Short Communication: Effects of Lactobacillus helveticus-Fermented Milk on the Differentiation of Cultured Normal Human Epidermal Keratinocytes

Abstract

Effects of Lactobacillus helveticus-fermented milk whey on the differentiation of normal human epidermal keratinocytes were studied. Analysis using real-time reverse transcription-polymerase chain reaction revealed that addition of Lactobacillus helveticus-fermented milk whey to the culture medium enhanced mRNA expression of keratin 10, an early differentiation marker, as well as involucrin, a late differentiation marker. Whey of artificially acidified milk, prepared by the addition of dl-lactic acid to milk instead of fermentation, also promoted expression of both markers, but Lactobacillus helveticus-fermented milk whey was more effective in increasing expression of those markers. These results indicate that milk whey has the potential to induce multiple stages of keratinocyte differentiation and that fermentation with Lactobacillus helveticus increases that activity. Furthermore, we examined the expression of profilaggrin, which increases with epidermal terminal differentiation, and found that Lactobacillus helveticus-fermented milk whey enhanced expression of profilaggrin mRNA in a dose-dependent manner. Expression also occurred to a greater extent than with artificially acidified milk whey or other whey samples prepared with several lactic acid bacterial species. Because the proteolytically processed form of profilaggrin, filaggrin, is very important for normal epidermal hydration and flexibility, our results indicate that Lactobacillus helveticus-fermented milk whey has the potential to enhance the production of filaggrin-related natural moisturizing factor, because of its effect on the induction of epidermal differentiation, and is expected to be a useful skin moisturizing agent.

Oral Intake ofLactobacillus helveticus-Fermented Milk Whey Decreased Transepidermal Water Loss and Prevented the Onset of Sodium Dodecylsulfate-Induced Dermatitis in Mice

“…[19][20][21] Hence we suppose that one or more biologically active peptides generated by the L. helveticus CM4-fermentation were responsible for our results. In this study, we did not examine the in vivo expression of filaggrin; furthermore, the relationship between the LHMW effects observed in this study and those observed in a previous in vitro study 4) remain unclear. In order to clarify the mechanisms and the effective substances involved, further investigation is required.…”

“…4) Further, L. helveticus CM4 strain showed very potent protease activity and efficiently produced several milk protein-derived biologically active peptides during fermentation. 6) Hence we expected that oral administration of LHMW, including a large amount of peptides, might have beneficial effects on the skin.…”

“…Fermented milk was prepared as reported previously. 4) In brief, reconstituted pasteurized 9% wt/wt skim milk (Yotsuba Milk Products, Sapporo, Japan) was fermented with the Lactobacillus helveticus CM4 strain 6) selected from a starter culture of Calpis sour milk (Calpis, Tokyo) at 37 C for 24 h. Artificially acidified milk was obtained by the addition of DL-lactic acid (Wako Pure Chemical Industries, Osaka, Japan) to reconstituted pasteurized 9% wt/wt skim milk to a final concentration of 2.29% wt/wt, to create the same acidity as that of the L. helveticus-fermented milk. These samples were separated into the whey fraction by centrifugation at 12;000 Â g for 20 min, and were named L. helveticus-fermented milk whey (LHMW) and artificially acidified milk whey (AAMW).…”

“…[1][2][3] With respect to the skin, fermented milk has been used in the clinical treatment of wounds and burns, 1) and is now being used widely as a cosmetic material. Recently, we reported that Lactobacillus helveticus-fermented milk whey (LHMW) promoted the differentiation of cultured normal human epidermal keratinocytes and increased the expression of profilaggrin, 4) which is further proteolytically processed and used in the generation of a natural moisturizing factor (NMF) believed to be crucial to epidermal hydration and flexibility. 5) However, little has been reported regarding the in vivo effects of ingestion or topical application of fermented milk on skin homeostasis, or on the in vitro effects of fermented milk components on cells constituting the skin.…”

Health-Promoting Properties of Lactobacillus helveticus

“…On the contrary, no significant effects were exerted by artificially acidified milk whey (Baba et al, 2010). In vitro experiments on human epidermal keratinocytes supplemented with the same fermented milk whey preparation identified an increase in profilaggrin mRNA, a precursor of filaggrin that is involved in epidermal hydration and flexibility and is a marker of epidermal terminal differentiation (Baba et al, 2006). …”