In vivo evaluation in man by two noninvasive methods of the stratum corneum barrier function after physical and chemical modifications

This study investigated the effect of different treatments on skin permeability, in vivo in man, by two noninvasive methods: transepidermal water loss (TEWL) determination measured with an evaporimeter, and Laser Doppler velocimetry (LDV) to measure the lag time before the vasodilatation induced by application of methyl and hexyl nicotinates.
Different treatments were performed on forearms of volunteers: 1. removal of the stratum corneum by stripping, 2. occlusion and hydration of the stratum corneum, 3. application of three surfactants in aqueous solution (Tween 60, sodium dodecyl sulphate and cetyl trimethylammonium bromide).
Increase in TEWL and decrease in lag time before the vasodilating effects of nicotinates measured with LDV after all treatments confirmed the modifications in skin permeability. The variations observed were more or less significant depending on the treatment, the investigative method, and the molecule used for the penetration study. With methyl (hydrophilic) nicotinate, the more TEWL increased, the more lag time decreased. A significant decrease in lag time after hexyl (lipophilic) nicotinate application was observed when there was a very high increase in TEWL. The lipophilic nature of the stratum corneum barrier was confirmed in vivo. Use of these two complementary methods, TEWL and LDV, allows the evaluation in vivo of the effect of different treatments on skin permeability in man.     

Surface distribution and depths profiling of particulate organic UV absorbers by Raman imaging and tape stripping

Tape stripping in conjunction with scanning Raman microscopy was used for assessing the lateral and vertical distribution of an organic particulate UV filter, methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT), in a sunscreen formulation. On the volar forearms of three volunteers, 1 mg cm(-2) formulation containing 10% MBBT was applied, and the average amount of MBBT was measured by Raman scanning microscopy in 15 consecutive tape strippings. The recovery of MBBT was 91.1% with 30.2% localizing on the skin surface (first strip), 42.5% in the upper stratum corneum (strips 2-5) and from 3.6 down to 0.8% in each of the 10 consecutive layers. The concentration of surface deposits of MBBT differed by a factor of 300 between folds, furrows and pores on the one hand and the interjacent ridges on the other hand. Seventy-five per cent of the applied particles occupied a fifth of the evaluated area - where concentrating in folds and furrows - as was confirmed by 3-D reconstruction. On interjacent ridges, 8.6% of MBBT distributed as very thin films preferentially. MBBT localized at sites not connected with the surface, such as in truncated pores or as potentially penetrated material amounted to 0.06% or to a twentieth of the 1.4% found in the lowest skin strippings. Scanning Raman microscopy in combination with tape stripping documented the lateral and vertical distribution quantitatively and at cellular (12.5 μm) lateral resolution. Our results confirmed an earlier report on the vertical distribution of organic particles applied to skin and was in line with similar reports on TiO(2) distribution.     

Influence of topically applied ceramide/phospholipid mixture on the barrier function of intact skin, atopic skin and experimentally induced barrier damage

The influence of a liposomal formulation of a ceramide/phospholipid mixture on the barrier function of the skin in atopic tests, in subjects with healthy skin, and after lipid extraction was tested. The transepidermal water loss and the stratum corneum hydration served as parameters for the evaluation of the influence of the ceramide/phospholipid-mixture treatment on the barrier function. Additionally a barrier-function test with nicotinic acid ester was performed after ceramide/phospholipid-mixture application. Stabilizing effects on the barrier function after application of ceramide/phospholipid-mixture were found only in atopic skin and after lipid extraction but not in healthy skin. The results lead us to expect positive effects of ceramide/phospholipid-mixture only if barrier damage is present.     

Non-contact laser-scanning confocal microscopy of the human cornea in vivo

PurposeTo investigate the utility of using non-contact laser-scanning confocal microscopy (NC-LSCM), compared with the more conventional contact laser-scanning confocal microscopy (C-LSCM), for examining corneal substructures in vivo.MethodsAn attempt was made to capture representative images from the tear film and all layers of the cornea of a healthy, 35 year old female, using both NC-LSCM and C-LSCM, on separate days.ResultsUsing NC-LSCM, good quality images were obtained of the tear film, stroma, and a section of endothelium, but the corneal depth of the images of these various substructures could not be ascertained. Using C-LSCM, good quality, full-field images were obtained of the epithelium, subbasal nerve plexus, stroma, and endothelium, and the corneal depth of each of the captured images could be ascertained.ConclusionsNC-LSCM may find general use for clinical examination of the tear film, stroma and endothelium, with the caveat that the depth of stromal images cannot be determined when using this technique. This technique also facilitates image capture of oblique sections of multiple corneal layers. The inability to clearly and consistently image thin corneal substructures – such as the tear film, subbasal nerve plexus and endothelium – is a key limitation of NC-LSCM.     

In vivo barrier challenge and long‐term recovery in human facial skin

Recently, we developed a biophysical approach to characterize in vivo facial cheek skin as a function of stratum corneum (SC) depth, barrier function and during a 24‐h recovery period. The current study extends this work and characterizes the human facial cheek after barrier challenge and, for the first time, facial SC barrier recovery over a 4‐week period. Changes in the corneocyte size over the 4‐week recovery period, and correlations with changes in Trans‐Epidermal Water Loss (TEWL) were monitored. This approach allows complete characterization of SC barrier function after a full biological regeneration of the SC barrier following tape stripping. The structural and compositional changes in facial cheek were investigated using Attenuated Total Reflectance‐Fourier Transform Infra Red (ATR‐FTIR) spectroscopy, tape stripping, TEWL measurements and image analysis combined with optical microscopy to characterize the SC depth profile during the tape stripping stress and over 4‐week recovery period. TEWL increased significantly from baseline after sequential tape stripping. Corneocyte size decreased with successive tape stripping. An inverse direct correlation was determined between TEWL and corneocyte surface area. After 4 weeks, the corneocyte size and TEWL for the facial cheek recovered 100% from the tape stripping procedure. The in vivo ATR‐FTIR data demonstrated that lipid and sebum components on the surface of the facial cheek SC recovered within 24 h post tape stripping, whereas protein (Amide II) and water components recovered after 1 week.     

Mobile connected dermatoscope and confocal laser scanning microscope: a useful combination applied in facial simple sensitive skin

Little is known as the effects of mobile connected dermatoscope services on diagnostic accuracy for sensitive skin. Confocal laser scanning microscope (CLSM) can non-invasively measure the thickness of epidermis. Combination of the two devices to observe sensitive skin may receive unexpected effects. To evaluate the application effect on sensitive skin with the combination of Handyscope and confocal laser scanning microscope. Twenty simple sensitive-skinned patients and 20 volunteers participated in the study. Cheek, typically, dermoscopic images were obtained from patients, and the changes in the skin texture were observed. Their epidermis thicknesses as well as the volunteers' were measured so that the thicknesses of the two groups were compared. Dermoscopic pictures of the skin texture obviously showed that dilated capillaries looked like earthworms with pigmented patches more or less floating above, and skin roughness as well as deepened dermatoglyph were also conspicuously present in some patients. The mean epidermal thickness of the patients was 79.01 μm and the volunteers' was 85.78 μm. The difference between the two groups reached 6.77 μm. There was a statistical significance (P = 0.001). Mobile connected dermatoscope and confocal laser scanning microscope might be the choice for simple sensitive skin investigation.     

Retrospective Shading Correction of Confocal Laser Scanning Microscopy Beef Images for Three-Dimensional Visualization

Because of the inherent imperfections of the image formation process, confocal laser scanning microscopy (CLSM) images are often corrupted by spurious intensity variations not present in the original scene, which is usually referred to as shading or intensity inhomogeneity. In this paper, a retrospective shading correction method for CLSM beef images was developed using a hybrid of image processing algorithms. A partial-differential-equation-based diffusion technique was applied, firstly, to reduce the additive shading component. To reduce the computational burden, a thresholding segmentation method was then presented to separate the muscle tissues from the background. After that, a robust, automatic, and fast method was applied for reduction of multiplicative shading component. The corrected images were finally used to construct a three-dimensional beef image, which could provide a valuable source of knowledge about beef microstructure that cannot be obtained via traditional two-dimensional images.     

In vivo confocal fluorescence imaging of skin surface cellular morphology: a pilot study of its potential as a clinical tool in skin research

The cellular morphology of the stratum corneum was studied in vivo using a novel imaging technique that uses confocal fluorescence microscopy in combination with topical application of a fluorescent contrast agent. Images obtained with this method show a strong variation in skin surface cellular morphology among healthy subjects. The results of several clinical studies suggest that cellular morphology is affected by the efficiency of the process of desquamation. As such, cellular morphology shows strong potential to serve as an indicator of skin health that yields mechanistic insight into the origins of skin ailments, such as xerosis, and the effectiveness of their treatments.     

An in vivo assessment of the biomechanical properties of human skin modifications under the influence of cosmetic products

A new device using a torsional technique has been developed to record the biomechanical properties of the skin.
After determination of the optimal experimental conditions, effects of cosmetic products on the stratum corneum biomechanical properties are described.     

Characterization of fibrillar collagen types using multi-dimensional multiphoton laser scanning microscopy

In dermal photodamage the ratio of the collagen types III to I changes. This makes the investigation of the fibrillar collagen type characteristics interesting for skin research. In this study collagen types were characterized using 5-dimensional multiphoton laser scanning microscopy (5D-IVT) that can be applied in vivo. Second harmonic generation (SHG) signals and fluorescence lifetimes of the collagen autofluorescence were analysed. Collagen type I generates a higher SHG intensity and a longer fluorescence lifetime compared to collagen type III. Thus, the SHG intensity decrease found in photodamaged skin might be explained by the increase in collagen type III. Calculating the in vivo relevant increase of collagen type III gives a negligible difference in fluorescence lifetime not qualifying this method for the determination of collagen type changes in dermal photodamage in vivo in human skin. However, for pathologies that exhibit higher differences in collagen types 5D-IVT analysis might be a suitable method.     

Microstructure and rheology of yogurt made with cultures differing only in their ability to produce exopolysaccharides

Yogurt was made using an exopolysaccharide-producing strain of Streptococcus thermophilus and its genetic variant that only differed from the mother strain in its inability to produce exopolysaccharides. The microstructure was investigated using confocal scanning laser microscopy, allowing observation of fully hydrated yogurt and the distribution of exopolysaccharide within the protein network. Yogurt made with the exopolysaccharide-producing culture exhibited increased consistency coefficients, but lower flow behavior index, yield stress, viscoelastic moduli and phase angle values than did yogurt made with the culture unable to produce exopolysaccharide. The exopolysaccharides, when present, were found in pores in the gel network separate from the aggregated protein. These effects could be explained by the incompatibility of the exopolysaccharides with the protein aggregates in the milk. Stirring affected the yogurt made with exopolysaccharide differently from yogurt without exopolysaccharide, as it did not exhibit immediate syneresis, although the structural breakdown was increased. The shear-induced microstructure in a yogurt made with exopolysaccharide-producing culture was shown to consist of compartmentalized protein aggregates between channels containing exopolysaccharide, hindering syneresis as well as the buildup of structure after stirring.     

Chemical and structural characterisation of almond oil bodies and bovine milk fat globules

Lipids in almonds are present as oil bodies in the nut. These oil bodies are surrounded by a membrane of proteins and phospholipids and are a delivery vehicle of energy in the form of triglycerides, similarly to the more studied bovine milk fat globule membrane. Chemical, physical and microscopic analyses revealed major differences in the composition and structure of almond oil bodies and bovine milk fat globules. The lipids of both natural emulsions differed in degree of unsaturation, chain length, and class. The almond oil body membrane does not contain any cholesterol or sphingomyelin unlike the bovine milk fat globule membrane. Therefore, the phospholipid distribution at the surface of the oil bodies did not present any liquid-ordered domains. The membranes, a monolayer around almond oil bodies and a trilayer around bovine fat globules, may affect the stability of the lipid droplets in a food matrix and the way the lipids are digested.     

油茶油体观察及其与含油量的相关性

为建立快速观察油茶种子油体的方法,揭示油茶种子含油量与其子叶贮藏细胞内油体之间的相关性,采用冰冻切片、荧光染料尼罗红染色和激光扫描共聚焦显微镜观察相结合技术,对不同含油量的7个油茶品系种子油体进行观察。研究表明:油茶油体为球形,直径平均为2.52μm,标准差为1.99μm,且不同含油量的油茶种子油体分布和数量存在明显差异,油茶种子油体截面积之和与其含油量呈显著正相关。研究结果可为油茶高油品系的选育提供理论依据。     

The influence of inulin addition on the morphological and structural properties of durum wheat pasta

The effect of the addition of different concentration of inulin fibre to durum wheat pasta was evaluated in terms of structure, texture and morphological characteristics. Inulin fibre fraction was incorporated into pasta at 5%, 10% and 15% inclusion rates. Changes to pasta quality were observed with inulin fibre fraction addition by scanning electron microscopy, X-ray diffraction and Raman spectroscopy methods. Investigation of pasta microstructure indicate a lowering in crystallinity increasing the inulin content and characterisation of starch gelatinisation events indicated that a combination of changes to the starch–protein matrix and the high water-binding capacity of inulin alters the physico-chemical properties and then the digestibility of the pastas.     

A study of the penetration of five novel synthetic steroids through hairless mouse skin in vitro

The change in the physiochemical properties of topical corticosteroids by addition of one or more chemical substituents is probably one of the most important factors affecting the activity and bioavailability of the steroid. During work on developing new steroids five synthetic steroids were tested for their ability to pass through freshly excised hairless mouse skin placed in diffusion cells at 33 1°C. Analysis of the compounds was made using high performance liquid chromatography. The presence of flourine at carbon 9, in the chemical grouping at carbon 17, or the addition of a pentyl group was found to increase the penetration of the steroid through the skin.     

自水解预处理对杨木聚木糖分布的影响

采用免疫荧光标记法结合共聚焦显微镜(CLSM)研究了自水解强度对杨木聚木糖分布的影响。结果表明,聚木糖荧光信号较均匀地分布于杨木纤维细胞壁中,随强度因子的增大细胞壁中心部位荧光信号下降逐渐增多,细胞壁边缘荧光信号下降较少,有些部位甚至会稍许增加。采用高效液相色谱(HPLC)检测法测得强度因子增强到3.72时,聚木糖含量从原料的14.64%下降到7.33%;对聚木糖荧光图谱进行区域强度统计,可得出与HPLC检测法结果相同的变化趋势。