Canine epidermal lipid sampling by skin scrub revealed variations between different body sites and normal and atopic dogs.

BACKGROUND: Previously, we evaluated a minimally invasive epidermal lipid sampling method called skin scrub, which achieved reproducible and comparable results to skin scraping. The present study aimed at investigating regional variations in canine epidermal lipid composition using the skin scrub technique and its suitability for collecting skin lipids in dogs suffering from certain skin diseases. Eight different body sites (5 highly and 3 lowly predisposed for atopic lesions) were sampled by skin scrub in 8 control dogs with normal skin. Additionally, lesional and non-lesional skin was sampled from 12 atopic dogs and 4 dogs with other skin diseases by skin scrub. Lipid fractions were separated by high performance thin layer chromatography and analysed densitometrically. RESULTS: No significant differences in total lipid content were found among the body sites tested in the control dogs. However, the pinna, lip and caudal back contained significantly lower concentrations of ceramides, whereas the palmar metacarpus and the axillary region contained significantly higher amounts of ceramides and cholesterol than most other body sites. The amount of total lipids and ceramides including all ceramide classes were significantly lower in both lesional and non-lesional skin of atopic dogs compared to normal skin, with the reduction being more pronounced in lesional skin. The sampling by skin scrub was relatively painless and caused only slight erythema at the sampled areas but no oedema. Histological examinations of skin biopsies at 2 skin scrubbed areas revealed a potential lipid extraction from the transition zone between stratum corneum and granulosum. CONCLUSIONS: The present study revealed regional variations in the epidermal lipid and ceramide composition in dogs without skin abnormalities but no connection between lipid composition and predilection sites for canine atopic dermatitis lesions. The skin scrub technique proved to be a practicable sampling method for canine epidermal lipids, revealed satisfying results regarding alterations of skin lipid composition in canine atopic dermatitis and might be suitable for epidermal lipid investigations of further canine skin diseases. Although the ceramide composition should be unaffected by the deeper lipid sampling of skin scrub compared to other sampling methods, further studies are required to determine methodological differences.

Comparison of three different sampling methods for canine skin lipids.

BACKGROUND: Epidermal lipids are of major interest in dermatological research, especially in canine atopic dermatitis. Owing to the existence of several sampling methods, the interpretation of study results is often complicated. OBJECTIVES: This study aimed to compare three different sampling methods and to establish a minimally invasive method for collecting canine epidermal lipids. ANIMALS AND METHODS: Skin samples from five dogs with no obvious skin abnormalities were taken from the caudal back and the inguinal region postmortem. Samples consisted of heat-separated epidermis of three skin biopsies, three scrapes and three skin scrubs. Lipids were analysed by high-performance thin-layer chromatography; the resulting bands were identified by using corresponding standards, retardation factors and mass spectrometry. The influences of the sampling method, the body site and the ceramide standards were investigated. RESULTS: Between body sites, significant differences were found for cholesterol sulphate, cholesteryl esters and triglycerides. Significant differences between sampling methods were detected for all lipid fractions except for cholesterol sulphate and glucosylceramides within the lipid profile, and for at least four ceramide classes within the ceramide profile. The most obvious discrepancies were found between heat-separated epidermis and skin scrub. The reproducibility was high for scraping and skin scrub, but was lowest for heat-separated epidermis. Furthermore, this study revealed a marked influence of ceramide standards on the results regarding the ceramide profile. CONCLUSIONS AND CLINICAL IMPORTANCE: Scraping and skin scrub are comparably suitable methods for skin lipid sampling, whereas the analysis of heat-separated epidermis may not be the method of first choice.