Evaluation of Effects on Skin Quality of a Centella asiatica Extracellular Vesicle-based SkinCare Formulation: A 28-Day Facial Skin Quality Study

Evaluation of Effects on Skin Quality of a Centella asiatica Extracellular Vesicle-based SkinCare Formulation: A 28-Day Facial Skin Quality Study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Evaluation of Effects on Skin Quality of a Centella asiatica Extracellular Vesicle-based SkinCare Formulation: A 28-Day Facial Skin Quality Study Tsong-Min Chang, Chung-Chin Wu, Huey-Chun Huang, Shr-Shiuan Wang, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6143379/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Skin damage results from the natural aging process, physical wear and tear, UV light, and environmental pollutants. Centella asiatica (Cica) is a widely used traditional medicinal herb and is known to have therapeutic effects on skin and wound healing. This study aims to evaluate the effects of Kristen Claire Supreme Rejuvenation Essence® with Cica EV as the main active ingredient, on facial appearance and skin quality in healthy participants. Twenty healthy participants (4 males and 16 females; average age 36.5) were enrolled and underwent a 24-hour skin patch test on the forearm to assess potential irritation or allergic reactions. The test product was applied twice daily for 28 days. Facial skin quality assessments were conducted before use and at 7, 14, 21, and 28 days of product application. Measured parameters included skin hydration, melanin content, skin elasticity, wrinkle percentage, redness area percentage, and pore percentage. After 28 days of test product treatment, significant improvements were observed in skin hydration and elasticity, wrinkle, redness, and pore distribution, while skin melanin content was reduced significantly. Kristen Claire Supreme Rejuvenation Essence® effectively improved skin hydration, elasticity, and texture, brightened the complexion, and showed potential in slowing skin aging, making it a promising candidate for skincare applications. Dermatology Centella asiatica Cica skincare cosmetics longevity extracellular vesicles skin quality skin hydration skin whitening skin elasticity wrinkle reduction Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction As the largest organ of the human body, skin is the main barrier of entry which protects the internal body from environmental damage and stress, maintains homeostasis, and provides key immune functions [ 1 ]. The skin consists of three main layers: the epidermis, which is the outermost protective layer composed of corneocytes, keratinocytes, melanocytes, and Langerhan’s cells; the dermis, which imparts the elasticity and structure of the skin by the extracellular matrix proteins such as collagen and elastin secreted by fibroblasts; and subcutaneous tissue, which is composed mostly of adipose tissue, containing adipocytes and stromal vascular cells [ 2 ]. Over time, skin undergoes aging due to intrinsic factors resulting in cell senescence characterized by reduced collagen production and decreased elasticity, as well as due to extrinsic factors like UV exposure, pollution, and oxidative stress [ 2 ]. To combat the effects of skin aging, modern skincare aims to restore skin hydration and elasticity, reduce pigmentation, and prevent or repair damage caused by aging and environmental factors. According to McKinsey, the global cosmetics market is a multi-billion industry which grew to $ 446 billion in 2023 and is estimated to reach $ 590 billion by 2028. It is characterized by a CAGR of 6%, with skincare representing the largest share of cosmetics, accounting for 44% of the market in 2023 [ 3 ]. There has been renewed interest in natural products for cosmetics, both from consumers and industry due to potential risks associated with synthetic compounds, as well as the purported benefits of natural products [ 4 ]. Medicinal plants have been used in many cultures since ancient times and contain multifaceted therapeutic properties, which can be attributed to a multitude of secondary metabolite phytochemicals [ 4 ]. Centella asiatica , also known as Cica, is commonly used in traditional herbal medicine such as Chinese traditional medicine and Indian Ayurveda therapy. The plant contains a rich assortment of phytochemicals such as triterpenoids, flavonoids, and polyphenols that contribute to its anti-inflammatory, wound-healing, and central nervous system protective properties [ 5 – 7 ]. Due to its broad spectrum benefits, commercial products containing Cica extracts are available for wound healing and skin care purposes [ 8 ]. A cursory glance at major cosmetics retailers in the United States and Japan revealed a wide variety of Cica-based skin care products, representing a sizeable section of natural skin care market [ 9 , 10 ]. Extracellular vesicles (EVs) are nanoscale lipid membrane-bound particles released by cells into the extracellular environment, carrying bioactive molecules that can modulate the physiological processes of target cells, even across disparate kingdoms of life [ 11 ]. Recent studies have shown that exosome-like EVs from plants, including Cica, promote skin improvement and wound healing more effectively than conventional plant extracts [ 12 – 14 ]. We have previously characterized ExoBella®, a proprietary preparation of EVs isolated from tissue culture Cica, EVs and confirmed their potent in-vitro antioxidant, anti-melanogenic, and anti-inflammatory properties [ 15 ]. Kristen Claire Supreme Rejuvenation Essence® is a skincare product formulated with ExoBella® Cica EVs as the main active ingredient. This clinical study aimed to evaluate skin improvement in volunteer users of Kristen Claire Supreme Rejuvenation Essence® over a period of 28 days. Materials and Methods Participants and Study Design Participants were healthy adults of any gender from the age of 18 to 60 years (inclusive). The inclusion criteria included the absence of chronic diseases, major illnesses, or allergies, and not currently using any medications or other skincare products. The participants were required to conduct a pre-test screening to confirm the absence of skin irritation or allergic reaction to the test product which consisted of applying a skin patch infused with the test product for 24 hours on the inner forearms. After screening, participants were asked to use the test product twice daily (morning and evening) after cleansing their face without using exfoliating products for 28 consecutive days. Each application involved 2 drops, evenly applied to the face using fingertips for absorption. Product usage was self-recorded, and the products were stored at room temperature. The skin quality tests were conducted at the investigation site Hungkuang University (Taichung City, Taiwan) onsite by the principal investigator. Each participant was to allocate 1 hour per test (including cleansing and waiting time). Skin quality tests were performed as a baseline test on day 0 (before using the test product), and subsequent tests at 7, 14, 21, and 28 days after test product use. Measurements of the skin quality tests included skin hydration, melanin content, and elasticity index using a C + K Multi Probe Adaptor MPA580 skin analysis device (Courage-Khazaka Electronic GmbH, Koln, Germany) equipped with probes for hydration, pigmentation, and elasticity measurements. Probes were placed 1 cm horizontally and 1 cm vertically from the outer corner of each eye (on the left and right cheeks) and 2 cm above the midpoint of the eyebrows (forehead). Additionally, the VISIA® Skin Analysis system (Canfield Imaging Systems, Fairfield, NJ, USA) was used to assess changes in wrinkle percentage, redness area percentage, and pore percentage by scanning full-face frontal and 45° angles on both sides of the face. The tests were conducted in a controlled environment at 20 ± 1°C and 50 ± 5% relative humidity, without direct sunlight or air conditioning airflow. All measurements were non-invasive. Probes contact the skin's surface to assess parameters based on electrical resistance, optical principles, and physical changes. Participants’ test results and photographic data were collected. Description of Test Product The test product (Kristen Claire Supreme Rejuvenation Essence®) is a proprietary water-based formulation containing the main active ingredient ExoBella®, a preparation of EVs derived from tissue culture C. asiatica . Other active ingredients include Rubus fruiticosus leaf extract, Glycyrrhiza glabrai root extract, Calendula officinalis flower extract, Aloe barbadensis leaf juice, Vaccinium angustifolium fruit extract, Morus nigra fruit extract, arbutin, hydrolyzed DNA, niacinamide, and tocopheryl acetate (Vitamin E). The appearance is clear with slightly gel-like in consistency. Assessment of Outcomes Skin hydration was measured using the Corneometer® CM825 (Courage-Khazaka Electronic GmbH, Koln, Germany), which evaluates the dielectric constant related to epidermal moisture at a depth of 60–100 µm. Mexameter® MX18 (Courage-Khazaka Electronic GmbH, Koln, Germany) was used to measure melanin content, which utilizes RGB light absorption (568 nm, 660 nm, and 880 nm) to measure melanin and hemoglobin content. Elasticity index was measured using the Cutometer® Dual MPA580 system (Courage-Khazaka Electronic GmbH, Koln, Germany), based on suction and stretching properties of the skin. VISIA® Skin Analysis System (Canfield Imaging Systems, Fairfield, NJ, USA) was used to evaluate the following parameters: Wrinkle percentage was measured by using standard white light to detect shadow variations to determine the distribution and number of wrinkles. Fewer wrinkles yield higher percentages. Redness percentage was measured by RBX polarized light to identify vascular or inflammatory issues. Smaller red zones result in higher percentages. Pore percentage was measured under white light to analyze shadowed pore depressions and darker areas relative to surrounding skin. Fewer pores yield higher percentages. Data Presentation and Statistical Analysis For each of the measured parameters, results are converted to skin improvement rate (%) calculated as follows: $$\:\frac{\left(Post‐treatment\:index\:-\:Pre‐treatment\:index\right)}{Pre‐treatment\:index}\:\times\:100\%$$ Where the pre-treatment index is the score of the parameter examined on Day 0, and the post-treatment index is the score of the parameter examined on Day 7, 14, 21, or 28. In skin hydration and elasticity, a positive skin improvement rate indicates higher hydration and high elasticity, respectively, when compared to Day 0. In skin melanin content, the skin improvement rate is negative because the parameter measures the decrease in melanin content and the lower skin improvement rate indicates greater decrease in melanin. In parameters measured using the VISIA® Skin Analysis System, a higher skin improvement rate indicates decreased wrinkles, redness, or pores. Results are presented as mean of the skin improvement rate with 95% confidence intervals shown as error bars. The analysis package in GraphPad Prism 6.01 (Boston, MA) was used for statistical analysis. A paired Student’s t-test was used to compare the difference between means of skin improvement rate on Days 7, 14, 21, or 28 against Day 0 (baseline). Results Participants From September 4, 2024, to October 4, 2024, 20 healthy participants (4 males and 16 females; average age 36.5) were enrolled and all were assessed negative for irritation or allergic reactions to the test product prior to the study. The participants ranged from the age of 23 to 54 years, and the mean age was 36.5 with a standard deviation of 9.2. Assessments of Participants The participants were assessed for changes in skin parameters, including skin hydration, melanin content, skin elasticity, wrinkle, redness (erythema), and skin pores. The raw data values of the assessment are presented in Tables S1 to S6. The evaluation was performed on three facial regions (left cheek, right cheek, and forehead) every 7 days over a 28-day period. In skin hydration, statistical significant increases in hydration percentage were observed across all facial regions (left cheek, right cheek, and forehead) over the course of 28 days compared to baseline, with the right cheek having slightly less improvement compared to other regions (Fig. 1 ). Skin hydration improved by 3.1–5.1%, 5.8–10.6%, 9.6–15.2%, and 14.6–21.2% on Days 7, 14, 21, and 28, respectively. The treatment also resulted in significantly lowered melanin content throughout the study period with melanin decreasing by 1.2–2.3%, 4.1–5.0%, 6.2–7.8%, and 9.2–11.3%, on Days 7, 14, 21, and 28, respectively (Fig. 2 ). Similarly, treatment resulted in significant improvement in skin elasticity over time uniformly across all regions tested (Fig. 3 ). The improvement in skin elasticity was 2.2–3.5%, 4.6–5.9%, 7.4–8.9%, and 12.0–12.5% on Days 7, 14, 21, and 28, respectively. The VISIA® Skin Analysis System was used to image and analyze skin wrinkles, redness, and pore distribution simultaneously. Skin wrinkle condition improved by 8.6–16.9%, 19.3–24.9%, 28.2–30.0%, and 32.9–34.8% on Days 7, 14, 21, and 28, respectively (Fig. 4 ). The forehead region lagged behind in wrinkle improvement at an earlier stage during the study but increased to approximately equal to that of other facial regions by Day 21 (Fig. 4 ). Skin redness was ameliorated during the study period by 5.6–7.9%, 13.1–14.6%, 19.9–22.7%, and 26.3–34.0% on Days 7, 14, 21, and 28, respectively (Fig. 5 ). Finally, skin pores reduction by treatment resulted in skin improvement by 9.3–12.4%, 19.8–21.8%, 28.5–32.1%, and 40.6–41.3% on Days 7, 14, 21, and 28, respectively (Fig. 6 ). In summary, the treatment had a significant improvement in skin hydration and elasticity, while reducing the distribution and area of melanin, wrinkles, redness, and pores. To investigate the effect of treatment in different age groups, the data was stratified according to the age of participants into two groups: > 40 years and < 40 years. The means of skin improvement rate of each parameter were compared between the two age groups at the three facial regions. We found no significant differences in skin hydration and elasticity between the two age groups (Figure S1). However, those aged above 40 years had significant less reduction in melanin in both cheek regions compared to participants aged less than 40 years on Day 28 of the study, while there were no significant differences between forehead region in both age groups (Figure S1). For parameters analyzed with VISIA® System, there were no significant differences between the two age groups in changes in skin wrinkles and pores (Figure S2). In redness skin area, the overall improvement in > 40 years age group was less than that of < 40 years age group, and the difference between two age groups was significant on Day 28 at the forehead region (Figure S2). Discussion In this small-scale clinical study, we observed positive improvements on the skin in participants treated with Cica EV-based test product. Skin integrity parameters such as hydration and elasticity increased after treatment with the test product. According to our previous study, Cica EVs could achieve improvement in skin hydration via upregulation of water retention genes such as aquaporin and filaggrin, and higher skin elasticity by increasing production of collagen and elastin [ 15 ]. The triterpene madecassoside in Cica has already been studied for its anti-melanogenic effect via anti-inflammatory effects [ 16 ]. Besides madecassoside, miRNAs within Cica EVs can also target melanogenic genes such as tyrosinase and other pathway inducers (WNT, CRTC1), as has been observed in our previous study and others [ 13 , 15 ]. Cica extracts have already been used in commercial skin care products due to their skin healing, antioxidant, and anti-melanogenic properties imparted by their bioactive compounds [ 6 – 8 ]. The mechanisms through which exosomes promote skin regeneration include angiogenesis, collagen synthesis, and regulation of inflammation [ 17 , 18 ]. A study discovered 11 novel miRNAs contained within Cica exosomes, which were predicated to inhibit genes involved in melanin production and dermatitis [ 13 ]. The study also found that Cica exosomes induced higher gene expression than C. asiatica extracts, possibly due to better penetration of skin cells by exosomes and the presence of packaged miRNAs [ 13 ]. In formulation of the test product, ExoBella® Cica EVs may work in concert with other herbal extracts and small molecule compounds (arbutin, niacinamide, vitamin E) to achieve the results seen in this study. The main limitation of the study is the small sample size, which may not include sufficient diversity of demographics such as different skin types and ethnicities due to the homogeneity of population in Taiwan. The small sample size also made it unfeasible to conduct a more robust analysis of differences between age groups, and stratification into smaller age groups (e.g. 16 ~ 25 years, 26 ~ 35 years, etc.). The sample population is also heavily gender-skewed toward female (80% of participants), which can also influence the generalizability of the results. The other limitation was the relatively short duration of the study (28 days). This may not be sufficient to capture the long-term effects of the test product as skin quality can be influenced by numerous factors over time including aging, UV exposure, and other environmental factors. A longer study period and follow-up would allow a more comprehensive understanding of the product's sustained efficacy and potential adverse effects. Lastly, as this was not a placebo-controlled nor double-blinded study, it would be difficult to determine whether the improvements observed were attributed to the use of test product, natural improvement of skin over time, or due to other external factors. In conclusion, the results of this study support the positive effects on skin quality of Kristen Claire Supreme Rejuvenation Essence® in improving skin hydration and elasticity while reducing melanin, wrinkles, redness, and pores. The differences observed between age groups and the three facial regions were minimal. Future studies should address the above mentioned limitations to provide clearer insights into the long-term benefits on skin improvement and application of the test product across more diverse set of population to address a global market. Declarations Author contributions Conceptualization, supervision and project administration: T.-M.C., C.C., T.-Y.K. Visualization and formal analysis: T.-M.C., L.T.-C.L. Investigation, data acquisition and curation: T.-M.C., H.-C.H., S.-S.W. Methodology: T.-M.C., C.-C.W., H.-C.H., S.-S.W., C.-H.C., P.-L.K., W.-H.T., W.-Y.Q. I.P., T.-Y.K. Resources: C.-C.W., C.-H.C., P.-L.K., W.-H.T., T.-Y.K. Writing—original draft preparation: T.-.M.C., L.T.-C.L. Writing—review and editing: T.-M.C., L.T.-C.L., W.-Y.Q., I.P., C.C., T.-Y.K. All authors reviewed the manuscript Funding Schweitzer Biotech Company provided the necessary funding for the study as well as provided test product Kristen Claire Supreme Rejuvenation Essence®, but had otherwise no role in enrollment, participation, data collection, data analysis, or completion of final report of the clinical study. Institutional Review Board Statement The final protocol, case report form, and informed consent form were reviewed and approved by the institutional review board of the Antai Tian-Sheng Memorial Hospital (IRB No. 24-064-A, approved May 16, 2024). All participants provided written informed consent prior to the start of the study. This study was conducted in compliance with Declaration of Helsinki and in compliance with all International Conference on Harmonization Good Clinical Practice Guidelines (ICH-GCP). Informed Consent Statement All participants provided written informed consent prior to the start of the study. Data Availability All data generated or analyzed during this study are included in this manuscript. Acknowledgements We thank Dr. Meei-Yun Lin of Schweitzer Biotech Company for reading and providing feedback for preparation of the manuscript. Conflicts of Interest C.-C. W., C.-H. C., P.-L. K., W.-H. T., L. T.-C. L., C. C., and T.-Y. K. are employees of Schweitzer Biotech Company. T.-M. C. and S.-S. W. have no competing interests to declare. This study and the test product were funded and provided by Schweitzer Biotech Company. References Rajkumar J., Chandan N., Lio P., Shi V.. The skin barrier and moisturization: function, disruption, and mechanisms of repair. Skin Pharmacol Physiol . 2023 , 36 , 174-85. Franco A.C., Aveleira C., Cavadas C.. Skin senescence: mechanisms and impact on whole-body aging. Trends Mol Med . 2022 , 28 , 97-109. The beauty boom and beyond: Can the industry maintain its growth? Available online: https://www.mckinsey.com/industries/consumer-packaged-goods/our-insights/the-beauty-boom-and-beyond-can-the-industry-maintain-its-growth. Accessed 10 December, 2024. Liu JK. Natural products in cosmetics. Nat Prod Bioprospect . 2022 , 12 , 40. Biswas D., Mandal S., Chatterjee Saha S., Tudu C.K., Nandy S., Batiha G.E., Shekhaway M.S., Pandey D.K., Dey A. Ethnobotany, phytochemistry, pharmacology, and toxicity of Centella asiatica (L.) Urban: A comprehensive review. Phytother Res . 2021 , 35 , 6624–6654. Bandopadhyay S., Mandal S., Ghorai M., Jha N.K., Kumar M., Radha, Ghosh A., Proćków J., Pérez de la Lastra J.M., Dey A. Therapeutic properties and pharmacological activities of asiaticoside and madecassoside: A review. J Cell Mol Med . 2023 , 27 , 593-608. Sun B., Wu L., Wu Y., Zhang C., Qin L., Hayashi M., Kudo M., Gao M., Liu T. Therapeutic potential of Centella asiatica and its triterpenes: a review. Front Pharmacol 2020 , 11 , 568032. Diniz L.R., Calado L.L., Duarte A.B., de Sousa D.P. Centella asiatica and its metabolite asiatic acid: wound healing effects and therapeutic potential. Metabolites . 2023 , 13 , 276. Sephora. Search results for “Cica”. Available online: https://www.sephora.com/search?keyword=cica. Accessed December 09, 2024 CosmeKitchen. Search results for “Cica”. Available online: https://www.cosmekitchen-webstore.jp/Form/Product/ProductList.aspx?shop=0&swrd=cica&disp=pro&dpcnt=42&fpfl=0&img=1&sort=16&pno=1. Accessed December 09, 2024. Jeppesen D.K., Zhang Q., Franklin J.L., Coffey R.J. Extracellular vesicles and nanoparticles: emerging complexities. Trends Cell Biol . 2023 , 33 , 667-81. Cho J.H., Hong Y.D., Kim D., Park S.J., Kim J.S., Kim H.M., Yoon E.J., Cho J.S. Confirmation of plant-derived exosomes as bioactive substances for skin application through comparative analysis of keratinocyte transcriptome. Appl Bio Chem . 2022 , 65 , 8. Kim M.H., Yoon E.J., Kim J.S., Park S.J., Lee H. Comparative analysis of the transcriptome and efficacy of bioactive Centella asiatica exosomes on skin cells. ResearchSquare (Preprint). 2023 . https://doi.org/10.21203/rs.3.rs-2787704/v1 Li A., Li D., Gu Y., Liu R., Tang X., Zhao Y., Qi F, Wei J, Liu J. Plant-derived nanovesicles: Further exploration of biomedical function and application potential. Acta Pharm Sin . 2023 , 13 , 3300-20. Chang T.M., Wu C.C., Huang H.C., Wang S.S., Chuang C.H., Kao P.L., Tang W.H., Liu L.T.C., Qiu W.Y., Percec I., Chen C., Kuo T.Y. Centella asiatica Tissue Culture-Derived Extracellular Vesicles: A Multifaceted Approach to Skincare Applications. bioRxiv (Preprint). 2024 . 12.03.624435. doi: https://doi.org/10.1101/2024.12.03.624435 Jung E., Lee J.A., Shin S., Roh K.B., Kim J.H., Park D. Madecassoside inhibits melanin synthesis by blocking ultraviolet-induced inflammation. Molecules . 2013 , 18 , 15724-36. Zhu Y., Zhao J., Ding H., Qiu M., Xue L., Ge D., Wen G., Ren H., Li P., Wang J. Applications of plant‐derived extracellular vesicles in medicine. MedComm . 2024 , 5 , e741. Kim Y.J., Yoo S.M., Park H.H., Lim H.J., Kim Y.L., Lee S., Seo K.W., Kang K.S. Exosomes derived from human umbilical cord blood mesenchymal stem cells stimulates rejuvenation of human skin. Biochem Biophys Res Commun. 2017 , 493 , 1102-8. Additional Declarations The authors declare potential competing interests as follows: C.-C. W., C.-H. C., P.-L. K., W.-H. T., L. T.-C. L., C. C., and T.-Y. K. are employees of Schweitzer Biotech Company. T.-M. C. and S.-S. W. have no competing interests to declare. 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Taipei City,Taiwan","correspondingAuthor":false,"prefix":"","firstName":"Charles","middleName":"","lastName":"Chen","suffix":""},{"id":423161054,"identity":"1ae326c1-d053-4f62-864c-b9a0fc8ac7e5","order_by":11,"name":"Tsun-Yung Kuo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYBACCWYGBsaGChsgk/kAiC9DpJYzaQw8DGwJID4PYS1AzNjYcgiohccAJEBYi2Q77+GXMxsOMNhL5Hx+daPGgoeB/fDRDfi0SDPzpVlu3HGHgUcid5t1zjGgw3jS0m7g0yLHzGNm+PDMMwYe6dxtxjlsQC0SPGZEaGk7DNSS88w45x8RWqSZeYwfboRoYX6c20aEFslmHjPGGWfSeHjuPzNjzu2T4GEj5BeJ82eMP/ZU2Mix9xx+/DnnW50cP/vhY3i1AAEbKG54YAwGNgLKQYD5AzpjFIyCUTAKRgEKAABY6UNPxbHxNgAAAABJRU5ErkJggg==","orcid":"","institution":"Schweitzer Biotech Company, Taipei City,Taiwan","correspondingAuthor":true,"prefix":"","firstName":"Tsun-Yung","middleName":"","lastName":"Kuo","suffix":""}],"badges":[],"createdAt":"2025-03-03 06:53:37","currentVersionCode":1,"declarations":{"humanSubjects":true,"vertebrateSubjects":false,"conflictsOfInterestStatement":true,"humanSubjectEthicalGuidelines":true,"humanSubjectConsent":true,"humanSubjectClinicalTrial":true,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-6143379/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6143379/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":77766646,"identity":"0235445e-9809-4de5-b168-a2ab8b346394","added_by":"auto","created_at":"2025-03-05 10:10:37","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":41536,"visible":true,"origin":"","legend":"\u003cp\u003eChange in skin hydration at 0, 7, 14, 21, and 28 days after treatment of test product. Higher values indicate more hydrated skin. Results are shown as bars representing means with error bars representing 95% confidence interval of the mean. Statistical significances on days 7, 14, 21, and 28 were calculated by paired t-test compared to value on day 0 (baseline). * P\u0026lt;0.05, ** P\u0026lt;0.01, *** P\u0026lt;0.001, *** P\u0026lt;0.0001\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/114c1fc40a34f13e04d9d5cf.png"},{"id":77766660,"identity":"a1372e88-d396-4958-9d22-3a85ac5f7029","added_by":"auto","created_at":"2025-03-05 10:10:37","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":43453,"visible":true,"origin":"","legend":"\u003cp\u003eChange in skin melanin content at 0, 7, 14, 21, and 28 days after treatment of test product. Lower values indicate decreasing melanin content. Results are shown as bars representing means with error bars representing 95% confidence interval of the mean. Statistical significances on days 7, 14, 21, and 28 were calculated by paired t-test compared to value on day 0 (baseline). * P\u0026lt;0.05, ** P\u0026lt;0.01, *** P\u0026lt;0.001, *** P\u0026lt;0.0001’\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/f0a059e501c7fc3c71ecac05.png"},{"id":77767478,"identity":"6c9a60ea-b584-45f4-9931-95bda717bbbc","added_by":"auto","created_at":"2025-03-05 10:18:37","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":42639,"visible":true,"origin":"","legend":"\u003cp\u003eChange in skin elasticity at 0, 7, 14, 21, and 28 days after treatment of test product. Higher values indicate increasing skin elasticity. Results are shown as bars representing means with error bars representing 95% confidence interval of the mean. Statistical significances on days 7, 14, 21, and 28 were calculated by paired t-test compared to value on day 0 (baseline). * P\u0026lt;0.05, ** P\u0026lt;0.01, *** P\u0026lt;0.001, *** P\u0026lt;0.0001\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/6570c388a6e83498f050f26a.png"},{"id":77768964,"identity":"e4e1424a-b701-47bd-beb9-a16b92e5e9ad","added_by":"auto","created_at":"2025-03-05 10:26:37","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":48091,"visible":true,"origin":"","legend":"\u003cp\u003eChange in skin wrinkles at 0, 7, 14, 21, and 28 days after treatment of test product. Higher values indicate decreasing distribution and number of wrinkles. Results are shown as bars representing means with error bars representing 95% confidence interval of the mean. Statistical significances on days 7, 14, 21, and 28 were calculated by paired t-test compared to value on day 0 (baseline). * P\u0026lt;0.05, ** P\u0026lt;0.01, *** P\u0026lt;0.001, *** P\u0026lt;0.0001\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/839bdcbe66f43cff55d8d8a6.png"},{"id":77770298,"identity":"9d3d9ecf-a559-4b97-b2ba-0b1c8eedc4c7","added_by":"auto","created_at":"2025-03-05 10:42:37","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":46686,"visible":true,"origin":"","legend":"\u003cp\u003eChange in skin redness at 0, 7, 14, 21, and 28 days after treatment of test product. Higher values indicate decreasing zone of redness. Results are shown as bars representing means with error bars representing 95% confidence interval of the mean. Statistical significances on days 7, 14, 21, and 28 were calculated by paired t-test compared to value on day 0 (baseline). * P\u0026lt;0.05, ** P\u0026lt;0.01, *** P\u0026lt;0.001, *** P\u0026lt;0.0001\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/13ed9bc4202e2934470da832.png"},{"id":77767482,"identity":"a3754d93-3871-42ea-84e6-7449df045354","added_by":"auto","created_at":"2025-03-05 10:18:37","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":47344,"visible":true,"origin":"","legend":"\u003cp\u003eChange in skin pores at 0, 7, 14, 21, and 28 days after treatment of test product. Higher values indicate decreasing number and area of pores. Results are shown as bars representing means with error bars representing 95% confidence interval of the mean. Statistical significances on days 7, 14, 21, and 28 were calculated by paired t-test compared to value on day 0 (baseline). * P\u0026lt;0.05, ** P\u0026lt;0.01, *** P\u0026lt;0.001, *** P\u0026lt;0.0001\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/11d24f321f14cc3d34040a79.png"},{"id":77770300,"identity":"86e53e88-6380-454e-85b2-bac2c01f1735","added_by":"auto","created_at":"2025-03-05 10:42:42","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":896224,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/51bddccd-9dda-462a-8e24-99278263329e.pdf"},{"id":77766647,"identity":"9adc0568-c002-4a1e-9b91-21c161e6f077","added_by":"auto","created_at":"2025-03-05 10:10:37","extension":"zip","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":170826,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary files\u003c/p\u003e","description":"","filename":"Supplementaryfiles.zip","url":"https://assets-eu.researchsquare.com/files/rs-6143379/v1/b7ee67a01fe550fbbcb8b77e.zip"}],"financialInterests":"The authors declare potential competing interests as follows: C.-C. W., C.-H. C., P.-L. K., W.-H. T., L. T.-C. L., C. C., and T.-Y. K. are employees of Schweitzer Biotech Company. T.-M. C. and S.-S. W. have no competing interests to declare. ","formattedTitle":"\u003cp\u003eEvaluation of Effects on Skin Quality of a \u003cem\u003eCentella asiatica\u003c/em\u003e Extracellular Vesicle-based SkinCare Formulation: A 28-Day Facial Skin Quality Study\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAs the largest organ of the human body, skin is the main barrier of entry which protects the internal body from environmental damage and stress, maintains homeostasis, and provides key immune functions [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The skin consists of three main layers: the epidermis, which is the outermost protective layer composed of corneocytes, keratinocytes, melanocytes, and Langerhan\u0026rsquo;s cells; the dermis, which imparts the elasticity and structure of the skin by the extracellular matrix proteins such as collagen and elastin secreted by fibroblasts; and subcutaneous tissue, which is composed mostly of adipose tissue, containing adipocytes and stromal vascular cells [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Over time, skin undergoes aging due to intrinsic factors resulting in cell senescence characterized by reduced collagen production and decreased elasticity, as well as due to extrinsic factors like UV exposure, pollution, and oxidative stress [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo combat the effects of skin aging, modern skincare aims to restore skin hydration and elasticity, reduce pigmentation, and prevent or repair damage caused by aging and environmental factors. According to McKinsey, the global cosmetics market is a multi-billion industry which grew to \u003cspan\u003e$\u003c/span\u003e446\u0026nbsp;billion in 2023 and is estimated to reach \u003cspan\u003e$\u003c/span\u003e590\u0026nbsp;billion by 2028. It is characterized by a CAGR of 6%, with skincare representing the largest share of cosmetics, accounting for 44% of the market in 2023 [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. There has been renewed interest in natural products for cosmetics, both from consumers and industry due to potential risks associated with synthetic compounds, as well as the purported benefits of natural products [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMedicinal plants have been used in many cultures since ancient times and contain multifaceted therapeutic properties, which can be attributed to a multitude of secondary metabolite phytochemicals [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. \u003cem\u003eCentella asiatica\u003c/em\u003e, also known as Cica, is commonly used in traditional herbal medicine such as Chinese traditional medicine and Indian Ayurveda therapy. The plant contains a rich assortment of phytochemicals such as triterpenoids, flavonoids, and polyphenols that contribute to its anti-inflammatory, wound-healing, and central nervous system protective properties [\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Due to its broad spectrum benefits, commercial products containing Cica extracts are available for wound healing and skin care purposes [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. A cursory glance at major cosmetics retailers in the United States and Japan revealed a wide variety of Cica-based skin care products, representing a sizeable section of natural skin care market [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eExtracellular vesicles (EVs) are nanoscale lipid membrane-bound particles released by cells into the extracellular environment, carrying bioactive molecules that can modulate the physiological processes of target cells, even across disparate kingdoms of life [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Recent studies have shown that exosome-like EVs from plants, including Cica, promote skin improvement and wound healing more effectively than conventional plant extracts [\u003cspan additionalcitationids=\"CR13\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. We have previously characterized ExoBella\u0026reg;, a proprietary preparation of EVs isolated from tissue culture Cica, EVs and confirmed their potent \u003cem\u003ein-vitro\u003c/em\u003e antioxidant, anti-melanogenic, and anti-inflammatory properties [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Kristen Claire Supreme Rejuvenation Essence\u0026reg; is a skincare product formulated with ExoBella\u0026reg; Cica EVs as the main active ingredient. This clinical study aimed to evaluate skin improvement in volunteer users of Kristen Claire Supreme Rejuvenation Essence\u0026reg; over a period of 28 days.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eParticipants and Study Design\u003c/h2\u003e \u003cp\u003eParticipants were healthy adults of any gender from the age of 18 to 60 years (inclusive). The inclusion criteria included the absence of chronic diseases, major illnesses, or allergies, and not currently using any medications or other skincare products. The participants were required to conduct a pre-test screening to confirm the absence of skin irritation or allergic reaction to the test product which consisted of applying a skin patch infused with the test product for 24 hours on the inner forearms.\u003c/p\u003e \u003cp\u003eAfter screening, participants were asked to use the test product twice daily (morning and evening) after cleansing their face without using exfoliating products for 28 consecutive days. Each application involved 2 drops, evenly applied to the face using fingertips for absorption. Product usage was self-recorded, and the products were stored at room temperature.\u003c/p\u003e \u003cp\u003eThe skin quality tests were conducted at the investigation site Hungkuang University (Taichung City, Taiwan) onsite by the principal investigator. Each participant was to allocate 1 hour per test (including cleansing and waiting time). Skin quality tests were performed as a baseline test on day 0 (before using the test product), and subsequent tests at 7, 14, 21, and 28 days after test product use.\u003c/p\u003e \u003cp\u003eMeasurements of the skin quality tests included skin hydration, melanin content, and elasticity index using a C\u0026thinsp;+\u0026thinsp;K Multi Probe Adaptor MPA580 skin analysis device (Courage-Khazaka Electronic GmbH, Koln, Germany) equipped with probes for hydration, pigmentation, and elasticity measurements. Probes were placed 1 cm horizontally and 1 cm vertically from the outer corner of each eye (on the left and right cheeks) and 2 cm above the midpoint of the eyebrows (forehead). Additionally, the VISIA\u0026reg; Skin Analysis system (Canfield Imaging Systems, Fairfield, NJ, USA) was used to assess changes in wrinkle percentage, redness area percentage, and pore percentage by scanning full-face frontal and 45\u0026deg; angles on both sides of the face.\u003c/p\u003e \u003cp\u003eThe tests were conducted in a controlled environment at 20\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C and 50\u0026thinsp;\u0026plusmn;\u0026thinsp;5% relative humidity, without direct sunlight or air conditioning airflow. All measurements were non-invasive. Probes contact the skin's surface to assess parameters based on electrical resistance, optical principles, and physical changes. Participants\u0026rsquo; test results and photographic data were collected.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eDescription of Test Product\u003c/h3\u003e\n\u003cp\u003eThe test product (Kristen Claire Supreme Rejuvenation Essence\u0026reg;) is a proprietary water-based formulation containing the main active ingredient ExoBella\u0026reg;, a preparation of EVs derived from tissue culture \u003cem\u003eC. asiatica\u003c/em\u003e. Other active ingredients include \u003cem\u003eRubus fruiticosus\u003c/em\u003e leaf extract, \u003cem\u003eGlycyrrhiza glabrai\u003c/em\u003e root extract, \u003cem\u003eCalendula officinalis\u003c/em\u003e flower extract, \u003cem\u003eAloe barbadensis\u003c/em\u003e leaf juice, \u003cem\u003eVaccinium angustifolium\u003c/em\u003e fruit extract, \u003cem\u003eMorus nigra\u003c/em\u003e fruit extract, arbutin, hydrolyzed DNA, niacinamide, and tocopheryl acetate (Vitamin E). The appearance is clear with slightly gel-like in consistency.\u003c/p\u003e\n\u003ch3\u003eAssessment of Outcomes\u003c/h3\u003e\n\u003cp\u003eSkin hydration was measured using the Corneometer\u0026reg; CM825 (Courage-Khazaka Electronic GmbH, Koln, Germany), which evaluates the dielectric constant related to epidermal moisture at a depth of 60\u0026ndash;100 \u0026micro;m. Mexameter\u0026reg; MX18 (Courage-Khazaka Electronic GmbH, Koln, Germany) was used to measure melanin content, which utilizes RGB light absorption (568 nm, 660 nm, and 880 nm) to measure melanin and hemoglobin content. Elasticity index was measured using the Cutometer\u0026reg; Dual MPA580 system (Courage-Khazaka Electronic GmbH, Koln, Germany), based on suction and stretching properties of the skin.\u003c/p\u003e \u003cp\u003eVISIA\u0026reg; Skin Analysis System (Canfield Imaging Systems, Fairfield, NJ, USA) was used to evaluate the following parameters:\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eWrinkle percentage was measured by using standard white light to detect shadow variations to determine the distribution and number of wrinkles. Fewer wrinkles yield higher percentages.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eRedness percentage was measured by RBX polarized light to identify vascular or inflammatory issues. Smaller red zones result in higher percentages.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003ePore percentage was measured under white light to analyze shadowed pore depressions and darker areas relative to surrounding skin. Fewer pores yield higher percentages.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e\n\u003ch3\u003eData Presentation and Statistical Analysis\u003c/h3\u003e\n\u003cp\u003eFor each of the measured parameters, results are converted to skin improvement rate (%) calculated as follows:\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:\\frac{\\left(Post‐treatment\\:index\\:-\\:Pre‐treatment\\:index\\right)}{Pre‐treatment\\:index}\\:\\times\\:100\\%$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eWhere the pre-treatment index is the score of the parameter examined on Day 0, and the post-treatment index is the score of the parameter examined on Day 7, 14, 21, or 28. In skin hydration and elasticity, a positive skin improvement rate indicates higher hydration and high elasticity, respectively, when compared to Day 0. In skin melanin content, the skin improvement rate is negative because the parameter measures the decrease in melanin content and the lower skin improvement rate indicates greater decrease in melanin. In parameters measured using the VISIA\u0026reg; Skin Analysis System, a higher skin improvement rate indicates decreased wrinkles, redness, or pores.\u003c/p\u003e \u003cp\u003eResults are presented as mean of the skin improvement rate with 95% confidence intervals shown as error bars. The analysis package in GraphPad Prism 6.01 (Boston, MA) was used for statistical analysis. A paired Student\u0026rsquo;s t-test was used to compare the difference between means of skin improvement rate on Days 7, 14, 21, or 28 against Day 0 (baseline).\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003eParticipants\u003c/h2\u003e\n \u003cp\u003eFrom September 4, 2024, to October 4, 2024, 20 healthy participants (4 males and 16 females; average age 36.5) were enrolled and all were assessed negative for irritation or allergic reactions to the test product prior to the study. The participants ranged from the age of 23 to 54 years, and the mean age was 36.5 with a standard deviation of 9.2.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eAssessments of Participants\u003c/h3\u003e\n\u003cp\u003eThe participants were assessed for changes in skin parameters, including skin hydration, melanin content, skin elasticity, wrinkle, redness (erythema), and skin pores. The raw data values of the assessment are presented in Tables S1 to S6. The evaluation was performed on three facial regions (left cheek, right cheek, and forehead) every 7 days over a 28-day period. In skin hydration, statistical significant increases in hydration percentage were observed across all facial regions (left cheek, right cheek, and forehead) over the course of 28 days compared to baseline, with the right cheek having slightly less improvement compared to other regions (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Skin hydration improved by 3.1\u0026ndash;5.1%, 5.8\u0026ndash;10.6%, 9.6\u0026ndash;15.2%, and 14.6\u0026ndash;21.2% on Days 7, 14, 21, and 28, respectively.\u003c/p\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003cp\u003eThe treatment also resulted in significantly lowered melanin content throughout the study period with melanin decreasing by 1.2\u0026ndash;2.3%, 4.1\u0026ndash;5.0%, 6.2\u0026ndash;7.8%, and 9.2\u0026ndash;11.3%, on Days 7, 14, 21, and 28, respectively (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003cp\u003eSimilarly, treatment resulted in significant improvement in skin elasticity over time uniformly across all regions tested (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). The improvement in skin elasticity was 2.2\u0026ndash;3.5%, 4.6\u0026ndash;5.9%, 7.4\u0026ndash;8.9%, and 12.0\u0026ndash;12.5% on Days 7, 14, 21, and 28, respectively.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003cp\u003eThe VISIA\u0026reg; Skin Analysis System was used to image and analyze skin wrinkles, redness, and pore distribution simultaneously. Skin wrinkle condition improved by 8.6\u0026ndash;16.9%, 19.3\u0026ndash;24.9%, 28.2\u0026ndash;30.0%, and 32.9\u0026ndash;34.8% on Days 7, 14, 21, and 28, respectively (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). The forehead region lagged behind in wrinkle improvement at an earlier stage during the study but increased to approximately equal to that of other facial regions by Day 21 (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003cp\u003eSkin redness was ameliorated during the study period by 5.6\u0026ndash;7.9%, 13.1\u0026ndash;14.6%, 19.9\u0026ndash;22.7%, and 26.3\u0026ndash;34.0% on Days 7, 14, 21, and 28, respectively (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003cp\u003eFinally, skin pores reduction by treatment resulted in skin improvement by 9.3\u0026ndash;12.4%, 19.8\u0026ndash;21.8%, 28.5\u0026ndash;32.1%, and 40.6\u0026ndash;41.3% on Days 7, 14, 21, and 28, respectively (Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e). In summary, the treatment had a significant improvement in skin hydration and elasticity, while reducing the distribution and area of melanin, wrinkles, redness, and pores.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003cp\u003eTo investigate the effect of treatment in different age groups, the data was stratified according to the age of participants into two groups: \u0026gt; 40 years and \u0026lt;\u0026thinsp;40 years. The means of skin improvement rate of each parameter were compared between the two age groups at the three facial regions. We found no significant differences in skin hydration and elasticity between the two age groups (Figure S1). However, those aged above 40 years had significant less reduction in melanin in both cheek regions compared to participants aged less than 40 years on Day 28 of the study, while there were no significant differences between forehead region in both age groups (Figure S1). For parameters analyzed with VISIA\u0026reg; System, there were no significant differences between the two age groups in changes in skin wrinkles and pores (Figure S2). In redness skin area, the overall improvement in \u0026gt;\u0026thinsp;40 years age group was less than that of \u0026lt;\u0026thinsp;40 years age group, and the difference between two age groups was significant on Day 28 at the forehead region (Figure S2).\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this small-scale clinical study, we observed positive improvements on the skin in participants treated with Cica EV-based test product. Skin integrity parameters such as hydration and elasticity increased after treatment with the test product. According to our previous study, Cica EVs could achieve improvement in skin hydration via upregulation of water retention genes such as aquaporin and filaggrin, and higher skin elasticity by increasing production of collagen and elastin [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. The triterpene madecassoside in Cica has already been studied for its anti-melanogenic effect via anti-inflammatory effects [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Besides madecassoside, miRNAs within Cica EVs can also target melanogenic genes such as tyrosinase and other pathway inducers (WNT, CRTC1), as has been observed in our previous study and others [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCica extracts have already been used in commercial skin care products due to their skin healing, antioxidant, and anti-melanogenic properties imparted by their bioactive compounds [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The mechanisms through which exosomes promote skin regeneration include angiogenesis, collagen synthesis, and regulation of inflammation [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. A study discovered 11 novel miRNAs contained within Cica exosomes, which were predicated to inhibit genes involved in melanin production and dermatitis [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The study also found that Cica exosomes induced higher gene expression than \u003cem\u003eC. asiatica\u003c/em\u003e extracts, possibly due to better penetration of skin cells by exosomes and the presence of packaged miRNAs [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. In formulation of the test product, ExoBella\u0026reg; Cica EVs may work in concert with other herbal extracts and small molecule compounds (arbutin, niacinamide, vitamin E) to achieve the results seen in this study.\u003c/p\u003e \u003cp\u003eThe main limitation of the study is the small sample size, which may not include sufficient diversity of demographics such as different skin types and ethnicities due to the homogeneity of population in Taiwan. The small sample size also made it unfeasible to conduct a more robust analysis of differences between age groups, and stratification into smaller age groups (e.g. 16\u0026thinsp;~\u0026thinsp;25 years, 26\u0026thinsp;~\u0026thinsp;35 years, etc.). The sample population is also heavily gender-skewed toward female (80% of participants), which can also influence the generalizability of the results. The other limitation was the relatively short duration of the study (28 days). This may not be sufficient to capture the long-term effects of the test product as skin quality can be influenced by numerous factors over time including aging, UV exposure, and other environmental factors. A longer study period and follow-up would allow a more comprehensive understanding of the product's sustained efficacy and potential adverse effects. Lastly, as this was not a placebo-controlled nor double-blinded study, it would be difficult to determine whether the improvements observed were attributed to the use of test product, natural improvement of skin over time, or due to other external factors.\u003c/p\u003e \u003cp\u003eIn conclusion, the results of this study support the positive effects on skin quality of Kristen Claire Supreme Rejuvenation Essence\u0026reg; in improving skin hydration and elasticity while reducing melanin, wrinkles, redness, and pores. The differences observed between age groups and the three facial regions were minimal. Future studies should address the above mentioned limitations to provide clearer insights into the long-term benefits on skin improvement and application of the test product across more diverse set of population to address a global market.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization, supervision and project administration: T.-M.C., C.C., T.-Y.K.\u003c/p\u003e\n\u003cp\u003eVisualization and formal analysis: T.-M.C., L.T.-C.L.\u003c/p\u003e\n\u003cp\u003eInvestigation, data acquisition and curation: T.-M.C., H.-C.H., S.-S.W.\u003c/p\u003e\n\u003cp\u003eMethodology: T.-M.C., C.-C.W., H.-C.H., S.-S.W., C.-H.C., P.-L.K., W.-H.T., W.-Y.Q. I.P., T.-Y.K.\u003c/p\u003e\n\u003cp\u003eResources: C.-C.W., C.-H.C., P.-L.K., W.-H.T., T.-Y.K.\u003c/p\u003e\n\u003cp\u003eWriting\u0026mdash;original draft preparation: T.-.M.C., L.T.-C.L.\u003c/p\u003e\n\u003cp\u003eWriting\u0026mdash;review and editing: T.-M.C., L.T.-C.L., W.-Y.Q., I.P., C.C., T.-Y.K.\u003c/p\u003e\n\u003cp\u003eAll authors reviewed the manuscript\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eSchweitzer Biotech Company provided the necessary funding for the study as well as provided test product Kristen Claire Supreme Rejuvenation Essence\u0026reg;, but had otherwise no role in enrollment, participation, data collection, data analysis, or completion of final report of the clinical study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstitutional Review Board Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe final protocol, case report form, and informed consent form were reviewed and approved by the institutional review board of the Antai Tian-Sheng Memorial Hospital (IRB No. 24-064-A, approved May 16, 2024). All participants provided written informed consent prior to the start of the study. This study was conducted in compliance with Declaration of Helsinki and in compliance with all International Conference on Harmonization Good Clinical Practice Guidelines (ICH-GCP).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll participants provided written informed consent prior to the start of the study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eAll data generated or analyzed during this study are included in this manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eWe thank Dr. Meei-Yun Lin of Schweitzer Biotech Company for reading and providing feedback for preparation of the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e C.-C. W., C.-H. C., P.-L. K., W.-H. T., L. T.-C. L., C. C., and T.-Y. K. are employees of Schweitzer Biotech Company. T.-M. C. and S.-S. W. have no competing interests to declare. This study and the test product were funded and provided by Schweitzer Biotech Company.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eRajkumar J., Chandan N., Lio P., Shi V.. The skin barrier and moisturization: function, disruption, and mechanisms of repair. \u003cem\u003eSkin Pharmacol Physiol\u003c/em\u003e. \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e36\u003c/em\u003e, 174-85. \u003c/li\u003e\n\u003cli\u003eFranco A.C., Aveleira C., Cavadas C.. Skin senescence: mechanisms and impact on whole-body aging. \u003cem\u003eTrends Mol Med\u003c/em\u003e. \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e28\u003c/em\u003e, 97-109. \u003c/li\u003e\n\u003cli\u003eThe beauty boom and beyond: Can the industry maintain its growth? Available online: https://www.mckinsey.com/industries/consumer-packaged-goods/our-insights/the-beauty-boom-and-beyond-can-the-industry-maintain-its-growth. Accessed 10 December, 2024.\u003c/li\u003e\n\u003cli\u003eLiu JK. Natural products in cosmetics. \u003cem\u003eNat Prod Bioprospect\u003c/em\u003e. \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e12\u003c/em\u003e, 40. \u003c/li\u003e\n\u003cli\u003eBiswas D., Mandal S., Chatterjee Saha S., Tudu C.K., Nandy S., Batiha G.E., Shekhaway M.S., Pandey D.K., Dey A. Ethnobotany, phytochemistry, pharmacology, and toxicity of \u003cem\u003eCentella asiatica\u003c/em\u003e (L.) Urban: A comprehensive review. \u003cem\u003ePhytother Res\u003c/em\u003e. \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e35\u003c/em\u003e, 6624\u0026ndash;6654. \u003c/li\u003e\n\u003cli\u003eBandopadhyay S., Mandal S., Ghorai M., Jha N.K., Kumar M., Radha, Ghosh A., Proćk\u0026oacute;w J., P\u0026eacute;rez de la Lastra J.M., Dey A. Therapeutic properties and pharmacological activities of asiaticoside and madecassoside: A review. \u003cem\u003eJ Cell Mol Med\u003c/em\u003e. \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e27\u003c/em\u003e, 593-608. \u003c/li\u003e\n\u003cli\u003eSun B., Wu L., Wu Y., Zhang C., Qin L., Hayashi M., Kudo M., Gao M., Liu T. Therapeutic potential of \u003cem\u003eCentella asiatica\u003c/em\u003e and its triterpenes: a review. \u003cem\u003eFront Pharmacol \u003c/em\u003e\u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e11\u003c/em\u003e, 568032. \u003c/li\u003e\n\u003cli\u003eDiniz L.R., Calado L.L., Duarte A.B., de Sousa D.P. \u003cem\u003eCentella asiatica\u003c/em\u003e and its metabolite asiatic acid: wound healing effects and therapeutic potential. \u003cem\u003eMetabolites\u003c/em\u003e. \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e13\u003c/em\u003e, 276. \u003c/li\u003e\n\u003cli\u003eSephora. Search results for \u0026ldquo;Cica\u0026rdquo;. Available online: https://www.sephora.com/search?keyword=cica. Accessed December 09, 2024\u003c/li\u003e\n\u003cli\u003eCosmeKitchen. Search results for \u0026ldquo;Cica\u0026rdquo;. Available online: https://www.cosmekitchen-webstore.jp/Form/Product/ProductList.aspx?shop=0\u0026amp;swrd=cica\u0026amp;disp=pro\u0026amp;dpcnt=42\u0026amp;fpfl=0\u0026amp;img=1\u0026amp;sort=16\u0026amp;pno=1. Accessed December 09, 2024.\u003c/li\u003e\n\u003cli\u003eJeppesen D.K., Zhang Q., Franklin J.L., Coffey R.J. Extracellular vesicles and nanoparticles: emerging complexities. \u003cem\u003eTrends Cell Biol\u003c/em\u003e. \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e33\u003c/em\u003e, 667-81.\u003c/li\u003e\n\u003cli\u003eCho J.H., Hong Y.D., Kim D., Park S.J., Kim J.S., Kim H.M., Yoon E.J., Cho J.S. Confirmation of plant-derived exosomes as bioactive substances for skin application through comparative analysis of keratinocyte transcriptome. \u003cem\u003eAppl Bio Chem\u003c/em\u003e. \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e65\u003c/em\u003e, 8. \u003c/li\u003e\n\u003cli\u003eKim M.H., Yoon E.J., Kim J.S., Park S.J., Lee H. Comparative analysis of the transcriptome and efficacy of bioactive \u003cem\u003eCentella asiatica\u003c/em\u003e exosomes on skin cells. \u003cem\u003eResearchSquare \u003c/em\u003e(Preprint). \u003cstrong\u003e2023\u003c/strong\u003e. https://doi.org/10.21203/rs.3.rs-2787704/v1\u003c/li\u003e\n\u003cli\u003eLi A., Li D., Gu Y., Liu R., Tang X., Zhao Y., Qi F, Wei J, Liu J. Plant-derived nanovesicles: Further exploration of biomedical function and application potential. \u003cem\u003eActa Pharm Sin\u003c/em\u003e. \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e13\u003c/em\u003e, 3300-20. \u003c/li\u003e\n\u003cli\u003eChang T.M., Wu C.C., Huang H.C., Wang S.S., Chuang C.H., Kao P.L., Tang W.H., Liu L.T.C., Qiu W.Y., Percec I., Chen C., Kuo T.Y. \u003cem\u003eCentella asiatica\u003c/em\u003e Tissue Culture-Derived Extracellular Vesicles: A Multifaceted Approach to Skincare Applications. bioRxiv (Preprint). \u003cstrong\u003e2024\u003c/strong\u003e. 12.03.624435. doi: https://doi.org/10.1101/2024.12.03.624435\u003c/li\u003e\n\u003cli\u003eJung E., Lee J.A., Shin S., Roh K.B., Kim J.H., Park D. Madecassoside inhibits melanin synthesis by blocking ultraviolet-induced inflammation. \u003cem\u003eMolecules\u003c/em\u003e. \u003cstrong\u003e2013\u003c/strong\u003e, \u003cem\u003e18\u003c/em\u003e, 15724-36. \u003c/li\u003e\n\u003cli\u003eZhu Y., Zhao J., Ding H., Qiu M., Xue L., Ge D., Wen G., Ren H., Li P., Wang J. Applications of plant‐derived extracellular vesicles in medicine. \u003cem\u003eMedComm\u003c/em\u003e. \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e5\u003c/em\u003e, e741. \u003c/li\u003e\n\u003cli\u003eKim Y.J., Yoo S.M., Park H.H., Lim H.J., Kim Y.L., Lee S., Seo K.W., Kang K.S. Exosomes derived from human umbilical cord blood mesenchymal stem cells stimulates rejuvenation of human skin. \u003cem\u003eBiochem Biophys Res \u003c/em\u003eCommun. \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e493\u003c/em\u003e, 1102-8. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Hungkuang University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Centella asiatica, Cica, skincare, cosmetics, longevity, extracellular vesicles, skin quality, skin hydration, skin whitening, skin elasticity, wrinkle reduction","lastPublishedDoi":"10.21203/rs.3.rs-6143379/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6143379/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSkin damage results from the natural aging process, physical wear and tear, UV light, and environmental pollutants. \u003cem\u003eCentella asiatica\u003c/em\u003e (Cica) is a widely used traditional medicinal herb and is known to have therapeutic effects on skin and wound healing. This study aims to evaluate the effects of Kristen Claire Supreme Rejuvenation Essence\u0026reg; with Cica EV as the main active ingredient, on facial appearance and skin quality in healthy participants. Twenty healthy participants (4 males and 16 females; average age 36.5) were enrolled and underwent a 24-hour skin patch test on the forearm to assess potential irritation or allergic reactions. The test product was applied twice daily for 28 days. Facial skin quality assessments were conducted before use and at 7, 14, 21, and 28 days of product application. Measured parameters included skin hydration, melanin content, skin elasticity, wrinkle percentage, redness area percentage, and pore percentage. After 28 days of test product treatment, significant improvements were observed in skin hydration and elasticity, wrinkle, redness, and pore distribution, while skin melanin content was reduced significantly. Kristen Claire Supreme Rejuvenation Essence\u0026reg; effectively improved skin hydration, elasticity, and texture, brightened the complexion, and showed potential in slowing skin aging, making it a promising candidate for skincare applications.\u003c/p\u003e","manuscriptTitle":"Evaluation of Effects on Skin Quality of a Centella asiatica Extracellular Vesicle-based SkinCare Formulation: A 28-Day Facial Skin Quality Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-05 10:10:32","doi":"10.21203/rs.3.rs-6143379/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"799bdade-7d70-4ed1-b7f3-6b963a16bfb9","owner":[],"postedDate":"March 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45096537,"name":"Dermatology"}],"tags":[],"updatedAt":"2025-03-05T10:10:32+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-05 10:10:32","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6143379","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6143379","identity":"rs-6143379","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}