Effect of fibroblasts on epidermal regeneration

El-Ghalbzouri, A.; Gibbs, Susan; Lamme, Evert N.; Blitterswijk, Clemens A. van; Ponec, Maria

doi:10.1046/j.1365-2133.2002.04871.x

Cited by 255 publications

(210 citation statements)

References 60 publications

(70 reference statements)

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“…It is generally believed that positional skin identity is regulated by epithelial (keratinocytes) and mesenchymal (fibroblasts) interactions. 1e5 Indeed, three-dimensional skin equivalent models and transplantation experiments show that epidermal stratification during the keratinocyte differentiation program is remarkably disrupted in the absence of fibroblasts, 2,4,5 demonstrating that epidermal development and homeostasis are regulated by extrinsic factors released by fibroblasts. Such interactions have been further clarified through an analysis of the differential localization of epidermal keratins in distinct epidermal layers and body sites.…”

mentioning

confidence: 99%

To Control Site-Specific Skin Gene Expression, Autocrine Mimics Paracrine Canonical Wnt Signaling and Is Activated Ectopically in Skin Disease

Kim¹,

Hossain²,

Nieves³

et al. 2016

The American Journal of Pathology

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Despite similar components, the heterogeneity of skin characteristics across the human body is enormous. It is classically believed that site-specific fibroblasts in the dermis control postnatal skin identity by modulating the behavior of the surface-overlying keratinocytes in the epidermis. To begin testing this hypothesis, we characterized the gene expression differences between volar (ventral; palmoplantar) and nonvolar (dorsal) human skin. We show that KERATIN 9 (KRT9) is the most uniquely enriched transcript in volar skin, consistent with its etiology in genetic diseases of the palms and soles. In addition, ectopic KRT9 expression is selectively activated by volar fibroblasts. However, KRT9 expression occurs in the absence of all fibroblasts, although not to the maximal levels induced by fibroblasts. Through gain-of-function and loss-of-function experiments, we demonstrate that the mechanism is through overlapping paracrine or autocrine canonical WNTeb-catenin signaling in each respective context. Finally, as an in vivo example of ectopic expression of KRT9 independent of volar fibroblasts, we demonstrate that in the human skin disease lichen simplex chronicus, WNT5a and KRT9 are robustly activated outside of volar sites. These results highlight the complexities of site-specific gene expression and its disruption in skin disease. Site-specific epidermal differentiation programs define the heterogeneity of skin identity across the human body. It is generally believed that positional skin identity is regulated by epithelial (keratinocytes) and mesenchymal (fibroblasts) interactions. 1e5 Indeed, three-dimensional skin equivalent models and transplantation experiments show that epidermal stratification during the keratinocyte differentiation program is remarkably disrupted in the absence of fibroblasts, 2,4,5 demonstrating that epidermal development and homeostasis are regulated by extrinsic factors released by fibroblasts. Such interactions have been further clarified through an analysis of the differential localization of epidermal keratins in distinct epidermal layers and body sites. Volar (palmoplantar) skin is characterized by thick epidermal layers, less pigmentation, and lack of hair. In addition to these features, cytoskeleton KERATIN (KRT) 9 6 has been thought to be almost exclusively localized to volar keratinocytes, 1,6e9 indicating that KRT9 can be a unique marker for volar skin. To date, many clinical studies have reported that KRT9 mutations cause epidermolytic palmoplantar keratoderma, characterized by compensatory thickened epidermal layers in the palms and soles. 10 Conditional deletion of Krt9 in a murine model demonstrated that Krt9 is responsible for maintaining mechanical integrity and terminal differentiation of volar skin.

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mentioning

confidence: 99%

To Control Site-Specific Skin Gene Expression, Autocrine Mimics Paracrine Canonical Wnt Signaling and Is Activated Ectopically in Skin Disease

Kim¹,

Hossain²,

Nieves³

et al. 2016

The American Journal of Pathology

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“…EGF is also able to activate PLSCR1 by inducing PLSCR1 to bind to the EGF receptor (EGFR) (40,41). Furthermore, human skin equivalents have been used because of the crucial role fibroblasts play in basement membrane formation (27,32). Factors released by fibroblasts resulted in the expression of basement membrane components (e.g.…”

Section: Discussionmentioning

confidence: 99%

“…Reconstruction of Human Skin Equivalents-Hydrated collagen gels were prepared using 4 mg/ml collagen solution isolated from rat tails, as described in detail elsewhere (27). Briefly, the collagen solution was used directly or mixed at 4°C with fibroblast suspension to reach a final density of 1 ϫ 10 5 cells/ml.…”

Section: Methodsmentioning

confidence: 99%

Phospholipid Scramblase 1 Is Secreted by a Lipid Raft-dependent Pathway and Interacts with the Extracellular Matrix Protein 1 in the Dermal Epidermal Junction Zone of Human Skin

Merregaert¹,

Langen²,

Hansen

et al. 2010

Journal of Biological Chemistry

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We examined the interaction of ECM1 (extracellular matrix protein 1) using yeast two-hybrid screening and identified the type II transmembrane protein, PLSCR1 (phospholipid scramblase 1), as a binding partner. This interaction was then confirmed by in vitro and in vivo co-immunoprecipitation experiments, and additional pull-down experiments with GST-tagged ECM1a fragments localized this interaction to occur within the tandem repeat region of ECM1a. Furthermore, immunohistochemical staining revealed a partial overlap of ECM1 and PLSCR1 in human skin at the basal epidermal cell layer. Moreover, in human skin equivalents, both proteins are expressed at the basal membrane in a dermal fibroblast-dependent manner. Next, immunogold electron microscopy of ultrathin human skin sections showed that ECM1 and PLSCR1 co-localize in the extracellular matrix, and using antibodies against ECM1 or PLSCR1 cross-linked to magnetic immunobeads, we were able to demonstrate PLSCR1-ECM1 interaction in human skin extracts. Furthermore, whereas ECM1 is secreted by the endoplasmic/Golgi-dependent pathway, PLSCR1 release from HaCaT keratinocytes occurs via a lipid raft-dependent mechanism, and is deposited in the extracellular matrix. In summary, we here demonstrate that PLSCR1 interacts with the tandem repeat region of ECM1a in the dermal epidermal junction zone of human skin and provide for the first time experimental evidence that PLSCR1 is secreted by an unconventional secretion pathway. These data suggest that PLSCR1 is a multifunctional protein that can function both inside and outside of the cell and together with ECM1 may play a regulatory role in human skin.The human ECM1 gene (11 exons) is located on chromosome 1q21.2 (1, 2) and encodes four splice variants. ECM1a (without exon 5a) is expressed in basal keratinocytes, dermal blood vessels, and adnexal epithelia, including hair follicles and glands, whereas ECM1b, which lacks exon 7, is expressed in the spinous and granular layers of the epidermis (3-5). ECM1c was found in the basal layer of the epidermis (6), and a fourth splice variant results in a truncated protein of 57 amino acids (7). The ECM1 protein contains a 19-amino acid signal peptide followed by four domains: a cysteine-free N-terminal segment, two tandem repeats, and a C-terminal segment. The two tandem repeats and the C-terminal domain contain cysteines in a typical CC-(X 7-10 )C arrangement that is capable of forming protein double loops that could be involved in protein-protein interactions (1, 8). More recently, a rudimentary three-dimensional model divided the ECM1a protein into four distinct domains: an NH 2 -terminal domain forming ␣-helical structures, followed by three domains, whose amino acid sequences were highly comparable with the third domain of human serum albumin: SASDL2 (serum albumin subdomain-like 2), SASDL3, and SASDL4 (9).The function of ECM1 has not yet been elucidated in detail; however, it has been reported that ECM1 could act as a novel paracrine factor involved in the regulation of endocho...

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“…In this reconstructed human skin model generated by culturing keratinocytes on fibroblast-populated collagen matrix, epidermal stratification is reproduced to a high extent, similar to that occurring in vivo. 20,23,35 Our recent study 26 showed that when HSE are grown in serum-free media and in the absence of growth factors, the number and the functional state of fibroblasts incorporated into the collagen matrix is crucial for the normalization of the epidermal differentiation program and of the deposition of various basement membrane and hemidesmosomal components. To examine the potential role of fibroblasts in the reepithelialization process, two types of wounds were introduced in our HSE: superficial wounds, in From the negative to a strong positive expression Patchy expression Human skin equivalents (HSE) were generated by seeding of keratinocytes onto the central region of fibroblast-populated collagen matrices and subsequent culturing for 2 weeks at the A/L interface in the absence of growth factors.…”

Section: Discussionmentioning

confidence: 99%

“…Fibroblasts have been found to exert profound effects on keratinocyte proliferation and differentiation and on the deposition of basement proteins. 20 Only in the presence of fibroblasts the normalization of the epidermal differentiation program and deposition of laminin 5, collagen type IV and VII at the dermal/epidermal junction occurred. We can speculate that also during wound-healing processes fibroblasts play an essential role, as it is well established that in vivo the closure of superficial wounds, in which only the epidermis is injured, is faster than in full-thickness wounds, where both the epidermal and dermal components are injured.…”

mentioning

confidence: 99%

Fibroblasts facilitate re-epithelialization in wounded human skin equivalents

et al. 2003

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The re-epithelialization of the wound involves the migration of keratinocytes from the edges of the wound. During this process, keratinocyte migration and proliferation will depend on the interaction of keratinocytes with dermal fibroblasts and the extracellular matrix. The present study aimed to investigate (1) the role of fibroblasts in the re-epithelialization process and on the reconstitution of the dermal-epidermal junction (DEJ) and (2) differential protein expression during re-epithelialization. For both purposes, three-dimensional human skin equivalents (HSE) were used. A full-thickness wound in HSE was introduced by freezing with liquid nitrogen and a superficial wound by linear incision with a scalpel. The closure of the wound in the absence or presence of exogenous growth factors was followed by monitoring the rate of re-epithelialization and regeneration of the DEJ. The results obtained in this study demonstrate that fibroblasts facilitate wound closure, but they differentially affected the deposition of various basement membrane components. The deposition of laminin 5 at the DEJ was delayed in superficial wounds as compared to the full-thickness wounds. During freeze injury, some basement membrane (BM) components remain associated with the dermal compartment and probably facilitate the BM reconstitution. The re-epithelialization process in full-thickness but not in superficial wounds was accelerated by the presence of keratinocyte growth factor and especially by epidermal growth factor. In addition, we have examined the deposition of various basement membrane components and the differences in protein expression in a laterally expanding epidermis in uninjured HSE. Laminin 5, type IV and VII collagen deposition was decreased in the laterally expanding epidermis, indicating that the presence of these proteins is not required for keratinocyte migration to occur in vitro. Using twodimensional polyacrylamide gel electrophoresis, we have identified DJ-1, a protein not earlier reported to be differently expressed during the epithelialization process of the skin.

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Effect of fibroblasts on epidermal regeneration

Abstract: Our results illustrate that numbers of fibroblasts in the collagen matrix and their functional state is a critical factor for establishment of normal epidermal morphogenesis.

Cited by 255 publications

References 60 publications

To Control Site-Specific Skin Gene Expression, Autocrine Mimics Paracrine Canonical Wnt Signaling and Is Activated Ectopically in Skin Disease

To Control Site-Specific Skin Gene Expression, Autocrine Mimics Paracrine Canonical Wnt Signaling and Is Activated Ectopically in Skin Disease

Phospholipid Scramblase 1 Is Secreted by a Lipid Raft-dependent Pathway and Interacts with the Extracellular Matrix Protein 1 in the Dermal Epidermal Junction Zone of Human Skin

Fibroblasts facilitate re-epithelialization in wounded human skin equivalents

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