Tomohiko Sano

The formation of wrinkles caused by transition of keratin intermediate filaments after repetitive UVB exposure

It has been reported that the formation of wrinkles involves changes in the elastic properties of the dermis due to the denaturation of elastic fibers. Several studies have shown that the hydration condition of the stratum corneum is also important in wrinkle formation. It is, however, still unclear how the stratum corneum contributes to wrinkle formation. Here we investigated the relationship between the formation of wrinkles and changes in the physical properties and condition of the skin after repetitive ultraviolet B (UVB) irradiation of hairless mice (HR/ICR). Repetitive UVB irradiation caused wrinkles on the dorsal skin of the mice. The elasticity (E′) of the stratum corneum of UVB-irradiated mice was significantly lower than that of age-matched control (unirradiated) mice. UVB exposure also caused a deterioration of the fibrous ultrastructure of keratin intermediate filaments (KIFs) in the skin. We conclude that the deterioration of KIFs in the stratum corneum caused by repetitive UVB irradiation decreases the elastic properties of the stratum corneum, resulting in the formation of wrinkles.

Structure and Rheology of a Self-Standing Nanoemulsion

A stable nanoemulsion consisting of nanometer-sized oil droplets in water having a self-standing capability was prepared by high-pressure emulsification. The nanoemulsion does not flow and has a yield stress. This nonfluidity is ascribed to the crystal-like lattice structure of nanodroplets. The lattice structure was observed by transmission electron microscopy of a freeze-fractured surface of the specimen. Small-angle neutron scattering (SANS) revealed the presence of an ordered structure in addition to spherical domains with a radius of 17 nm. This long-range order is, in principle, due to electrostatic repulsive interaction between charged nanodroplets. Dynamic light scattering (DLS) showed two relaxation modes, one for the collective motion of the lattice and the other for the translational diffusion of the nanodroplets. Dilution of the nanoemulsion resulted in a transition from a crystal-like structure to a typical colloidal solution.

Evaluation of photoaging in facial skin by multiphoton laser scanning microscopy.

Background/purpose: It has been reported that autofluorescence (AF) and second harmonic generation (SHG) generated in the upper dermis are related with skin photoaging. In this study, we assessed the photoaging of facial skin exposed to daily sunlight using in vivo multiphoton laser microscopy to measure AF and SHG.

Hyperbaric Oxygen Exposure Reduces Age-Related Decrease in Oxidative Capacity of the Tibialis Anterior Muscle in Mice

The effects of exposure to hyperbaric oxygen on the oxidative capacity of the skeletal muscles in mice at different ages were investigated. We exposed 5-, 34-, 55-, and 88-week-old mice to 36% oxygen at 950 mmHg for 6 hours per day for 2 weeks. The activities of succinate dehydrogenase (SDH), which is a mitochondrial marker enzyme, of the tibialis anterior muscle in hyperbaric mice were compared with those in age-matched mice under normobaric conditions (21% oxygen at 760 mmHg). Furthermore, the SDH activities of type IIA and type IIB fibers in the muscle were determined using quantitative histochemical analysis. The SDH activity of the muscle in normobaric mice decreased with age. Similar results were observed in both type IIA and type IIB fibers in the muscle. The decrease in the SDH activity of the muscle was reduced in hyperbaric mice at 57 and 90 weeks. The decreased SDH activities of type IIA and type IIB fibers were reduced in hyperbaric mice at 90 weeks and at 57 and 90 weeks, respectively. We conclude that exposure to hyperbaric oxygen used in this study reduces the age-related decrease in the oxidative capacity of skeletal muscles.

Life cycle of human melanocytes is regulated by endothelin-1 and stem cell factor in synergy with cyclic AMP and basic fibroblast growth factor.

BACKGROUND Although the function of human melanocytes is well characterized at cellular and molecular levels, the mechanism of the regulation of the life cycle (proliferation, differentiation, and cell death) of human melanocytes is not fully understood. OBJECTIVE This study aims to clarify what factors are involved in regulating the life cycle of human melanocytes using serum-free culture system. METHODS Human epidermal melanocytes were cultured in a serum-free growth medium supplemented with several kinds of growth factors, cytokines, and hormones and the effects of these factors on the life cycle of melanocytes were investigated in detail. RESULTS Of the factors tested, endothelin-1 (ET-1) stimulated the proliferation of melanoblasts and melanocytes in the presence of cyclic AMP (cAMP)-elevating factor such as dibutyryl cAMP (DBcAMP) and of basic fibroblast growth factor (bFGF). ET-1 also stimulated the proliferation and differentiation of human melanocytes in the presence of DBcAMP. Moreover, stem cell factor (SCF) stimulated the proliferation of melanoblasts and melanocytes synergistically with ET-1. The removal of ET-1 and SCF from the culture medium greatly inhibited the proliferation of melanocytes followed by apoptotic cell death. CONCLUSION These results suggest that the life cycle of human melanocytes is regulated by ET-1 and SCF in synergy with cAMP and bFGF.

In situ small-angle neutron scattering and rheological measurements of shear-induced gelation.

The microscopic structure of shear-induced gels for a mixed solution of 2-hydroxyethyl cellulose and nanometer-size spherical droplets has been investigated by in situ small-angle neutron scattering (SANS) with a Couette geometry as a function of shear rate gamma. With increasing gamma, the viscosity increased rapidly at gamma approximately 4.0 s(-1), followed by a shear thinning. After cessation of shear, the system exhibited an extraordinarily large steady viscosity. This phenomenon was observed as a shear-induced sol-gel transition. Real-time SANS measurements showed an increase in the scattering intensity exclusively at low scattering angle region. However, neither orientation of polymer chains nor droplet deformation was detected and the SANS patterns remained isotropic irrespective of gamma. It took about a few days for the gel to recover its original sol state. A possible mechanism of gelation is proposed from the viewpoint of shear-induced percolation transition.

Hyperbaric oxygen improves ultraviolet B irradiation-induced melanin pigmentation and diminishes senile spot size.

Background: The effects of exposure to hyperbaric oxygen on ultraviolet B (UVB) irradiation‐induced melanin pigmentations of skins and on senile spot sizes of faces were investigated.

Evaluation of unique elastic aggregates (elastic globes) in normal facial skin by multiphoton laser scanning tomography

BackgroundThere is no reliable marker to estimate the degree of skin aging in vivo. It now has become possible to quantitatively determine the dermal characteristics of the extracellular matrix (ECM) in vivo using multiphoton laser tomography (MLT).MethodsFifty-seven healthy Japanese female volunteers, aged from 20 to 60 years old, were examined using multiphoton depth-resolved measurements of autofluorescence (AF) and second harmonic generation (SHG) at three sites on their right cheek. Paraffin-embedded skin specimens obtained from the faces of 12 normal individuals aged 38-68 years old were stained with Elastica van Gieson (EVG).ResultsWe found unique elastic aggregates at a 20µm depth from the dermo-epidermal junction (DEJ) in vivo which increased in size with aging of subjects from 20 to 60 years old. SHG fibers seemed to surround those elastic aggregates. Histological examination of specimens from normal individuals stained with EVG confirmed the occurrence of elastic aggregates with varied sizes just beneath the epidermis or hair follicles.ConclusionsThe elastic aggregates are morphologically similar to previously described ‘elastic globes’ and can serve as a marker of the early stage of photoaging. MLT will contribute to determine age-related dermal changes using a non-invasive technique.

Relationship between the echogenicity of subcutaneous tissue and the depth of eye corner wrinkles.

Background/purpose: A close relationship was found between the internal structure of subcutaneous tissue and wrinkle depth in our previous study of human forehead. The present study examined the relationship in the eye corner.

Relationship between the echogenicity of subcutaneous tissue and the depth of forehead wrinkles: Forehead wrinkle profile by ultrasonography

Background/purpose: Subcutaneous tissue is rarely studied in research on wrinkles. We used diagnostic ultrasonography to produce images of subcutaneous tissue of the forehead, one of the areas where age‐related wrinkles form. We quantified the resulting echogenicity and investigated its relationship with wrinkle depth.