Ubiquinone (coenzyme Q10, Q10) represents an endogenously synthesized lipid‐soluble antioxidant which is crucial for cellular energy production but is diminished with age and under the influence of external stress factors in human skin. Here, it is shown that topical Q10 treatment is beneficial with regard to effective Q10 replenishment, augmentation of cellular energy metabolism, and antioxidant effects. Application of Q10‐containing formulas significantly increased the levels of this quinone on the skin surface. In the deeper layers of the epidermis the ubiquinone level was significantly augmented indicating effective supplementation. Concurrent elevation of ubiquinol levels suggested metabolic transformation of ubiquinone resulting from increased energy metabolism. Incubation of cultured human keratinocytes with Q10 concentrations equivalent to treated skin showed a significant augmentation of energy metabolism. Moreover, the results demonstrated that stressed skin benefits from the topical Q10 treatment by reduction of free radicals and an increase in antioxidant capacity. © 2015 BioFactors, 41(6):383–390, 2015

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Preservation of skin in a "close-to -native state" without the loss of lipids can be achieved by high-pressure freezing, followed by cryo-sectioning and investigation in a Cryo-TEM (sections) or Cryo-SEM (remaining "bloc-face"). Therefore, vitrified samples were cryo sectioned at 100 K and investigated in the frozen-hydrated state at 103 K . An alternative way to minimize loss of lipids and ultra structural changes, as determined by mass spectroscopy, is freeze-substitution in the presence of acetone saturated with uranyl acetate and embedding at low temperatures followed by sectioning and investigation at room temperature (1). We compared the skin ultrastructure in cryosections ( Fig. 3C ~70 nm thickness) with that of skin in resin sections ( Fig. 3A&B) to determine the structural changes occurring with each procedure. We found ultrastructural details e.g. corneodesmosomes and lamellar bodies, in the process of fusion with the plasma membrane at the SC/S.granulosum interface in both types of sections. Cryosections show a strong deformation in the direction of the sectioning process. These deformations render study of the fine structure to a difficult task, and interpretation of fine details can only be performed in selected areas of a cryosection whereas no deformation of the structure could be found on the corresponding "bloc-face" observed by Cryo-SEM (2). Our experience shows, nevertheless, that it is possible to investigate the ultrastructure of skin without the use of any chemical fixatives or dehydration techniques. But it also clearly reveals a significantly different lipid and protein (e.g. filaggrin) distribution when samples were chemically fixed prior to high pressure freezing (Fig.1&2). In general lipids are redistributed over the entire epidermis during or after the chemical fixation process (Fig. 1A), whereas protein e.g. filaggrin are masked by the chemical fixative ( Fig. 2A)! Consequently, a routine comparison of the lipid structure and a better understanding of the ultrastructure of the viable skin and mainly the skin barrier are only possible by comparing EM and LM data (3) of skin biopsies, which have been prepared by "close-to-native state" preserving preparation protocols and approved by cryosectioning for cryo-TEM. Figure 1-3: High pressure frozen human skin biopsie; 1-3 A) chemical fixation prior to high pressure freezing followed by identical freeze substitution in Acetone/UrAc as for 1-3 B). 3C Frozen hydrated TEM section. Labeling for filaggrin (1A-B) and glucosylceramid 3 (2A-B).