Six Amino Acids among Natural Moisturizing Factors Responsible for Skin Hydration: Improvement and Anti-Aging of Skin by <i>Galactomyces</i> Ferment Filtrate-Pitera^TM Containing Skin Moisturizer

Abstract

<b>Background:</b> Natural moisturizing factors (NMFs) are filaggrin-derived components in the cornified layer that are critical for maintaining healthy skin moisturization and barrier function. However, studies have reported conflicting findings on the relationship between NMF levels and aging, while few studies have investigated this relationship clinically. To fill this research gap, we determined the levels of major NMF components such as free amino acids, pyrrolidone carboxylic acid, and urocanic acids, and individually verified their relationships with skin hydration, barrier function, age, and skin aging. <b>Purpose:</b> The objective of this study was to clinically investigate the relationship between NMF components&#8217; levels and skin aging in facial skin. The main NMF components were obtained from facial skin and quantified. We then selected NMF components showing strong relationships to skin hydration, and analyzed the relationships of the levels of these selected NMF components with signs of skin aging, namely, texture, pores, wrinkles, and dullness (L-value). We also examined the efficacy of treatment with a skin care formula (SK-II Facial Treatment Essence, called SK-II FTE hereafter) including <i>G</i><i>a</i><i>lactomyces</i> ferment filtrate (GFF, Pitera^TM) on the selected NMF component levels associated with skin hydration and barrier function, and the signs of skin aging of texture, pores, wrinkles, and dullness (L-value). <b>Method:</b> We conducted two clinical trials in this research. In Study 1, we measured 23 NMF components using tape-stripped cornified layer to quantify them via an HPLC method in 196 Asian females aged 20 to 59 (mean &#177; S.D., 38.6 &#177; 9.4). Facial visual aging parameters [texture, pores, wrinkles, and dullness (L-value)], as well as elasticity (R7), skin hydration, and TEWL, were quantified using facial skin imaging and skin physical property measurement devices.<b> </b>Study 2 was performed to evaluate whether the facial application of SK-II FTE affects the NMF levels and skin aging parameters in 63 Asian female volunteers aged 20 to 55 (38.4 &#177; 9.03). During the course of Study 2, 0.6 mL of SK-II FTE was applied to the face twice daily in the morning and afternoon. Skin measurements were performed at the start of the day (baseline) and at week 8. <b>R</b><b>e</b><b>sults:</b> In Study 1, we examined the stratum corneum levels of 23 NMF components comparing to the skin hydration status in 196 female subjects. The subjects were divided into two groups using the median of each measured NMF component. Skin hydration values were compared between the two groups defined for each NMF component. The results showed that subjects with higher levels of six amino acids, alanine, arginine, asparagine, glutamine, glycine, and histidine, exhibited significantly higher skin hydration than those with lower amino acid levels. No significant differences in skin hydration values were found for the other 17 NMF components. We then analyzed whether the sum of these six amino acid NMF components (called 6-AA-NMFs, hereafter) is affected by aging. The 6-AA-NMF level peaked in the subjects aged 25-29, and then gradually and significantly decreased with age. Interestingly, the 6-AA-NMF level was significantly correlated with the skin hydration value, but not with TEWL. In addition, the 6-AA-NMF level demonstrated significant correlations with the signs of skin aging of texture, pores, wrinkles, and dullness (L-value). Then, in Study 2, we examined whether the daily application of SK-II FTE affects the 6-AA-NMF level and visual aging parameters in 63 females. SK-II FTE demonstrated significant increases of the levels of 6-AA-NMFs and each of its components associated with hydration and barrier function, and improvements of skin texture, pores, wrinkles, and dullness (L-value) during the 8 weeks of treatment of facial skin. <b>Conclusion</b>: These clinical studies with large numbers of subjects across a wide age range revealed that six amino acids as NMF components were highly correlated with facial skin hydration in the stratum corneum. The levels of these six NMF components were also found to decrease at ages after the 30 s and were significantly correlated with major signs of skin aging. Notably, these six NMF components (6-AA-NMFs) were increased by SK-II FTE treatment associated with improvements of skin hydration and signs of skin aging, namely, texture, pores, wrinkles, and dullness (L-value). These studies were limited by the lack of investigation of why some NMF components were not associated with skin hydration. More clinical trials examining various NMF components and their relationship with aging are anticipated.