 12 Examples of ultrastructure observed when comparing the treated versus untreated side.
Notes: Examples of ultrastructure of the papillary dermis observed when comparing the treated versus untreated side. Untreated side (A): Presence of an amorphous
substance (*) in the spaces between fragmented EFs. Treated side (B, C, D, and E). (B) Restructured EFs in the vicinity of Fb with clear cytoplasm and developed
mitochondria. Fb express well-developed rough endoplasmic network (C) and plasma membrane–attached small secretory vesicles often facing the EFs (D). (E) Immunogold
labeling of α-smooth muscle actin in the Fb of the papillary dermis.
Abbreviations: EFs, elastic bers; Fb, broblasts.
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and the increase of fibroblasts migration suggest a beneficial
early action of mechanical stimulation on the synthesis of
extracellular matrix components. Mécano-Stimulation stimu-
lates the decreased cell activity found in aged skin through
cellular mechanotransduction.
The results suggest that a prolonged action of Mécano-
Stimulation might provide a significant difference in the
synthesis of extracellular matrix components such as collagen
I, elastin, and hyaluronic acid. The expected changes include
improvement of contraction forces and the components of
the extracellular matrix.
Ultrastructural histopathologic positive changes observed
in four out of ten study subjects correlated well with the
clinical and biochemical findings. One of the most striking
features observed with electron microscopy was apparent
restructuring of the fragmented elastin fibers. Although it is
difficult to differentiate them from newly synthesized elastic
fibers or from the degradation of the fiber remnants, there is
some evidence confirming that the mechanically stimulated
skin could recycle this functionally important component of
the extracellular matrix. However, the restructuring effects
were less remarkable than those observed after long-term
treatment with topical cream containing vitamin C.25,26
Formation of stress fibers and their attachment to focal
adhesions via vinculin have been demonstrated as hallmarks
of fibroblast organization in collagen lattices subjected to
tension.6 Accordingly, the overall increase in α-SM-actin
expression on the treated side in half of the cases might
support our clinical and biochemical results pointing to the
morphological improvement. Yet, expression of actin stress
fibers by dermal fibroblasts appeared to be a less reliable
feature in the evaluation of the tissue response to mechanical
constrains. Labeling of the α-SM-actin was observed in all
biopsies, either from the treated or control sides, although the
level of expression varied among the cells within the same
biopsy and/or between the subjects.
Our results confirm then the restructuring potential of the
physical stimulation procedure.
Conclusion
In conclusion, this original study allowed observing and
demonstrating not only the clinical and visible immediate
results but also the internal mechanisms of the skin through
an external, noninvasive, and painless stimulation.
The study provides the evidence that the use of
Mécano-Stimulation, a technique based on the application
of mechanical stimuli of the cutaneous and subcutaneous
tissue by delivering microbeats to the skin’s surface, is
able to induce clinically identifiable improvement in facial
skin appearance. This improvement is associated with a
tendency to increase the synthesis of extracellular matrix
components.
Acknowledgments
This research was made possible by the help and support
of the volunteers, technicians, and research engineers at
our center; we gratefully acknowledge Mrs Aurélie Durai,
Vanessa Ecarnot, and Celine Thiebaud for their enthusiasm
in carrying out the sensitive and accurate care and evalua-
tion and Mrs Clélia Monteux and Mr Christian Gagnière for
providing us with the device.
Disclosure
The authors certify that there is no conflict of interest with
any financial organization regarding the research described
in the article. The study was realized under the university
auspices.
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