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The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product(

CLS02021) Versus Placebo in a 4-week Application Period

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DOI: 10.5455/medarh.2022.76.108-114

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The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product( CLS02021) Versus Placebo in a 4-week Application Period
ORIGINAL PAPER
doi: 10.5455/medarh.2022.76.108-114
MED ARCH. 2022 APR; 76(2): 108-114
RECEIVED: FEB 25, 2022
ACCEPTED: APR 15, 2022
1
Department of Pharmacy at Medical
Faculty, Univertsity Sarajevo School of
Science and Technology, Sarajevo, Bosnia
and Herzgovina
2
Department of Pharmacology and
Clinical Pharmacy, Faculty of Pharmacy,
University of Sarajevo, Sarajevo, Bosnia
and Herzegovina
Corresponding author: Tarik Catic, PhD.
Pharm. University Sarajevo School of
Science and Technology Medical School,
71000 Sarajevo, Bosnia and Herzegovina.
E-mail: tarikcatic@bih.net.ba. ORDID ID:
http//www.orcid.org/0000-0002-0240-
8558.
© 2022 Tarik Catic, Belma Pehlivanovic,
Nejla Pljakic, Amina Balicevac
This is an Open Access article distributed under
the terms of the Creative Commons Attribution
Non-Commercial License (http://creative-
commons.org/licenses/by-nc/4.0/) which
permits unrestricted non-commercial use,
distribution, and reproduction in any medium,
provided the original work is properly cited.
108
The Moisturizing Efficacy of a
Proprietary Dermo-Cosmetic
Product( CLS02021) Versus Placebo
in a 4-week Application Period
Tarik Catic1, Belma Pehlivanovic2, Nejla Pljakic1, Amina Balicevac1
ABSTRACT
Background: Studies suggest that applying probiotic skincare products may have benefi-
cial effects on the skin due to bacteria competitiveness and specific metabolites produced
by probiotics. The cream CLS02021 is a postbiotic blend of metabolites, including organic
acids, enzymes, and peptides that are a result of the co-fermentation of three proprietary
probiotic strains and is specific for its high concentration of the enzyme Sphingomyeli-
nase which plays a significant role in cell signaling pathways and ceramide production.
Objective: The aim of this study is to evaluate the cosmetic effects of probiotic-derived
Ingredient CLS02021 and its safety in healthy volunteers. Methods: Fifty healthy volunteers
between 18 and 69 years old were recruited to participate in this multicentric, randomized,
intra-individual, double-blind group study. Following a face-split design, volunteers applied
both face cream containing CLS02021 on one side of their face and a placebo cream on
the other side of their face for a total duration of four weeks, two times a day. Evaluation of
the cosmetic effects included instrument measurements of moisturizing, elasticity, wrin-
kle depth, sebum production, pore size, melanin production, sensitivity, and side effects.
Results: A significant difference of CLS02021 over the placebo group was observed for
moisture and elasticity increase (both p<0.001), pore size (p<0.01), and a wrinkle depth de-
crease (p<0.05). No significant differences were noted in sebum production, melanin level,
skin cleanliness, and sensitivity between groups. Conclusion: CLS02021, as a probiotic-de-
rived ingredient, showed strong beneficial effects on skin moisture and elasticity increase,
wrinkle depth, and pore size reduction. Face cream containing CLS02021 is well tolerated
and could be an organic choice for cosmetic hydrating and antiaging skincare treatment.
Keywords: probiotics, skin care, cosmetic product, antiaging.
1. BACKGROUND
The human skin is an essential defense barrier against environmental fac-
tors such as various toxins and microbial pathogens. To achieve this function,
our skin hosts a variety of protective microbiota. In healthy skin, the micro-
biota is in homeostasis and balance. Due to physical, chemical, and other ex-
ternal and internal factors, our microbiota can be easily disrupted, leading to
skin conditions including acne, atopic dermatitis, psoriasis, rosacea, or even
premature aging (1-3). Beneficial skin properties of probiotic bacteria can be
assigned to bacteria competitiveness with harmful bacteria and bacteria-spe-
cific metabolite production. In more simple terms, beneficial bacteria drive
the harmful bacteria from the skin and restore its balanced environment
while producing various organic acids, polysaccharides, and proteins - most-
ly enzymes that participate in the normal metabolic processes of the skin
such as hyaluronic acid production, ceramide synthesis, melanocyte stimu-
lation, and many others. A further emphasis is attributed to Lactic Acid Bac-
teria (LAB) compounds such as cell wall fragments and their metabolites (4).
Different studies have shown that LAB metabolites such as hyaluronic acid,
lipoteichoic acid, sphingomyelinase, and lactic acid have promising cosmetic
and therapeutic effects on the skin (5-8). The proprietary cream CLS02021
is a postbiotic blend of metabolites, including organic acids, enzymes, and
peptides, that are a result of the co-fermentation of three proprietary probi-
otic strains. These strains are lactic acid bacteria (LAB) strains isolated from
native fermented dairy products from the Balkan region. Genomic tools
were used to fully characterize and denominate the strains as Lactobacillus
ORIGINAL PAPER | Med Arch. 2022 APR; 76(2): 108-114
 The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product( CLS02021) Versus Placebo in a 4-week Application Period
plantarum (AN057), Lactobacillus casei (AN177), and
Streptococcus thermophilus (AN157). The absence of
antimicrobial and virulence factors, a toxicological re-
view, and the traditional utilization of these strains in
dairy products have substantiated enough evidence to
testify to the safety of these strains. The beneficial effects
of probiotic strains are based on particular metabolites
which probiotic bacteria naturally produce during their
fermentation (4). The strains of CLS02021 were careful-
ly selected based on their potential to produce specific
metabolites in certain amounts. Metabolites that were
identified as most promising were Lipoteichoic Acid
(LTA), Hyaluronic Acid (HA), Lactic Acid (LA), and
Sphingomyelinase (SMase). The scientific details of the
ingredients of the cream CLS2021 are as follows:
Lipoteichoic Acid (LTA)
LTA is a structural component of gram-positive cell
wall bacteria with antigenic properties stimulating spe-
cific immune responses. LTA isolated from probiot-
ic bacteria, such as LAB, has been reported to protect
against the overproduction of proinflammatory cyto-
kines and stimulate skin defense against microbial infec-
tion (9). Recently, scientists have demonstrated that LTA
isolated from L. Plantarum is also an effective agent in
skin protection from UV-induced skin damage and pho-
to-aging by inhibiting the expression of collagen degrad-
ing enzyme: matrix metalloproteinase-1 (MMP-1) (10).
Suppression of MMP-1 by LTA is mediated through
MAPK-1 signaling cascade proteins, which exert influ-
ence on AP-1 and NF-kb transcription factors. Consid-
ering proteins as proto-oncogene products involved in
tumorigenesis, these findings also indicate the potential
ability of LTA as an agent for the prevention of skin can-
cer (10).
Hyaluronic Acid (HA)
HA consists of a basic unit of two sugars, glucuronic
acid, and N-acetylglucosamine, polymerized into large
macromolecules of 2000–25 000 repeating units (11).
HA is essential for maintaining the normal structure
of the stratum corneum and conserving epidermal bar-
rier functions. Topical application of HA for 60 days
showed significant improvement in skin hydration and
elasticity. Application of low-molecular-weight HA re-
sulted in a significant reduction in wrinkle depth, which
was attributed to better penetration (12). Furthermore,
low-molecular-weight HA has also been reported to
increase epithelial defense by inducing one of the most
common types of antimicrobial peptides, β-defensin-2,
via Toll-like receptors (TLR), participating in the host
response against bacterial infections (13). HA is com-
mercially obtained by extraction from rooster combs
and by fermentation by LAB (14).
Lactic Acid
Lactic acid is an organic acid with one hydroxyl group
attached to the alpha position of the acid. It has been
widely used for many years in cosmetic regimens and
skincare products such as moisturizers, exfoliants, and
emollients (15, 16). The use of lactic acid in skincare in
different concentrations leads to the improvements in
both the epidermal and dermal components providing
ORIGINAL PAPER | Med Arch. 2022 APR; 76(2): 108-114
useful treatment for rejuvenating photo-damaged skin
(17). Lactic acid is regularly produced through microbial
fermentation and is a predominant end-product of LAB
carbohydrate metabolism.
Sphingomyelinase (SMase)
Sphingomyelinase (SMase) is an enzyme that is re-
sponsible for ceramide production. Ceramides are lip-
ids (fats) found naturally in high concentrations in the
uppermost layers of skin, making over 50% of the skin’s
composition. Ceramides help retain the skin together by
forming a protective layer that limits moisture loss and
protects against visible damage from pollution and oth-
er environmental stressors. A decrease in ceramide in
the stratum corneum causes water loss and barrier dys-
function in the epidermis, including a loss of protection
against antigens and bacteria (18).
SMase is detected in bacteria, with great variations in
SMase activity among different bacterial strains. Bac-
terial SMase is a secretory protein released from cells
into the media. In vitro studies have confirmed that
keratinocyte cell lines treated with sonicated S. ther-
mophilus significantly increase ceramide generation.
Further examinations proved that increased levels of
ceramide production were due to SMase (>0.1 mU/mL)
obtained from sonicated cells of S. thermophilus. These
results were further confirmed using in vivo testing on
17 healthy Caucasian volunteers with a cream formula-
tion containing sonicated S. thermophilus applied twice
daily. After seven days of treatment, S. thermophilus in-
duced a very significant increase in ceramide levels as
compared to controls (19). Based on the mechanisms of
action of these specific metabolites, it can be hypoth-
esized that the primary effect of daily use of a topical
product with CLS02021 is an increase in overall skin
moisture levels.
The chemical composition of Metabiotic Complex
provides a promising base for other beneficial effects
on the skin such as anti-inflammatory effects, stimula-
tion of collagen production, increase in skin elasticity,
brightening of skin complexion, reduction of fine lines
and wrinkles, and general improvement of skin appear-
ance.
There is a growing body of evidence, and ongoing re-
search focus on the benefits of technologies based on
probiotics for cosmetic purposes (19). However, there
is a lack of well-designed Randomized Controlled Tri-
als (RCTs) for the study of cosmetic products and their
measurable effects on the skin.
The aim of this randomized and controlled study is to
explore the cosmetic effects of the ingredient “Metabiot-
ic Complex,” developed by Anbiome Labs. It is hypothe-
sized that Metabiotic Complex-based formulations may
have beneficiary effects on a wide array of cosmetic ap-
pliances.
2. OBJECTIVE
The aim of this study is to evaluate the cosmetic effects
of probiotic-derived Ingredient CLS02021 and its safety
in healthy volunteers.
109
The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product( CLS02021) Versus Placebo in a 4-week Application Period

3. SUBJECTS AND METHODS Statistical analysis

Study subjects Data analysis is conducted using SPSS Statistics for
Fifty healthy volunteers without any prior skin-related Windows, Version 23.0 (IBM SPSS Statistics for Win-
or health-related conditions, between 18 and 69 years dows, Version 23.0. Armonk, NY: IBM Corp). Descrip-
old, were recruited to participate in this study. The ex- tive statistics were performed for the characterization
clusion criteria included pregnancy, nursing, and plan- of demographic characteristics of the volunteers and
ning a pregnancy. All volunteers signed a written in- included frequency (n), mean, a standard deviation of
formed consent in accordance with the study procedure mean, minimum, median, and maximum values. Fre-
approved by the Bioethical Committee of the Sarajevo quency and percentages were provided for categorical
Medical School at the Sarajevo School of Science and variables. Mann-Whitney U Wilcoxon test was used
Technology and were free to discontinue participation to compare the efficiency of test and control products
at any time without any obligation. Volunteers were re- between measurements. All results were evaluated at a
quired to follow specific instructions for four weeks ac- confidence level of 95%, and the significance level was
cording to the study design. set at p<0.05. The per-protocol (PP) population included
Study design all randomized volunteers who fulfilled inclusion crite-
This was a randomized, intra-individual, double-blind n (%) Mean STD Min Max
controlled study performed between July 2021 and Sep-
Male 7 (14,9) 35,57 13,28 18,0 59,0
tember 2021 in Sarajevo, Bosnia, and Herzegovina. The
Female 40 (85,1) 35,73 11,28 20,0 60,0
study was performed in agreement with the principles
Age 35,70 11,44 18,0 60,0
of the Declaration of Helsinki and recommendations of
Good Clinical Practice ICHE6(R2). Furthermore, the Table 1. Distribution of study subjects
study was applied and registered at ClinicalTrials.gov,
receiving an Identifier code: NCT05048121. Parameter Active Placebo p
All volunteers were assigned a Subject Identification Moisture
code for easier monitoring and to maintain subject an- W0-W2 ΔMean±STD 12,96±16,57 1,94±20,72 0,001**
onymity. Each volunteer was given a set of two identi- ΔMedian 13 4
cal tubes, one tube containing CLS02021 and another Δ Min-Max -30-76 -48-80
one containing a placebo, and they were instructed to W0-W4 ΔMean±STD 11,49±19,09 -4,45±21,47 0,000**
apply a product cream in an amount of a hazelnut size ΔMedian 10 0
while avoiding the eye region. The study products were ΔMin-Max -32-56 -76-38
applied twice a day, for four weeks, to the left and right W2-W4 ΔMean±STD -1,47±16,06 -6,38±17,48 0,199
sides of the face using the indication on the tubes shown Δ Median 0 -4
as “Left” and “Right,” a method widely known as split- Δ Min-Max -36-35 -76-22
face testing. Figure 1. briefly provides an overview of the Elasticity
study design. W0-W2 ΔMean±STD 11,87±15,64 1,91±19,45 0,001**
Study points and parameters ΔMedian 11 3
During the four-week study, a total of three measure- Δ Min-Max -28-74 -45-78
ments were performed at the following points: baseline W0-W4 ΔMean±STD 11,04±17,62 -3,51±18,32 0,000**
point (W0), point Week 2 (W2), and endpoint Week 4 ΔMedian 10 0
(W4). Initial measurement prior to the study beginning ΔMin-Max -28-55 -47-42
served as a baseline point. After the measurement of
W2-W4 ΔMean±STD -0,83±14,88 -5,43±14,71 0,134
endpoint W4, the study was concluded. At every study
Δ Median 0 -4
point, with the aim to assess the safety profile, volunteers
Δ Min-Max -39-38 -53-24
were prompted to report any side effects that might have
Sebum U zone
stemmed from the product use. All of the measurements
W0-W4 ΔMean±STD 0,79±25,23 6,79±28,9 0,421
were carried out by an independent cosmetologist, com-
ΔMedian 0 0
pletely blinded to the procedure.
The measurement process was performed in two ΔMin-Max -57-97 -49-97
designated pharmacies. The API Skin 100 instrument Sebum T zone    
(ARAM HUVIS, South Korea) was used for measure- W0-W4 ΔMean±STD 4,79±36,56 6,21±36,34 0,814
ment and evaluation of the following skin parameters: ΔMedian 0 2
moisturizing and elasticity using corneometry (Bioelec- ΔMin-Max -97-86 -97-89
trical Impedance Analysis); wrinkle depth by photom- Pore
etry (shadow method and picture processing); sebum 0,850
W0-W2 ΔMean±STD 6,11±18,17 7,94±21,11
production by photometry; pore size by photometry
(polarized light and picture processing); melanin pro- ΔMedian 3 3
duction by photometry (light absorption); sensitivity by Δ Min-Max -35-50 -36-61
photometry (light reflection); cleanliness by photometry 0,008**
W0-W4 ΔMean±STD -3,94±14,65 3,74±16,29
(polarized light and picture processing).
ΔMedian -5 5

110 ORIGINAL PAPER | Med Arch. 2022 APR; 76(2): 108-114

 The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product( CLS02021) Versus Placebo in a 4-week Application Period

ΔMin-Max -31-29 -44-36 ipants withdrew from the study; therefore, forty-seven
W2-W4 ΔMean±STD -10,04±22,34 -4,19±19,79 0,235 participants completed the study (Figure 1). Baseline
Δ Median -8 -4 demographic data of volunteers are presented in Table
Δ Min-Max -51-32 -49-51 1. All of the volunteers reported the use of facial creams
Melanin in the past and no prior skin-related or health-related
W0-W2 ΔMean±STD -5,68±35,98 -6,09±31,99 0,689 conditions (Table 2). During the study, three side effects
ΔMedian -9 -2
were reported; two volunteers reported pimple break-
Δ Min-Max -75-90 -70-89
down, which occurred in the first week of product ap-
plication, and one volunteer reported redness on the
W0-W4 ΔMean±STD -12,57±34,24 -8,87±36,33 0,586
forehead during the first 3-4 days of use. Further anal-
ΔMedian -11 -9
ysis shows that reported redness occurred on the side
ΔMin-Max -89-68 -78-71
of the face where the active product was applied for one
W2-W4 ΔMean±STD -6,89±27,82 -2,79±31,72 0,553
subject and in the other subject where the placebo was
Δ Median -5 0
applied, which may be caused by the base cream for-
Δ Min-Max -69-59 -71-70 mulation and not the CLS02021. Overall, the safety and
Cleanliness tolerability results of CLS02021 were comparable with
W0-W2 ΔMean±STD -1,32±33,26 -0,77±34,06 0,907 those of the placebo and local tolerability was good for
ΔMedian 0 0 both treatments. A comparison of treatment groups,
Δ Min-Max -85-66 -75-89 based on a difference in measured scores (Delta Δ) at
W0-W4 ΔMean±STD 3,38±30,61 3,47±27,46 0,626 baseline point W0, point W2, and after endpoint W4, is
ΔMedian 3 0 presented in Table 3. Sebum T zone and Sebum U zone
ΔMin-Max -74-61 -59-77 parameters were measured not at point W2. Our results
W2-W4 ΔMean±STD 4,7±33,18 4,23±30,77 0,838 indicate a significant difference between CLS02021 and
Δ Median 0 0 the placebo groups in the following parameters: Mois-
Δ Min-Max -78-72 -87-83 ture (p<0.0001), Elasticity (p<0.0001), Pore Size (p<0.01),
Wrinkle and Wrinkle Depth (p<0.05). However, the change was
W0-W2 ΔMean±STD 0,83±19,86 5,7±25,08 0,181 not statistically significant from the placebo group in
ΔMedian -1 2 the following parameters: Sebum U and T zone, Mela-
Δ Min-Max -52-47 -80-65 nin, Cleanliness, and Sensitivity. Statistically significant
W0-W4 ΔMean±STD -5,68±14,84 0,4±17,21 0,047* increases in moisture (p<0.0001) are observed in the
ΔMedian -4 0 CLS02021 group at W0 to W4 (11,49±19,09) compared
ΔMin-Max -52-27 -61-64 to the Placebo group (-4,45±21,47). A similar effect has
W2-W4 ΔMean±STD -6,51±16,63 -5,3±22,63 0,666 been observed in the CLS02021 group with parameter
Δ Median -3 -1 Elasticity (p<0.0001), where the mean improvement at
Δ Min-Max -63-23 -72-58 W0 was 11,04±17,62, while the Placebo group recorded
Sensitivity a mean decrease of -3,51±18,32 at W4 (Table 3). Statis-
0,889
tically significant reduction of pore size and decrease in
W0-W2 ΔMean±STD 3,49±26,8 3,13±23,63 wrinkles depth (p<0.05) was observed from W4 to W0,
ΔMedian 0 0 while no significant difference was observed between
Δ Min-Max -70-71 -65-63 W2 and W0. Obtained results suggest improvement of
0,922
the following skin parameters: moisture, elasticity, and
W0-W4 ΔMean±STD -5,94±26,95 -2,83±25,66 wrinkle depth over a period of four weeks. Furthermore,
ΔMedian 0 -4 a comparison of mean values at points W0, W2, and W4
ΔMin-Max -76-62 -86-64 demonstrates a positive effect on moisture, elasticity,
W2-W4 ΔMean±STD -9,43±31,12 -5,96±24,15 0,994
pore size, and wrinkle depth of the tested product ver-
sus the placebo (Graph 1). Although there was a positive
Δ Median -2 -4
trend, no statistically significant difference between the
Δ Min-Max -81-71 -68-64
CLS02021 group and the placebo group was observed
Table 2. Comparison of treatment groups based on difference in the following measured parameters: sebum, melanin,
of scores measures on baseline, W 2 and W 4. Mann-Whitney U cleanliness, and sensitivity. However, volunteers using
Test; *p<0,05; **p<0,01
test tubes containing CLS02021 showed a rapid and
ria and were assessed at W4. The primary efficacy crite- constant decrease in melanin levels as well as sensitivity
rion was the mean change in the instrument measured improvement. Regarding sebum levels in the U zone, the
moisture and elasticity from W0 to W4. Other effica- CLS02021 group shows a lower and slower increase in
cy variables included the mean change in instrumental sebum levels and lower levels of sebum in the T zone.
measured scores from W0 to W4.
5. DISCUSSION
4. RESULTS Recent studies have shown that the intestinal microbi-
A total of fifty healthy volunteers were enrolled in this ome plays an essential role in modulating system inflam-
study. However, during the first two weeks, three partic- mation and skin diseases by modulating the host im-

ORIGINAL PAPER | Med Arch. 2022 APR; 76(2): 108-114 111

The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product( CLS02021) Versus Placebo in a 4-week Application Period

Moisture - mean value W0, Elasticity - mean value W0, W2,

W2, W4 W4
39,49 42,55 35,04 45
45 34,4 37,55 30,89
41,09
36,72
41,43
29,6 36,32
25 25 25,68
W0 W2 W4 W0 W2 W4

CLS02021 Placebo CLS02021 Placebo

A B

Pore size - mean value W0, Wrinkle depth - mean value

W2, W4 W0, W2, W4
55 32,64
45,57 35
41,38 28,96
45 38,57 30 28,13 27,34
44,68
35 37,64 34,64 25 26,94
22,45
25 20
W0 W2 W4 W0 W2 W4

CLS02021 Placebo CLS02021 Placebo

C D
Graph 1. Comparison of mean values at point Wo, W2 and W4 for moisture (A), elasticity (B), pore size (C) and Wrinkle depth (D)
Graph 1. Comparison of mean values at point Wo, W2 and W4 for moisture (A), elasticity
mune system (20, 21). It is suggested that oral probiotics
can treat certain skin conditions, such as acne, atopic
(B), porephotoaging,
dermatitis, size (C) and Wrinkle
psoriasis, and depth
wound(D)healing
(22). Findings from this study suggest that the der-
mo-cosmetic product CLS02021, with our proprietary
cosmetic ingredient “Metabiotic Complex,” has statisti-
cally significant beneficial effects on the skin regarding
skin moisturizing, elasticity, and wrinkle improvement.
These findings are in accordance with the recently re-
ported beneficial effects of probiotics and their metabo-
lites (23). Ceramides are key components in skin barrier
integrity maintenance because of their ability to lock in
moisture. Furthermore, ceramides contribute to skin
elasticity giving it a vibrant, healthy look (24). Studies
have shown that topical preparations of precursors can
lead to an increase in ceramide levels of Sphingomyelin-
ase, which is naturally produced during the probiotic life
cycle and is responsible for the conversion of native skin
sphingomyelin into ceramides (25). Metabiotic Com-
plex contains a significant amount of naturally produced
Sphingomyelinase, which can be attributed to facilitat-
ing the production of ceramides, leading to a measur-
able increase in skin moisture levels and improvement
of skin elasticity. Due to the growing evidence, it is be-
coming more popular to use topical preparations of pro-
biotics for the treatment of atopic dermatitis, psoriasis,
and wound healing but also for photo and skin aging.
Recent studies have classified probiotic skincare as one
of the latest skincare trends for fighting photoaging and
skin aging (26). Probiotics have been shown to have a
preventative effect on skin photoaging induced by short-
term UV irradiation. Satoh et al. reported that the use
of probiotic Bifidobacterium breve B-3 showed its effect
on suppressing changes in transepidermal water loss,
skin hydration, epidermal thickening, and attenuated
the damage to the tight junction structure and basement
membrane induced by chronic UVB irradiation (27). In
our study, a cream consisting of “Metabiotic Complex,”
a probiotic derived ingredient, showed beneficial effects
on the main potential risks contributing to the skin ag-
ing process. Fast improvement and increase of facial skin
moisture, which persists, are one of the effects on overall
long-term effects on skin condition. Sharma et al. ex-
plain that probiotics can restore acidic skin pH, alleviate
oxidative stress, attenuate photoaging, and improve skin
barrier function (28). In vitro study of the effects of plant
extracts fermented with Lactobacillus buchneri (PELB)
on ultraviolet light (UVB)-induced photoaging process
reported that PELB could be useful in the cosmetic
industry due to its protective effects against UVB-in-
duced photoaging. PELB works by decreasing elastase
activity and increasing type I collagen expression, re-
ducing collagenase activity, and promoting the expres-
sion of moisture factor and antioxidant enzymes (29).

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 The Moisturizing Efficacy of a Proprietary Dermo-Cosmetic Product( CLS02021) Versus Placebo in a 4-week Application Period
Our study product showed a significant and persisting
increase in skin elasticity which was expressed in the
first two weeks of product application. Notay et al. in-
vestigated the use of topical Nitrosomonas eutropha as
a cosmetic ingredient for the reduction of facial wrinkles
and found a significant improvement in wrinkle depth
severity, hyperpigmentation of the forehead, and glabel-
la in the group receiving high topical concentrations of
the probiotic formula (30). Hyaluronic acid is a humec-
tant; it binds large amounts of water, more precisely 100
times its weight (31). Its ability to tie water helps rehy-
drate the skin, thereby filling in fine lines and reducing
the appearance of wrinkles (32). Along with lactic acid,
hyaluronic acid is a large constituent of a Metabiotic
Complex, leading to a reduction of wrinkle depth during
four weeks of product use. Topical application of lactic
acid has been shown to lead to an increase in skin hy-
dration and elasticity, a decrease in transepidermal wa-
ter loss, and a decrease in pore size (33). A statistically
significant reduction in pore size (p<0.01), was shown in
healthy volunteers in the area where the tested product,
CLS2021, was applied.
LTA is a natural constituent of lactic acid bacteria cell
walls. Through its inhibition of expression of collagen
degrading enzyme matrix metalloproteinase-1 (MMP-
1), it can be attributed to being an effective agent in skin
protection from UV-induced skin damage and pho-
to-aging (34). Four weeks of topical use of a cosmetic
product containing Metabiotic Complex infused with
LTA has not shown a statistically significant difference
in melanin production decrease. However, a melanin
production increase was also not observed, which gives
cause to hypothesize that prolonged product use could
lead to positive results. Considering that the study was
performed during the warmest summer months with
high UV exposure, melanin production levels could
have been significantly influenced. Therefore, further
research should be carried out during a different sea-
son. In order to achieve more significant scientific data,
further evaluations should be performed in more con-
trolled conditions.
6. CONCLUSION
The use of oral and topical probiotics for skincare
and the treatment of skin problems has grown in pop-
ularity over the last decade. Ongoing research studies
attempt to evaluate the efficacy, mechanism of action,
safety, and indications of new products when they are
introduced to the market. Developed by Anbiome Labs
cream CLS02021 contains “Metabiotic Complex,” a pro-
biotic-derived ingredient, which is a blend of several or-
ganic acids, enzymes, and peptides that are a result of a
co-fermentation of three proprietary probiotic strains.
Our findings have demonstrated strong beneficial ef-
fects of face cream containing “Metabiotic Complex” on
skin moisture and elasticity increase, wrinkle depth, and
pore size reduction during a four-week-long study. Fur-
thermore, face cream containing “Metabiotic Complex”
is well tolerated and could be the organic choice for cos-
metic moisturizing and antiaging skincare treatment.
ORIGINAL PAPER | Med Arch. 2022 APR; 76(2): 108-114
However, there is a need to conduct further research
with an extended study duration on a larger sample of
study subjects.
• Acknowledgment: The authors would like to thank Dr. Oguz Akbas (Monitor CRO) for
his ample contributions to study design and motivating discussion.
• Authors contribution: All authors were involved in all steps of preparation this article. Final
proofreading was made by the first author.
• Conflict of interest: None declared.
• Financial support and sponsorship: Nil.
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