Skin care
Skin care
peer-reviewed
Next-generation resveratrol from plant-derived exosomes: exosomic resveratrol and biopeptides as a novel strategy for skin longevity
ÒSCAR EXPÓSITO
CEO, CSO and co-founder, Vytrus Biotech S.A.
ABSTRACT:Inspired by intermittent fasting, a Vitis vinifera derived secretome was developed under nutrient-deprivation conditions to emulate cellular hormesis for skin longevity. These conditions yield a metabolically enriched secretome containing bioactive compounds known as Plant Fasting Factors, resveratrol, signaling peptides, and polyamines, encapsulated within plant-derived exosomes, suitable for targeted skin delivery. Studies showed the secretome significantly enhanced autophagy (e.g., a 138% increase in LC3 fluorescence in aged fibroblasts) and reduced lipofuscin and melanin accumulations in vitro. Clinical trials revealed a +38.3% increase in skin antioxidant potential in Caucasian volunteers and a –5.2% reduction in global skin pigmentation in Asian volunteers. This groundbreaking plant-based solution offers a potent, next-generation cosmetic bioactive for healthy aging skin.
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“A study in healthy women providing probiotic yogurt for four weeks showed an improvement in emotional responses as measured by brain scans”
Figure 1. Skin Section with Microbiome. Most microorganisms live in the superficial layers of the stratum corneum and in the upper parts of the hair follicles. Some reside in the deeper areas of the hair follicles and are beyond the reach of ordinary disinfection procedures. There bacteria are a reservoir for recolonization after the surface bacteria are removed.
Materials and methods
Studies of major depressive disorder have been correlated with reduced Lactobacillus and Bifidobacteria and symptom severity has been correlated to changes in Firmicutes, Actinobacteria, and Bacteriodes. Gut microbiota that contain more butyrate producers have been correlated with improved quality of life (1).
A study in healthy women providing probiotic yogurt for four weeks showed an improvement in emotional responses as measured by brain scans (2). A subsequent study by Mohammadi et al. (3) investigated the impacts of probiotic yogurt and probiotic capsules over 6 weeks and found a significant improvement in depression-anxiety-stress scores in subjects taking the specific strains of probiotics contained in the yogurt or capsules. Other studies with probiotics have indicated improvements in depression scores, anxiety, postpartum depression and mood rating in an elderly population (4-7).
Other studies have indicated a benefit of probiotic supplementation in alleviating symptoms of stress. In particular, researchers have looked at stress in students as they prepared for exams, while also evaluating other health indicators such as flu and cold symptoms (1). In healthy people, there is an indication that probiotic supplementation may help to maintain memory function under conditions of acute stress.
Introduction
Unveiling the benefits of intermittent fasting at a cellular level
Intermittent fasting has emerged as a promising strategy to enhance health and longevity by modulating key metabolic pathways (1,2). Nutrient scarcity activates adaptive mechanisms and induces at a cellular level:
- Hormesis, stimulating stress response pathways and DNA repair, including via autophagy stimulation (LC3II) telomere protection (shelterin complex stabilization), and modulation of the IGF-1 signaling pathway, all of which support cellular longevity and homeostasis (3, 4).
- Antioxidant defenses, including increased expression of catalase, peroxiredoxins, and superoxide dismutase, with protective effects on telomere integrity (5),
- Autophagy, facilitating the recycling of damaged cellular components and maintenance of homeostasis (6, 7, 8),
- Prevention of senescence, by reducing pigment accumulation (lipofuscin, melanin) and oxidative burden (9, 10).
Fasting on Plants and Hormesis
Like humans, plants exhibit adaptive responses under nutrient-deprived conditions. Due to their sessile nature, they have evolved complex pathways, such as the optimization of enzymatic processes and the production of antioxidant secondary metabolites, especially phenolic metabolites, so that energy metabolism is optimized, mutation accumulations are reduced, and plant longevity is prolonged (11, 12, 13, 14, 15, 16).
Drawing from cellular responses to nutrient restriction mechanisms and inherent stress resilience of perennial plants, Vytrus Biotech has developed a novel cosmetic active derived from Vitis vinifera stem cells—an emblematic species known for its exceptional longevity, with vines documented to live over 400 years (25). Utilizing its proprietary Phyto-Peptidic Fractions (PPF) platform, Vytrus cultured these cells under controlled nutrient-deprivation to mimic a fasting state, thereby inducing a hormetic response. This process yields a metabolically enriched secretome containing signaling peptides, antioxidants such as polyphenols, and bioactive-loaded plant-derived exosomes (17, 18).
A New Type of Resveratrol: Exosomic Resveratrol
Emerging research confirms that Vitis vinifera can release extracellular vesicles loaded with polyphenols, lipids, and plant microRNAs, including those with structural homology to human inflammation-regulating miRNAs (19). Among the vesicular contents, resveratrol is of particular interest for its dual role as both an antioxidant and a signaling molecule (20, 21).
When encapsulated within exosomes under fasting-mimicking conditions, exosomic resveratrol may influence caloric restriction–like pathways in skin cells, as suggested by literatura on including SIRT1 activation, mitochondrial enhancement, and inflammatory modulation—mimicking the systemic benefits of fasting at a cellular level (22, 23, 24).
In this study, we examine the molecular profile and skin-related effects of this Vitis vinifera fasting-induced secretome, demonstrating that exosomic resveratrol functions as a potent, next-generation cosmetic bioactive with both preventive and reparative capacities.
Experimental
Vitis vinifera cell cultures
To replicate the effects of intermittent fasting at the cellular level, a Vitis vinifera cell cultures two-phase cultivation protocol was developed. In the non-fasting protocol, cells were cultured for 8 days under nutrient-sufficient conditions. In the fasting-mimicking protocol, the culture period was extended by an additional 9 days following carbon source depletion, imposing a controlled caloric restriction.
Extracellular vesicles isolation
After mixing the fruit extracts, samples were centrifuged at 500× g for 10 min; the supernatants were filtered with 100 µm filters and serially centrifugated at 2000× g for 20 min to eliminate cell debris and then at 15,000× g for 30 min to collect the fraction enriched in microvesicles (25).
Exosomes visualization
The isolated PDEVs were resuspended in PBS and, after a proper dilution, placed on 200-mesh carbon-coated copper grids. TEM images were captured using a Philips CM 100 Electron Microscope (26).
Exosomes characterization
Size distribution and concentration of extracellular vesicles in liquid suspension were analyzed through Nanoparticle Tracking Analysis (NTA) from Malvern (NanoSight NS300, Salisbury, UK). While proprietary constraints limit full disclosure, internal validation included particle size (NTA), TEM morphology, and surface marker consistency with known EV profiles.
Peptidomics analysis
The sample preparation and Instrumental analysis were performed by Creative Proteomics services (Creative Proteomics, NY, USA).
Sample preparation
CA and s secretomes were diluted in 0.1% TFA and loaded onto the ziptip column twice, followed by desalting with 0.1% TFA and elution with 60% acetonitrile. The eluates were combined, lyophilized to near dryness, and resuspended in 20 μ L of 0.1% formic acid for LC-MS/MS analysis.
Instrumental analysis
The nano LC-MS/MS analysis utilized an Ultimate 3000 nano UHPLC system coupled with an Orbitrap Q Exactive HF mass spectrometer (Thermo Scientific, Waltham, MA, USA). Peptides were separated on a PepMap C18 column and analyzed using a linear gradient of mobile phase A (0.1% formic acid in water) and mobile phase B (0.1% formic acid in acetonitrile) with a flow rate of 250 nL / min.
Peptidome characterization
Raw LC-MS / MS files were analyzed based on the species of the samples using PEAKS STUDIO 8.5 (Bioinformatics Solutions Inc., Ontario, Canada). MS data was used to decipher the sequence based on fragmentation patterns and mass differences between ions.
Metabolomic analysis
The polyphenol content of exosomes was also analyzed to assess their potential bioactive properties. The methodology employed included ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UPLC/MS/MS - ESP/EScl) using a photodiode array (PDA) detector and a triple quadrupole analyzer.
In vitrostudies
Evaluation of fasting-mimicking effect and cell longevity phases
To emulate fasting conditions at the cellular level, a two-phase cultivation protocol was implemented for Vitis vinifera stem cells. In the first phase, cells were cultured under nutrient-sufficient conditions for eight days, during which exponential growth was promoted and basal metabolic activity established. Throughout this phase, daily monitoring included sugar consumption assays, pH stability checks, and visual morphological assessment to ensure optimal culture health. Upon reaching the nutrient plateau, the culture transitioned to the second phase—a nine-day nutrient-restriction period, characterized by the depletion of carbon sources to trigger a fasting-mimicking stress response. During this phase, secondary metabolite biosynthesis was stimulated and tracked through metabolomic profiling and optical density measurements, allowing the identification of hormesis-associated shifts in the secretome. This metabolic conditioning is fundamental for the enrichment of bioactive compounds such as resveratrol, peptides, and polyamines that comprise the core efficacy profile of Vitis vinifera secretome.
Evaluation of longevity markers in Vitis vinifera secretome
A novel in vitro model was established to investigate the effects of the Vitis vinifera fasting-induced secretome on cellular fasting pathways and longevity-associated biomarkers in human skin cells. Human dermal fibroblasts (HDFs) were cultured for two months across eight passages (P2–P8) under four experimental conditions: untreated control, metformin as a positive control, and the secretome at 0.65% and 2.5% concentrations. These doses were selected based on internal screening for efficacy and cytocompatibility in HDF cultures.
Enhancement of autophagy processes
Autophagy activation in human dermal fibroblasts was assessed by quantifying LC3 expression through both immunofluorescence. For immunostaining, cells were fixed, permeabilized, and incubated with anti-LC3 and FITC-conjugated secondary antibodies, followed by Hoechst nuclear staining.
Melanin and lipofuscin reduction
Melanin levels were quantified in dark melanocytes exposed to 2.5 and 0.65 % vol/vol of Vitis vinifera or the positive control (Metformin at 100 µM) or kojic acid (500µM) for 7 days. Lipofuscin levels were quantified in the human dermal fibroblasts exposed to 2.5 and 0.65 % vol/vol of Vitis vinifera secretome or the positive control (Metformin at 100 µM) for the indicated times (days 28 and 56) by Sudan black staining.
In vivostudies
- Fasting-mimicking effect potential of the secretome of Vitis vinifera
Tested on Caucasian volunteer panel: Skin antioxidant potential
A clinical study was conducted on 66 Caucasian volunteers (aged 40–65), divided into three groups (n=22 each): non-fasting placebo, fasting placebo (12–14 h), and Vitis vinifera secretome (1%, applied twice daily without fasting). Assessments were performed at baseline, day 28, and day 56 using non-invasive techniques: PrimosCR SF (profilometry), Mexameter (pigmentation), and Corneofix® skin stripping (stratum corneum analysis).
- Radiance and depigmentation benefits of the secretome of Vitis vinifera
Tested on Asian volunteer panel: Skin global pigmentation
A second in vivo study was conducted on 22 Asian volunteers (aged 40–65) to assess the efficacy of Vitis vinifera secretome across diverse skin types. A 1% Vitis vinifera secretome formulation was applied twice daily for 56 days using a hemifacial design (Vitis vinifera secretome vs. placebo). Assessments were performed at baseline, day 28, and day 56 using CM-700D spectrophotometry, Mexameter (melanin index), and Primos (skin profilometry).
Results
A step forward in exosomics: analysis of the fasting-mimicking exosomes
The Vitis vinifera secretome exhibited an average exosome concentration of 1.47 × 10⁹ particles/mL, with a mean vesicle diameter of 124.2 nm. Following isolation, comprehensive peptidomic and metabolomic analyses were performed to characterize the bioactive cargo—referred to as Plant Fasting Factors—naturally selected and encapsulated by the cells.
Identification of peptides in the fasting-mimicking exosomes of Vitis vinifera secretome
These peptides ranged from 11 to 26 amino acids—predominantly between 14 and 20 residues—were identified within the Vitis vinifera exosomes. The distribution of peptide lengths is non-random, with a significant prevalence of 14, 17, and 20 amino acid sequences, suggesting a selective cellular mechanism for exosomal loading. Comparison with Vytrus' proprietary PEPVYT database revealed no significant matches, underscoring the novelty of these peptides in the cosmetic landscape.
Identification of metabolites in the exosomes of Vitis vinifera secretome
The characterization of secondary metabolites within Vitis vinifera exosomes revealed a targeted selective molecular cargo, despite the inherently low metabolite concentrations. Through advanced analytical methods, Vytrus identified four polyphenols, with resveratrol and isorhamnetin being the most abundant. These compounds are not only known for their antioxidant activity but also for their capacity to activate cellular pathways involved in skin longevity. Specifically, resveratrol demonstrated potential to mimic fasting effects by activating autophagy and promoting cellular cleanup mechanisms. Specifically, the fasting-mimicking conditions trigger cellular adaptation mechanisms such as autophagy induction (via LC3II upregulation), reidelays the onset of senescence through ß-Galactosidase pathway activation, and the potential modulation of telomere maintenance systems, all contributing to improved cellular resilience and skin longevity. (20, 4).
Encapsulated within plant exosomes, resveratrol exhibited a novel role as a signaling molecule, capable of triggering SIRT1 activation, improved mitochondrial efficiency, reduced inflammation, and modulation of gene expression (1, 2).
In vitro assays: Vitis vinifera secretome activities
Evaluation of longevity markers
- Autophagy pathways activation
The impact of Vitis vinifera secretome on autophagy—a crucial cellular mechanism for maintaining homeostasis and recycling damaged components—was assessed through LC3 immunofluorescence (Figure 1) analysis in human dermal fibroblasts (HDFs) at early (P4) and later (P8) passages. LC3-II accumulation, indicative of autophagosome formation, was significantly enhanced in secretome-treated cells, particularly at 2.5% concentration. At P8, aged fibroblasts exhibited a 138% increase in LC3 fluorescence compared to untreated controls, surpassing even the positive control (metformin at 100 µM), highlighting a dose-dependent enhancement of autophagy.
Figure 1. LC3 immunofluorescence in HDFs showing autophagic activity. Hoechst (blue) marks nuclei; LC3 (green) indicates autophagosomes. Values represent mean of 3 (P4) and 6 (P8) biological replicates. Average diameter of exosomes = 124.2 nm
2. Inhibition of lipofuscin accumulations
Lipofuscin, a hallmark of cellular ageing and oxidative stress, was assessed in HDFs as an indicator of waste accumulation and declining autophagic efficiency. In untreated controls, lipofuscin levels increased markedly from P4 to P8, reflecting progressive cellular ageing (Fig. 1 A–B). In contrast, treatment with Vitis vinifera secretome (0.16% and 2.5%) or metformin at P4 significantly reduced lipofuscin accumulation, as seen by fewer dark spots (Fig. 1 C–E).
3. Inhibition of melanin accumulations
Melanin, while essential for photoprotection, can accumulate excessively with age, contributing to cellular dysfunction. In this study, senescent melanocytes were used to evaluate the depigmenting potential of Vitis vinifera secretome. As shown in Figure 20, the secretome reduced melanin content in a dose-dependent manner—by 2% at 2.5%, matching metformin (100 µM), and by 4% at 0.65%, similar to kojic acid (500 µM) (Figure 2).
Figure 2. Melanin reduction in senescent melanocytes. Data represents the mean of three biological replicates.
In vivoassays
In vivo 1: Assessment of the Fasting-Mimicking Potential of Vitis vinifera Secretome in a Caucasian Volunteer Panel
Skin antioxidant potential
The antioxidant capacity of Vitis vinifera secretome was assessed in vivo by measuring the reductive power of stratum corneum samples via the FRAP assay. While the placebo and fasting groups showed minimal and statistically non-significant changes, the secretome group exhibited a significant increase in skin antioxidant potential—+29.3% at day 28 and +38.3% at day 56 (p < 0.05) (Figure 3).
Figure 3. Skin antioxidant potential assessed in the first skin layers. Data represent the mean percentage change in antioxidant potential at T28 and T56 relative to T0. Significant differences compared to placebo and fasting are indicated by (p < 0.05).
In vivo 2: Evaluation of Radiance and Depigmentation Effects of Vitis vinifera Secretome in an Asian Volunteer Pane
- Skin global pigmentation
Skin global pigmentation was assessed in an Asian volunteer panel using hemifacial application and VISIA® imaging (Figure 4). Treatment with 1% Vitis vinifera secretome resulted in a statistically significant reduction in pigmentation: –4.1% at day 28 versus –1.1% with placebo (3-fold greater, p < 0.05), and –5.2% at day 56 versus –3.8% with placebo (1.4-fold greater). These results confirm a sustained depigmenting effect, supporting the ingredient’s efficacy in evening skin tone and enhancing radiance—contributing meaningfully to the luminous, uniform appearance characteristic of the glass skin effect.
Figure 4. Hemifacial comparison versus placebo under controlled conditions. Images captured with VISIA® system.
Conclusion
The biotechnological development of the Vitis vinifera fasting-mimicking secretome offers a novel, plant-based solution for skin longevity. By applying a controlled nutrient deprivation protocol, a hormetic cellular response is triggered in the plant, leading to the production of bioactive compounds known as Plant Fasting Factors—resveratrol, signaling peptides, and polyamines—naturally encapsulated within plant-derived exosomes for targeted skin delivery.
Although the present study primarily focused on the biological efficacy of Vitis vinifera secretome, the formulation’s physicochemical stability is a key factor in preserving its bioactivity. Drawing on Vytrus Biotech's methodology used in previous actives, stability efforts are oriented toward monitoring the integrity of the most functionally relevant molecular fractions, including proteins and peptides. In that case, targeted stability studies evaluated the shelf life by quantifying specific protein and peptide content under both refrigerated and room temperature conditions over extended periods, ensuring consistent performance of the active ingredient. Following a similar rationale, Vitis vinifera secretome future stability assessments will focus on its key bioactive compounds—particularly resveratrol, biopeptides, and exosome-associated molecules—under recommended storage conditions. These efforts will support long-term performance and guide formulation and packaging optimization without compromising the integrity of the active system.
Furthermore, this study provides relevant insights into the cosmetic potential of fasting-mimicking plant secretomes, highlighting their capacity to engage cellular longevity pathways such as autophagy, oxidative stress resistance, and pigment modulation. The results support Vitis vinifera secretome as an active for skin tone uniformity and vitality. Continued research focusing on deeper molecular characterization, formulation optimization, and broader clinical validation will help to further substantiate and refine this innovative cosmetic strategy.
Conclusion
The future of cosmetics lies in the continued evolution of holistic approaches which represents a transformative shift in the industry, merging scientific advancements, natural ingredients, and wellness principles. By understanding and embracing the interconnectedness of these elements, the cosmetics industry can cultivate products that not only enhance external beauty but also contribute to the overall well-being of individuals and the planet.
The interplay between beauty from within and topical cosmetics is the key for future products. The integration of biotechnology and green chemistry is revolutionizing cosmetic formulations, offering sustainable and biocompatible alternatives.
Developers can implement blockchain to trace the journey of ingredients from source to product. Nevertheless, the efficacy of the natural products should be scientifically proven. Marketers can communicate transparency as a brand value, and parallelly educate consumers by highlighting how specific ingredients contribute to radiant and healthy skin.
By embracing the synergy between these approaches and leveraging scientific advancements, the cosmetics industry can provide consumers with comprehensive beauty solutions that cater to both internal and external dimensions of beauty.
Surfactant Applications
The application area lends itself particularly well to the use of AI. Active today in this area is the US company Potion AI (6). The company provides AI-powered formulation tools for beauty and personal care R&D. Their offerings include Potion GPT, next generation ingredient and formula databases and AI document processing. Potion’s work could have a significant impact on the entire surfactant value chain, from raw material suppliers to end consumers. By using their GPT technology, they can help target work toward novel surfactant molecules that have optimal properties for specific applications. By using their ingredient and formula databases, they can access and analyze a vast amount of data on surfactant performance, safety, and sustainability. By using their AI document processing, they can extract and organize relevant information from patents, scientific papers, and regulatory documents. These capabilities could enable Potion AI's customers to design and optimize surfactant formulations that are more effective, eco-friendly, and cost-efficient. A particularly interesting application for this type of capability is deformulation.
Deformulation is the process of reverse engineering a product's formulation by identifying and quantifying its ingredients. Deformulation can be used for various purposes, such as quality control, competitive analysis, patent infringement, or product improvement. However, deformulation can be challenging, time-consuming, and costly, as it requires sophisticated analytical techniques, expert knowledge, and access to large databases of ingredients and formulas.
AI can potentially enhance and simplify the deformulation process by using data-driven methods to infer the composition and structure of a product from its properties and performance. For example, AI can use machine learning to learn the relationships between ingredients and their effects on the product's characteristics, such as color, texture, fragrance, stability, or efficacy. AI can also use natural language processing to extract and analyze information from various sources, such as labels, patents, literature, or online reviews, to identify the possible ingredients and their concentrations in a product.
Figure 2. Skin Section with Microbiome. Most microorganisms live in the superficial layers of the stratum corneum and in the upper parts of the hair follicles. Some reside in the deeper areas of the hair follicles and are beyond the reach of ordinary disinfection procedures. There bacteria are a reservoir for recolonization after the surface bacteria are removed.
About the Author
Òscar Expósito
Òscar Expósito - Biologist with a PhD in Plant Biotechnology, and cofounder of Vytrus Biotech. Author of more than 15 scientific articles and 6 patents in pharmaceutical and cosmetic sectors. Speaker in more than 35 international academic congresses, professor in entrepreneurship and biotechnology fields. Member of associations such as SEQC, SCS and IFSCC. With Vytrus, he won international awards from the cosmetic sector.
Òscar Expósito
CEO, CSO and co-founder, Vytrus Biotech S.A.
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