Culturing Keratinocytes and Fibroblasts in a Three-Dimensional Mesh Results in Epidermal Differentiation and Formation of a Basal Lamina-Anchoring Zone

Contard, Paul; Bartel, Ronnda L.; Jacobs, Lloydstone; Perlish, Jerome S.; MacDonald, Ewan; Handler, Lionel; Cone, Diana; Fleischmajer, Raúl

doi:10.1111/1523-1747.ep12349952

Cited by 107 publications

(56 citation statements)

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“…Formation of a regular epidermal tissue architecture had been achieved in vitro in an organotypic three-dimensional coculture system (3D-coculture) by growing NEK at the air-liquid interface on top of type I collagen gels populated with skin fibroblasts (Asselineau et al, 1985;Kopan et al, 1989;Smola et al, 1994;Limat et al, 1996;Ponec et al, 1997). These results corresponded well with those of other reports on similar models, but using fibroblast-produced extracellular matrices instead of collagen gels (Bouvard et al, 1992;Contard et al, 1993). However, in the latter cases these processes occurred in a wider time frame, which severely restricts the choice of external manipulations.…”

Section: Introductionsupporting

confidence: 79%

“…However, in the latter cases these processes occurred in a wider time frame, which severely restricts the choice of external manipulations. It was also confirmed in such coculture systems that the BM constituents were produced in a cooperative manner (Contard et al, 1993;Limat et al, 1996;Smola et al, 1998;Fleischmajer et al, 2000), nidogen being exclusively expressed by fibroblasts and laminin-5 by keratinocytes, presumably providing a 'nucleation site' for BM assembly. With expression of the BM components in the right sequence an ultrastructurally defined BM zone was also developed (Contard et al, 1993;Tsunenaga et al, 1998;Stark et al, 1999), which corresponded to the findings in surface transplants of NEK on athymic nude mice (Breitkreutz et al, 1997).…”

Section: Introductionmentioning

confidence: 91%

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Inhibition of basement membrane formation by a nidogen-binding laminin γ1-chain fragment in human skin-organotypic cocultures

Breitkreutz

Mirancea

Schmidt

et al. 2004

Journal of Cell Science

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Basement membranes generally determine different tissue compartments in complex organs, such as skin, playing not only an important structural but also a regulatory role. We have previously demonstrated the formation of a regular basement membrane in organotypic three-dimensional (3D)-cocultures of human skin keratinocytes and fibroblasts by indirect immunofluorescence and transmission electron microscopy. In this assembly process, cross-linking of type IV collagen and the laminin γ1 chain by nidogen is considered a crucial step. For a functional proof, we have now competitively inhibited nidogen binding to laminin in 3D-cocultures with a recombinant laminin γ1 fragment (γ1III3-5 module) spanning this binding site. Repeated treatment abolished the deposition of nidogen at the epithelial-matrix interface but also greatly perturbed the presence of other matrix constituents such as laminin and perlecan. This effect persisted over the entire observation period of 10 to 21 days. In contrast, some components of the basement membrane zone were only moderately affected, with the laminin-5 isoform (γ2 chain), type IV collagen and integrin α6ß4 still showing a distinct staining at their regular position, when seen by light microscopy. Furthermore, epidermal morphology and differentiation remained largely normal as indicated by the regular location of keratins K1/K10 and also of late differentiation markers. Ultrastructural examination demonstrated that the γ1 fragment completely suppressed any formation of basement membrane structures (lamina densa) and also of hemidesmosomal adhesion complexes. As a consequence of hemidesmosome deficiency, keratin filament bundles were not attached to the ventral basal cell aspect. These findings were further substantiated by immuno-electron microscopy, revealing either loss or drastic reduction and dislocation of basement membrane and hemidesmosomal components. Taken together, in this simplified human skin model (representing a `closed system') a functional link has been demonstrated between compound structures of the extra- and intracellular space at the junctional zone providing a basis to interfere at distinct points and in a controlled fashion.

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Section: Introductionsupporting

confidence: 79%

Section: Introductionmentioning

confidence: 91%

Inhibition of basement membrane formation by a nidogen-binding laminin γ1-chain fragment in human skin-organotypic cocultures

Breitkreutz

Mirancea

Schmidt

et al. 2004

Journal of Cell Science

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“…One of the major arachidonic acid metabolites in UVB-irradiated epidermal cells is PGE 2 which induces edema and erythema in skin [2, 4]. PGE 2 release can be detected as early as 6 h and peaks between 24 and 48 h following UVB exposure.…”

Section: Resultsmentioning

confidence: 99%

“…An acute UVB exposure of skin causes ‘sunburn’ with histopathological changes including tissue necrosis, hemorrhage, edema and infiltration of leukocytes [2]. Arachidonic acid metabolites such as leukotrienes and prostaglandins contribute to these histopathological changes [6, 28, 29].…”

Section: Resultsmentioning

confidence: 99%

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Anti-Inflammatory Activity of 2,4,6-Trihydroxy-α-<i>p</i>-Methoxyphenyl- acetophenone (Compound D-58)

Malaviya

Uckun³

2000

Dermatology

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Background: Active oxygen radicals as well as a variety of cytosolic protein tyrosine kinases play a role in the regulation of prostaglandin E₂ (PGE₂), a key inflammatory mediator, released by skin cells in response to irradiation with ultraviolet B light (UVB). Identification of chemical compounds that can interrupt such events may provide a basis for the development of potent anti-inflammatory agents. Objective: To investigate the effect of a novel genistein analog, 2,4,6-trihydroxy-α-p-methoxyphenylacetophenone, with antioxidant property (compound D-58), on UVB-induced inflammatory responses. Methods: Epidermal cell cultures were irradiated with UVB both in the presence and absence of compound D-58 and the PGE₂ released in the medium was determined by ELISA. For in vivo studies, skin inflammation was induced in mice either by carrageenan challenge of the air pouch or by an acute exposure of skin to UVB radiation. The resulting inflammatory mediator release, skin edema and the histological changes of the skin were determined both in the presence and absence of compound D-58. Results: Compound D-58 treatment effectively inhibited the development of edema and histological changes in the skin of UVB-irradiated mice as well as the release of PGE₂ in vitro as well as in vivo. Conclusion: Compound D-58 (2,4,6-trihydroxy-α-p-methoxyphenylacetophenone) has potent anti-inflammatory properties.

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Comparison of an in vitro skin model to normal human skin for dermatological research

Monteiro‐Riviere

Inman

Snider

et al. 1997

Microsc. Res. Tech.

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Culturing Keratinocytes and Fibroblasts in a Three-Dimensional Mesh Results in Epidermal Differentiation and Formation of a Basal Lamina-Anchoring Zone

Cited by 107 publications

References 10 publications

Inhibition of basement membrane formation by a nidogen-binding laminin γ1-chain fragment in human skin-organotypic cocultures

Inhibition of basement membrane formation by a nidogen-binding laminin γ1-chain fragment in human skin-organotypic cocultures

Anti-Inflammatory Activity of 2,4,6-Trihydroxy-α-<i>p</i>-Methoxyphenyl- acetophenone (Compound D-58)

Comparison of an in vitro skin model to normal human skin for dermatological research

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