Continuous Skin Rejuvenation by Combining Nonablative Fractional Laser With Daily Application of a Multibeneficial Composition Formulation: A Blinded Randomized Clinical Trial Study

Abstract

Background and Aims
Skin aging is a common concern among individuals, and laser treatments are recognized as one of the most effective approaches to mitigate the aging process. The study aims to compare a multibeneficial formula serum versus a blank formulation in achieving maximum efficacy following a single treatment of nonablative fractional laser for facial skin rejuvenation.

Methods
This study was a double‐blind, split‐face, monocentric, randomized clinical trial in China (September 24, 2023–March 07, 2024), and 37 patients seeking the Fotona 4D laser treatment for aging‐related facial changes were enrolled. After one full‐face laser treatment, each patient applied the test serum to one side and the blank formulation to the other, randomly, twice daily for 28 days. Two dermatologists assessed facial skin quality and aging signs at baseline and Day 0 (D0, immediately after the laser treatment), D3, D7, D14, and D28. Noninvasive measurement and self‐assessment questionnaires were also administered at each visit. According to the types of variables, appropriate statistical tests, including the Friedman test, ANOVA test, and Wilcoxon signed‐rank test, were used to examine the within‐groups or between‐groups differences.

Results
Thirty‐three women, aged 35–49 years, completed the study. After 28 days of the test serum application, the visual clinical scores rated by investigators showed more significantly beneficial changes on the test side than those on the control. More significant improvements in index parameters for the test sides were also found both in wrinkles with a 21.14% decrease of SEw value from the baseline and in elasticity with a 14.99% decrease of R2 value, while the corresponding reductions were 3.83% for SEw and 4.10% for R2 found on the control sides. The reduction of the nasolabial folds area proportion, analyzed by Primos, was 10.61% on the test sides and 3.39% on the control. No adverse events were reported.

Conclusion
The serum with a multi‐beneficial composition can contribute to achieving a more significant and sustainable efficacy after the Fotona 4D treatment in skin rejuvenation improvement.
ClinicalTrials.gov ID: NCT06140628.

Full text

Health Science Reports
ORIGINAL RESEARCH
Continuous Skin Rejuvenation by Combining
Nonablative Fractional Laser With Daily Application of a
Multibeneficial Composition Formulation: A Blinded
Randomized Clinical Trial Study
Xinxuan Zhang
1
| Manru Ning
2
| Mengqing Lin
1
| Qi Tang
1
| Yihuai Liang
3
| Feifei Wang
2
| Xiaoke Xu
1
1
Xiaoke BeauCare Clinic, Shenzhen, Guangdong, China |
2
Yunnan Characteristic Plant Extraction Laboratory Co. Ltd., Kunming, Yunnan, China |
3
Yunnan
Botanee Bio‐Technology Group Co. Ltd., Kunming, Yunnan, China
Correspondence: Feifei Wang (wangfeifei@botanee.com) | Xiaoke Xu (xxk782@qq.com)
Received: 17 July 2024 | Revised: 23 December 2024 | Accepted: 16 January 2025
Funding: The study was supported by the Yunnan Characteristic Plant Extraction Laboratory (2022YKZY002 and 2022YKZY006).
Keywords: anti‐aging | formulation | non‐ablative fractional laser | skin barrier | skin physiology | skin wrinkles
ABSTRACT
Background and Aims: Skin aging is a common concern among individuals, and laser treatments are recognized as one of the
most effective approaches to mitigate the aging process. The study aims to compare a multibeneficial formula serum versus a
blank formulation in achieving maximum efficacy following a single treatment of nonablative fractional laser for facial skin
rejuvenation.
Methods: This study was a double‐blind, split‐face, monocentric, randomized clinical trial in China (September 24,
2023–March 07, 2024), and 37 patients seeking the Fotona 4D laser treatment for aging‐related facial changes were enrolled.
After one full‐face laser treatment, each patient applied the test serum to one side and the blank formulation to the other,
randomly, twice daily for 28 days. Two dermatologists assessed facial skin quality and aging signs at baseline and Day 0 (D0,
immediately after the laser treatment), D3, D7, D14, and D28. Noninvasive measurement and self‐assessment questionnaires
were also administered at each visit. According to the types of variables, appropriate statistical tests, including the Friedman
test, ANOVA test, and Wilcoxon signed‐rank test, were used to examine the within‐groups or between‐groups differences.
Results: Thirty‐three women, aged 35–49 years, completed the study. After 28 days of the test serum application, the visual
clinical scores rated by investigators showed more significantly beneficial changes on the test side than those on the control.
More significant improvements in index parameters for the test sides were also found both in wrinkles with a 21.14% decrease of
SEw value from the baseline and in elasticity with a 14.99% decrease of R2 value, while the corresponding reductions were
3.83% for SEw and 4.10% for R2 found on the control sides. The reduction of the nasolabial folds area proportion, analyzed by
Primos, was 10.61% on the test sides and 3.39% on the control. No adverse events were reported.
Conclusion: The serum with a multi‐beneficial composition can contribute to achieving a more significant and sustainable
efficacy after the Fotona 4D treatment in skin rejuvenation improvement.
ClinicalTrials.gov ID: NCT06140628.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly
cited.
© 2025 The Author(s). Health Science Reports published by Wiley Periodicals LLC.
Zhang and Ning equally contributed to this work.
1of10Health Science Reports, 2025; 8:e70423
https://doi.org/10.1002/hsr2.70423

1 | Introduction
The skin, as the body's largest organ, provides a physical barrier
against environmental factors and is exposed to damage
throughout life [1]. Skin aging is influenced by both intrinsic
and extrinsic factors [2]. Intrinsic aging is primarily determined
by genetics and age, while extrinsic aging is characterized by
fine lines, pigmentation, and roughness [3]. Since facial skin
visibly shows signs of aging, it is of significant concern to many
people. Although intrinsic aging is irreversible, photoaging can
be mitigated through various protective measures. Therapies for
photodamage [4], such as non‐ablative fractional laser (NAFL),
intense pulsed light (IPL), and radio‐frequency light (RFL),
have become increasingly popular. Additionally, incorporating
skincare products into daily routines, including cleaning,
moisturizing, and sun protection, can effectively protect the
skin and slow the aging process.
Fotona 4D, an NAFL with two complementary wavelengths
(2940 nm and 1064nm), treats four distinct dimensions of the
skin to rejuvenate it from the inside out. It is particularly effective
for tightening lines around the eyes, though temporary pain,
redness, and swelling may occur, affecting postoperative comfort.
Because of the desire for improved appearance with shorter
recovery times and longer‐lasting effects, the concept of “inte-
grated skincare”has emerged [5]. This approach combines
clinically proven skincare products with professional medical
esthetics to enhance treatment benefits. Studies have shown
that “integrated skincare”can synergistically reduce adverse
reactions and promote skin barrier repair [6]. Numerous
botanical extracts with antiaging properties have been identi-
fied, targeting various aspects of aging. For example, Cordyceps
sinensis extract [7], Paeonia suffruticose extract [8], and er-
gothioneine [9] regulate the extracellular matrix; Kappaphycus
alvarezii extract [10] limits the telomere shortening and slows
the senescence of dermal fibroblasts; Acmella oleracea extract
[11] relaxes muscle to recuperation of contractile activity; and
Morus alba extract inhibits mitogen‐activated protein kinases
(MAPKs). Furthermore, a recent preliminary study demon-
strated that daily use of a multi‐beneficial composition formu-
lation can promote facial rejuvenation [12]. Based on the
efficacy of Fotona 4D for facial rejuvenation and the concept of
“integrated skincare,”this study aimed to systematically eval-
uate the short‐and long‐term effects of Fotona 4D treatment
alone and in combination with a multi‐component antiaging
composition formulation. The assessment was conducted using
clinical evaluations, noninvasive methods, and self‐assessment
questionnaires. The short‐term effect on the epidermal barrier
was measured by transepidermal water loss (TEWL), while the
long‐term effects on facial skin rejuvenation were evaluated
across five parameters including wrinkles, firmness, smooth-
ness, roughness, and elasticity.
2 | Methods
2.1 | Ethics
The study was approved by Shanghai Ethics Committee for
Clinical Research (Approval Number: SECCR/2023‐101‐01).
Before any study‐related procedures or measurements, written
informed consent was obtained from all participants. Partici-
pants were informed about the purpose of the study, procedures
involved, potential risks, and their right to withdraw at any time
without any consequences.
2.2 | Study Design
This study was a double‐blind, split‐face, monocentric, random-
ized clinical trial. Thirty‐seven participants were enrolled and
underwent a 2‐week washout period using basic skincare prod-
ucts, and 33 of them received the Fotona 4D laser treatment and
completed the study. The skin biophysical parameters, objective
assessments, and self‐assessments were evaluated on D0, D3, D7,
D14, and D28 use after the Fotona 4D laser treatment.
2.3 | Participants
Participants meeting all inclusion criteria and none of the ex-
clusion criteria were enrolled and provided informed consent.
Inclusion criteria: (1) Healthy female aged 30–50; (2) subjects
with under‐eye wrinkles, pore appearance, and forehead wrin-
kles (scores are all > 2 from the Asian Skin Aging Atlas, Volume
2[13]; (3) subjects with nonsensitive skin; (4) subjects are
willing to use the test serum on split‐face for 28 days; (5) sub-
jects with no history of antibiotics in the past 3 months; and (6)
subjects understand the nature of the study and sign the in-
formed consent form (ICF). Exclusion criteria: (1) Subjects with
known intolerance or hypersensitivity to skincare ingredients;
(2) female subjects who are pregnant, lactating or plan to
become pregnant; (3) subjects who have received esthetic
medical treatment (photoelectric, tightening, filling, etc.) within
the past 6 months; (4) subjects with erythema, rash, flaking and
edema on faces; (5) subjects who participated in clinical trials
within 1 month; (6) subjects with antiallergy medication history
within 1 month; and (7) any subjects those the investigator
considers ineligible.
2.4 | Laser System
Each participant received a single laser treatment (Fotona 4D Pro,
Fotona d.o.o., Slovenija) on D0 using the following three modes:
FRAC3 Mode: 4 mm spot size, 20–25 J/cm² fluence, 1.6 s pulse
width, a frequency of 4.0 Hz, and 1500 rounds at each session.
PIANO Mode: 9 mm spot size, 120–150 J/cm² fluence, 5 s pulse
width, a frequency of 4.0 Hz, and 1500 rounds at each session.
Superficial Mode: 5 mm spot size of 5 mm, 1.0–1.5 J/cm² flu-
ence, and a frequency of 2.0 Hz.
2.5 | Treatment Protocol
All enrolled participants were instructed to discontinue their
usual skincare products for 2 weeks before the treatment. Basic
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skincare products, that is, sunscreen, cleansing foam, and
moisturizing emulsion, were provided for daily use throughout
the study.
On D0, all participants' facial skin was cleansed with a neutral
lotion before the session, and no topical anesthesia was applied.
Each of them lying in a horizontal position, received the three
modes of laser therapy, successively. Immediately after the laser
treatment, they applied the antiaging formulation on the test
side and the blank formulation on the control side, maintaining
this regimen twice daily for 28 days. The facial midline, defined
by anatomical landmarks such as the glabella, nasal bridge,
philtrum, and chin, served as the physical boundary between
the two sides.
2.6 | Formulation
The test item is a commercially available antiaging serum,
produced by Yunnan Botanee Co. Ltd in China, containing a
proprietary complex of C. sinensis extract, P. suffruticosa extract,
K. alvarezii extract, ergothioneine, A. oleracea extract, M. alba
extract, β‐alanyl hydroxyprolyldiaminobutyroyl benzylamide,
and palmitoyl hexapeptide‐12. The placebo control is a base
formulation without the mentioned complex.
2.7 | Instrumental Evaluation
All outcomes were assessed under controlled environmental
conditions (21°C ± 1°C with 50% ± 10% relative humidity).
Participants acclimated for 30 min before any evaluation.
Skin surface microstructure and wrinkles were investigated using
PRIMOS Clinical Research System (Canfield Scientific), measuring
depth and area percentage of crow's feet wrinkles and nasolabial
foldsateachvisit.SkintexturewasrecordedusingVisioScanVC20
plus (Courage & Khazaka), analyzing parameters such as skin
smoothness (SEsm) and wrinkles (SEw). Skin elasticity was deter-
mined by the Cutometer MPA 580 (Courage & Khazaka), focusing
on R2 (total elasticity) and F4 (firmness). Trans‐epidermal water
loss (TEWL) was measured using Tewameter (Courage & Kha-
zaka). The VISIA system (Canfield Scientific) was used for captur-
ing facial images, and skin thickness was determined by Ultrascan
UC22 (Courage & Khazaka).
2.8 | Clinical Evaluation
Clinical evaluations were conducted during all visits by two
independent dermatologists using the Evaluation Criteria for
Human Skin Aging and Asian Skin Aging Atlas, Volume 2.The
Evaluation Criteria for Human Skin Aging scores forehead
wrinkles from 0 to 8, crow's feet wrinkles from 0 to 6, under‐
eye wrinkles from 0 to 9, and nasolabial folds from 0 to 7. The
AsianSkinAgingAtlas,Volume2scores cheek sebaceous pores
from 0 to 5.
Clinical evaluations also included a 10‐point visual analog scale
(VAS, 0–9) for attributes such as elasticity, roughness, firmness,
and global face wrinkles, conducted by the two dermatologists.
The Global Esthetic Improvement Scale (GAIS) was used at
each visit after the Fotona 4D treatment to assess facial
improvement on a 5‐point scale: 3 = improved completely;
2 = improved significantly; 1 = improved slightly; 0 = no dif-
ference; −1 = worsened. All participants were asked to com-
plete a satisfaction questionnaire regarding the antiaging effects
of the treatment for both face sides, with answers recorded on a
9‐point scale (1 = completely disagree, 9 = totally agree).
2.9 | Statistical Analysis
Statistical analysis was performed by using SPSS version 22.0.
The categorical variables were reported as counts and percent-
ages, while the continuous variables were reported in the form of
mean ± standard deviation (SD) or as the minimum, maximum,
and median. The Shapiro–Wilk test was used to assess the nor-
mality of the data set. The intra‐group comparison was per-
formed between the baselines and the other visits, using the
analysis of variance (ANOVA) test for data normally distributed,
otherwise, the Friedman test would be used. Intergroup com-
parisons were performed between the treatment and control
sides. Independent samples t‐test was used for data with normal
distribution, and the Mann–Whitney Utest was used for the non‐
normally distributed data. Additionally, the Mann–Whitney U
test and the Friedman test were applied to the categorical vari-
ables for comparing the between‐group differences and the
within‐group ones, respectively. A two‐sided p‐value less than
0.05 was considered to indicate statistically significant.
3 | Result
3.1 | Demographic Characteristics of Subjects
A total of 37 females were included and 33 of them finished the
study, with two withdrawing during the washout period and
another two withdrawing after the Fotona 4D treatment. The
mean age of the 33 participants was (43.45 ± 3.94) years (range:
35–49 years).
3.2 | Clinical Evaluation of Overall
Improvement (GAIS)
During the study period, the GAIS scores gradually increased on
both facial sides. The distribution of GAIS scores indicated that
overall improvement in facial skin was more prominent on the test
side than the control side, with statistical significance observed from
D4 to D28 (Figure 1). On the test side, all participants achieved at
least an “Improved”rating by D7, whereas the same effect was
observed starting from D14 on the control side. At the final follow‐
up visit on D28, 27 cases (82%) had a GAIS score of 2, and six cases
(18%) scored 1 on the test side; correspondingly, 13 cases (39%)
scored 2, and 20 cases (61%) scored 1 on the control side.
3.3 | Improvement in Skin Barrier
The measurement of TEWL on the skin surface provides in-
sights into the barrier function of the stratum corneum.
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Figure 2illustrates the TEWL on the cheek before and after
28 days of applying the test or control items. TEWL values on
the control side were significantly higher than the baseline on
D3 (p< 0.02), indicating an imperfect skin barrier, and showed
only a 1.58% decrease from the baseline on D28. However, on
the test side, the TEWL values gradually decreased to baseline
levels by D7 and further decreased by 9.51% at the end of the
study (p< 0.001), which suggested the test item may help
accelerate skin barrier recovery and enhance skin barrier
integrity.
3.4 | Improvement in Antiwrinkle
3.4.1 | Clinical Evaluation Based on a 10‐Point
VAS (0–9)
The VAS evaluation, where a higher score indicates a worse
condition, revealed a decreasing trend in scores on both
sides with each successive visit. A bilateral comparison of
the VAS scores for roughness and global face wrinkles at
each visit relative to the baselines is shown in Figure 3.
Statistically significant differences in score changes between
the test and control sides were observed in tactile roughness
and global wrinkles starting from D7, while in visual
roughness from D14.
3.4.2 | Clinical Evaluation Based on Atlas
The distribution of scores across different parameters was
similar, with a median of about 3 points (Figure 4). Therefore,
the analysis compared the improvement on both sides using a
cutoff of 3 points. The proportion of scores related to wrinkle
parameters indicated a trend of improvement on both sides
from D7 to D28.
In comparison to their baselines, statistically significant chan-
ges in each clinical efficacy indicator were demonstrated on
both sides by D28, except for the nasolabial folds of the control
side. Daily use of the test serum resulted in improvements in
under‐eye wrinkles, crow's feet, forehead wrinkles, and
FIGURE 1 | Global Esthetic Improvement Scale (GAIS) scores for
skin aging on the test and control sides at baseline, on D7, D14,
and D28.
FIGURE 2 | Changes in transepidermal water loss (TEWL) over
time among participants using the test or control items. Data were
analyzed using the Friedman test; *p< 0.05, compared with the
baseline.
FIGURE 3 | Bilateral comparison of the clinical evaluation scores at each visit relative to baseline. (a) Mean change in visual roughness. (b) Mean
change in tactile roughness. (c) Mean change in global face wrinkles. Data were analyzed using the Wilcoxon Rank Sum Test test; *p< 0.05,
compared with both sides.
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nasolabial folds as early as D7. On the control side, significant
improvement in under‐eye wrinkles was observed by D14,
while crow's feet and forehead wrinkles showed significant
improvement only by D28.
3.4.3 | Measurement of Skin Texture
The treatment with the test serum contributed to a significant
reduction in the facial wrinkle parameter (SEw). From the
FIGURE 4 | Clinical evaluation based on atlas scores on D7, D14, and D28 showing the percentage of scores on each side. (a) Forehead wrinkles.
(b) Nasolabial fold wrinkles. (c) Underneath eye wrinkles. (d) Crow's feet wrinkles.
FIGURE 5 | Evolution of skin wrinkle parameters by skin biophysics analysis after 28 days of using test or control products. (a) Mean changes in
SEw (mean ± SD). (b) Mean changes in SEsm (mean ± SD). Data were analyzed using the Friedman test; *p< 0.05, compared with the baseline. SD,
standard deviation.
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baseline, the test side achieved a reduction by 21.14% over
28 days, whereas the control side showed only a slight anti-
wrinkle effect, with a decrease by 3.83%. The mean SEw
improvement of the test side was more significant than that of
the control side on both D14 and D28 (p< 0.002) (Figure 5).
Especially, the mean SEsm of the test side decreased by 20.68%
on D28, that is, from 229.98 at baseline to 182.41, while the
control side showed a decline from 208.73 to 189.68, a reduction
by 9.13% (Figure 5).
3.4.4 | Measurement of Skin Wrinkles
The area proportion of the nasolabial folds on the test side
decreased significantly by D3 and this decline trend was sus-
tained through D28, with a decrease by 7.03% from 18.00% at
baseline to 16.73% on D28. However, a slight decrease by 3.36%
occurred to the control side, from 18.47% to 17.85% (Table 1and
Figure 6). Additionally, a similar pattern was found for changes
over time in the area proportion of crow's feet (Table 1and
Figure 6).
Wrinkle depth is another indicator of evaluating the anti-
wrinkle effect. Both crow's feet and nasolabial folds showed a
decreasing mean depth over 28 days, although not statistically
significant compared to the baseline (Table 1). As illustrated in
Figure 7, representative examples of 3D images exhibited
wrinkle changes within the dotted circles.
3.5 | Improvement in Skin Firmness
3.5.1 | Clinical Evaluation Based on a 10‐point
VAS (0–9)
The scores of firmness and elasticity graded visually by
dermatologists were falling with each successive visit for
both sides (Figure 8). A bilateral comparison of the scores
relative to the baseline showed statistically significant lower
scores were observed on the test side than the control side
early as on D7.
3.5.2 | Measurement of Skin Mechanical Properties
The increasing values in R2 and decreasing ones in F4 from
baselines were demonstrated on both sides (Figure 9). Notably,
the test sides suggested a more pronounced improvement in the
two measured indices than their counterparts.
3.5.3 | Measurement of Dermal Thickness
The dermis of the cheeks, indicated by the ultrasonographic
measurement, becomes thicker on D14 and D28 for both sides
(Table 1). There was a greater average magnitude of thickness
elevated from baselines on the test sides compared with the
control ones. The representative examples of ultrasonographic
skin images are shown in Figure 10.
TABLE 1 | Changes associated with skin wrinkles and skin thickness parameters during the test.
Side Baseline D3 (△1) D7 (△2) D14 (△3) D28 (△4)
Percentage of
wrinkle area (%)
Nasolabial
folds
Test 18.00% ± 0.33% 17.95% ± 0.16% (−0.27%) 17.39% ± 0.25% (−3.36%) 17.32% ± 0.31% (−3.79%) 16.73% ± 0.27% (−7.03%)
Control 18.47% ± 0.13% 18.34% ± 0.12% (−0.70%) 17.95% ± 0.14% (−2.81%) 17.94% ± 0.14% (−2.88%) 17.85% ± 0.18% (−3.36%)
Crow's feet Test 17.42% ± 0.38% 17.33% ± 0.39% (−0.47%) 16.69% ± 0.41% (−4.15%) 15.98% ± 0.42% (−8.24%) 15.43% ± 0.49% (−11.41%)
Control 17.56% ± 0.26% 17.85% ± 0.22% (+1.68%) 17.06% ± 0.22% (−2.81%) 16.94% ± 0.21% (−3.54%) 16.98% ± 0.24% (−3.28%)
Wrinkle
depth (μm)
Nasolabial
folds
Test 77.36 ± 4.05 74.36 ± 2.90 (−3.88%) 74.24 ± 3.26 (−4.03%) 73.30 ± 2.89 (−5.25%) 72.39 ± 2.64 (−6.42%)
Control 76.52 ± 4.78 76.12 ± 4.11 (−0.51%) 75.03 ± 3.26 (−1.94%) 73.64 ± 3.43 (−3.76%) 73.21 ± 3.01 (−4.32%)
Crow's feet Test 56.18 ± 3.72 53.15 ± 3.37 (−5.39%) 52.48 ± 3.05 (−6.58%) 51.58 ± 3.01 (−8.20%) 51.48 ± 2.85 (−8.36%)
Control 59.88 ± 6.11 56.58 ± 5.07 (−5.52%) 57.09 ± 5.14 (−4.66%) 56.76 ± 5.68 (−5.21%) 57.30 ± 5.91 (−4.30%)
Dermal thickness Test 1794.42 ± 40.78 1748.82 ± 48.93 (−2.54%) 1885.97 ± 42.96 (+5.10%) 1925.85 ± 49.62 (+7.32%) 2009.45 ± 46.26 (+11.98%)
Control 1725.21 ± 44.75 1680.76 ± 50.30 (−2.58%) 1845.15 ± 46.02 (+6.95%) 1879.67 ± 42.66 (+8.95%) 1938.79 ± 49.16 (+12.38)
Note: Data at baseline, Day (D)3, D7, D14, and D28 were indicated as mean ± standard deviation (SD); Δ1, the percentage changes on D3 (vs. baseline); Δ2, the percentage changes on D7 (vs. baseline); Δ3, the percentage changes on D14
(vs. baseline); Δ4, the percentage changes on D28 (vs. baseline). Data related to wrinkles were analyzed using the Friedman test, and data related to dermal thickness were analyzed using the analysis of variance (ANOVA) test.
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3.5.4 | Evaluation of Sebaceous Pores
The changes in sebaceous pores, as reductions in pore size and pore
area percentage reflect firmer skin, canbeanalternativeindicatorof
skin firmness. Low sebaceous‐poresscores(≤3) based on the atlas
rose in proportion for both sides throughout the study (Figure 11).
Additionally, larger proportions of low scores appeared on the test
side than on the control one from D7 on.
FIGURE 6 | Evolution of skin wrinkle parameters by skin biophysics analysis after 28 days of using test or control products. (a) Changes in the
distribution of data in the nasolabial folds wrinkles area. (b) Changes in the distribution of data in the crow's feet wrinkles area. Data were analyzed
using the Friedman test; *p< 0.05, compared with the baseline.
FIGURE 7 | Representative three‐dimensional images of wrinkle changes at baseline and on D28. (a) Three‐dimensional images for crow's feet
wrinkles on the test and control sides in Subject 18. (b) Three‐dimensional images for nasolabial folds wrinkles on the test and control sides in
Subject 33.
FIGURE 8 | Bilateral comparison of clinical evaluation scores at each visit relative to the baseline. (a) Mean change in firmness. (b) Mean change
in elasticity. Data were analyzed using the Friedman test; *p< 0.05, compared with both sides.
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The area percentages of sebaceous pores produced by PRIMOS
were depicted in Figure 9. After 28 days, the test side exhibited a
reduction in mean area percentage by 22.49%, while the control
one did by 11.65% only. Signs of improvement were observed on
the test sides at visits of D14 (p< 0.02) and D28 (p< 0.001). The
representative example images of the sebaceous pores are
shown in Figure 12.
3.6 | Self‐Assessment
Each participant completed a questionnaire on D7, D14, and
D28, and revealed self‐perceived benefits brought by the test
serum consistent with expert visual grading and noninvasive
biophysics measurement.
3.7 | Adverse Events
No serious adverse events were observed during the entire
study. None of the participants suffered erythema or papules
due to the laser treatment and daily use of test serum. Only one
case of very mild edema was reported, and the symptom was
considered secondary to the laser treatment and relieved
spontaneously within 3 days.
4 | Discussion
Participants enrolled in this study sought the Fotona 4D laser
treatment due to concerns about aging‐related appearance chan-
ges. Therefore, optimizing the efficacy of skin rejuvenation and
minimizing adverse effects from cosmetic laser therapy is crucial.
Compared with traditional fractional lasers, NAFLs can en-
hance skin quality with fewer adverse effects and reduced
downtime. Nevertheless, the three modes employed in this
FIGURE 9 | Evolution of skin firmness parameters by skin biophysics analysis after 28 days of using test or control products. (a) Changes in R2.
(b) Changes in F4. (c) Changes in the proportion of sebaceous pores area. Data for R2 and sebaceous pore area distribution were analyzed using
analysis of variance (ANOVA), while F4 data and the percentage of sebaceous pores area were analyzed using the Friedman test; *p< 0.05, compared
with the baseline.
FIGURE 10 | Images of skin condition at baseline and on D28. Ultrasonographic skin aspect on the test and control sides in Subject 33, the white
circle represents the dermis.
FIGURE 11 | Clinical evaluation based on atlas related to seba-
ceous pores at D7, D14, and D28 showing the percentage of score at
each side.
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study may still induce mild erythema, edema, flushing, pimples,
and desquamation. The active ingredients, including C. sinensis
extract [7], M. alba extract [14], and ergothioneine [15], incor-
porated into the test serum, can counteract oxidative stress and
stimulate collagen renewal. This efficacy was confirmed by the
TEWL values and tolerability assessments.
Furthermore, penetration of the epidermal barrier has
always been a significant challenge for active ingredients in
drugs and cosmetics, with the stratum corneum being the
primary rate‐limiting step [16, 17]. Multiple studies showed
that laser pretreatment can enhance the permeability and
depth of molecule penetration [18–20]. The superficial mode
used in this study is a type of micro‐ablative fractional laser
that may facilitate deeper penetration, allowing active
ingredients to reach deeper skin layers through ablated
channels extending into the dermis.
Previous research has shown that dermal density and the
thickness of the epidermis and dermis are key parameters in
demonstrating the cutaneous regeneration process [21]. The
test serum significantly improved most measured aging
parameters, with these improvements sustained over 28 days
and progressively enhanced from baseline on the test sides
compared to the control sides. The gradual increase in dermal
density and thickness is probably facilitated by the addition of
Cortex moutan extract [8], which induces fibroblast synthesis
and sustains the effects of the Fotona 4D laser, thereby pro-
longing the benefits of facial rejuvenation achieved through
medical esthetics.
However, the study's limitations include a small sample size and
a relatively short follow‐up period. A larger sample size could
enhance the reliability of the study and strengthen the statistical
power of the analysis. Extending the follow‐up period is also
helpful in exploring the sustainable benefits of product use on
skin rejuvenation. In addition, common adverse events associ-
ated with lasers, such as erythema, desquamation, and burning,
were infrequent in this study. Some anti‐inflammatory ingredi-
ents like Portulaca oleracer extract, Helichrysum stoechas [22]
extract, and M. alba extract [23–25] are included in the formu-
lation, but whether the test serum can reduce inflammation is
still unclear and needs to be proved in the future research.
Overall, the serum with a multi‐component antiaging compo-
sition enhances the efficacy of Fotona 4D in skin rejuvenation.
However, more solid evidence originating from well‐designed
clinical trials is necessary.
Author Contributions
Xinxuan Zhang: conceptualization, investigation, writing –review and
editing. Manru Ning: conceptualization, supervision, writing –original
draft. Mengqing Lin: investigation. Qi Tang: investigation. Yihuai
Liang: conceptualization, methodology, writing –review and editing.
Feifei Wang: funding acquisition, writing –review and editing, project
administration. Xiaoke Xu: conceptualization, investigation, method-
ology, project administration, writing –review and editing.
Acknowledgments
All authors have read and approved the final version of the manuscript.
Feifei Wang and Xiaoke Xu had full access to all of the data in this study
and takes complete responsibility for the integrity of the data and the
accuracy of the data analysis. This study was supported by the inde-
pendent research fund of Yunnan Characteristic Plant Extraction Lab-
oratory (2022YKZY004 and 2022YKZY006). We are thankful for the
financial support, which was used for the study design, data collection,
and data analysis.
Conflicts of Interest
The authors declare no conflicts of interest.
Data Availability Statement
The data that support the findings of this study are available on request
from the corresponding author. Study Protocol and all of the individual
participant data collected during the trial, after deidentification can be
shared, immediately following publication.
Transparency Statement
The lead author Feifei Wang, Xiaoke Xu affirms that this manuscript is
an honest, accurate, and transparent account of the study being
reported; that no important aspects of the study have been omitted; and
that any discrepancies from the study as planned (and, if relevant, re-
gistered) have been explained.
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