Elucidating the molecular landscape of the stratum corneum

Characterization of the molecular structure of skin, especially the barrier layer, the stra-tum corneum, is a key research priority for generating understanding to improve diag-nostics, aid pharmaceutical delivery, and prevent environmental damage. Our study uses the recently developed 3D OrbiSIMS technique to conduct in situ analysis of ex vivo human skin tissue and reveals the molecular chemistry of skin in unprecedented detail, as a result of the step change in high mass resolving power compared with previ-ous studies. This characterization exposes the nonhomogeneity of the stratum corneum, both laterally and as a function of depth. Chemical variations relating to fundamental biological processes, such as the epidermal cholesterol sulfate cycle, are visualized using in situ analysis. We are able to resolve the debate around the chemical gradients present within the epidermis, for example, whether palmitic acid is of sebaceous origin or a true component of the stratum corneum. Through in situ depth analysis of cryogenically preserved samples, we are able to propose that it is actually a component of both surface sebum and the intrinsic lipid matrix. This approach also suggests similarity between the epidermis compounds found in human and porcine skin as a function of depth. Since porcine skin is a widely used model for permeation testing this result has clinical rele-vance. In addition to using this technique for endogenous species, we have used it to demonstrate the permeation of a commercially important antiaging peptide into the human stratum corneum. Due to its chemical similarity to native skin components and exceptionally low effective concentration, this information was previously unattainable.