Preparation of Model Intercellular Membrane Using Low-Toxic Solvent Ethanol.

The stratum corneum, the outermost layer of the skin, is composed of keratinized cells known as corneocytes and intercellular lipids (IC lipids) such as ceramides (Cers), cholesterol (Chol), and free fatty acids (FFAs). These lipids form highly ordered multilamellar membranes in the intercellular spaces, serving as a key regulator of the skin barrier function. To investigate the mechanistic relation between the membrane structure and the skin barrier function, model membranes composed of Cers/Chol/FFA mixtures have been widely employed. In most of those studies, highly toxic solvents such as chloroform (CHCl3) and methanol (MeOH) were used to dissolve and mix the IC lipids. However, growing concerns regarding safety and environmental hygiene have driven the selection of safer alternative solvents. In this study, we developed a new protocol for preparing model membranes using less toxic ethanol (EtOH). In the EtOH-based protocol, IC lipids were dissolved in near-boiling EtOH, diluted by a fivefold large volume of buffer, and cooled to 30°C. After 18 h of equilibration at 30°C, the model intercellular lamellae were collected by centrifugation. In addition, we demonstrated that the lamellar sample exhibited thermodynamic and structural properties comparable to those of the lamellae prepared by the conventional CHCl3/MeOH-based protocols. Moreover, the developed EtOH-based protocol enhanced the miscibility of IC lipids to some extent. Therefore, the EtOH-based protocol would be a useful methodology for preparing model intercellular membranes, being well-suited for environmental hygiene.

• Publication year

  2025

• Venue

  Chemical and pharmaceutical bulletin

• Publication date

  2025-10-16

• Fields of study

  Medicine, Materials Science, Chemistry

• Identifiers
  DOI 10.1248/cpb.c25-00332PMID 41093595
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Biomimetic Stratum Corneum Liposome Models: Lamellar Organization and Permeability Studies

  2023cited by this paper
• Distribution of Domains Formed by Lateral Packing of Intercellular Lipid in the Stratum Corneum.

  2023cited by this paper
• Investigating the nanostructure of a CER[NP]/CER[AP]-based stratum corneum lipid matrix model: A combined neutron diffraction & molecular dynamics simulations approach.

  2022cited by this paper
• Phase Separation in Lipid Lamellae Result from Ceramide Conformations and Lateral Packing Structure.

  2021cited by this paper
• The Importance of Free Fatty Chain Length on the Lipid Organization in the Long Periodicity Phase

  2021cited by this paper
• Roles of Lipids in the Permeability Barriers of Skin and Oral Mucosa

  2021cited by this paper
• Dual-functional liposomes for curcumin delivery and accelerating silk fibroin hydrogel formation.

  2020cited by this paper
• New insight into phase behavior and permeability of skin lipid models based on sphingosine and phytosphingosine ceramides.

  2019cited by this paper
• Molecular Interaction between Intercellular Lipids in the Stratum Corneum and l-Menthol, as Analyzed by Synchrotron X-Ray Diffraction.

  2017cited by this paper
• Potential of Stratum Corneum Lipid Liposomes for Screening of Chemical Skin Penetration Enhancers.

  2017cited by this paper
• Analysis of the pretreatment effect of ethanol on the stratum corneum- and hair follicular-penetration of drugs using the hair follicle-plugging method.

  2014cited by this paper
• Ceramides in the skin lipid membranes: length matters.

  2013cited by this paper
• SC lipid model membranes designed for studying impact of ceramide species on drug diffusion and permeation, part III: influence of penetration enhancer on diffusion and permeation of model drugs.

  2012cited by this paper
• Ethanol perturbs lipid organization in models of stratum corneum membranes: An investigation combining differential scanning calorimetry, infrared and (2)H NMR spectroscopy.

  2012cited by this paper
• The structure and function of the stratum corneum.

  2012cited by this paper
• A possible regulation mechanism of water content in human stratum corneum via intercellular lipid matrix.

  2012cited by this paper
• The thermotropism and prototropism of ternary mixtures of ceramide C16, cholesterol and palmitic acid. An exploratory study.

  2011cited by this paper
• Crystalline lipid domains: characterization by X-ray diffraction and their relation to biology.

  2011cited by this paper
• Model membranes prepared with ceramide EOS, cholesterol and free fatty acids form a unique lamellar phase.

  2010cited by this paper
• Novel preparation of intercellular lipid models of the stratum corneum containing stereoactive ceramide.

  2010cited by this paper
• Preparation and characterization of a stratum corneum substitute for in vitro percutaneous penetration studies.

  2006cited by this paper
• Coexistence of two domains in intercellular lipid matrix of stratum corneum.

  2006cited by this paper
• Aberrant lipid organization in stratum corneum of patients with atopic dermatitis and lamellar ichthyosis.

  2001cited by this paper
• Determination of stratum corneum lipid profile by tape stripping in combination with high-performance thin-layer chromatography

  2001cited by this paper
• Lipids and barrier function of the skin.

  2000cited by this paper
• The lipid organisation in the skin barrier.

  2000cited by this paper
• Electron diffraction provides new information on human stratum corneum lipid organization studied in relation to depth and temperature.

  1999cited by this paper
• Preparation of liposomes from stratum corneum lipids.

  1986cited by this paper

• No citing papers are available for this paper.