ArticlePDF Available

Mecano-Stimulation (TM) of the skin improves sagging score and induces beneficial functional modification of the fibroblasts: clinical, biological, and histological evaluations

Authors:

Abstract and Figures

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.
Content may be subject to copyright.
© 2015 Humbert et al. This work is published by Dove Medical Press Limited, and licensed under Creative Commons Attribution – Non Commercial (unported, v3.0)
License. The full terms of the License are available at http://creativecommons.org/licenses/by-nc/3.0/. Non-commercial uses of the work are permitted without any further
permission from Dove Medical Press Limited, provided the work is properly attributed. Permissions beyond the scope of the License are administered by Dove Medical Press Limited. Information on
how to request permission may be found at: http://www.dovepress.com/permissions.php
Clinical Interventions in Aging 2015:10 387–403
Clinical Interventions in Aging Dovepress
submit your manuscript | www.dovepress.com
Dovepress 387
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 benecial functional
modication 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
Number of times this article has been viewed
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
388
Humbert et al
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
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
389
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 specic areas of interest dening 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 specically with the sequential aspiration.
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
390
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
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
391
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
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
392
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 efcient) was observed in 70% of
treated hemiface of the subjects.
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
393
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 signicantly improved after treatment on the treated hemiface (P0.05); at T1, the
color scores as well as skin transparency, brightness, and luminosity were signicantly improved on the treated compared to the nontreated hemiface. All the parameters presented
a signicant improvement in terms of time/treatment evolution. *Signicantly different from T0 within each hemiface. §Signicantly 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 signicant 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:
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
394
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 signicant
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.
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
395
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 signicant
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 dened as the sum of the previous scores (the lower the score, the better the
relief is). A signicant improvement in terms of time/treatment evolution was observed for all the evaluated parameters. *Signicantly different from T0 within each hemiface.
§Signicantly different from treated hemiface at T1.
Abbreviations: SD, standard deviation; T, time.
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
396
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 signicantly improved (decrease)
on the treated side (blue) compared to the nontreated hemiface (red); a signicant
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 dened 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 signicant improvement in terms of time/treatment evolution was observed for skin complexion
heterogeneity, under-eye dark circles, redness, and color score. *Signicantly different from T0 within each hemiface. §Signicantly different from treated hemiface at T1.
Abbreviation: T, time.
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
397
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)
Signicant difference
at T1 between treated
and untreated
Signicant 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 signicant for seven of the ten parameters, and trend for one. Signicant 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.
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
398
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
Figure 8 Effect of Mécano-Stimulation™ on broblast retraction according to
time.
Notes: Fibroblasts capacity to retract free-oating collagen lattices reects their
migration, and is presented to time. The retraction of collagen lattices obtained with
broblasts exposed to Mécano-Stimulation™ was signicantly increased (decreasing
of the diameter of retracted collagen lattices) compared to nonstimulated broblasts
(P0.001).
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
399
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 Quantication 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 signicant 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 quantication 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, nonsignicant 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.
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
400
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 quantication of MMP-1, MMP-9, and TIMP-1 was obtained using an enzyme-linked immunosorbent assay kit. Results are expressed as % respective control. No
signicant change was observed in the synthesis of MMP-1 (A) and TIMP-1 (C). At T1, a signicant 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.
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
401
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.
Clinical Interventions in Aging 2015:10
submit your manuscript | www.dovepress.com
Dovepress
Dovepress
402
Humbert et al
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.
References
1. Courderot-Masuyer C, Robin S, Tauzin H, Humbert P. Evaluation
of the behaviour of wrinkles fibroblasts and normal aged fibroblasts
in the presence of poly-l-lactic acid. J Cosmet Dermatol Sci Appl.
2012;2:20–27.
2. Jacquet L. Le massage plastique à double action dans le traitement
des dermatoses [The double action plastic massage in the treatment
of dermatosis]. Paris Médical: la semaine du clinicien. 1911;01:
355–358. French.
3. Eastwood M, MacGrouther DA, Brown RA. Fibroblasts responses to
mechanical forces. Proc Inst Mech Eng. 1998;212:85–92.
4. Chiquet M. Regulation of extracellular matrix gene expression by
mechanical stress. Matrix Biol. 1999;18:417–426.
5. Harris AK, Stopak D, Wild P. Fibroblast traction as a mechanism for
collagen morphogenesis. Nature. 1981;290:249–251.
6. Kessler D, Dethlefsen S, Haase I, et al. Fibroblasts in mechanically
stressed collagen lattices assume a “synthetic” phenotype. J Biol Chem.
2001;276:36575–36585.
7. Varani J, Dame MK, Rittie L, et al. Decreased collagen production
in chronologically aged skin: roles of age-dependent alteration in
fibroblast function and defective mechanical stimulation. Am J Pathol.
2006;168(6):1861–1868.
8. Hu JJ, Humphrey JD, Yeh AT. Characterization of engineered tissue
development under biaxial stretch using nonlinear optical microscopy.
Tissue Eng Part A. 2009;15(7):1553–1564.
9. Revuz J, Adhoute H, Cesarini JP, Poli F, Lacarriere C, Emiliozzi C.
Clinical and histological effects of the Lift6® device used on facial skin
ageing. Nouv. Dermatol. 2002;21:335–342.
10. Fitzpatrick TB. The validity and practicality of sun-reactive skin type
I through VI (Editorial). Arch Dermatol. 1988;124:869–871.
11. Ezure T, Hoshoi J, Amano S, Tsuchiya T. Sagging of the cheek is
related to skin elasticity, fat mass and mimetic muscle function. Skin
Res Technol. 2009;15(3):299–305.
12. Musnier C, Piquemal P, Beau P, Pitter J. Visual evaluation in vivo of
“complexion radiance” using the C.L.B.T. sensory methodology. Skin
Res Technol. 2004;10(1):50–56.
Clinical Interventions in Aging
Publish your work in this journal
Submit your manuscript here: http://www.dovepress.com/clinical-interventions-in-aging-journal
Clinical Interventions in Aging is an international, peer-reviewed journal
focusing on evidence-based reports on the value or lack thereof of treatments
intended to prevent or delay the onset of maladaptive correlates of aging
in human beings. This journal is indexed on PubMed Central, MedLine,
CAS, Scopus and the Elsevier Bibliographic databases. The manuscript
management system is completely online and includes a very quick and fair
peer-review system, which is all easy to use. Visit http://www.dovepress.
com/testimonials.php to read real quotes from published authors.
Clinical Interventions in Aging 2015:10 submit your manuscript | www.dovepress.com
Dovepress
Dovepress
Dovepress
403
Clinical, biological, and histological evaluations of Mécano-Stimulation™
13. Bell E, Ivarsson G, Merrill C. Production of tissue-like structure by con-
traction of collagen lattices by human fibroblasts of different proliferative
potential in vitro. Proc Natl Acad Sci U S A. 1979;76:1274–1278.
14. Mac Mary S, Sainthillier JM, Jeudy A, et al. Assessment of cumulative
exposure to UVA through the study of asymmetrical facial skin aging.
Clin Interv Aging. 2010;5:277–284.
15. Gilchrest BA. Skin aging 2003: recent advances and current concepts.
Cutis. 2003;72:5–10.
16. Fisher GJ, Varani J, Voorhees JJ. Fibroblast collapse and therapeutic
implication. Arch Dermatol. 2008;144(5):666–672.
17. Fisher GJ, Quan T, Purohit T, et al. Collagen fragmentation promotes
oxidative stress and elevates matrix metalloproteinase-1 in fibroblasts
in aged human skin. Am J Pathol. 2009;174:101–114.
18. Qiu H, Long X, Ye JC, et al. Influence of season on some skin proper-
ties: winter vs summer, as experienced by 354 Shanghainese women
of various ages. Int J Cosmet Sci. 2011;33(4):377–383.
19. Fanian F, Mac-Mary S, Jeudy A, et al. Efficacy of micronutrient supple-
mentation on skin aging and seasonal variation: a randomized, placebo-
controlled, double-blind study. Clin Interv Aging. 2013;8:1527–1537.
20. Watson RE, Bowden JJ, Bastrilles JY, Long SP, Griffiths CE. A cos-
metic “anti-ageing” product improves photoaged skin: a double-blind,
randomized controlled trial. Br J Dermatol. 2009;161:419–426.
21. Haftek M, Mac-Mary S, Le Bitoux MA, et al. Clinical, biometric and
structural evaluation of the long-term effects of a topical treatment
with ascorbic acid and madecassoside in photoaged human skin. Exp
Dermatol. 2008;17:946–952.
22. Chen W, Fu X, Ge S, Sun T, Sheng Z. Differential expression of matrix
metalloproteinases and tissue-derived inhibitors of metalloproteinase in
fetal and adult skins. Int J Biochem Cell Biol. 2007;39(5):997–1005.
23. Dasu MR, Spies M, Barrow RE, Herndon DN. Matrix metalloprotei-
nases and their tissue inhibitors in severely burned children. Wound
Repair Regen. 2003;11(3):177–180.
24. Mohan R, Chintala SK, Jung JC, et al. Matrix metalloproteinase
gelatinase B (MMP-9) coordinates and effects epithelial regeneration.
J Biol Chem. 2002;277(3):2065–2072.
25. Nusgens BV, Humbert P, Rougier A, et al. Topically applied vitamin
C enhances the mRNA level of collagens I and III, their processing
enzymes and tissue inhibitor of matrix metalloproteinase in the human
dermis. J Invest Dermatol. 2001;116(6):853–859.
26. Humbert P, Haftek M, Creidi P, et al. Topical ascorbic acid on photoaged
skin. Clinical, topographical and ultrastructural evaluation: double-blind
study vs. placebo. Exp Dermatol. 2003;12(3):237–244.
... These viscous biomaterials show hydrating capabilities and provide a temporary scaffolding effect for dermal tissue reconstruction through mechanical stimulation of fibroblasts. Therefore, they allow the restoration of a hydrated, supple, and dense skin [5]. Natural polysaccharides are particularly interesting for designing new intradermal devices since they represent structural analogues of living tissues (i.e. the glycosaminoglycans [GAGs]) [6]. ...
Article
Injectable soft-tissue devices are increasingly used for improving skin defects and deficiencies related to ageing. To assess the safety and efficacy of KIO015, a new injectable soft-tissue device formulated with carboxymethyl chitosan for the intradermal treatment of skin defects associated with ageing. Twenty-two subjects (40-65 years) were randomized to receive injections in the neckline of KIO015 and a non-cross-linked HA-based device, and were followed for up to 10 months. Injection site reactions (ISRs) and adverse events (AEs) were documented. Skin improvement was assessed instrumentally and clinically. Skin biopsies at injection zones in the lower back were taken at Day 28 for histopathology and immunohistochemistry analyses, to further assess product performance. Histomorphometric analyses on rabbits and in vitro assessment of KIO015 antioxidant capacity were also conducted. KIO015 was very well tolerated. Only expected and transient ISRs were observed; mainly erythema and hematoma. No adverse local effects or foreign body granuloma were observed histologically. Both clinical and instrumental evaluations confirmed the performance of KIO015. The skin was firmer and more elastic. Skin hydration showed significant improvement three days after injection. KIO015 exhibited superior overall maintenance of skin hydration after 10 months as compared to HA. These clinical results were supported by in vitro trials and implantation tests in the rabbit. The results from this pilot study support the use of KIO015 as an innovative alternative to HA-based devices for intradermal treatment of skin disorders.
... The results are set out in . [2,10,13,15,17] An improvement in blood perfusion was noticed in four studies. Fibroblast proliferation was enhanced together with an increase in collagen content. ...
Chapter
Full-text available
Vacuum massage is a noninvasive mechanical massage technique performed with a mechanical device that lifts the skin by means of suction to create and mobilize a skin fold. It was invented by a French engineer suffering from burn scars after a car accident and has since then been frequently used for the treatment of burn scars. The two most reported physical effects of vacuum massage were improvement of the tissue hardness and the elasticity of the skin. Besides physical effects, a variety of physiological effects are reported in the literature, for example, an increased number of fibroblasts and collagen fibers accompanied by an alteration of fibroblast phenotype and collagen orientation. Little information was found on the decrease of pain and itch due to vacuum massage. Although vacuum massage initially had been developed for the treatment of burn scars, a literature review found little evidence for the efficacy of this treatment. Variations in duration, amplitude, or frequency of the treatment have a substantial influence on collagen restructuring and reorientation, thus implying possible beneficial influences on the healing potential by mechanotransduction pathways. Vacuum massage may release the mechanical tension associated with scar retraction and thus induce apoptosis of myofibroblasts. Suggestions for future research include upscaling the study design, investigating the molecular pathways and dose dependency, comparing effects in different stages of repair, including evolutive parameters and the use of more objective assessment tools.
... This therapy was introduced to treat traumatic or burn scars. [14][15][16][17] Here, we report our experience using endermology to prepare irradiated skin for autologous fat injection. We illustrate the high efficiency and low morbidity of our minimally invasive approach combining peri-operative skin preparation with prepectoral lipofilling associated with prosthesis-based reconstruction. ...
Article
Full-text available
Background: Post-mastectomy irradiation severely impairs skin trophicity resulting in poor prosthetic implant outcome. Autologous fat grafting improves skin quality allowing minimally invasive approach with prosthetic reconstruction. Here, we report our pilot experience of preoperative mechanotherapy to optimize lipofilling and subsequent prosthetic reconstruction outcome. Methods: We retrospectively included 65 women that had breast reconstruction using autologous fat grafting and implant placement from 2012 to 2018 benefiting or not from mechanotherapy before the reconstructive procedure. Demographic and surgical outcomes were recorded. Results: The volume of fat injected was significantly superior in the mechanotherapy group compared with the controls for the first and second lipofilling (259.3 mL vs 150.6 mL and 251.8 mL vs 154 mL, respectively). Sixteen patients among controls required a pre-expansion prosthesis compared with none in the endermology group. The prosthesis volume was smaller in the endermology group. Six patients in the endermology group had a reconstruction without prosthesis. The aesthetic score evaluated by patients was 4.8 with no statistically significant difference between the two groups. Conclusion: Preoperative skin mechanotherapy and postoperative skin mechanotherapy increase skin compliance. It is associated with a higher volume of fat injection and lower prosthesis volume. If confirmed in a prospective study, endermology could become a standard in patients' preparation for lipofilling-based reconstruction.
... The relative improvement observed in nearly half of patient area receiving placebo cream probably results of the beneficial mechanical effects of massage since skin mechanical stimulation as it was recently shown stimulates fibroblast functions, i.e. enhances collagen, hyaluronic acid and elastin production. 10 Thus, a 5% vitamin C cream has shown for the first time beneficial effects on Bateman purpura. ...
Article
Background: Bateman purpura is characterized by diffuse senile skin atrophy, senile purpura and spontaneous stellar pseudo-cicatrices. Cutaneous changes in the course of aging have been related to lower levels of ascorbic acid into the dermis of elderly people. Objective: In the present study, we postulate that senile purpura could be linked to dermal vitamin C deficiency and could be corrected by topical administration of this vitamin. Methods: A 12-weeks, hemi-member (forearm or leg), randomized double-blind comparative study, was conducted in 18 patients with Bateman purpura aged over than 60 years. At each visit, clinical assessment and biometrological measurements were performed. Clinical examination and scoring by experts showed a significant improvement on the vitamin C-treated side compared with the control, with reduction of hemorrhage areas, increase of dermal thickness. Results: Twice-daily application of 5% topical vitamin C led to a clinically apparent improvement of the skin symptoms and allows beneficial effects on skin elasticity and thickness. Bateman purpura, a classical sign of photoaging whose origin has not clearly been recognized could be improved by vitamin C applied on to the skin. Conclusion: These results confirm the hypothesis of underlying of role vitamin C deficiency in the determinism of Bateman purpura. This article is protected by copyright. All rights reserved.
... The ex vivo study presented here demonstrates that an oscillatory mechanical stimulus exerted upon the skin surface can induce changes in the expression of some structural proteins, as previously shown in cellular models [1,8,40,41]. When the stimulation protocol is repeated twice year old donor, after 5 days of massaging for all markers, with the exception of type VII collagen and procollagen 1, which were sampled after 10 days of treatment from the same donor. ...
Article
Full-text available
Mechanical and geometrical cues influence cell behaviour. At the tissue level, almost all organs exhibit immediate mechanical responsiveness, in particular by increasing their stiffness in direct proportion to an applied mechanical stress. It was recently shown in cultured-cell models, in particular with fibroblasts, that the frequency of the applied stress is a fundamental stimulating parameter. However, the influence of the stimulus frequency at the tissue level has remained elusive. Using a device to deliver an oscillating torque that generates cyclic strain at different frequencies, we studied the effect(s) of mild skin massage in an ex vivo model and in vivo. Skin explants were maintained ex vivo for 10 days and massaged twice daily for one minute at various frequencies within the range of 65–85 Hz. Biopsies were analysed at D0, D5 and D10 and processed for immuno-histological staining specific to various dermal proteins. As compared to untreated skin explants, the massaging procedure clearly led to higher rates of expression, in particular for decorin, fibrillin, tropoelastin, and procollagen-1. The mechanical stimulus thus evoked an anti-aging response. Strikingly, the expression was found to depend on the stimulus frequency with maximum expression at 75Hz. We then tested whether this mechanical stimulus had an anti-aging effect in vivo. Twenty Caucasian women (aged 65-75y) applied a commercial anti-aging cream to the face and neck, followed by daily treatments using the anti-aging massage device for 8 weeks. A control group of twenty-two women, with similar ages to the first group, applied the cream alone. At W0, W4 and W8, a blinded evaluator assessed the global facial wrinkles, skin texture, lip area, cheek wrinkles, neck sagging and neck texture using a clinical grading scale. We found that combining the massaging device with a skin anti-aging formulation amplified the beneficial effects of the cream.
Article
Skin wound healing aims to repair and restore tissue through a multistage process that involves different cells and signalling molecules that regulate the cellular response and the dynamic remodelling of the extracellular matrix. Nowadays, several therapies that combine biomolecule signals (growth factors and cytokines) and cells are being proposed. However, a lack of reliable evidence of their efficacy, together with associated issues such as high costs, a lack of standardization, no scalable processes, and storage and regulatory issues, are hampering their application. In situ tissue regeneration appears to be a feasible strategy that uses the body's own capacity for regeneration by mobilizing host endogenous stem cells or tissue-specific progenitor cells to the wound site to promote repair and regeneration. The aim is to engineer instructive systems to regulate the spatio-temporal delivery of proper signalling based on the biological mechanisms of the different events that occur in the host microenvironment. This review describes the current state of the different signal cues used in wound healing and skin regeneration, and their combination with biomaterial supports to create instructive microenvironments for wound healing.
Article
This study aimed to evaluate the protective effect of artocarpin-enriched (Artocarpus altilis) heartwood extract on the mechanical properties of UVB-irradiated fibroblasts. Human skin fibroblasts were pretreated with 50 μg/mL(-1) extract and later irradiated with UVB (200 mJ/cm(-2) ). They were then cultured within three-dimensional of free-floating and tense collagen lattices. The pretreatment of fibroblasts with the extract prior to UVB radiation showed cells protection against UVB-induced suppression of α-SMA expression, fibroblast migration and contraction. These results reveal that the extract prevents mechanical damages induced by UVB irradiation in fibroblast-embedded collagen lattices, and therefore, has a potential as a natural photo-protectant.
Article
Full-text available
Background: Several studies have confirmed dramatic changes in skin surface parameters during the winter months. Although there are many studies supporting the positive effects of topical treatment, there are no published studies demonstrating the effects of oral supplementation in the prevention of negative skin changes during winter. The purpose of this study was to evaluate the efficacy of an oral micronutrient supplement in preventing the negative effects of winter weather on skin quality using noninvasive biometrologic instruments. Methods: This study included 80 healthy female volunteers aged 35-55 years with phototype II-IV skin. Randomization was balanced. Two tablets of a micronutrient supplement (Perfectil® Platinum) or placebo were administered once daily for 4 months. The volunteers were examined at baseline, after 4 months, and 6 weeks after termination of treatment (month 5.5). The evaluation included skin microrelief by Visioscan® as the main outcome, and the secondary outcomes were results on standard macrophotography, skin tension by Reviscometer®, skin high-frequency ultrasound, and self-assessment. Results: For all pseudoroughness and microrelief indicators, there was a significant increase from baseline to month 4 in the placebo group (P<0.05) but no change in the active group. Descriptive statistics for the mean minimum, mean maximum, and minimum to maximum ratio on the nonexposed study zone showed a significant and dramatic difference between baseline and month 4 and between baseline and month 5.5 (P<0.05) in the active group, indicating decreasing anisotropy of the skin. High-frequency ultrasound on the exposed study zone revealed that skin thickness was significantly decreased in the placebo group during winter but was stable in the treated group (P<0.01). The photography scaling and self-assessment questionnaire revealed no significant changes in either group. Conclusion: These results indicate that the skin is prone to seasonal changes during winter, particularly in exposed areas. The data also indicate that oral supplementation can be a safe treatment, with no serious side effects, and may prevent or even eliminate the negative effects of winter on the skin.
Article
Full-text available
Background: Wrinkles are characterized by changes in the organization and structure of the dermis. Human wrinkle fibroblasts (WF) have a different functional behaviour in comparison with normal-aged fibroblasts (NF). Decreases in migration capacities and collagen I synthesis are observed. Mitochondrial function is impaired with an increase in lac-tate production during aging. Sculptra® (poly-L-lactic acid: PLLA), a biodegradable synthetic polymer, is used for sub-cutaneous volume restoration. Thus we decided to investigate different fibroblast functions when placed in contact with PLLA. Objectives: The potential of PLLA to compensate for the reduction of metabolic activity, to restore the migra-tion capacity of WF and to inhibit the lactate production, was investigated and compared to NF. Methods: Two differ-ent skin samples were used from each of the three women’s facelift (one inside a face wrinkle and one from normal aged skin). Collagen I, lactate productions and proliferation capacities were investigated on monolayer cultures. Migra-tion properties were evaluated using three-dimensional collagen lattices. Results: PLLA increased collagen I synthesis, restored migration capacities and tended to decrease lactate production in WF, whereas PPLA stimulated proliferation in NF and tended to improve the migration of NF. Conclusion: These results suggested that PLLA from Sculptra® acted as a stimulus for collagen production in WF and that it is suitable for correcting skin depressions, such as wrinkles.
Article
Full-text available
The facial skin of 354 women, aged 18–80, living in Shanghai, was investigated over two successive 6 month periods, summer and winter. Results from clinical assessments indicate that aging signs, such as wrinkling and sagging, are unaffected over such period. However, physical measurements revealed alterations in some functional criteria of the skin, such as sebum output, skin colour, melanin content of pigmented spots, skin hydration, all being increased during summer. The relationships between all criteria, as well as technical or applied inferences/consequences from this study, are discussed. La peau du visage de 354 femmes résidant à Shanghai, âgées de 18 à 80 ans a été observée à deux périodes successives, été et hiver. Les résultats des évaluations cliniques montrent que Les signes liés au vieillissement, tels que rides et affaissement des traits, ne sont pas modifiés durant ces périodes. En revanche, des modifications de certains critères fonctionnels cutanés ont été observées par les mesures physiques, telle la production sébacée, la couleur de la peau, le contenu mélanique des taches pigmentaires, l’hydratation, tous étant augmentés au cours de l’été. Les relations entre ces critères, ainsi que les conséquences techniques ou appliquées résultant de cette étude, sont discutées.
Article
Full-text available
Published studies assessing whether asymmetrical facial ultraviolet light exposure leads to underlying differences in skin physiology and morphology report only clinical observations. The aim of this study was to assess the visual impact on the skin of repeated ultraviolet-A (UVA) exposure through a window. Eight women and two men presenting with asymmetrical signs of photoaging due to overexposure of one side of their face to the sun through a window over a long period of time were enrolled in the study. Split-face biometrologic assessments were performed (clinical scoring, hydration with Corneometer, mechanical properties with Cutometer, transepidermal water loss with AquaFlux, skin relief with fringe projection, photography, stripping, and then lipid peroxidation analysis). Significant differences were observed in clinical scores for wrinkles, skin roughness assessed by fringe projection on the cheek, and skin heterogeneity assessed with spectrocolorimetry on the cheekbone. Other differences were observed for skin hydration, as well as skin laxity, which tended towards significance. This study suggests the potential benefit of daily UVA protection during nondeliberate exposure indoors as well as outside.
Conference Paper
In developed countries, interest in cutaneous aging is in large part the result of a progressive, dramatic rise over the past century in the absolute number and proportion of the population who are elderly. The psychosocial, as well as physiologic, effects of skin aging on older persons have created a demand for a better understanding of the aging process and particularly for effective interventions. Skin aging is a complex process determined by the genetic endowment of the individual and by environmental factors. The appearance of old skin and the clinical consequences of skin aging have been well-known for centuries, but it is only in the past 50 years that mechanisms and mediators have been pursued systematically. Still, within a relatively short time, there has been tremendous progress, a progress greatly enhanced by basic gerontologic research using immunologic, biochemical, and in particular, molecular biologic approaches.
Article
Mechanical stimulation of fibroblasts induces extracellular matrix modifications. The aim of this study is to evaluate the effects on face of a "firming" treatment, dispensed with the mechanical device, Lift 6®, during 8 weeks. Efficacy on ageing skin has been measured by cutometry, stereophotogrammetry, histology, simple blind clinical exam by dermatologist and self-evaluation questionnaire. Cutometry could not detect any significant effect, but, an the contrary, the study could evaluate significant differences with the parameters of stereophotogrammetry, clinical exam, self evaluation questionnaire and on histological analysis. Although some parameters became improved as early as S4, (cutaneous laxity, smoother skin, skin improvment) the beneficial effect is mainly observed after 8 weeks of treatment and is maintained 2 weeks after the end of the treatment. Tolerance of the product was found excellent all along the study and 100% of the volonteers were satisfied with the treatment. The results observed argue in favor of a firming and remodeling action of the Lift 6®.
Article
Ascorbic acid (vitamin C) is a cofactor required for the function of several hydroxylases and monooxygenases. It is not synthesized in humans and some other animal species and has to be provided by diet or pharmacologic means. Its absence is responsible for scurvy, a condition related in its initial phases to a defective synthesis of collagen by the reduced function of prolylhydroxylase and production of collagen polypeptides lacking hydroxyproline, therefore, they are unable to assemble into stable triple-helical collagen molecules. In fibroblast cultures, vitamin C also stimulates collagen production by increasing the steady-state level of mRNA of collagen types I and III through enhanced transcription and prolonged half-life of the transcripts. The aim of the experimental work has been to evaluate the effect on dermal cells of a preparation of vitamin C topically applied on one side vs placebo on the other side of the dorsal face of the upper forearm of postmenopausal women. Biopsies were collected on both sides and the level of mRNA measured by non competitive reverse transcription–polymerase chain reaction made quantitative by the simultaneous transcription and amplification of synthetic RNA used as internal standards. The mRNA of collagen type I and type III were increased to a similar extent by vitamin C and that of three post-translational enzymes, the carboxy- and amino-procollagen proteinases and lysyloxidase similarly increased. The mRNA of decorin was also stimulated, but elastin, and fibrillin 1 and 2 were not modified by the vitamin. The expression of matrix metalloproteinases 1, 2, and 9 was not significantly changed, but an increased level of tissue inhibitor of matrix metalloproteinase 1 mRNA was observed without modification of tissue inhibitor of matrix metalloproteinase 2 mRNA. The stimulating activity of topical vitamin C was most conspicuous in the women with the lowest dietary intake of the vitamin and unrelated to the level of actinic damage. The results indicate that the functional activity of the dermal cells is not maximal in postmenopausal women and can be increased.Keywords: ADAMTS2, BMP1, decorin, elastin, matrix metalloproteinases
Article
Facial sagging is associated with aging, although the mechanism remains unclear. The aim of this study was to investigate the mechanism of facial sagging by examining the relationship of sagging severity to changes of skin elasticity, fat mass and facial muscle function at the cheek. Faces of 108 healthy Japanese female volunteers, aged 20-60 years were photographed at an angle of 45 degrees . Standard scores of sagging severity were established by analyzing the photographs. We examined the correlations of scored sagging levels with skin elasticity measured with a Cutometer MPA 580, fat content estimated by bioelectrical impedance analysis and facial muscle function (lip sealing force and occlusal force) in middle-aged female volunteers (30-40 years) with a wide range of sagging scores. Because the upper, lower and lateral areas in the cheek may show severe sagging, a six-grade score of sagging severity was separately established for each area. Each score was significantly correlated positively with age (20-60 years). In middle-aged volunteers, the sagging scores in all three areas of the cheek were significantly and negatively associated with skin elasticity. Body fat percentage was significantly and positively correlated with the sagging scores in the lower and lateral areas, although the correlation was only weakly positive in the upper area. Mimetic muscle function, measured in terms of lip sealing pressure, was significantly and negatively correlated with the sagging score only at the upper area of the cheek, but masticatory muscle function, measured in terms of occlusal force pressure, was not associated with the sagging score. Sagging may be associated with the reduction of skin elasticity and mimetic muscle function and increase of fat mass, but the relationships are different in different areas of the cheek.
Article
Very few over-the-counter cosmetic 'anti-ageing' products have been subjected to a rigorous double-blind, vehicle-controlled trial of efficacy. Previously we have shown that application of a cosmetic 'anti-ageing' product to photoaged skin under occlusion for 12 days can stimulate the deposition of fibrillin-1. This observation infers potential to repair and perhaps clinically improve photoaged skin. We examined another similar over-the-counter cosmetic 'anti-ageing' product using both the patch test assay and a 6-month double-blind, randomized controlled trial (RCT), with a further 6-month open phase to assess clinical efficacy in photoaged skin. For the patch test, commercially [corrected] available test product and its vehicle were applied occluded for 12-days to photoaged forearm skin (n = 10) prior to biopsy and immunohistochemical assessment of fibrillin-1; all-transretinoic acid (RA) [corrected] was used as a positive control. Sixty photoaged subjects were recruited to the RCT (test product, n = 30 vs. vehicle, n = 30; once daily for 6-months; face & hands) [corrected] with clinical assessments performed at recruitment and following 1-, 3- & 6-months of use [corrected]. Twenty-eight subjects had skin biopsies (dorsal wrist) at baseline and at 6 months of treatment for immunohistochemical assessment of fibrillin-1 (test product, n = 15; vehicle, n = 13). All subjects [corrected] received test product for a further 6-months. Final clinical assessments were performed at the end of this open period; 27 subjects received test product for 12-months [corrected]. In the 12-day patch test assay, we observed significant immunohistological deposition of fibrillin-1 in skin treated by test product and RA as compared to untreated baseline (P = 0.005 and 0.015 respectively). In the clinical RCT, at 6 months, compared to baseline assessment, 43% of subjects on test product had an improvement in facial wrinkles (P = 0.013), whereas only 22% of subjects using vehicle had clinical improvement (P = ns). Between group comparison of test product and vehicle was non-significant (P = 0.10). After 12 months, there was a significant benefit of test product over that projected for vehicle (70% vs. 33% of subjects improving; combined Wilcoxon rank tests, P = 0.026). There was significant deposition of fibrillin-1 in skin treated for 6 months with test product (mean +/- SE; vehicle, 1.84 +/- 0.23; test product, 2.57 +/- 0.19; P = 0.019). An over-the-counter cosmetic 'anti-ageing' product demonstrated clear benefit over vehicle in fibrillin-1 deposition over a 6-month trial period. There was a corresponding but non-significant trend towards clinical improvement in facial wrinkles. Clinical improvements in the treated group were increased after a further 6-months of use. This study demonstrates that a cosmetic may improve the appearance of wrinkles and further supports the use of fibrillin-1 as a robust biomarker for repair of photoaged dermis.