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Article

Wrinkle Improvement in HanDam (Twist) on Ultraviolet B Irradiation-Induced Skin Photoaging in Hairless Mice

by
Inbong Song
1,2,
Judong Song
2,
Ilseok Jang
2,
Dayoung Noh
2,
Chaemyeong Lee
2 and
Jungkee Kwon
1,*
1
Department of Laboratory Animal Medicine, College of Veterinary Medicine, Jeonbuk National University, 79 Gobong-ro, Iksan-si 54596, Republic of Korea
2
Plastic Material R&D Center, OSSTEM IMPLANT, Magokjungang 12-ro 3, Gangseo-gu, Seoul 07522, Republic of Korea
*
Author to whom correspondence should be addressed.
Appl. Sci. 2025, 15(9), 4879; https://doi.org/10.3390/app15094879
Submission received: 25 March 2025 / Revised: 24 April 2025 / Accepted: 25 April 2025 / Published: 28 April 2025
Browse Figures
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Abstract

:
Background: Skin photoaging caused by ultraviolet B (UV-B) irradiation leads to the formation of wrinkles. A method to lessen wrinkles is the application of Polydioxanone (PDO) lifting threads. HanDam (Twist) is a PDO lifting thread with a unique morphological twisted shape. Objective: The aim of this study was to evaluate the wrinkle improvement ability of HanDam (Twist) on UV-B irradiation-induced skin photoaging in a hairless mouse model. Methods: Wrinkles were induced by UV-B irradiation to the backs of female hairless mice for six weeks. After induction, the wrinkles were treated with threads, and the mice were monitored for six weeks post-treatment. Results: Our results showed that treatment with HanDam (Twist) effectively ameliorated UV-B irradiation-induced wrinkle depth and significantly increased collagen density by 13% compared to HanDam (non-Twist) in the histological analysis. In measuring protein expression related to collagen production, HanDam (Twist) significantly increased transforming growth factor beta (TGF-β) and collagen type 1 (COL1) by 46% and 67% compared to HanDam (non-Twist). Matrix metalloproteinase-1 (MMP-1) protein expression showed similar density and no significance compared to HanDam (non-Twist). Conclusions: These findings suggest that HanDam (Twist) improves the effectiveness of lifting threads for skin care compared to that of existing products.

1. Introduction

Skin aging is classified into intrinsic aging, which occurs with age, and extrinsic aging due to the influence of ultraviolet B (UV-B) rays, tobacco use, and environmental pollution factors. Photoaging is caused by extrinsic aging, and UV-B irradiation is one of the important causes of photoaging [1]. Wrinkle formation is a major sign of photoaging [2]. Although the mechanism of wrinkle production is not completely known, one cause is an imbalance in collagen production within the skin tissue.
The wavelength range of UV-B is between 280 and 320 nm, which allows UV-B to penetrate the epidermis and UV-B-induced damage to be made to the dermis [3,4]. UV-B-induced damage causes photoaging and internal and external changes in the skin [5,6]. The internal changes associated with UV-B-induced damage include the inhibition of collagen production; the external changes include wrinkles, pigmentation, and dryness [7,8]. Wrinkle formation in the skin is the main characteristic of photoaging, and the reduction in collagen type 1 (COL1) expression and the increase in matrix metalloprotease-1 (MMP-1) expression are key mechanisms of wrinkle formation [9]. According to a study of aberrant collagen synthesis in the skin caused by photoaging, UV-B irradiation induces the activation of MMPs, which act to break down elastic collagen in the dermis and connective tissue components [10,11]. MMP-1 activity plays a key role in collagen degradation, and collagen synthesis is inhibited by fragments of decomposed collagen. This reaction is a continuous process in photoaging conditions and is a major cause of wrinkles due to the decrease in collagen content in skin tissue [12].
Multiple strategies have been developed to delay the aging process, including filler implants, botulinum toxin injections, and laser treatments [13,14,15]. Rhytidectomy, or facelift surgery, has been a recognized procedure since the early 20th century [16]. The incision facelift method has disadvantages such as high cost, long surgery and recovery times, deep scars, and patient anxiety [17]. Using anti-wrinkle drugs such as retinoids [18] can avoid the disadvantages of surgery, but it takes a long time for the wrinkle improvement to take effect. Therefore, other face-lifting surgical techniques that can overcome the disadvantages are being developed. Face-lifting using lifting threads made of Polydioxanone (PDO) material is particularly safe and effective [19]. The recently developed PDO lifting thread is designed with a barb-like “cog”, and the surgical approach involves implanting this thread directly above the superficial muscular aponeurotic system (SMAS) of the facial skin [20]. The PDO lifting thread is a biocompatible, absorbable thread, and recent research has shown that this thread decreases wrinkle formation through both a physical lifting effect and the promotion of collagen synthesis [21].
HanDam (Twist) is a PDO lifting thread with a cog shape. Common lifting threads do not involve twisting, but HanDam (Twist) applies a unique and characteristic Twist technique. We expect that lifting threads with a Twist shape will further increase the positive effect of the cog shape. Previous research has been conducted on the improvement in materials and cog shape [22,23]. However, the effectiveness of Twist-type lifting threads has not been studied in vivo. Thus, we examined the beneficial effects of HanDam (Twist) for the promotion of collagen synthesis and relief from wrinkles in a UV-induced photoaging hairless mouse model.

2. Materials and Methods

2.1. Animals and Treatment

Eight-week-old female SKH-1 hairless mice were purchased from Orientbio (Seongnam, Republic of Korea) with a weight ranging from 22 to 25 g. The mice were housed in the animal housing facility of the Medicine and Health Sciences Department of the OSSTEM IMPLANT institute. All mice were managed in accordance with institutional ethical guidelines for the care and use of experimental animals at OSSTEM IMPLANT (approval number OST-2103). The mice were housed with 12 h cycles of dark and light, were fed a commercial diet and allowed tap water ad libitum, and were maintained at a constant environmental temperature (25 ± 2 °C) and humidity (about 60%) throughout the study.
After the mice entered the facility, the mice were acclimated for 1 week, and then the experiment was conducted. All mice were randomly divided into 4 groups: normal, UV-B (UV-B irradiated, no treatment), HanDam (non-Twist) (UV-B irradiated + non-Twist thread-treated mice), and HanDam (Twist) (UV-B irradiated + Twist thread-treated mice). Mice were divided into 12 mice in each group. In total, 6 mice were measured for wrinkle measurement throughout the experimental period, and 6 other mice were sacrificed at week 4 for histological and Western blotting analysis. A total of 48 mice were used in the experiment, and euthanasia was performed using carbon dioxide (CO2) gas.

2.2. UV-B Irradiation

UV-B irradiation was performed for 6 weeks. A UV-B lamp manufactured by Sankyodenki, Tokyo, Japan, was used as the light source. Irradiation intensity was measured with a UV radiometer (Solartech Inc., Harrison Township, MI, USA). The mice were irradiated for 6 weeks, and the irradiation dose varied by week as follows: the radiation dose was set to 1 MED = 100 mJ/cm2 and gradually increased to the minimum erythema dose (MED) from 1 minimal erythema dose (MED) 3 times a week for the first week, 2 MED 3 times a week for the next 2 weeks, and 4 MED 2 times a week for the final 3 weeks (Figure 1).

2.3. Thread Treatment

The HanDam (Twist) sample was created by applying a physical process to HanDam (non-Twist) to implement the Twist form (Figure 2). The thread was inserted between the back and legs of the mice and in a wrinkle-prone area due to photoaging. For thread treatment, the mouse was anesthetized by inhalation, and the 2 sides of the back skin were lifted to place the thread in as straight a line as possible in the subcutaneous layer. The length of the suture was 50 mm. Thread-treated mice were monitored daily for abnormalities in the treated area [9].

2.4. Wrinkle Measurement

Replicas of the upper and lower skin surfaces were created using a silicon-based impression material (Courage & Khazaka Electronic GmbH, Koln, Germany) at 4 time points (complete UV-B irradiation, thread treatment, 4 and 6 weeks post-treatment). The replicas were cut into circular sections with a diameter of 1 cm, and the backs of the sections were flattened. Images of the negative replicas were observed using a wrinkle analysis system, Visionline VL 650 (Courage & Khazaka Electronic GmbH, Koln, Germany).

2.5. Histological Analysis

The dorsal skin containing the test materials was sampled 4 weeks after thread treatment and fixed in 10% formaldehyde solution for 24 h. Individual samples were fixed again in a fresh 10% formaldehyde solution for 24 h. After paraffin embedding, transverse sections (5 μm) were obtained along the thread axis. Deparaffinization and dehydration were performed using xylene and alcohol, respectively. Masson’s trichrome staining (Abcam, Cambridge, MA, USA) was performed according to the manufacturer’s manual. Images were visualized using a light microscope (Olympus BX51; Olympus Co., Tokyo, Japan) and digital imaging system (Olympus DP71; Olympus Co., Tokyo, Japan). The images were observed at 200× magnification, and the investigators involved in the histological evaluation were blinded to the study group. Images of the stained sections were analyzed with the Image-Pro Plus 6.0 software (Media Cybernetics, Rockville, MD, USA). The dermal collagen density was calculated as the collagen area/total area in the dermis.

2.6. Western Blotting Analysis

For Western blotting, the dorsal skin containing the test materials was sampled 4 weeks after thread treatment. The protein in tissue lysates was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis using 8–12% SDS–polyacrylamide gel. Separated proteins were transferred to polyvinylidene difluoride membranes (Bio-Rad Laboratories, Hercules, CA, USA). To evaluate protein activity, membranes were blocked with 5% skim milk in PBS and then incubated with primary antibodies to transforming growth factor beta (TGF-β), COL1, MMP-1 (Abcam, Cambridge, MA, USA), and β-actin (Cell Signaling, Danvers, MA, USA) in 1% skim milk and then washed with PBS overnight in the dark. After the blots were washed, the membranes were incubated with goat anti-rabbit and goat anti-mouse IgG antibodies (1:5000 v/v, Millipore, Bedford, CA, USA) for 1 h at room temperature. Each antigen–antibody complex was confirmed by Super Signal West Dura Extended Duration Substrate (Thermo Scientific, San Jose, CA, USA) on the Chemi Imager system (Alpha Innotech, San Leandro, CA, USA) [24,25].

2.7. Statistical Analysis

Data are presented as the mean ± SEM. Significant differences between groups were analyzed using a one-way ANOVA in GraphPad Prism version 5.0 (GraphPad Software, San Diego, CA, USA), and the differences were considered significant at p < 0.05 in Tukey’s multiple range test [26].

3. Results

3.1. Effects of HanDam (Twist) on Wrinkle Formation in UV-B-Irradiated Hairless Mice

We used dorsal skin silicon replicas to investigate wrinkle formation from the pretreatment of thread to 6 weeks after treatment (Figure 3). The wrinkles of the UV-B-irradiated group were deep and thick compared to those in the normal group. Light and thin wrinkle formation was observed in the thread-treated groups (HanDam [non-Twist] and HanDam [Twist]). Changes in wrinkle formation due to thread treatment were maintained for 6 weeks. There was no significance between thread-treated groups (HanDam [non-Twist] and HanDam [Twist]).

3.2. Effects of HanDam (Twist) on Wrinkle Depth in UV-B-Irradiated Hairless Mice

We established the effects on wrinkle depth from the pretreatment of thread treatment to 6 weeks after treatment (Figure 4). Before treatment, all groups irradiated with UV-B had deep wrinkles (>80 μm). After treatment, the thread groups showed a significant reduction in wrinkle depth to similar to that of the normal group. This reduction in wrinkles was maintained for 6 weeks, and the HanDam (Twist) group showed a greater reduction in wrinkles compared to the HanDam (non-Twist) group.

3.3. HanDam (Twist) Ameliorated Collagen Loss in UV-B-Irradiated Hairless Mice

Histological evaluation using Masson’s trichrome stain showed significant differences among the experimental groups (Figure 5a). UV-B irradiation caused a significant decrease in collagen compared to the normal group, and this reduction was ameliorated in both groups of UV-B-irradiated and thread-treated mice and the HanDam (non-Twist) and HanDam (Twist) groups. Collagen density was evaluated histologically by visualizing Masson’s trichrome stain as an index of collagen accumulation. As shown in Figure 5b, in the UV-B-irradiated mice, the density of collagen in the dermis decreased; however, an increase in collagen density was confirmed in the thread-treated groups, and the HanDam (Twist) group showed the highest collagen density.

3.4. HanDam (Twist)-Induced Collagen Synthesis Protein Expression in UV-B-Irradiated Hairless Mice

TGF-β, which is involved in collagen synthesis by physical stimulation, increased in thread-treated mice compared to normal and UVB-irradiated group mice. In addition, COL1 was significantly increased in the thread-treated groups. The HanDam (Twist) group had the greatest increase in TGF-β and COL1 expression. The collagen-degrading enzyme MMP-1 was highest in the UV-B-irradiated mice. However, the thread treatment of irradiated mice significantly counteracted the effects of UV-B irradiation on the levels of both COL1 and MMP-1. These results suggest that HanDam (Twist) protects the skin against UV-B irradiation-induced collagen loss by increasing the production of COL1 and decreasing the production of MMP-1 (Figure 6a,b).

4. Discussion

Skin photoaging induced by UV-B irradiation is characterized by wrinkle formation and the inhibition of collagen synthesis. We conducted an experiment using a photoaging model that induces wrinkles in irradiated mice through UV-B exposure. UV-B-induced photoaging inhibits collagen synthesis in the skin, and decreased collagen synthesis impedes skin elasticity, resulting in wrinkle formation [27]. In the current study, we confirmed that mice irradiated with UV-B developed wrinkles. The HanDam (non-Twist) and HanDam (Twist) groups treated with PDO thread showed wrinkle improvement to the normal state. Wrinkle improvement was maintained for up to 4 weeks after treatment (Figure 3). The observed wrinkle improvement is associated with the presence of cogs on PDO threads and is attributed to skin attraction induced by physical stimulation [28].
Wrinkles induced by UV-B irradiation are graded according to depth. The more severe the decline in collagen synthesis and the ease of skin folding, the deeper the wrinkles are formed, and depth is used as an indicator of wrinkles [29]. We confirmed the depth of wrinkles created by UV irradiation. HanDam (non-Twist) and HanDam (Twist) reduced wrinkle depth to normal levels after treatment. In addition, the wrinkle depth increased at 6 weeks compared to immediately post-treatment in the thread-treated groups, and HanDam (Twist) improved wrinkle depth compared to HanDam (non-Twist) (Figure 4). The 6-week post-treatment results confirmed that HanDam (Twist) had an improved ability to maintain wrinkle depth reduction compared with HanDam (non-Twist).
Photoaging induced by UV-B irradiation in the skin leads to lower collagen density in the dermis [23]. Masson’s trichrome staining is a representative method to selectively stain collagen and is suitable for examining collagen density in histological analysis [30,31]. Histological analysis in the present study confirmed that the decrease in collagen density due to UV-B was recovered through HanDam (non-Twist) and HanDam (Twist) treatment. The HanDam (non-Twist) and HanDam (Twist) groups showed greater collagen production than the normal group collagen due to the physical stimulation of the skin by the lifting thread (Figure 5).
The increase in TGF-β and the synthesis of COL1 induced by physical stimulation have been shown to be active at 4 weeks post-treatment [32]. Therefore, we evaluated the correlation between TGF-β and collagen production induced by physical stimulation through protein density analysis 4 weeks after PDO thread treatment. TGF-β plays a key role in the wound-healing process related to collagen synthesis caused by mechanical stimulation by PDO threads [33,34]. Our experimental results confirmed that TGF-β and COL1 increased equally in the HanDam (Twist) group and that compared to the HanDam (non-Twist) group, the production of these two proteins was higher in the HanDam (Twist) group (Figure 6). This result is based on the physical characteristics of the Twist form. The non-Twist form is a shape where the protrusions are arranged in a straight line, whereas the Twist form is a shape where the cogs are arranged in all directions. The PDO lifting thread is known to have a lifting effect due to its mechanical effect and collagen production by fibroblast stimulation by increasing the tension between tissues [35,36,37,38].
Therefore, the Twist shape added to PDO thread is likely to induce greater physical stimulation for collagen production than the non-Twist shape, and stronger physical stimulation causes the increase in collagen synthesis. According to the results of this experiment, the Twist shape of PDO thread is thought to be able to alleviate photoaging of the skin by increasing collagen synthesis. The physical stimulation of this Twist-shaped PDO immediately improves wrinkles after treatment, and the increase in collagen synthesis is expected to maintain skin health even after thread metabolism. In addition, since skin characteristics such as the thickness of mice are different from those of humans, we plan to overcome these limitations through future animal and clinical studies.

5. Conclusions

Our results suggest that HanDam (Twist) has beneficial effects on skin photoaging induced by UV-B in hairless mice. HanDam (Twist) effectively reduced wrinkles induced by UV-B irradiation and promoted collagen synthesis in this mouse model. The collagen synthesis increase induced by HanDam (Twist) appears to be mediated through TGF-β activation. Our results provide evidence that a Twist shape of PDO thread is effective in wrinkle reduction by increasing physical stimulation compared to conventional PDO thread shapes. This study is significant in that it suggests the possibility of becoming a model for the shape of the next generation lifting thread by applying the Twist shape of the lifting thread. In future studies, we plan to confirm the wrinkle improvement effect of the Twist-shaped PDO thread through animal and clinical evaluations with skin thickness similar to humans.

Author Contributions

Conceptualization, I.S. and J.K.; methodology, J.S. and I.J.; validation, D.N. and C.L.; resources, J.S., I.J., and D.N.; writing—original draft preparation, I.S.; writing—review and editing, I.S. and J.K.; visualization, I.S. and C.L.; supervision, J.S. and J.K. project administration, I.J. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

The animal study protocol was approved by the Institutional Ethics Committee of OSSTEM IMPLANT (protocol code: OST2103 and date of approval: 10 August 2021).

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author. The data are not publicly available due to ethical restrictions.

Acknowledgments

This research was supported by OSSTEM IMPLANT Co., Ltd.

Conflicts of Interest

Author Inbong Song, Judong Song, Ilseok Jang, Dayoung Noh, Chaemyeong Lee was employed by the company OSSTEM IMPLANT. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Abbreviations

The following abbreviations are used in this manuscript:
UV-BUltraviolet B
PDOPolydioxanone
TGF-βTransforming growth factor beta
COL1Collagen type 1
MMP-1Matrix metalloproteinase-1

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Figure 1. Experimental design of the study.
Figure 1. Experimental design of the study.
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Figure 2. Photograph of HanDam (non-Twist) and HanDam (Twist) threads. Photographs of the threads at 50× magnification.
Figure 2. Photograph of HanDam (non-Twist) and HanDam (Twist) threads. Photographs of the threads at 50× magnification.
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Figure 3. Morphological observation of skin. A representative of each group is shown from immediately post-UV irradiation to 6 weeks after thread treatment. After UV-B irradiation, before thread treatment; thread treatment, immediately after thread treatment; 4w, 4 weeks after thread treatment; 6w, 6 weeks after thread treatment. Photographs of the skin at 50× magnification. Arrows indicate wrinkles created by UV-B.
Figure 3. Morphological observation of skin. A representative of each group is shown from immediately post-UV irradiation to 6 weeks after thread treatment. After UV-B irradiation, before thread treatment; thread treatment, immediately after thread treatment; 4w, 4 weeks after thread treatment; 6w, 6 weeks after thread treatment. Photographs of the skin at 50× magnification. Arrows indicate wrinkles created by UV-B.
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Figure 4. Evaluation of wrinkle depth from skin samples. The wrinkle depth of each group is shown from immediately post-UV irradiation to 6 weeks after thread treatment. UV-B irradiation, before thread treatment; Thread Treatment, immediately after thread treatment; 4w, 4 weeks after thread treatment; 6w, 6 weeks after thread treatment. Photographs of the skin at 50× magnification. Arrows indicate wrinkles created by UV-B. (a–d) Mean values with different letters differed significantly (p < 0.05) among groups by Tukey’s test.
Figure 4. Evaluation of wrinkle depth from skin samples. The wrinkle depth of each group is shown from immediately post-UV irradiation to 6 weeks after thread treatment. UV-B irradiation, before thread treatment; Thread Treatment, immediately after thread treatment; 4w, 4 weeks after thread treatment; 6w, 6 weeks after thread treatment. Photographs of the skin at 50× magnification. Arrows indicate wrinkles created by UV-B. (a–d) Mean values with different letters differed significantly (p < 0.05) among groups by Tukey’s test.
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Figure 5. Histological evaluations of collagen fibers in dorsal skin of mice against UV-B-induced skin damage. Four weeks post-thread treatment, (a) Masson’s trichrome stain (magnification 200×), and (b) histogram of estimated relative collagen density. (a–d) Mean values with different letters differed significantly (p < 0.05) among groups by Tukey’s test. Scale bars = 200 μm.
Figure 5. Histological evaluations of collagen fibers in dorsal skin of mice against UV-B-induced skin damage. Four weeks post-thread treatment, (a) Masson’s trichrome stain (magnification 200×), and (b) histogram of estimated relative collagen density. (a–d) Mean values with different letters differed significantly (p < 0.05) among groups by Tukey’s test. Scale bars = 200 μm.
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Figure 6. Effects of HanDam (Twist) on activation of TGF-β, COL1, and MMP-1 in UV-B irradiation-induced skin aging in hairless mice. Four weeks post-thread treatment, (a) the protein expression related to collagen synthesis through physical stimulation was evaluated using Western blot analysis, and (b) displays a histogram showing TGF-β, COL1, and MMP-1 relative densities. (a–c) Mean values with different letters differed significantly (p < 0.05) among groups, as shown by Tukey’s test.
Figure 6. Effects of HanDam (Twist) on activation of TGF-β, COL1, and MMP-1 in UV-B irradiation-induced skin aging in hairless mice. Four weeks post-thread treatment, (a) the protein expression related to collagen synthesis through physical stimulation was evaluated using Western blot analysis, and (b) displays a histogram showing TGF-β, COL1, and MMP-1 relative densities. (a–c) Mean values with different letters differed significantly (p < 0.05) among groups, as shown by Tukey’s test.
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MDPI and ACS Style

Song, I.; Song, J.; Jang, I.; Noh, D.; Lee, C.; Kwon, J. Wrinkle Improvement in HanDam (Twist) on Ultraviolet B Irradiation-Induced Skin Photoaging in Hairless Mice. Appl. Sci. 2025, 15, 4879. https://doi.org/10.3390/app15094879

AMA Style

Song I, Song J, Jang I, Noh D, Lee C, Kwon J. Wrinkle Improvement in HanDam (Twist) on Ultraviolet B Irradiation-Induced Skin Photoaging in Hairless Mice. Applied Sciences. 2025; 15(9):4879. https://doi.org/10.3390/app15094879

Chicago/Turabian Style

Song, Inbong, Judong Song, Ilseok Jang, Dayoung Noh, Chaemyeong Lee, and Jungkee Kwon. 2025. "Wrinkle Improvement in HanDam (Twist) on Ultraviolet B Irradiation-Induced Skin Photoaging in Hairless Mice" Applied Sciences 15, no. 9: 4879. https://doi.org/10.3390/app15094879

APA Style

Song, I., Song, J., Jang, I., Noh, D., Lee, C., & Kwon, J. (2025). Wrinkle Improvement in HanDam (Twist) on Ultraviolet B Irradiation-Induced Skin Photoaging in Hairless Mice. Applied Sciences, 15(9), 4879. https://doi.org/10.3390/app15094879

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