GALACTOMYCES FERMENT FILTRATE

Interleukin (IL)-37 suppresses systemic and local inflammation. It is expressed in the epidermis, the external layer of the skin, and is decreased in inflammatory skin diseases including atopic dermatitis (AD) and psoriasis. Therefore, an agent applied topically on the skin that can increase IL-37 could be promising for treating AD and psoriasis; however, the mechanism regulating IL-37 remains largely unknown. Given that IL-37 expression is induced in differentiated keratinocytes, a major component of the epidermis, and that activation of aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor, promotes keratinocyte differentiation, we hypothesized that AHR might be involved in the IL-37 expression in human keratinocytes. We analyzed normal epidermal human keratinocytes (NHEKs) treated with tapinarof and Galactomyces ferment filtrate (GFF), which are potent AHR modulators. We found that tapinarof and GFF upregulated IL-37 in NHEKs, which was canceled by the knockdown of AHR using siRNA transfection, indicating that AHR mediates IL-37 expression in NHEKs. Furthermore, we found that the knockdown of IL-37 resulted in the upregulation of IL-33, an alarmin cytokine with crucial roles in the pathogenesis of AD and psoriasis. These findings suggest that IL-37 negatively regulates IL-33 expression in NHEKs. Finally, we examined whether tapinarof and GFF treatment modulates IL-33 expression in NHEKs. Such treatment inhibited IL-33 expression, which was partially reversed by the knockdown of either AHR or IL-37. Taken together, our findings provide the first evidence that tapinarof and GFF could have potential to prevent IL-33-overexpressing disorders such as AD and psoriasis via the AHR/IL-37 axis.

Galactomyces ferment filtrate (GFF, Pitera™) is a cosmetic ingredient known to have multiple skin care benefits, such as reducing redness and pore size via the topical application of its moisturizer form. Although GFF is known to act partly as an antioxidative agonist for the aryl hydrocarbon receptor (AHR), its significance in keratinocyte biology is not fully understood. In this study, we conducted a transcriptomic analysis of GFF-treated human keratinocytes. Three different lots of GFF consistently modulated 99 (22 upregulated and 77 downregulated) genes, including upregulating cytochrome P450 1A1 ( CYP1A1 ), a specific downstream gene for AHR activation. GFF also enhanced the expression of epidermal differentiation/barrier-related genes, such as small proline-rich proteins 1A and 1B ( SPRR1A and SPRR1B ), as well as wound healing-related genes such as serpin B2 ( SERPINB2 ). Genes encoding components of tight junctions claudin-1 ( CLDN1 ) and claudin-4 ( CLDN4 ) were also target genes upregulated in the GFF-treated keratinocytes. In contrast, the three lots of GFF consistently downregulated the expression of inflammation-related genes such as chemokine (C-X-C motif) ligand 14 ( CXCL14 ) and interleukin-6 receptor ( IL6R ). These results highlight the beneficial properties of GFF in maintaining keratinocyte homeostasis.

Facial skin aging is an important psychophysical and social concern, especially in women. We compared facial parameters reflecting aging of the skin in 1999 and 2010 in 86 female volunteers. Then, all subjects applied three Galactomyces ferment filtrate-containing skin care products (G3 products; SK-II Facial Treatment Essence, SK-II Cellumination Essence, and SK-II Skin Signature Cream) twice daily for 12 months (M), with the skin parameters being measured at 2 M, 8 M, and 12 M during this period. Facial skin aging parameters such as wrinkles, hyperpigmented spots, and roughness significantly deteriorated during the 11-year interval. This 11-year aging process was associated with reduced hydration and increased transepidermal water loss (TEWL). Notably, treatment with G3 products significantly and cumulatively increased skin hydration with a correlated reduction of TEWL during the 12 M treatment period. Such treatment also significantly and cumulatively reversed the 11-year facial skin aging in the three parameters of wrinkles, spots, and roughness. These results suggest that facial skin retains the potential to recover from the aging process when it is applied with appropriate cosmetic agents.

Post-acne hyperpigmentation (PAH) occurs secondary to acne vulgaris and may cause significant adverse effects. Although may occur in any skin types, PAH has been found to be more common and severe in people with colored skin. This study aimed to assess the effectiveness of combination serum containing galactomyces ferment filtrate (GFF), dexpanthenol, and Centella asiatica for treating PAH. This randomized controlled clinical trial involved Fitzpatrick skin type (FST) IV and V patients with PAH. Subjects were equally divided into treatment group, who received three drops of combination serum twice daily for 8 weeks, and placebo group. The melanin index (MI) and Lightness (L*) score were assessed every 2 weeks. Out of 51 subjects, the L* score of the treatment group in subjects with FST V was significantly higher on the 4th and 6th week compared to the placebo group ( P ˂ 0.05). The MI of subjects with FST IV was significantly lower compared to the placebo group after 8 weeks ( P ˂ 0.05). The treatment group showed consistent increasing and decreasing trend in L* score and MI, respectively ( r ˃ 0.9, P ˂ 0.05). Combination serum containing GFF, dexpanthenol, and C. asiatica may be effective in treating PAH in subjects with colored skin by accelerating lessening of PAH.

Skin is exposed to multiple external factors such as solar radiation and environmental oxidative stressors. Ageing and the accumulation of stress damage lead to reduced skin health and loss of skin firmness, which contribute to wrinkling and sagging. We investigated if Galactomyces ferment filtrate (GFF) could modulate skin cell mechanical properties through changes to the actin cytoskeleton and anchoring junctions. Proteomic analysis was performed to identify age-associated changes in cellular components in skin biopsies and GFF-mediated changes in full thickness 3D skin equivalent models. Nanoindentation was used to measure the Young's modulus (resistance to elastic deformation) of keratinocyte cells under Latrunculin B (LatB)-induced destabilization of the actin cytoskeleton. Immunofluorescence and confocal microscopy were performed on skin equivalents to validate the GFF-induced changes observed in proteomic analysis and identify expression profiles of key structural proteins within the skin. Aged skin had decreased expression of internal cellular components 'actin cytoskeleton' and 'focal adhesion'. Destabilization of actin cytoskeleton by LatB significantly reduced the Young's modulus of keratinocyte cells. Pre-treatment of keratinocytes with GFF was able to prevent LatB-induced decline of skin cell mechanical properties. Proteomic analysis of full thickness 3D skin equivalent models revealed that GFF increased the expression of 'actin cytoskeleton' and 'anchoring junctions', especially the actin-binding focal adhesion proteins. Immunofluorescence staining confirmed that GFF upregulated expression of the actin stabilizer, calponin 2 and integrin α2 in basal keratinocytes. GFF increased integrin localisation to the dermal-epidermal junction (DEJ) and colocalised with collagen IV in vitro, suggesting increased linkage between the epidermis and dermis. Our data establish the actin cytoskeleton and anchoring junctions which bind it as important cellular structures which decline with age. We also identified that the cellular structures are required for the maintenance of keratinocyte Young's modulus, which could be an important property impacting skin firmness. Furthermore, we have identified that GFF promotes actin stability and builds a rigid holding structure of the anchoring junctions to increase cell-cell and epidermis-dermis anchoring in skin equivalent models. Therefore, GFF has the potential to help maintain and restore skin firmness in young and old skin, respectively. La peau est exposée à de multiples facteurs externes tels que le rayonnement solaire et les facteurs de stress oxydatifs environnementaux. Le vieillissement et l'accumulation de dommages causés par le stress entraînent une diminution de la santé et de la fermeté de la peau, ce qui contribue à l'apparition de rides et au relâchement cutané. Nous avons étudié si le filtrat de ferment de Galactomyces (FFG) pouvait moduler les propriétés mécaniques des cellules cutanées par des modifications du cytosquelette d'actine et des jonctions d'ancrage. MÉTHODES: Une analyse protéomique a été réalisée pour identifier les changements associés à l'âge des composants cellulaires dans des biopsies cutanées et les changements médiés par le FFG dans des modèles 3D équivalents à la peau de pleine épaisseur. La nano‐indentation a été utilisée pour mesurer le module de Young (résistance à la déformation élastique) des cellules kératinocytaires sous déstabilisation induite par la latrunculine B (LatB) du cytosquelette d'actine. Une immunofluorescence et une microscopie confocale ont été réalisées sur des équivalents cutanés pour valider les changements induits par le FFG observés dans l'analyse protéomique et identifier l

Human skin is the first line of defence from environmental factors such as solar radiation and is susceptible to premature ageing, including a disruption in epidermal differentiation and homeostasis. We evaluated the impact of a Galactomyces Ferment Filtrate (GFF) on epidermal differentiation and response to oxidative stress. We used transcriptomics, both spatial and traditional, to assess the impact of GFF on epidermal biology and homeostasis in keratinocytes (primary or immortalized) and in ex vivo skin explant tissue. The effect of GFF on cell adhesion rates, cellular ATP levels and proliferation rates were quantitated. Oxidative phosphorylation and glycolytic rates were measured under normal and stress-induced conditions. Transcriptomics from keratinocytes and ex vivo skin explants from multiple donors show GFF induces keratinocyte differentiation, skin barrier development and cell adhesion while simultaneously repressing cellular stress and inflammatory related processes. Spatial transcriptomics profiling of ex vivo skin indicated basal keratinocytes at the epidermal-dermal junction and cornifying keratinocytes in the top layer of the epidermis as the primary cell types influenced by GFF treatment. Additionally, GFF significantly increases crosstalk between suprabasal and basal keratinocytes. To support these findings, we show that GFF can significantly increase cell adhesion and proliferation in keratinocytes. GFF also protected overall cellular bioenergetics under metabolic or oxidative stress conditions. Our findings provide novel insights into cellular differences and epidermal spatial localization in response to GFF, supporting previous findings that this filtrate has a significant impact on epidermal biology and homeostasis, particularly on spatially defined crosstalk. We propose that GFF can help maintain epidermal health by enhancing keratinocyte crosstalk and differentiation/proliferation balance as well as promoting an enhanced response to stress. La peau humaine constitue la première ligne de défense contre les facteurs environnementaux tels que le rayonnement solaire et est susceptible de vieillir prématurément, notamment d’une perturbation de la différenciation épidermique et de l’homéostasie. Nous avons évalué l'impact d'un filtrat de ferment Galactomyces (GFF) sur la différenciation épidermique et la réponse au stress oxydatif. MÉTHODES: Nous avons utilisé la transcriptomique, à la fois spatiale et traditionnelle, pour évaluer l'impact du GFF sur la biologie épidermique et l'homéostasie des kératinocytes (primaires ou immortalisés) et des explants cutanés ex vivo. L'effet du GFF sur les taux d'adhésion cellulaire, les niveaux d'ATP cellulaire et les taux de prolifération ont été quantifiés. La phosphorylation oxydative et les taux de glycolytique ont été mesurés dans des conditions normales et induites par le stress. RÉSULTATS: La transcriptomique des kératinocytes et des explants cutanés ex vivo provenant de plusieurs donneurs montrent que le GFF induit la différenciation des kératinocytes, le développement de la barrière cutanée et l'adhésion cellulaire tout en réprimant simultanément le stress cellulaire et les processus inflammatoires associés. Le profilage transcriptomique spatial de la peau ex vivo a indiqué que les kératinocytes basaux à la jonction épidermique‐dermique et les kératinocytes cornifiants dans la couche supérieure de l'épiderme étaient les principaux types de cellules influencés par le traitement GFF. De plus, le GFF augmente considérablement la diaphonie entre les kératinocytes suprabasaux et basaux. Pour étayer ces résultats, nous montrons que le GFF peut augmenter cons

Mask wearing is described as one of the main public health measures against COVID-19. Mask wearing induces various types of subjective and objective facial skin damage, such as hair pore dilatation and redness. Facial pore size and redness show morning-to-evening intra-day fluctuations. It remains unknown whether mask usage affects fluctuations in pore size and redness. We measured facial skin hydration, transepidermal water loss (TEWL), pore size, and redness four times a day for 6 weeks in 20 healthy young women. After a 2-week no-mask-usage period (baseline period), all subjects wore unwoven masks for 2 weeks; then, for the following 2 weeks, they applied masks after the topical application of a moisturizer containing a Galactomyces ferment filtrate (GFF) skin care formula (Pitera™). We demonstrated that mask wearing significantly increased the intra-day fluctuations of pore size, redness, and TEWL. In addition, significant correlations were evident among these three parameters. Notably, these mask-induced skin changes were significantly improved, achieving a return to baseline levels, by the application of a GFF-containing moisturizer. In conclusion, mask wearing aggravates intra-day fluctuations in pore size and redness. Appropriate moisturization can minimize this mask-related skin damage, most likely by normalizing the elevated TEWL.

To compare the effectiveness of tranexamic acid (TA) combination serum with hydroquinone, the gold standard in whitening agents for healthy populations. This was a three-arm randomized controlled trial. The subjects were divided into three groups: the first group received 3% TA combination serum (3% TA, 4% galactomyces ferment filtrate, 2% niacinamide, and 4% alpha arbutin), the second group received 2% TA combination serum, and the third group received 4% hydroquinone. One milliliter of each serum was applied on three holes: Hole A, which was located 4 cm from the left cubital fossa, Hole B, which was located 4 cm from the first hole, and Hole C, which was located 4 cm from the right cubital fossa. The skin brightness and pigmentation intensity were evaluated each week for 4 weeks using a chromameter. A total of 44 subjects were recruited for this study. All groups showed a significant improvement in skin brightness and pigmentation intensity after 4 weeks (p < .001). There were no differences between the treatment groups and hydroquinone (p > .05). TA serum (2 and 3%) combined with 4% galactomyces ferment filtrate, niacinamide, and alpha arbutin is an effective depigmenting agent.

Caspase-14 is a cysteinyl-aspartate-specific proteinase that is specifically expressed in epidermal keratinocytes. Dysregulation of caspase-14 expression is implicated in impaired skin barrier formation. To elucidate the regulation of caspase-14 in differentiated keratinocytes, we characterized the expression of caspase-14 in normal human epidermal keratinocytes (NHEKs) and two types of three-dimensional (3D) human epidermis culture models, EPI-200 and EPI-201, via RT-PCR and immunoblot analyses. Caspase-14 expression was absent in subconfluent NHEKs, but was present in confluent NHEKs as well as those induced to differentiate by calcium. Caspase-14 expression levels in the 3D epidermis models were almost equal to that in the Ca(2+)-treated differentiated NHEKs. Despite the presence of caspase-14 expression in these models, caspase-14 activity was found only in the mature 3D skin model, EPI-200. This was confirmed by detection of a 17 kDa cleaved fragment of caspase-14 present only in the EPI-200 model. Since glucocorticoid (GC) receptor is required for skin barrier competence, we investigated whether the GC dexamethasone (Dex) and various natural components of common skin moisturizers affect caspase-14 expression in keratinocytes. Dex decreased caspase-14 expression in undifferentiated, but not differentiated, NHEKs. Conversely, Dex increased caspase-14 expression in both 3D skin models, although it did not alter caspase protease activity. Similar to treatment with Dex, treatment of the premature 3D skin mode, EPI-201 with a Galactomyces ferment filtrate markedly increased expression of caspase-14. Further, these results suggest that the effect of Dex, or lack thereof, on caspase-14 expression is dependent on the stage of keratinocyte differentiation.

Skincare products play a crucial role in preventing the dry skin induced by various causes. Certain ingredients can help to improve the efficacy of skincare products. Galactomyces ferment filtrate (GFF) is such a functional ingredient. Its use originated from the empirical observation that the hands of sake brewers who deal with yeast fermentation retain a beautiful and youthful appearance. Consequently, skincare products based on GFF are widely used throughout the world. Recent studies have demonstrated that GFF activates an aryl hydrocarbon receptor (AHR) and upregulates the expression of filaggrin, a pivotal endogenous source of natural moisturizing factors, in epidermal keratinocytes. It also activates nuclear factor erythroid-2-related factor 2 (NRF2), the antioxidative master transcription factor, and exhibits potent antioxidative activity against oxidative stress induced by ultraviolet irradiation and proinflammatory cytokines, which also accelerate inflammaging. GFF-mediated NRF2 activation downregulates the expression of CDKN2A, which is known to be overexpressed in senescent keratinocytes. Moreover, GFF enhances epidermal terminal differentiation by upregulating the expression of caspase-14, claudin-1, and claudin-4. It also promotes the synthesis of the antiinflammatory cytokine IL-37 and downregulates the expression of proallergic cytokine IL-33 in keratinocytes. In addition, GFF downregulates the expression of the CXCL14 and IL6R genes, which are involved in inflammaging. These beneficial properties might underpin the potent barrier-protecting and anti-inflammaging effects of GFF-containing skin formulae.

Young women often complain about the daily fluctuation of their facial skin conditions. However, no objective study has been carried out on such changes. This study is aimed at quantitatively elucidating daily skin fluctuation and evaluating the efficacy of cosmetic skin care treatment. We developed the first portable and self-guided facial skin imaging device (eMR Pro) to reproducibly capture facial images at home. Two 8 week clinical studies were then conducted to analyze daily skin fluctuation of facial pore areas, roughness and redness in young Japanese women ( n = 47 in study 1 and n = 57 in study 2) by collecting facial images three times a day, during the morning after wake-up, during the morning after face wash, and during the evening after face wash. After a 4 week baseline measurement period (week -4 to week -1), all subjects applied Galactomyces ferment filtrate (GFF, Pitera ® ) skin care formula twice a day for 4 weeks (week 1 to week 4). These three skin conditions did exhibit different fluctuation patterns. The pore area and roughness showed the "morning after wake-up"-largest fluctuation pattern, whereas redness showed the "evening after face wash"-largest fluctuation pattern. GFF treatment significantly reduced the net values and delta fluctuation of pore area, roughness, and redness, which were consistently observed in two studies. In conclusion, the daily fluctuation of facial skin conditions is potentially a new target field for investigating healthy skin maintenance.