Extracellular matrix proteins induce changes in intracellular calcium and cyclic AMP signalling systems in cultured human keratinocytes

Goberdhan, N.J.; Edgecombe, Mandy; Freedlander, E.; MacNeil, Sheila

doi:10.1016/s0305-4179(96)00087-3

Cited by 16 publications

(6 citation statements)

References 30 publications

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“…Even allowing for the possibility of crosstalk between these two pathways, perhaps mediated by mitogen-activated protein kinase, a control group of islets embedded into a solid matrix in the absence of CT failed to transform to ducts. This is in keeping with the report of Goberdhan et al (1997) who showed that RGD peptide did not have an effect on intracellular cAMP in keratinocytes. Conversion of a solid to a hollow structure is a morphogenetic process observed frequently during vertebrate embryogenesis (Coucouvanis & Martin 1995).…”

Section: Discussionsupporting

confidence: 93%

Factors mediating the transdifferentiation of islets of Langerhans to duct epithelial-like structures

Wang

Li²,

Rosenberg

2001

Journal of Endocrinology

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We have previously shown that isolated islets embedded in type 1 collagen gel in the presence of a defined medium undergo transdifferentiation within 96 h to duct epithelial structures. The aim of this study was to identify the factors implicated in this process. Freshly isolated canine islets were embedded in type 1 collagen gel, Matrigel or agarose for up to 120 h and cultured in (i) Dulbecco's modified Eagle's medium (DMEM)/F12 plus cholera toxin (CT), (ii) medium CMRL1066 plus CT, (iii) CMRL1066 plus forskolin and (iv) CMRL1066 alone. At 16 h, intracellular levels of cAMP (fmol/10 3 islets) were increased in groups i-iii (642 17, 338 48, 1128 221) compared with group iv (106 19, P<0·01). Epithelial differentiation correlated with the total amount of intracellular cAMP measured over 120 h. Islet-epithelial transformation during the initial 36 h was associated with a wave of apoptosis which was followed by a wave of cell proliferation. During epithelial differentiation there was a progressive loss of all islet hormones and the concomitant expression of cytoskeletal proteins characteristic of duct epithelial cells. Islets in collagen and Matrigel demonstrated high rates of epithelial differentiation (63 2% and 71 4% respectively) compared with those in agarose gel (0 0%, P<0·001). Islets suspended in DMEM/F12 plus CT supplemented with soluble laminin or fibronectin did not undergo transformation. Prior incubation of freshly isolated islets with an integrin-binding arginine-glycineaspartate motif-presenting synthetic peptide also reduced islet transformation. These studies confirm the biological potential of islets of Langerhans to differentiate to duct epithelial structures. cAMP-mediated signal transduction and an appropriate integrin-matrix interaction are necessary for this process to proceed.

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Section: Discussionsupporting

confidence: 93%

Factors mediating the transdifferentiation of islets of Langerhans to duct epithelial-like structures

Wang

Li²,

Rosenberg

2001

Journal of Endocrinology

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“…Keratinocytes were obtained from four consenting adult patients undergoing routine cosmetic surgery, resulting in a surgical discard, with institutional ethics committee approval, and in adherence to the Declaration of Helsinki Principles. Cells were isolated and cultured using a modification of the Rheinwald and Green method 32 as described previously 33 . In brief, pieces of skin were incubated in 0.125% trypsin (Invitrogen, Mulgrave, Vic, Australia) overnight at 4°C to promote the separation of the epidermal and dermal layers.…”

Section: Methodsmentioning

confidence: 99%

Hyperbaric oxygen stimulates epidermal reconstruction in human skin equivalents

Kairuz

Upton

Dawson

et al. 2007

Wound Repair Regeneration

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The crucial role of oxygen during the complex process of wound healing has been extensively described. In chronic or nonhealing wounds, much evidence has been reported indicating that a lack of oxygen is a major contributing factor. Although still controversial, the therapeutic application of hyperbaric oxygen (HBO) therapy can aid the healing of chronic wounds. However, how HBO affects reepithelization, involving processes such as keratinocyte proliferation and differentiation, remains unclear. We therefore used a three-dimensional human skin-equivalent (HSE) model to investigate the effects of daily 90-minute HBO treatments on the reconstruction of an epidermis. Epidermal markers of proliferation, differentiation, and basement membrane components associated with a developing epidermis, including p63, collagen type IV, and cytokeratins 6, 10, and 14, were evaluated. Morphometric analysis of hematoxylin and eosin-stained cross sections revealed that HBO treatments significantly accelerated cornification of the stratum corneum compared with controls. Protein expression as determined by immunohistochemical analysis confirmed the accelerated epidermal maturation. In addition, early keratinocyte migration was enhanced by HBO. Thus, HBO treatments stimulate epidermal reconstruction in an HSE. These results further support the importance of oxygen during the process of wound healing and the potential role of HBO therapy in cutaneous wound healing.

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“…Normal human adult keratinocytes (obtained from breast reductions and abdominoplasties) were isolated from the dermal/epidermal junction, as previously described 4. Cells were cultured in complete Green's media (Hams F10) that included cholera toxin (0.1 n M ), hydrocortisone (0.4 μg/mL), adenine (1.8 × 10 −4 M ), triiodo‐1‐thyronine (2 × 10 −7 M ), insulin (5 mg/mL), transferrin (5 μg/mL), glutamine ( 2× 10 −3 M ), fungizone (0.625 μg/mL), penicillin (1000 IU/mL), streptomycin (1000 μg/mL), and 10% fetal calf serum.…”

Section: Methodsmentioning

confidence: 99%

Plasma‐polymerized surfaces for culture of human keratinocytes and transfer of cells to an in vitro wound‐bed model

Haddow

Steele

Short

et al. 2002

J Biomedical Materials Res

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The aim of this study was to develop plasma-polymerized surfaces suitable for the attachment and culture of human keratinocytes and that would allow their subsequent transfer to a wound-bed model. Keratinocyte attachment has been assessed on a carrier polymer, either untreated or treated with a hydrocarbon plasma polymer, collagen I, or carboxylic-acid-containing plasma copolymers. Cell attachment was poor on the "bare" carrier polymer and hydrocarbon plasma polymer (PP) surfaces. Cell attachment was good and comparable on collagen I-coated carrier polymer and carrier polymer plasma coated with carboxylic acid functionalities. After 24 h of cell culture, surfaces were inverted so that cells were adjacent to a de-epidermalized dermis (DED) for 4 days. After 4 days in contact with DED, the surfaces were removed and the level of residual cells and cells transferred to DED were assessed using a cell viability assay. Cell transfer from the collagen I-coated surface was on the order of 90%. Transfer from the carrier polymer surface and the hydrocarbon-coated surface was poor while cells cultured on acid-containing surfaces showed high levels of transfer. Cell transfer was greatest from those surfaces containing the highest level of acid functionality (ca. 21%). Cell transfer was not significantly affected by the choice of carrier polymer material although some sample-to-sample variation was seen. To determine that plasma-polymerized surfaces could be used clinically, selected samples were sterilized with ethylene oxide. Subsequent analysis and cell culture indicated that the surface chemistry and cell-transfer capability of these plasma-polymerized surfaces were unaffected by the sterilization procedure. Plasma-polymerized carboxylic-acid-containing surfaces show great promise in the field of wound healing, encouraging keratinocyte attachment and permitting keratinocyte transfer to a wound bed.

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Extracellular matrix proteins induce changes in intracellular calcium and cyclic AMP signalling systems in cultured human keratinocytes

Cited by 16 publications

References 30 publications

Factors mediating the transdifferentiation of islets of Langerhans to duct epithelial-like structures

Factors mediating the transdifferentiation of islets of Langerhans to duct epithelial-like structures

Hyperbaric oxygen stimulates epidermal reconstruction in human skin equivalents

Plasma‐polymerized surfaces for culture of human keratinocytes and transfer of cells to an in vitro wound‐bed model

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