Improved epidermal barrier formation in human skin models by chitosan modulated dermal matrices

Abstract

Full thickness human skin models (FTMs) contain an epidermal and a dermal equivalent. The latter is composed of a collagen dermal matrix which harbours fibroblasts. Current epidermal barrier properties of FTMs do not fully resemble that of native human skin (NHS), which makes these human skin models less suitable for barrier related studies. To further enhance the resemblance of NHS for epidermal morphogenesis and barrier formation, we modulated the collagen dermal matrix with the biocompatible polymer chitosan. Herein, we report that these collagen-chitosan FTMs (CC-FTMs) possess a well-organized epidermis and maintain both the early and late differentiation programs as in FTMs. Distinctively, the epidermal cell activation is reduced in CC-FTMs to levels observed in NHS. Dermal-epidermal interactions are functional in both FTM types, based on the formation of the basement membrane. Evaluation of the barrier structure by the organization of the extracellular lipid matrix of the stratum corneum revealed an elongated repeat distance of the long periodicity phase. The ceramide composition exhibited a higher resemblance of the NHS, based on the carbon chain-length distribution and subclass profile. The inside-out barrier functionality indicated by the transepidermal water loss is significantly improved in the CC-FTMs. The expression of epidermal barrier lipid processing enzymes is marginally affected, although more restricted to a single granular layer. The novel CC-FTM resembles the NHS more closely, which makes them a promising tool for epidermal barrier related studies.

Characterization of human skin equivalents developed at body's core and surface temperatures

“…FTMs were snap frozen in liquid nitrogen using gelatin capsules filled with Tissue‐Tek® O.C.T.™ Compound (Sakura Finetek Europe B.V., Alphen aan den Rijn, The Netherlands) or fixated with 4% formaldehyde (Added Pharma, Oss, The Netherlands), dehydrated and paraffin embedded (Mieremet et al, ).…”

“…SC was isolated from the skin, air‐dried, and stored under Argon gas over silica until further use as described before (Bouwstra, Gooris, van der Spek, & Bras, ; Mieremet, et al, ). Small‐angle X‐ray diffraction measurements were performed at the European Synchrotron Radiation Facility (Grenoble, France) at station BM26B for a period of 2 × 150 s as described elsewhere (Mieremet, et al, ; Mojumdar, Kariman, et al, ). From the positions of a series of equidistant diffraction peaks (located at q n ), the repeat distance (d) of a lamellar phase was calculated using the equation d = n · 2π/q n , where n is the order number of the diffraction peak.…”

3D skin models for 3R research: The potential of 3D reconstructed skin models to study skin barrier function

“…Recent evidence revealed the influence of reducing the relative humidity . Recently, chitosan was introduced in the collagen matrix, which resulted in an improved skin barrier as evaluated by measurement of transepidermal water loss . Optimization of culture conditions will be topic of future studies.…”

“…[89][90][91] Recently, chitosan was introduced in the collagen matrix, which resulted in an improved skin barrier as evaluated by measurement of transepidermal water loss. [92] Optimization of culture conditions will be topic of future studies. Medium composition is also prone to optimization, as multiple studies revealed improved barrier formation after topical application of metabolic active molecules improving the barrier formation in vivo.…”

Hemostatic, anti-inflammatory and antibacterial effects of Sanqixiantao dressing <i>in vivo</i> and <i>in vitro</i>

“…The skin is the largest organ of human body and provides physiological barrier against infections [1,2]. Acute skin wounds usually occur in field battles, traffic accidents and in daily life [3].…”