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ORIGINAL RESEARCH
Skin protective effects of an antipollution,
antioxidant serum containing Deschampsia
antartica extract, ferulic acid and vitamin C:
a controlled single-blind, prospective trial in
women living in urbanized, high air pollution area
This article was published in the following Dove Press journal:
Clinical, Cosmetic and Investigational Dermatology
Massimo Milani
1
Bita Hashtroody
2
Marco Piacentini
3
Leonardo Celleno
3
1
Medical Department, Cantabria Labs
Difa Cooper, Caronno P, VA, Italy;
2
Cantabria Labs Medical Department,
Madrid, Spain;
3
Eurofins Cosmetics &
Personal Care, Italy
Introduction: Air pollution causes skin damage and favors skin aging processes such as
dark spots and wrinkles, through oxidative stress. Pollutant substances accelerate skin aging
through a specific activation of intracellular receptors called AhR (aryl-hydrocarbon recep-
tors). Deschampsia antartica aqueous extract (DAE) has shown to counteract the pollutant-
induced AhR activation. Ferulic acid (FA) and vitamin C (VC) are potent antioxidant
substances. A serum containing DAE/FA/VC has been recently developed. So far, no clinical
data are available regarding the protective actions of this serum against the detrimental
effects of air pollution on the skin.
Objective: We conducted a prospective, single-blind, 28-day study to assess efficacy and
protective effects against air pollution skin damage of a new serum containing Deschampsia
antartica extract.
Materials and methods: Twenty, photo type I–III, women (mean age 42 years) with at least
three dark spots on the face, living in a homogenous urbanized, high pollution area (Rome)
were evaluated. The objectives of the study were to evaluate the effects of treatment on skin
barrier function, assessed by transepidermal water loss (TEWL) measurement (Tewameter),
the effect on dark spots, evaluated by means of colorimetry (Colorimeter CL 400), and the
effect on squalene peroxide (SQOOH)/squalene (SQ) skin ratio assessed with face swabs.
Results: The trial was conducted between November 20 and December 19, 2018. In
comparison with baseline, the product induced a significant improvement of skin hydration
(−19% of TEWL), a significant improvement of dark spots (+7%) and a significant improve-
ment of SQOOH/SQ ratio (−16%). The product was evaluated very well by >90% of the
treated subjects regarding cosmetic acceptability.
Discussion: A serum containing DAE/FA/VC has shown to improve skin barrier function,
to reduce dark spots and to counteract the skin oxidative stress in women living in high
pollution urban area.
Keywords: Deschampsia antartica aqueous extract (DAE), air pollution, skin aging, Squalene
Introduction
The skin is an organ in direct contact with various air pollutants.
1
Air pollution
causes skin damage and favors skin aging processes such as dark spots and
wrinkles, through oxidative stress.
2
In particular, air pollution causes alteration
Correspondence: Massimo Milani
Medical Department, Cantabria Labs Difa
Cooper, Via Milano 160, Caronno P., VA,
Italy
Tel +3 902 965 9031
Email massimo.milani@difacooper.com
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http://doi.org/10.2147/CCID.S204905
of skin barrier functions, oxidative stress, and
inflammation.
3
Skin exposed to pollutants reduces its
antioxidative potential.
4
Furthermore, pollutant sub-
stances, mainly particulate matter (PM10, PM2.5),
accelerate skin aging through specific activation of intra-
cellular receptors called AhRs (aryl-hydrocarbon
receptors).
5
Several studies have demonstrated that
AhRs are involved in the pathophysiology of skin,
including skin hyperpigmentation, photocarcinogenesis
andskininflammation.
6
Deschampsia antartica (DA)
is a polyextremophile plant native to Antarctica.
7
This
plant lives under high solar irradiation, salinity and
oxygen concentration, low temperature and extreme
dryness.
8
DA aqueous extract (DAE) has shown,
in vitro, to counteract the pollutant-induced AhR
activation.
9
Ferulic acid (FA) and vitamin C (VC) are
potent antioxidant substances.
10,11
The standardized
Crypthomphalus aspersa (a snail of the Helicidae
family) secretion (SCA), rich in proteins, glycosamino-
glycans and antioxidant, is extensively used in cosmetic
products as an antiaging and skin regenerating
substance.
12
A serum containing DAE/FA, VC and
SCA (10%) has been recently developed (EC serum).
So far, no clinical data are available regarding the pro-
tective actions of this serum in vivo against the detri-
mental effects of air pollution on the skin.
Trial objectives
We conducted an explanatory, experimental, prospective,
pilot, single-blind, controlled, 28-day study to evaluate the
skin protective effects of EC-serum in women living in
urbanized area.
Materials and methods
Subjects
We enrolled 20 women (mean age 42 years) living in
a homogenous urbanized, high pollution area (Rome) compel-
ling to spend at least 2 hrs outdoor. The main inclusion criteria
were:womenagedfrom35to45years,withaphototype
(Fitzpatrick): I–III, residents in a homogeneous urban area,
compelled to spend at least 2 hrs a day outdoors, with at least
three dark spots on the face without any other facial skin
diseases. During the entire study duration, the volunteers
were requested not to use on face any topical products (cos-
metics/drugs/medical devices) or sunscreen or any products
with an SPF. Table 1 reports subjects’characteristics at
baseline.
Ethical aspects
All the enrolled women gave their written informed con-
sent. The study was performed according to: 1) the general
principles of medical ethics in clinical research coming
from the Declaration of Helsinki (June 1964) and its suc-
cessive amendments;
13
2) the international recommenda-
tions relating to Good Clinical Practices for conducting
clinical trials for drugs ICH E6(R1) of 10/06/1996
(CPMP/ICH/135/95);,
14
3) the Directive of the European
Parliament and Council 2001/20/EC concerning the harmo-
nization of legislative, statutory and administrative provi-
sions of the member States relating to the application of
good clinical practices when conducting clinical trials for
drugs for human use –OJ/EC of 01/05/2001 and 4) the
recommendations of Colipa guidelines, August 1997.
15
The
study protocol was approved by the Local Ethical
Committee. The study was performed at the Institute of
Eurofins BioPharma Product Testing, certified ISO
9001:2015 and equipped with material and technical
means suitable for noninvasive clinical researches, compa-
tible with the safety requirements for human subjects. The
independent Ethical Committee of Eurofins Institute
approved the study protocol in October 2018.
Application of DAE/FC/VC serum
The subjects were instructed to apply the tested product
(Endocare-C Edafence serum; Cantabria Labs, Madrid,
Spain) at home under the normal conditions of use, for
28 consecutive days, on the entire face. The product was
applied on the face twice daily (in the morning and in
the evening). In particular, specific instructions were
given to each subject for the correct application of the
tested product: 1) do not wash the face after the appli-
cation of the serum; 2) do not apply any other skincare
products and 3) do not use any make-up products during
the study period.
Trial outcomes
The objectives of the study were to evaluate the effects of
treatment on skin barrier function, assessed by transepi-
dermal water loss (TEWL) measurements, the effect on
dark spots, evaluated by means of colorimetry and the
antioxidative protective skin effect evaluating the squalene
peroxide (SQOOH)/squalene (SQ) skin ratio assessed by
face swabs. TEWL measurements were performed on the
face by means of Tewameter TM 300 (Courage &
Khazaka, Koln, Germany ) and expressed as g/m
2
/hr.
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The evaluation of the dark spots was performed using the
Colorimeter CL 400 (Courage & Khazaka) device. This
instrument allows evaluating the skin color using the chro-
matic space L*a*b*. Those data represent the luminance
(L*), the quantity of red (a*: red–green axis) and the color
intensity (b*: yellow–blue axis) and allow the exact loca-
tion of each color in the chromatic tridimensional diagram.
In this study, to evaluate the intensity of the color of the
dark spots, the individual typology angle (ITA°) was con-
sidered: ITA°= [arc tangent ((L* - 50)/b*)] 180/3.14159. If
the ITA° value increases, the dark spots are lighter. To
evaluate the effect on dark spots, high-resolution photo-
graphs of the dark spots were performed under standar-
dized lighting with Visioface Quick (Courage & Khazaka).
Standardized digital photographs of the full face were
taken at D0 and at D28 using the VISIA-CR photo station
(Canfield Imaging System, Fairfield, NJ, USA). All these
measurements were performed by the same technician,
before any application (D0/T0) and then after 28 consecu-
tive days of product use (D28). Serum samples were taken
from forehead with a swabbing method at D0 and at D28.
Swab homogenates were centrifuged at 10,000 gand sub-
sequently analyzed for squalene monohydroperoxide
(SQOOH) and SQ content with liquid chromatography
system according to Jourdain et al.
16
A specific subjects’
cosmetic qualities and efficacy evaluation questionnaire
was also performed at day 28 to evaluate the perceived
efficacy by the subjects and the cosmetic acceptability.
Statistical methods
This was a pilot study and therefore a formal calculation of
sample size was not performed. Statistical analysis was done
using GraphPad statistical software ver. 13.0 (La Jolla, CA,
USA). Continuous variables were expressed as mean±SD.
The paired Student's t-test and the Wilcoxon tests were used
for the analysis of the study outcomes comparing baseline
(D0) and D28 values. We calculated also the 95% CIs of the
difference in all the variables evaluated. A p-value of <0.05
was considered significant.
Results
Tab le 1 summarizes the main demographic and clinical char-
acteristics of enrolled subjects. All the enrolled women con-
cluded the 28-day study period. No clinical sign imputable to
the investigational product was observed by the investigator
during the study. No sensation of discomfort was reported by
the subjects during the study. The trial was conducted
between November 20 and December 19, 2018. During this
period, elevated PM10 concentrations (>20 µg/m
3
)inthe
respective urban areas were registered (ARPA official data-
base, Agenzia regionale per la protezione ambientale)in23
of 29 days. The PM10 average during the study period 35±9
µg/m
3
. At baseline, the TEWL values were 10.4±2 g/m
2
/hr.
After treatment, TEWL was reduced significantly (P=0.002;
Wilcoxon Test) to 8.3±1 g/m
2
/hr, with an absolute difference
of 2.1 (95% CI: 1.3–2.9) (Figure 1). In comparison with
baseline, this represents a reduction of −19% in the TEWL
mean values. A significant (P=0.01; Wilcoxon test) improve-
ment (decrease in intensity of pigmentation) of dark spots
(+7%) was observed after treatment. The ITA° angle at base-
line was 23.8±4.4, and it increased significantly to 25.6±5.6
at D28 (P=0.0021; Wilcoxon test), with an absolute differ-
ence of 1.7 (95% CI: 0.80–2.7) (Figure 2). A significant
improvement of SQOOH/SQ ratio was observed at day 28
in comparison with baseline data (−16%). The SQOOH/SQ
ratio at baseline was 12.7±7.6% and 9.0±5.6% after treat-
ment (P=0.017; Wilcoxon test) with an absolute difference of
3.6 (95% CI: 0.40–7.7) (Figure 3). Table 2 reports numerical
data analysis for each instrumental parameter. In Figure 4 are
presented four high definition pictures of two subjects at
baseline and after treatment,l demonstrating the whitening
Table 1 Subjects’characteristics at baseline
Number 20
Mean age, years, mean±SD 42±7
Fitzpatrick photo type, % I: 0%
II:40%
III:60%
Subjects with sensitive skin, n(%) 10 (50%)
Transepidermal water loss, (TEWL) g/m
2
/hr, mean±SD 10.4±2
TEWL
*
D0
0
5
10
15
g/m2/h
D28
Figure 1 Evolution of transepidermal water loss (TEWL). *P=0.002; Wilcoxon test.
Abbreviations: TEWL, Trans Epidermal Water Loss; D0, Day zero; D28, Day 28.
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effect on dark spots of the tested serum. These subjects have
provided written informed consent for their images to be
published. The product was evaluated very well by >90%
of the treated subjects regarding cosmetic acceptability. In
more detail, 80% of the subjects judged that the product was
able to reduce hyperpigmentation; 95% stated that the pro-
duct made the skin brighter, smoother and softer; 90% of the
subjects judged that the product has a good texture and it was
easy to apply. No side effects were reported.
Discussion
According to the WHO,
17
93% of the population world-
wide is exposed to an excessive level of outdoor and/or
indoor pollution. Air pollution exposure is associated
with premature skin aging, wrinkle formation, pigmen-
ted spots, urticaria and eczema.
3
Pollutant substances,
mainly particulate matter (PM10, PM2.5), cause oxida-
tive stress via the production of ROS and the secretion
of proinflammatory cytokines such as TNF, IL-1 and IL-
8.
18
Furthermore, exposure to air pollutants increases
matrix metalloproteinases (MMP) including MMP-1,
MMP-2 and MMP-9, with an increase of dermal col-
lagen degradation.
19
PM accelerates skin aging through
specific activation of intracellular receptors called
AhR.
20
Air pollution causes alteration of skin barrier
functions, oxidative stress and inflammation.
21
Exposure to traffic air pollution is related to skin aging
in light-skinned women.
22,23
Subjects living in high
pollution area have a reduced content of SQ in sebum,
increased lactic acid levels and altered cohesion of stra-
tum corneum.
24
The pollutant substance benzopyrene
increases the skin amount of SQOOHs, and this process
contributes to skin hyperpigmentation and dark spot
formation.
25
Pham et al have demonstrated that oxidized
SQ is a reliable marker of environmental pollution.
26
In
our study, we have documented that the use of DAE/FA/
VC serum decreases the formation of SQOOHs at skin
level, reducing the ratio SQOOX/SQ. SQOOX is con-
sidered a secondary mediator of environmental induced
stress on the skin.
27
SQOOHs induce hyperpigmentation
in experimental animal models.
28
Several cosmetic pro-
ducts present on the market declare antipollution
effects.
29
However, it is important that these claims
should be validated by specificinvitroandinvivo
testing.
30
The serum we have evaluated in this trial
contains DAE, FA and VC. DAE has shown, in vitro,
to reduce the pollutant-induced AhR activation.
9
This
extract is able also to counteract the detrimental effect
of tobacco smoke, heavy metals and oxidant substances
on cell viability of cultivated human dermal
fibroblasts.
31
FA is a potent ubiquitous plant antioxidant,
and it can improve the chemical stability of vitamins
C.
32
Topical VC has a relevant antioxidant and light-
ening effect.
33
In our study, we observed that the 28-day
use of DAE/FA/VC serum improves skin barrier
Colorimetry of dark spot
ITA angle D0
ITA angle D28
0
20
10
30
ITA angle value
*
Figure 2 Evolution of darkspots evaluated withcolorimetry;*P=0.0021; Wilcoxon test.
Abbreviation: ITA, Individual Typology Angle.
0
5
10
SQOOH/SQ %
*
Ratio
15
20
SQOOH/SQ D0
SQOOH/SQ D28
Figure 3 Evolution of SQOOH/SQ ratio. *P=0.017; Wilcoxon test.
Abbreviations: SQOOH, Squalene peroxide; SQ, Squalene.
Table 2 Data analysis of outcomes variables
Baseline Day 28 Variation
and P-value
Transepidermal water loss
(TEWL: g/m
2
/hr; mean±SD
10.4±1.99 8.3±1.3 −19.1%
(<0.05)
Colorimetry(individual typology
angle (ITA°); mean±SD
23.8±4.4 25.5±5.6 +7%
(<0.05)
Squalene peroxided SQOOH/
Squalene SQ %
12.7±7.6 9.0±5.6 −16%
(<0.05)
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function, has a relevant whitening effect on dark spots
and reduces the oxidative stress of the skin. Study
limitations should be taken into account in evaluating
the results of our study. The trial was single-blind;
however, the main endpoints of the study were evalu-
ated in an objective investigator-independent manner
(TEWL, colorimetry, sebum analysis). In our trial, we
did not perform any evaluation of the effect on AhR
activity which is problematic in a clinical setting. In
addition, the present study was performed during
a winter period only, when the main air pollutant sub-
stances are PMs. During the summer period, a relevant
pollutant is ozone.
34
Ozone is an important pro-
oxidative substance able to damage skin barrier.
35
Future trials, conducted during the summer period,
assessing the clinical effects of the tested product
could be useful to evaluate the skin protective effect
also against this dangerous pollutant substance. The
tested serum contains several active compounds
(DAEFA, VC and SCA), and therefore it is not possible
to discriminate the clinical effects we have observed in
this study. The main goal was to evaluate the capability
of the serum to protect the skin of subjects living in
a high pollution environmental, during a time frame
(winter season) characterized by high outdoor pollution
(especially PM 10). During the clinical evaluation, high
pollution days (PM10 exceeding 20 µ/m
3
) represented
80% of the entire study period. This further supports the
fact that we tested the skin protective effect of this
serum under an appropriate time frame for assessing
its real antipollution effect.
Conclusion
Short-term use of a serum containing DAE/FA VC and
SCA has shown to improve skin barrier function, to reduce
Figure 4 High-resolution pictures of two subjects evaluating dark spots (Aand A1: baseline, D0; Band B1: after treatment D28).
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dark spots and to counteract the skin oxidative stress in
women living in high pollution urban area.
Acknowledgments
Difa Cooper Spa, Cantabria Labs Group, supported this
trial with an unrestricted grant.
Author contributions
MP and LC conducted the trial performing visits and
instrumental evaluations. MM was involved in study pro-
tocol design. All the authors contributed toward data ana-
lysis, drafting and critically revising the paper, gave final
approval of the version to be published and agree to be
accountable for all aspects of the work.
Disclosure
Dr Massimo Milani and Dr Bita Hashtroody are employees of
Cantabria Labs Difa Cooper and they, as well as Dr Marco
Piacentini, report personal fees from Cantabria Labs Difa
Cooper, during the conduct of the study. The authors report
no other conflicts of interest in this work.
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2133.2005.06939.x
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