In vitro reconstitution of stratum corneum lipid lamellae

In the final stages of differentiation in the epidermis of terrestrial mammals, lipids are extruded into the intercellular spaces. The initially extruded lipid becomes transformed into broad, multilamellar sheets that are found in the intercellular spaces throughout the stratum corneum. These lamellae display an unusual alternating broad-narrow-broad pattern of lucent bands as revealed by transmission electron microscopy (TEM). This arrangement results in two periodicities that can be measured from electron micrographs and are also evident in X-ray diffraction-5 nm (broad) and 13 nm (broad-narrow-broad). The goal of the present study was to reconstitute these lamellae in vitro. Porcine stratum corneum lipids were applied to Millipore filters. The disks were placed in water and heated to 80 degrees C for 1 h. After cooling, the disks were stored over desiccant. At each stage, the disks were prepared for TEM. TEM revealed that the application of the lipid solutions onto the disks resulted to deposition of mostly amorphous material. Heating in water resulted in the formation of many lamellae. The width of the lamellae was uniform and in the range of 5 to 6 nm with no broad-narrow-broad pattern; however, after storage under desiccating conditions, the broad-narrow-broad pattern was reproduced.