The Water Permeability of Primary Mouse Keratinocyte Cultures Grown at The Air-Liquid Interface

Cumpstone, Michelle B.; Kennedy, Alane H.; Harmon, Charles S.; Potts, Russell O.

doi:10.1111/1523-1747.ep12712043

Cited by 27 publications

(8 citation statements)

References 14 publications

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“…This is not suitable in our study, as the weight of the Ti disc could damage the interface if it were placed vertically. Some studies have reported performing the permeability test directly on the insert of the cell culture plate, as used in the present study, in which the sample was placed horizontally on the insert without disturbing the interface [24,25].…”

Section: Discussionmentioning

confidence: 99%

The biological seal of the implant–soft tissue interface evaluated in a tissue-engineered oral mucosal model

Chai

Brook

Palmquist

et al. 2012

J. R. Soc. Interface.

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For dental implants, it is vital that an initial soft tissue seal is achieved as this helps to stabilize and preserve the peri-implant tissues during the restorative stages following placement. The study of the implant -soft tissue interface is usually undertaken in animal models. We have developed an in vitro three-dimensional tissue-engineered oral mucosal model (3D OMM), which lends itself to the study of the implant -soft tissue interface as it has been shown that cells from the three-dimensional OMM attach onto titanium (Ti) surfaces forming a biological seal (BS). This study compares the quality of the BS achieved using the threedimensional OMM for four types of Ti surfaces: polished, machined, sandblasted and anodized (TiUnite). The BS was evaluated quantitatively by permeability and cell attachment tests. Tritiated water (HTO) was used as the tracing agent for the permeability test. At the end of the permeability test, the Ti discs were removed from the three-dimensional OMM and an Alamar Blue assay was used for the measurement of residual cells attached to the Ti discs. The penetration of the HTO through the BS for the four types of Ti surfaces was not significantly different, and there was no significant difference in the viability of residual cells that attached to the Ti surfaces. The BS of the tissue-engineered oral mucosa around the four types of Ti surface topographies was not significantly different.

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Section: Discussionmentioning

confidence: 99%

The biological seal of the implant–soft tissue interface evaluated in a tissue-engineered oral mucosal model

Chai

Brook

Palmquist

et al. 2012

J. R. Soc. Interface.

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“…However, this REp is still more permeable to water than normal skin incubated under the same condi tions. Cumpstone et al [21] measured the flux of water through mouse kératinocyte layers cultured on collagen-coated filters which were maintained at the air-liquid inter face. In their experiments, the penetration of water through the REp, when expressed rela tive to normal skin, was much higher than in our assays, most likely because they used fresh untreated skin as the reference.…”

Section: Influence O F the Age O F The Culturesmentioning

confidence: 99%

Reconstructed Human Epidermis: A Model to Study in vitro the Barrier Function of the Skin

Régnier

Caron

Reichert

et al. 1992

Skin Pharmacol Physiol

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In previous studies, we have shown that architecture and differentiation of human epidermis reconstructed by culturing dissociated keratinocytes on deepidermized dermis at the air-liquid interface resembles very much those of epidermis in vivo. The various types of epidermal layers are present with the appropriate differentiation markers, such as the bullous pemphigoid antigen, suprabasal keratins, involucrin, membrane-bound transglutaminase and filaggrin, positioned almost like in normal epidermis. At the ultrastructural level, heterogeneous keratohyalin granules and intra- and extracellular membrane-coating granules are observed. After 7 days of culture, a compact stratum corneum covers the reconstructed epidermis. In the present work, the barrier function of the reconstructed epidermis was evaluated by measuring fluxes of ³H-water. Our results show that under the various culture conditions tested, the presence of the reconstructed epidermis reduces dramatically the permeability of the deepidermized dermis although the skin equivalent exhibits a higher permeability than normal skin. Conditions that favor terminal differentiation of the keratinocytes such as maintaining the cultures in delipidized serum improve the barrier function. Reduction of the relative humidity has a similar effect, whereas the age of the culture is of no influence. Experiments are in progress to reconstruct human epidermis with a barrier function as efficient as in normal skin.

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“…However, since the permeability barrier is a very complex structure, properly assembled in vivo only during undisturbed keratinocyte differentiation, it has been difficult to produce an organotypic culture model with a barrier function comparable with that of intact skin (Cumpstone et al 1989;Ponec et al 1990;Mak et al 1991;Nolte et al 1993;Regnier et al 1993). The high permeability exhibited by organotypic keratinocyte cultures probably reflects abnormalities in the stratum corneum lipid profile (Ponec 1991;Vicanova et al 1996Vicanova et al , 1998b as well as the intercellular organization of lamellar body contents (Boyce and Williams 1993;Fartasch and Ponec 1994), the main determinants of the permeability barrier (Elias and Menon 1991).…”

Section: Introductionmentioning

confidence: 99%

Vitamin C enhances differentiation of a continuous keratinocyte cell line (REK) into epidermis with normal stratum corneum ultrastructure and functional permeability barrier

Pasonen‐Seppänen

Suhonen

Kirjavainen

et al. 2001

Histochem Cell Biol

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A continuous rat epidermal cell line (rat epidermal keratinocyte; REK) formed a morphologically well-organized epidermis in the absence of feeder cells when grown for 3 weeks on a collagen gel in culture inserts at an air-liquid interface, and developed a permeability barrier resembling that of human skin. By 2 weeks, an orthokeratinized epidermis evolved with the suprabasal layers exhibiting the differentiation markers keratin 10, involucrin, and filaggrin. Granular cells with keratohyalin granules and lamellar bodies, and corneocytes with cornified envelopes and tightly packed keratin filaments were present. Morphologically, vitamin C supplementation of the culture further enhanced the normal wavy pattern of the stratum corneum, the number of keratohyalin granules present, and the quantity and organization of intercellular lipid lamellae in the interstices of the stratum corneum. The morphological enhancements observed with vitamin C correlated with improved epidermal barrier function, as indicated by reduction of the permeation rates of tritiated corticosterone and mannitol, and transepidermal water loss, with values close to those of human skin. Moreover, filaggrin mRNA was increased by vitamin C, and western blots confirmed higher levels of profilaggrin and filaggrin, suggesting that vitamin C also influences keratinocyte differentiation in aspects other than the synthesis and organization of barrier lipids. The unique REK cell line in organotypic culture thus provides an easily maintained and reproducible model for studies on epidermal differentiation and transepidermal permeation.

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The Water Permeability of Primary Mouse Keratinocyte Cultures Grown at The Air-Liquid Interface

Cited by 27 publications

References 14 publications

The biological seal of the implant–soft tissue interface evaluated in a tissue-engineered oral mucosal model

The biological seal of the implant–soft tissue interface evaluated in a tissue-engineered oral mucosal model

Reconstructed Human Epidermis: A Model to Study in vitro the Barrier Function of the Skin

Vitamin C enhances differentiation of a continuous keratinocyte cell line (REK) into epidermis with normal stratum corneum ultrastructure and functional permeability barrier

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