Effects of a cosmetic ‘anti-ageing’ product improves photoaged skin

Article (PDF Available)inBritish Journal of Dermatology 161(2):419-26 · May 2009with116 Reads
DOI: 10.1111/j.1365-2133.2009.09216.x · Source: PubMed
Abstract
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.
THERAPEUTICS BJD British Journal of Dermatology
A cosmetic ‘anti-ageing’ product improves photoaged skin:
a double-blind, randomized controlled trial
R.E.B. Watson, S. Ogden, L.F. Cotterell, J.J. Bowden, J.Y. Bastrilles, S.P. Long* and C.E.M. Griffiths
Dermatological Sciences Research Group, School of Translational Medicine, Faculty of Medical and Human Sciences, The University of Manchester, Oxford Road,
Manchester M13 9PT, U.K.
*Alliance Boots Ltd, Nottingham NG2 3AA, U.K.
Note added after online publication:
Since the publication of this article online on 28 April 2009, the authors wish to recognize the following changes to the
article:
Title: Effects of a cosmetic ‘anti-ageing’ product on photoaged skin
Conflicts of Interest: This study was funded by Alliance Boots Ltd.
Summary
Methods For the patch test, commercially 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-trans
retinoic acid (RA) was used as a positive control. Sixty photoaged subjects were recruited to the RCT (test pro-
duct, n = 30 vs. vehicle, n = 30; once daily for 6-months; face & hands) with clinical assessments performed at
recruitment and following 1-, 3- & 6-months of use. 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 received test product for a further 6-months. Final clinical assessments were per-
formed at the end of this open period; 27 subjects received test product for 12-months.
Results 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 =0Æ005 and 0Æ015 respectively). In the clini-
cal RCT, at 6 months, compared to baseline assessment, 43% of subjects on test product had an improvement in
facial wrinkles (P =0Æ013), whereas only 22% of subjects using vehicle had clinical improvement (P = ns).
Between group comparison of test product and vehicle was non-significant (P =0Æ10). After 12 months, there
was a significant benefit of test product over that projected for vehicle (70% vs. 33% of subjects improving; com-
bined Wilcoxon rank tests, P =0Æ026). There was significant deposition of fibrillin-1 in skin treated for 6 months
with test product (mean ± SE; vehicle, 1Æ84 ± 0Æ23; test product, 2Æ57 ± 0Æ19; P =0Æ019).
Conclusion 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.
2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426 419
Correspondence
R.E.B. Watson.
E-mail: rachel.watson@manchester.ac.uk
Accepted for publication
3 April 2009
Key words
fibrillin-1, patch-test assay, randomized controlled
trial, wrinkles
Conflicts of interest
S.P.L. is employed by Alliance Boots Ltd., the
manufacturer of the commercially available
preparation tested in this study.
Re-use of this article is permitted in accordance
with the Creative Commons Deed, Attribution
2Æ5, which does not permit commercial
exploitation.
DOI 10.1111/j.1365-2133.2009.09216.x
Summary
Background Very few over-the-counter cosmetic ‘anti-ageing’ products have been
subjected to a rigorous double-blind, vehicle-controlled trial of efficacy. Previ-
ously 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.
Objective 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 effi-
cacy in photoaged skin.
Methods For the patch test, a commercially 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-trans retinoic acid
(RA) 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
and hands) with clinical assessments performed at recruitment and following 1,
3 and 6 months of use. Twenty-eight volunteers had skin biopsies (dorsal wrist)
at baseline and at 6 months treatment for immunohistochemical assessment of
fibrillin-1 (test product, n = 15; vehicle, n = 13). All volunteers received the test
product for a further 6 months. Final clinical assessments were performed at the
end of this open period.
Results In the 12-day patch test assay, we observed significant immuno-
histological deposition of fibrillin-1 in skin treated with the test product and
RA compared with the untreated baseline (P =0Æ005 and 0Æ015, respectively).
In the clinical RCT, at 6 months, the test product produced statistically
significant improvement in facial wrinkles as compared to baseline assessment
(P =0Æ013), whereas vehicle-treated skin was not significantly improved
(P =0Æ11). After 12 months, there was a significant benefit of the test
product over that projected for the vehicle (70% vs. 33% of subjects
improving; combined Wilcoxon rank tests, P =0Æ026). There was significant
deposition of fibrillin-1 in skin treated for 6 months with the test product
[(mean ± SE) vehicle 1Æ84 ± 0Æ23; test product 2Æ57 ± 0Æ19;
ANCOVA
P =0Æ019).
Conclusions In a double-blind RCT, an over-the-counter cosmetic ‘anti-ageing’
product resulted in significant clinical improvement in facial wrinkles, which was
associated with fibrillin-1 deposition in treated skin. This study demonstrates that
a cosmetic product can produce significant improvement in the appearance of
wrinkles and further supports the use of fibrillin-1 as a robust biomarker for the
repair of photoaged dermis.
All tissues, regardless of body site, are subject to intrinsic age-
ing, the result of the passage of time. Few clinically apparent
changes occur in intrinsically aged skin until the individual is
over 70 years of age at which point fine wrinkles become
apparent.
1
Skin, more than any other organ, is also subject
to environmental influences which can lead to extrinsic
ageing. One such environmental factor is chronic exposure to
sunlight which results in phenotypic changes termed photo-
ageing—inevitably a combination of intrinsic ageing and
photodamage. By comparison with intrinsic ageing, photoaged
skin is rough, dyspigmented and exhibits both fine and deep
wrinkles.
2,3
Histological examination of intrinsically aged skin
reveals atrophy of the dermal extracellular matrix (ECM), with
reduced levels of collagen and elastin.
4
Photoaged skin has a
different ECM morphology with solar elastosis—the deposition
of dystrophic elastic fibres in the dermis—being a prominent
histological feature.
5
Photoaged dermis contains significantly
reduced levels of collagen types I and III,
6
fewer anchoring
2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426
420 Clinical efficacy of a cosmetic ‘anti-ageing’ product, R.E.B. Watson et al.
fibrils at the dermal–epidermal junction (DEJ; collagen VII)
7
and loss of the fibrillin-rich microfibrillar architecture in the
papillary dermis.
8
These remodelled ageing phenotypes are
thought in part to be due to increased cutaneous expression
of matrix metalloproteinases (MMPs).
9–11
Topical retinoids are used as the clinical, evidence-based ‘gold
standard’ for the treatment of photoaged skin.
12
Numerous
studies have shown the reparative effects of topical application
of all-trans retinoic acid (RA), which includes the partial restora-
tion of collagens I, III
13
and VII
14
and restoration of the fibril-
lin-rich microfibrillar network.
15
These ECM changes, together
with reduced MMP expression may in part explain the clinical
improvement of photoaged skin produced by topical retin-
oids.
16–18
We showed previously, in a 12-day occluded patch
test assay, that a specific cosmetic ‘anti-ageing’ product also has
the ability to stimulate the accumulation of fibrillin-1.
19
Although prescription retinoids can affect these significant
clinical and histological changes in photoaged skin there is
scant evidence that any of the plethora of cosmetic ‘anti-
ageing’ products can produce similar effects. We firstly
examined whether another, similar cosmetic ‘anti-ageing’
product can induce accumulation of fibrillin-1 in photoaged
skin using the patch test protocol. We then investigated the
same product in a rigorous double-blind, randomized
controlled trial (RCT) to ascertain whether or not its use
results in a clinically detectable benefit.
Methods
Test products
A commercially available product provided by Alliance Boots
Ltd (No7 Protect & Perfect Intense Beauty Serum; Alliance
Boots Ltd, Nottingham, UK) was investigated in these studies,
together with a vehicle formulation. The product is a water in
silicone emulsion with glycerine and other emollients and a
complex of ‘anti-ageing’ ingredients comprising natural
extracts and peptides: sodium ascorbyl phosphate, Panax ginseng,
Morus alba, Lupinus alba, tocopherol, palmitoyl oligopeptide,
palmitoyl tetrapeptide-7, Medicago sativa and retinyl palmitate.
The vehicle was of identical composition, but without the
complex of ‘anti-ageing’ ingredients.
In vivo patch test study
Ten healthy but photoaged volunteers were recruited (four
men, six women; age range 61–76 years) and subjected
to an extended 12-day patch test assay.
19
Test substances
(vehicle and test product 20 lL) were applied separately to the
extensor photoaged aspect of the forearm under standard 6-mm
diameter Finn chambers (Scanpore, Tuulsula, Finland). In add-
ition, an area was left untreated but was occluded to provide a
baseline control sample. Test products were applied to clean skin
on days 1, 4 and 8 of the assay. RA (0Æ025%; Retin-A
cream;
Janssen-Cilag Ltd, Beerse, Belgium; 20 lL) was applied to an
untreated site on day 8 and left in situ for 4 days to avoid
potential complications of irritancy caused by extended occlu-
sion. On day 12, the Finn chambers were removed and 3-mm
punch biopsies were obtained under 1% lignocaine local
anaesthesia from each test site. Biopsied tissue was embedded
in optimal cutting temperature compound (Tissue-Tek
;Miles
Laboratories, Elkhart, IN, U.S.A.), snap frozen in liquid nitrogen
and stored at )70 C prior to immunohistochemical analyses.
The North Manchester Local Research Ethics Committee approved
the study and all subjects gave written, informed consent.
Slide preparation
Frozen sections were prepared at a thickness of 10 lm (OTF
cryostat; Bright Instruments Ltd, Cambridge, U.K.) and moun-
ted onto gelatin-coated slides prior to histological analysis.
Immunohistochemistry
Immunohistochemistry was performed as previously des-
cribed
19
to identify a panel of ECM molecules or remodelling
enzymes in frozen sections from the 12-day patch test assay
and from the RCT. Primary antibodies were applied overnight
at 4 C. These were: mouse antihuman fibrillin-1 (clone
11C1.3; Neomarkers, Union City, CA, U.S.A.) diluted 1 : 100;
rat antihuman procollagen-1 (pCI) (clone M-58; Chemicon
International Inc., Temecula, CA, U.S.A.) diluted 1 : 1000; or
mouse antihuman MMP-1 (Oncogene Research Products, Bos-
ton, MA, U.S.A.) diluted 1 : 100. Negative controls were by
incubation of isotype sera at the appropriate concentration or
omission of the primary antibody. Sections were washed in
TBS prior to incubation with the appropriate biotinylated sec-
ondary antibody for 30 min. Antibody staining was visualized
using a well-characterized immunoperoxidase reaction (Vecta-
Stain
Elite ABC system; Vector Laboratories, Burlingame, CA,
U.S.A.) utilizing Vector SG
as chromogen. Following light
counterstaining with nuclear fast red, sections were serially
dehydrated and permanently mounted. Stained sections were
randomized, blinded and examined on a Nikon OPTIPHOT
microscope (Tokyo, Japan). The degree of immunostaining for
fibrillin-1 and pCI was assessed as previously described.
8,15,19
In brief, a five-point semiquantitative scale was used where
0 = no staining and 4 = maximal staining within the experi-
ment. The numbers of epidermal keratinocytes positive for
MMP-1 were quantified per high-power field (hpf; · 400).
Four sections (including control) were examined per subject,
per site, per treatment and the average score calculated.
Randomized controlled trial
Sixty healthy but photoaged volunteers were recruited to this
study (11 men, 49 women; age range 45–80 years). All test
products were supplied in identically packaged, coded con-
tainers so that the investigators and subjects were unaware as
to the treatment. Subjects were randomly allocated to self-
treatment with either the vehicle formulation or the test prod-
uct as described by a randomization programme (StatsDirect
2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426
Clinical efficacy of a cosmetic ‘anti-ageing’ product, R.E.B. Watson et al. 421
Ltd, Altrincham, U.K.) and instructed on the use of their allot-
ted cream—daily evening application to the entire face and
dorsa of the hands, including the wrists and extensor forearm,
for 6 months. Clinical assessments of the skin of the face and
dorsal hands were performed for all participants at baseline
and following 1, 3 and 6 months of product use. The follow-
ing four parameters were assessed at each visit: fine lines and
wrinkles, dyspigmentation, overall clinical grade of photoage-
ing and tactile roughness. The degree of fine lines and wrin-
kles, dyspigmentation and the overall level of photoageing
were scored according to the well-characterized Griffiths
photonumeric scale for photoaged skin.
20
The scale ranges
from 0 to 8, where 0 represents no evidence of photoageing
and 8 represents the most severe photoageing. Pigment was
assessed on a similar 0–8 scale, where 0 denotes a uniform
coloration of the skin with absence of photoageing-related
colour change and 8 represents severe dyspigmentation. Like-
wise, tactile roughness was scored on the treated areas from 0
to 8, where 0 represents totally smooth skin with no rough
patches and 8 represents very roughened skin.
In addition, 28 subjects provided 3-mm skin biopsies from
the dorsal wrist at the beginning and end of the 6-month
study period (vehicle formulation, n = 13; test product,
n = 15). These biopsies were evaluated for the expression of
fibrillin-1 in the papillary dermis, as previously described.
19
All the subjects were monitored for the occurrence of serious
adverse events up to, and including, 28 days after their
involvement with this study. The Salford and Trafford Local
Research Ethics Committee approved the study and all the sub-
jects gave written, informed consent.
Statistical analyses
In vivo patch test study
Differences in the amount of fibrillin-1 immunostaining
produced by the vehicle and test product were assessed for
significance using the repeated measures analysis of variance
(
ANOVA). Results were considered significant if P <0Æ05 (95%
confidence level) and were calculated using SPSS+ v 11.5 soft-
ware (SPSS Inc., Chicago, IL, U.S.A.).
Randomized controlled trial
Clinical assessment. Analysis of covariance (
ANCOVA), using the
baseline as covariate, was used to assess the 6-month data.
Linear regression analysis was used to extrapolate the vehicle
response to 12 months, thus allowing direct comparison with
the test product, validity confirmed by the Monte Carlo simu-
lation.
21
As all volunteers used the test product in the final
6 months, the 12-month clinical assessment data were analy-
sed using a combination of Wilcoxon’s matched pairs signed
rank and rank sum tests, to give an overall P-value.
In-use biopsy samples.
ANCOVA was performed, using the baseline
as covariate to assess changes in the deposition of fibrillin-1 in
the papillary dermis, with significance taken at the 95% confi-
dence level (SAS 9.1; SAS Institute Inc., Cary, NC, U.S.A.).
Results
In vivo patch test study
RA (the clinical ‘gold standard’) produced a significant deposi-
tion of fibrillin-1 in the papillary dermis compared with that
observed at baseline (P =0Æ015). Application of the vehicle,
following the 12-day patch test assay, produced little effect on
fibrillin-1 deposition (P >0Æ05). However, application of the
test product resulted in a significant deposition of fibrillin-1,
the accumulation being at a similar level to that observed
using RA (mean ± SE) (baseline 1Æ27 ± 0Æ11; vehicle formu-
lation 1Æ70 ± 0Æ17; test product 2Æ64 ± 0Æ22, P =0Æ005; RA
2Æ51 ± 0Æ28, P =0Æ015; Fig. 1e). As in previous studies,
15
treatment with RA had little effect on deposition of pCI or on
the expression of MMP-1 in the epidermis (Table 1).
Randomized controlled trial
Clinical assessment
At 6 months, the test product produced statistically significant
improvement in facial wrinkles as compared to baseline assess-
ment (P =0Æ013), whereas vehicle-treated skin was not sig-
nificantly improved (P =0Æ11). 43% of the subjects who had
received the test product showed an improvement in facial
wrinkles compared with the baseline assessment, whereas only
22% of the subjects receiving the vehicle showed improve-
ment compared with baseline (Fig. 2). Use of the test product
produced a clinically significant improvement in facial wrin-
kles after 12 months of use, with a statistically significant
between-groups benefit of test product vs. the vehicle (test
product, 70% of subjects improving compared with vehicle,
33% improving; combined Wilcoxon rank tests, P =0Æ026)
(Fig. 2). No benefits of the test product were seen
for improvement in mottled dyspigmentation. Use of either
formulation produced an improvement in skin texture
over that recorded at baseline (vehicle, P =0Æ001; test prod-
uct, P =0Æ001), but the test formulation did not perform sig-
nificantly better than the vehicle (data not shown; P =0Æ72).
In-use biopsy samples
The test product produced a significant accumulation of fibril-
lin-1 in the papillary dermis of photoaged skin at 6 months
compared with the vehicle (mean ± SE) (vehicle formulation
1Æ84 ± 0Æ23; test product 2Æ57 ± 0Æ19;
ANCOVA, P =0Æ019;
Fig. 3).
Discussion
We show here, for the first time, that a commercially avail-
able over-the-counter ‘anti-ageing’ product improves the
2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426
422 Clinical efficacy of a cosmetic ‘anti-ageing’ product, R.E.B. Watson et al.
appearance of facial wrinkles when used in the long term.
This improvement is associated with restoration of fibrillin-1,
the major component of fibrillin-rich microfibrils, in product-
treated skin.
In these studies, we performed a double-blind RCT
to assess whether or not an over-the-counter cosmetic
‘anti-ageing’ product can produce clinically significant
improvement in the appearance of photoaged facial skin.
The trial was executed to the highest standards, with study
creams coded and randomized at source, and with the
volunteers, investigators and independent statistician ‘blind’
to the coding until after study completion and initial data
analysis. We did not observe any benefit of the vehicle or
test product on the appearance of mottled dyspigmentation
4
3
2
1
0
Baseline Vehicle
*
**
Test product All-trans RA
Treatment
Fibrillin staining, ordinal scale
(a)
(b)
(c)
(e)
(d)
Fig 1. Fibrillin-rich microfibrils are deposited in test product- and all-trans retinoic acid (RA)-treated human skin in a short-term patch test assay.
Representative photomicrographs showing fibrillin-rich microfibrils (arrows) in photoaged extensor forearm following the extended 12-day
patch test protocol: (a) baseline; (b) vehicle; (c) test product. (d) The positive control (0Æ025% RA) was applied for 4 days to avoid harmful
side-effects. Original magnification · 400. (e) Quantification of fibrillin-rich microfibrils following the occluded patch test assay with vehicle, test
product or the positive control (RA), compared with the baseline. We identified significantly more fibrillin-rich microfibrils with both the test
product (**P =0Æ005) and RA (*P =0Æ015).
2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426
Clinical efficacy of a cosmetic ‘anti-ageing’ product, R.E.B. Watson et al. 423
or actinic lentigines associated with photoageing. Expert
clinical assessments (S.O., J.Y.B.) showed that subjects
treated with the vehicle formulation had improved skin
texture compared with that observed at their recruitment,
but did not exhibit any change in the appearance of their
facial wrinkles. Those treated with the test product showed
improvements in both skin texture (compared with their
baseline clinical assessment) and, more importantly, in the
appearance of facial wrinkles. This improvement in the
appearance of facial wrinkles became significant only after
12 months of daily product use comparing between groups.
It is interesting to note, however, that when compared with
the baseline, the test product did lead to a noticeable clini-
cal improvement in facial wrinkles (P =0Æ013) in 43% of
treated individuals after 6 months, compared with only 22%
(a)
(b)
(c)
100
90
80
70
60
50
40
30
20
10
0
Vehicle Test product
Treatment
Subjects showing clinical
improvement in facial wrinkles
at 12-months (%)
Fig 2. Application of the test product results in clinical improvement
in facial wrinkles. (a) Clinical assessment identified a significant
improvement (P =0Æ026) in wrinkle grade following treatment
with the test product compared with the vehicle at 12 months, as
ascertained by using a Monte Carlo simulation. Standardized
photographs of the face of a 56-year-old woman at (b) baseline and
(c) after 6 months’ application of the test product show an
improvement in periorbital fine lines and wrinkles.
(a)
(b)
4
3
2
1
0
Baseline
Vehicle
*
Test product
Treatment
Mean fibrillin staining,
ordinal scale
(c)
Fig 3. Fibrillin-rich microfibrils were assessed by immunohisto-
chemistry at recruitment (baseline, a) and after treatment with either
vehicle (data not shown) or test product (b). (c) Significant
deposition of fibrillin-rich microfibrils was observed in the test
product-treated skin. *P =0Æ 019.
Table 1 Expression of extracellular matrix
(ECM) molecules in photoaged skin
following 12-day occlusion with a cosmetic
‘anti-ageing’ product
ECM component Baseline Vehicle Test product 0Æ05% RA
Fibrillin-1 1Æ27 ± 0Æ11 1Æ70 ± 0Æ17 2Æ64 ± 0Æ22** 2Æ51 ± 0Æ28*
pCI 2Æ65 ± 0Æ15 2Æ64 ± 0Æ15 2Æ70 ± 0Æ13 2Æ63 ± 0Æ21
MMP-1 (cells hpf) 115Æ8±7Æ9 140Æ3±13Æ6 127Æ2±14Æ1 143Æ7±12Æ5
RA, all-trans retinoic acid; pCI, procollagen-1; MMP, matrix metalloproteinase; hpf, high-
power field. *P =0Æ015; **P =0Æ005.
2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426
424 Clinical efficacy of a cosmetic ‘anti-ageing’ product, R.E.B. Watson et al.
of those treated with the vehicle where there was no signi-
ficant improvement in appearance (P =0Æ11). In a compari-
son between groups, this improvement was not statistically
significant but does indicate that larger clinical trials of cos-
metic products might be expected to show useful clinical
improvement after 6 months’ use. The data from this study
are indicative that cosmetic ‘anti-ageing’ products can result
in noticeable clinical improvement in facial wrinkles. To our
knowledge, this is the first time such benefits have been
reported for a commercially available cosmetic ‘anti-ageing’
product and paves the way for larger studies with more
statistical power.
In addition, we examined the distribution of a key bio-
marker of photoageing—loss of fibrillin-1 in the papillary
dermis—in skin samples obtained during the RCT. Fibrillin-1
is the major glycoprotein component of fibrillin-rich micro-
fibrils (oxytalan fibres). Skin treated with the test product
contained significantly more fibrillin-rich microfibrils in the
papillary dermis than either the baseline samples or those
treated with the vehicle (P =0Æ019). Our previous research,
which demonstrated that an over-the-counter cosmetic
‘anti-ageing’ product can bring about increased fibrillin-1
deposition in the papillary dermis, used a short-term, exagger-
ated-use patch test assay.
19
In the current study, we sought to
determine whether results obtained using such an experimen-
tal system were predictive of clinical improvement following
long-term use of such a product.
Consumers purchasing cosmetic skin care products—partic-
ularly those purporting ‘anti-ageing’ properties—are presented
with a broad choice of available products and only limited
data regarding their efficacy. The situation is further compli-
cated for consumers by the use of trademarked proprietary
names for ingredients and the relative lack of published long-
term studies to demonstrate product performance. We previ-
ously sought to investigate if a cosmetic ‘anti-ageing’ product
can have a measurable effect on fibrillin-1 and established that
histological improvement could be achieved in a short-term
exaggerated-use assay.
19
The results from that study were
indicative of some degree of structural change in the skin
following use of a cosmetic but provided no evidence for
actual clinical improvement. To address this question, the
current study compared the use of a commercially available
cosmetic ‘anti-ageing’ skincare serum to its vehicle and has
shown that such a product is capable of bringing about a clin-
ical improvement in the appearance of photoaged skin when
used for 12 months.
The difference in efficacy between the vehicle and the test
product demonstrates that a correctly formulated skincare
product can deliver clinically relevant skin improvement,
above that delivered by the vehicle base. The studied product
contains the retinol ester, retinyl palmitate, together with nat-
ural plant extracts, peptides and lipopeptides and antioxidants.
Other authors have shown evidence for the role of many of
these cosmetic ingredients in protecting against mechanisms
that lead to dermal degradation, such as increased MMP activ-
ity
22
and stimulating repair of dermal components.
23–25
It
was our belief that a combination of ingredients with activi-
ties known to address the multiple changes which occur in
photoaged skin (degradation of collagen and elastin, the
appearance of surface wrinkling and textural changes) may be
beneficial in a cosmetic product when used long term. Specif-
ically, the retinyl palmitate, palmitoyl peptides and Medicago
sativa extracts have been shown in vitro to lead to deposition
of collagen 1 in model skin systems and the extract of white
lupin has been shown to inhibit MMP-1 (S.P. Long, unpub-
lished data). In our previous study using the 12-day assay, a
similar product induced deposition of pCI as well as fibrillin,
together with inhibition of MMPs.
19
These data suggest that
the long-term benefits described here may be due to a com-
bination of actions. The product does not contain sunscreens,
so the visible skin improvements cannot be attributed to
photoprotection of the skin. Further work is underway to elu-
cidate the relative contributions of the individual ingredients.
The finding that a clinically relevant improvement in the
appearance of photoaged skin was demonstrated with long-term
use of a commercially available cosmetic ‘anti-ageing’ product
may cause some to question whether such effects are within the
definition of a cosmetic. Improvement in the appearance of
wrinkles is considered to be a cosmetic action, as the effect is
localized to the skin and is not concerned with treatment or cor-
rection of a disease condition. Several other authors report that
application of cosmetic products leads to changes in skin physi-
ology, including changes in barrier function,
26,27
stratum
corneum thickness
28
and lipidogenesis
29
and that cumulative
effects of cosmetic products are possible, a principle demon-
strated for skin moisturization
30
and recognized by regulatory
authorities.
31
When compared with the long-term clinical
effects of topical RA, it can be seen that the degree of improve-
ment offered by a cosmetic product is still markedly less than
that which is achievable with a prescribed medicine.
32–38
In conclusion, these studies provide evidence that use of an
over-the-counter cosmetic ‘anti-ageing’ product is able to
induce clinically identifiable improvement in the appearance
of facial wrinkles following long-term use. This improvement
is associated with deposition of fibrillin-rich microfibrils in
the papillary dermis of treated skin. The study further supports
the use of fibrillin-1 in a short-term assay as a biomarker for
assessing efficacy of potential photoageing repair products.
Acknowledgments
Independent statistical analysis of the RCT phase of this study
was provided by Chirostat Statistical Consulting, Nottingham,
U.K. The authors would also like to acknowledge the helpful
and insightful comments by Mr I. Marlow (Alliance Boots
Ltd.) during the preparation of this manuscript.
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2009 Boots Company Plc British Journal of Dermatology 2009 161, pp419–426
426 Clinical efficacy of a cosmetic ‘anti-ageing’ product, R.E.B. Watson et al.
    • "Signs of ageing are accordingly more prominent on photo-exposed areas such as the face. Facial ageing obviously differs between individuals depending on their age, lifestyle and ethnicity, but can be summarized through a combination of different grades of severity of dry skin, visible fines line and wrinkles, uneven skin pigmentation, loss of elasticity and skin sagging1234. The recently published case of a truck driver with unilaterally photodamaged skin, known as dermatoheliosis [5], resulting in thickening of the epidermis as well as the destruction of elastic fibres in the dermal extracellular matrix (ECM), shows the consequences of chronic exposure to ultraviolet A. It demonstrated the dramatic impact of photoageing, that is extrinsic vs. intrinsic ageing, on the skin appearance, thus strengthening the need for using sun protection and preventative or reparative skincare solutions. "
    [Show abstract] [Hide abstract] ABSTRACT: Like all organs, the skin is subject to intrinsic aging. In addition to being an external organ, the skin is in direct contact with the environment thus making it particularly exposed to extrinsic damage, mainly ultraviolet radiation (UVR), which accelerates the aging process, called photoageing (1). Signs of ageing are accordingly more prominent on photo-exposed areas such as the face. Facial ageing obviously differs between individuals depending on their age, lifestyle and ethnicity, but can be summarized through a combination of different grades of severity of dry skin, visible fines line and wrinkles, uneven skin pigmentation, loss of elasticity and skin sagging (1-4). This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Full-text · Article · Jun 2015
    • "The findings of this study have now been extended to test a new formulation of the product under both an occluded patch test protocol and a 6-month randomised, controlled trial [83]. Again, deposition of fibrillin-1 at the DEJ was noted following application of the anti-ageing product for both 12 days and 6 months [83]. This study supported the use of fibrillin-1 as an informative and robust biomarker for assessing efficacy of potential photoageing repair products. "
    [Show abstract] [Hide abstract] ABSTRACT: Cutaneous ageing is the result of two distinct, biological processes which may occur concurrently: (i) the passage of time, termed intrinsic ageing and (ii) environmental influences, termed extrinsic ageing. Intrinsic ageing of the skin is a slow process which causes changes in tissue structure and impairs function in the absence of additional biological, chemical and physical factors. The clinical features of intrinsically aged skin are not usually evident until old age when, although smooth and unblemished, the skin surface appears pale and is characterized by fine wrinkles with occasional exaggerated expression lines. Functionally, intrinsically aged skin is dry and less elastic than more youthful skin. In contrast, extrinsically aged skin is exemplified by deep, coarse wrinkles, mottled hyperpigmentation and a marked loss of elasticity and recoil. The two major environmental influences which induce extrinsic ageing are: (i) chronic exposure to solar ultraviolet (UV) irradiation (termed photoageing) and (ii) smoking. This review discusses the changes associated with the ageing process in the skin, with particular emphasis on the role played by the elastic fibre network in maintaining dermal function. The review concludes with a discussion of a short-term assay for independent assessment of the efficacy of anti-ageing cosmetic products using the elastic fibre component fibrillin-1 as a biomarker of extracellular matrix repair. © 2010 Society of Cosmetic Scientists and the Société Française de Cosmétologie.
    Full-text · Article · Sep 2010
  • Article · Sep 2009
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