• Nutrition Research and Practice
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• v.5 no.5
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• pp.396-403
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• 2011

Ceramides (Cer) comprise the major constituent of sphingolipids in the epidermis and are known to play diverse roles in the outermost layers of the skin including water retention and provision of a physical barrier. In addition, they can be hydrolyzed into free sphingoid bases such as $C_{18}$ sphingosine (SO) and $C_{18}$ sphinganine (SA) or can be further metabolized to $C_{18}$ So-1-phosphate (S1P) and $C_{18}$ Sa-1-phosphate (Sa1P) in keratinocytes. The significance of ceramide metabolites emerged from studies reporting altered levels of SO and SA in skin disorders and the role of S1P and Sa1P as signaling lipids. However, the overall metabolism of sphingoid bases and their phosphates during keratinocyte differentiation remains not fully understood. Therefore, in this study, we analyzed these Cer metabolites in the process of keratinocyte differentiation. Three distinct keratinocyte differentiation stages were prepared using 0.07 mM calcium (Ca$^{2+}$) (proliferation stage), 1.2 mM Ca$^{2+}$ (early differentiation stage) in serum-free medium, or serum-containing medium with vitamin C (50 ${\mu}L$/mL) (late differentiation stage). Serum-containing medium was also used to determine whether vitamin C increases the concentrations of sphingoid bases and their phosphates. The production of sphingoid bases and their phosphates after hydrolysis by alkaline phosphatase was determined using high-performance liquid chromatography. Compared to cells treated with 0.07 mM Ca$^{2+}$, levels of SO, SA, S1P, and SA1P were not altered after treatment with 1.2 mM Ca$^{2+}$. However, in keratinocytes cultured in serum-containing medium with vitamin C, levels of SO, SA, S1P, and SA1P were dramatically higher than those in 0.07- and l.2-mM Ca$^{2+}$-treated cells; however, compared to serum-containing medium alone, vitamin C did not significantly enhance their production. Taken together, we demonstrate that late differentiation induced by vitamin C and serum was accompanied by dramatic increases in the concentration of sphingoid bases and their phosphates, although vitamin C alone had no effect on their production.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.32 no.1 s.55
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• pp.17-22
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• 2006

Phosphatidyiserine (PS) is a phospholipid which plays the structural role in membranes and serves as a cofactor of signaling enzymes for diverse cellular functions. In this study, we observed that topical treatment with PS significantly decreased trans-epidermal water loss (TEWL) induced by tape-stripping in hairless mice. Also, ceramides in epidermis were increased in PS-treated group compared to vehicle-treated one in vivo. the amounts of non-hydroxyl ceramide (NHCER) (1.4 fold) and glucosylceramide (glucosylCER) (1.6 fold), in the skin of hairless mice, were increased by topical treament with PS. Also, we demonstrated that PS stimulated keratinocyte differentiation. We observed that PS treatment morphologically altered normal human keratinocyte (NHK) from the proliferating phase to the differentiating one, suggesting that PS stimulated epidermal differentiation in NHK. We also showed that the expressions of the specific markers for epidermal differentiation, involucrin (INV) (3.5 fold up) and transglutaminase 1 (TG'ase 1) (3 fold up), were significantly increased by PS treatment, compared to untreated control in vitro. In addition, topical treatment with PS resulted in a progressive increase in INV and loricrin protein levels in vivo. In conclusion, we provide the first evidence for the physiological activities of PS in skin, and we suggest that PS strengthen the epidermal permeability harrier by stimulation of keratinocyte differentiation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.1
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• pp.31-37
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• 2018

Oil/water (O/W) nanoemulsions are effective vehicles to change the permeability of the skin. In this study, we focused on the preparation and characterization of nanoemulsion which serve as colloidal carriers for the dermal application of ceramide IIIB (CIIIB) and stratum corneum (SC) lipids such as cholesterol, and palmitic acid. In order to optimize the nanoemulsions, emulsification process conditions were conducted with regard to droplet size, nanoemulsion stability, and solubility of CIIIB. A decrease in droplet size was observed through emulsification temperature of $80^{\circ}C$ and phase inversion composition (PIC) method. CIIIB has low solubility in oil and water. When the concentration of CIIIB was increased, the droplet size of nanoemulsion was increased. When Lipoid S75-3 was added to the oil phase, the solubility of CIIIB increased, indicating some interactions shown in DSC measurements. CIIIB and SC lipids could be successfully incorporated in nanoemulsions without crystallization or physical instability. In conclusion, a stable nanoemulsion containing the SC lipids could be effective as an efficient moisturizing system for skin.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.2
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• pp.119-126
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• 2016

Skin is the largest organ that protects the body from the external environmental factors such as smog, cigarette smoke, UV. Protective skin barrier is composed with keratinizational keratinocytes and intercellular lipids such as ceramides, cholesterols and fatty acids combined by the lamellar liquid crystal structure. In this research, we confirmed that the Jeju wild ginseng (JWG) extracts dose-dependently increased the expression of serine-palmitoyltransferase (SPT) protein which is associated with ceramide biosynthesis. In addition, emulsion containing 5% JWG extract was applied on skin of human volunteers for 2 weeks and then significantly reduced transepidermal water loss (TEWL) compared to that of control group. As a results, JWG extract increased the biosynthesis of ceramides that is the key components of the skin lipid through enhancing expression of SPT. In addition, JWG extract reduced TEWL resulting in improvement of skin barrier function. In this context, we suggest that JWG extract could be used as a skin barrier enhancer and moisturing agents in cosmetic fileds.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.61-72
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• 2003

In cosmetic area, retinol is prominent ingredient for anti-wrinkle but unstable against light, heat, oxygen and so on. Therefore the stabilization of retinol is required. Here, we capsulated doubly retinol in the SLM(Skin Lipid Matrix) that makes three dimensional lamellar structure similar to skin, after formation of primary liposome (retinol-nanoemulsion). First, we make primary liposome from retinol / hydrogenated lecithin / polysorbate20 / caprylic ＆ capric triglyceride / ethanol / and so on, and the mean diameter to 70 nm, using microfluidizer passed three times at 800 Bar, repeatedly. Then we produce DC-liposome (doubly capsulated-liposome) that was encapsulated primary liposome with SLM made of hydrogenated phosphatidyl choline / caprylic ＆ capric triglyceride / 1, 3-butylene glycol / ceramide3 / cholesterol /etc. We measured for color stability against light and heat with chromameter. As a result of this experiment, we observed DC-liposome was more than from 1.5 to 3 times as stable as general liposome. Livability of retinol has improved from 2 to 6 times when we analyzed it by HPLC. Also, penetration effect of DC-liposome has improved.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.570-577
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• 2003

The importance of cornified envelope (CE)-bound lipids to epidermal barrier function is increasingly being recognized. In the present study, we intentionally damaged the cornified layer of hairless mice by ultraviolet irradiation and sodium dodecyl sulfate (SDS) treatment, and assessed the changes in epidermal barrier function by measuring Trans Epidermal Water Loss (TEWL). We also measured changes in the amount of CE-bound lipids using thin layer chromatography (TLC). The results showed that both treatments increased TEWL and decreased CE-bound lipids (omega-hydroxy cerami de and omega-hydroxy acid). In addition, investigation of the chronological changes in TEWL revealed an inverse relationship between TEWL and CE-bound lipids, and a correlation between CE-bound lipids and epidermal barrier function. We then measured the amount of CE-bound lipids in the cheek and the medial side of the upper arm in humans. The results showed that because the cheek receives external stimulation on a daily basis, the amount of CE-bound lipids was significantly lower, while the level of TEWL was higher. These observations, together with those from the animal study, indicate that CE-bound lipids are related to epidermal barrier function.

• Molecules and Cells
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• v.25 no.2
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• pp.224-230
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• 2008

Ceramides are well-known second messengers that induce apoptosis in various kinds of cancer cells, and their effects are closely related to radiation sensitivity. Phytoceramides, the yeast counterparts of the mammalian ceramides, are also reported to induce apoptosis. We investigated the effect of a novel ceramide derivative, N-acetylphytosphingosine (NAPS), on the radiosensitivity of NCI-H460 human lung carcinoma cells and its differential cytotoxicity in tumor and normal cells. The combination of NAPS with radiation significantly increased clonogenic cell death and caspase-dependent apoptosis. The combined treatment greatly increased Bax expression and Bid cleavage, but not Bcl-2 expression. However, there was no effect on radiosensitivity and apoptosis in BEAS2B cells, which derive from normal human bronchial epithelium. Cell proliferation and DNA synthesis were significantly inhibited by NAPS in both NCI-H460 and BEAS2B cells, but only the BEAS2B cells recovered by 48h after removal of the NAPS. Furthermore, the NCI-H460 cells underwent more DNA fragmentation than the BEAS2B cells in response to NAPS. Our results indicate that NAPS may be a potential radiosensitizing agent with differential effects on tumor vs. normal cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.1
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• pp.53-72
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• 2001

Dry skin is caused mainly by the perturbation of stratum corneum lipids which affected by ageing, change of season, excess use of surfactant and the effect of disease like atopic dermatitis and psoriasis. Intercellular lipid structures in stratum corneum are responsible for the barrier function of mammalian skin. The major lipd classes that can be extracted from stratum corneum are ceramides, cholesterol and fatty acid, which make up approximately 50, 25, 10 percent of the stratum corneum lipid mass, respectively. Small amount of cholesterol sulfate, phospholipids, glycosylceramide and cholesterol esters are also present. Recent studies have shown that application of one or two these lipids to the perturbed skin delays barrier recovery; only equimolar mixtures allow normal recovery. We observed that barrier recovery rate was improved in hairless mouse by topical application of single neutral lipids (ceramide, free fatty acid, cholesterol) and lipid mixtures. Whereas the application of single lipid didn’t allows a significant enhancement comparing with normal barrier repair, the equimolar mixtures of 3 components(including synthetic pseudoceramide PC104) improved barrier repair, as assessed by the transepidermal water loss. At clinical study to the volunteers aged over sixty, skin dryness recuperated by the increase of moisture(capacitance) and the reduction of scaling. Utilization of physiologic lipid mixture containing natural ceramides or synthetic pseudoceramide could lead to new forms of topical therapy for the dryness and dermatoses(e.g., psoriasis, atopic dermatitis and irritant dermatitis).

• Biomolecules & Therapeutics
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• v.19 no.1
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• pp.134-139
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• 2011

Sphingosine kinase (SPHK) has a central role to control cell death and cell proliferation, which is suggested as a sphingolipid rheostat by regulating the levels between ceramide and sphingosine 1-phosphate (S1P). Therefore, physiological regulators of SPHK will be a good candidate to develop a new targeted drug. For this purpose, a series of synthetic pseudoceramides were tested by SPHK assay either cell-based or cell-free system. K10PC-5 strongly inhibited SPHK, while K6PC-5 activated SPHK in cell-free system. Specifically, K6PC-5 activated SPHK under the co-treatment with $50\;{\mu}M$ dimethylsphingosine (DMS), a SPHK inhibitor. Collectively, we developed a simple SPHK assay system to find SPHK regulatory pseudoceramide compounds, K10PC-5 and K6PC-5 which may be useful to cancer treatment or immune regulation like FTY720, a synthetic sphingolipid mimetic compound.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.6
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• pp.590-595
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• 1991

In order to investigate polar lipid ingredients and fat쇼 acid compositions in corn embryo, lipids were extracted with n-hexane (HX), pet. ether (PE), chloroform-methanol (2:1, v/v) (CM), dichoromethane-methanol (2:1, v/v DM) and hexane-diethyl ether (5:1, v/v HD). Of the glycolipid in polar lipids were separated by thin layer chormatogarphy (TLC), monoglycosyl diacyglycerol was most efficient with CM, HD, and monoglycosy sterol and monoglycosyl ceramide were similar to five solvents, but HX, PE and DM, HD were somewhat superior, respectively. Of the phospholipid, phosphatidyl inositol was most efficient with CM, DM, and phosphatidyl choline was similar to five solvents as well. Phosphatidyl serine was superior PE, HD, CM to HX, and phosphatidyl ethanolamine was inferior CM to HX. The major fatty acid in the glycolipid was linoleic acid, and it was most efficient with CM the same as plamitic acid, but oleic acid was superior in using HX, PE. The major fatty acids in the phospholipid were palmitic, heptadecanoic aicds, and they were superior in using HX and PE, respectively. Also oleic acid was most efficient with HX and CM, but HD was somewhat inferior.