Arrangement of Ceramides in the Skin: Sphingosine Chains Localize at a Single Position in Stratum Corneum Lipid Matrix Models.

Understanding the structure of the stratum corneum (SC) is essential to understand the skin barrier process. The long periodicity phase (LPP) is a unique trilayer lamellar structure located in the SC. Adjustments in the composition of the lipid matrix, as in many skin abnormalities, can have severe effects on the lipid organization and barrier function. Although the location of individual lipid subclasses has been identified, the lipid conformation at these locations remains uncertain. Contrast variation experiments via small-angle neutron diffraction were used to investigate the conformation of ceramide (CER) N-(tetracosanoyl)-sphingosine (NS) within both simplistic and porcine mimicking LPP models. To identify the lipid conformation of the twin chain CER NS, the chains were individually deuterated, and their scattering length profiles were calculated to identify their locations in the LPP unit cell. In the repeating trilayer unit of the LPP, the acyl chain of CER NS was located in the central and outer layers, while the sphingosine chain was located exclusively in the middle of the outer layers. Thus, for the CER NS with the acyl chain in the central layer, this demonstrates an extended conformation. Electron density distribution profiles identified that the lipid structure remains consistent regardless of the lipid's lateral packing phase, this may be partially due to the anchoring of the extended CER NS. The presented results provide a more detailed insight on the internal arrangement of the LPP lipids and how they are expected to be arranged in healthy skin.

Recombinant COVID-19 vaccine based on recombinant RBD/Nucleoprotein and saponin adjuvant induces long-lasting neutralizing antibodies and cellular immunity.

SARS-CoV-2 has caused a global pandemic, infecting millions of people. An effective preventive vaccine against this virus is urgently needed. Here, we designed and developed a novel formulated recombinant receptor-binding domain (RBD) nucleocapsid (N) recombinant vaccine candidates. The RBD and N were separately expressed in E. coli and purified using column chromatography. The female Balb/c mice were immunized subcutaneously with the combination of purified RBD and N alone or formulated with saponin adjuvant in a two-week interval in three doses. Neutralization antibody (Nabs) titers against the SARS-CoV-2 were detected by a Surrogate Virus Neutralization (sVNT) Test. Also, total IgG and IgG1, and IgG2a isotypes and the balance of cytokines in the spleen (IFN-γ, Granzyme B, IL-4, and IL-12) were measured by ELISA. The percentages of CD4+ and CD8+ T cells were quantified by flow cytometry. The lymphoproliferative activity of restimulated spleen cells was also determined. The findings showed that the combination of RBD and N proteins formulated with saponin significantly promoted specific total IgG and neutralization antibodies, elicited robust specific lymphoproliferative and T cell response responses. Moreover, marked increase in CD4+ and CD8+ T cells were observed in the adjuvanted RBD and N vaccine group compared with other groups. The results suggest that the formulations are able to elicit a specific long-lasting mixed Th1/Th2 balanced immune response. Our data indicate the significance of the saponin-adjuvanted RBD/N vaccine in the design of SARS-CoV-2 vaccines and provide a rationale for the development of a protective long-lasting and strong vaccine.

Supramolecular architecture of a multi-component biomimetic lipid barrier formulation.

HYPOTHESIS: Biomimetic liquid crystalline systems are widely used in skin care cosmetics and topical pharmaceutical preparations. Our ability to rationally design such formulations, however, is hampered by our incomplete understanding of their structure on the nanoscale. EXPERIMENTS: Using polarized light microscopy and small-angle and wide-angle X-ray scattering, the molecular architecture and properties of a barrier formulation prepared from distearoylphosphatidylcholine mixed with long chain fatty acid and alcohols, with and without antimicrobial pentanediols are directly probed. The nature and composition of the phases identified are determined through small-angle neutron scattering studies using chain-deuterated components, and the detailed structure and dynamics of the gel network lamellae are determined through molecular dynamics simulations. FINDINGS: The formulations show molecular ordering with long and short periodicity lamellar phases and there is little change in these structures caused by changes in temperature, drying, or the application of shear stress. The diol-free formulation is demonstrated to be self-preserving, and the added pentanediols are shown to distribute within the interlamellar regions where they limit availability of water for microbial growth. In culmination of these studies, we develop a more complete picture of these complex biomimetic preparations, and thereby enable their structure-based design.

The Influence of Co-Surfactants on Lamellar Liquid Crystal Structures Formed in Creams.

It is well-established that oil-in-water creams can be stabilised through the formation of lamellar liquid crystal structures in the continuous phase, achieved by adding (emulsifier) mixtures comprising surfactant(s) combined (of necessity) with one or more co-surfactants. There is little molecular-level understanding, however, of how the microstructure of a cream is modulated by changes in co-surfactant and of the ramifications of such changes on cream properties. We investigate here the molecular architectures of oil-free, ternary formulations of water and emulsifiers comprising sodium dodecyl sulfate and one or both of the co-surfactants hexadecanol and octadecanol, using microscopy, small-angle and wide-angle X-ray scattering and small-angle neutron scattering. We then deploy these techniques to determine how the structures of the systems change when liquid paraffin oil is added to convert them to creams, and establish how the structure, rheology, and stability of the creams is modified by changing the co-surfactant. The ternary systems and their corresponding creams are shown to contain co-surfactant lamellae that are subtly different and exhibit different thermotropic behaviours. The lamellae within the creams and the layers surrounding their oil droplets are shown to vary with co-surfactant chain length. Those containing a single fatty alcohol co-surfactant are found to contain crystallites, and by comparison with the cream containing both alcohols suffer adverse changes in their rheology and stability.

Revealing the Hidden Details of Nanostructure in a Pharmaceutical Cream.

Creams are multi-component semi-solid emulsions that find widespread utility across a wide range of pharmaceutical, cosmetic, and personal care products, and they also feature prominently in veterinary preparations and processed foodstuffs. The internal architectures of these systems, however, have to date been inferred largely through macroscopic and/or indirect experimental observations and so they are not well-characterized at the molecular level. Moreover, while their long-term stability and shelf-life, and their aesthetics and functional utility are critically dependent upon their molecular structure, there is no real understanding yet of the structural mechanisms that underlie the potential destabilizing effects of additives like drugs, anti-oxidants or preservatives, and no structure-based rationale to guide product formulation. In the research reported here we sought to address these deficiencies, making particular use of small-angle neutron scattering and exploiting the device of H/D contrast variation, with complementary studies also performed using bright-field and polarised light microscopy, small-angle and wide-angle X-ray scattering, and steady-state shear rheology measurements. Through the convolved findings from these studies we have secured a finely detailed picture of the molecular structure of creams based on Aqueous Cream BP, and our findings reveal that the structure is quite different from the generic picture of cream structure that is widely accepted and reproduced in textbooks.