Comparisons of invasion scores in wounded melanoma TE models.

<p>Comparisons of invasion scores in wounded melanoma TE models.</p

Quantitation of the effects of wounding and Ibuprofen on melanoma cell invasion in TE skin constructs.

<p>(A) with HBL cells, (B) with A375SM cells and (C) with C8161 cells. To the constructs were added W=Wound, Fn=Fibrin clots, TNF=300U/ml TNF-α, NaIbu=Ibuprofen sodium salt as shown. Values (bars) are expressed as Mean+SEM of invasion score. * indicates p<0.05; ** indicates p<0.01.</p

Effects of Melanoma Cells on Wound Healing in TE Skin.

<p>Effects of Melanoma Cells on Wound Healing in TE Skin.</p

Effects of Ibuprofen on wounded melanoma skin construct model.

<p>All constructs contained fibroblasts, keratinocytes and A375SM cells and fibrin clots plus 300 U/ml TNF-α (as in A and C). To (E) and (G) was added unloaded gel. To (B), (D), (F) and (H) were added 100μM NaIbu hydrogel. (B), (C), (D), (E), (G) and (H) = 4x magnification; (A) and (F) = 10x magnification. The site of the wound is indicated by an arrow. DED=De-Epidermised acellular Dermis, F=Fibroblasts, K=Keratinocytes, W=Wound.</p

Effect of mechanical wounding and addition of fibrin and TNF-α on wound healing in TE skin.

<p>(A) DED+Wound (4x magnification); (B) DED+Wound+Fibrin (4x magnification); (C) DED+Fibroblasts+Wound (10x magnification); (D) DED+Fibroblasts+Wound+Fibrin (10x magnification); Wounded standard skin constructs (E, F and G) (4x, 10x and 10x magnifications respectively). Wounded standard skin constructs with the addition of fibrin and TNF-α (H and I) (10x magnification). The site of the wound is indicated by an arrow. DED = De-Epidermised acellular Dermis, F = Fibroblasts, W = Wound.</p

Effect of fibrin clot, TNF-α and skin cells on wound healing.

<p>Effect of fibrin clot, TNF-α and skin cells on wound healing.</p

Development of a mini 3D cell culture system using well defined nickel grids for the investigation of cell scaffold interactions

A bioreactor system was developed using a series of fine mesh nickel grids as free standing scaffolds to investigate the behaviours of fibroblasts and keratinocytes in tissue culture. It was found that the mesh size of the suspended grids, but not of the grids that attached to tissue culture surface, had significant influences on cell behaviour and there was a maximum size for fibroblast to span within the defined culture period. Time lapse video microscopy demonstrated fibroblasts cultured on these grids initially migrated onto the struts but then worked together to fill in the voids between struts with a membranous sheet of tissue. In contrast keratinocytes barely migrated from the initial site of cell deposition and when they moved (to a modest extent) it was as an integrated sheet of cells. Similar results were observed when both types of cells were co-cultured in the system

Effects of NSAID on melanoma invasion in wounded skin constructs.

<p>Effects of NSAID on melanoma invasion in wounded skin constructs.</p

Invasion of melanoma cells into tissue engineered skin.

<p>(A) Control TE skin constructs to which fibroblasts and keratinocytes were added to the DED (4x magnification); (B) Skin cells (fibroblasts and keratinocytes) and HBL melanoma cells in DED stained with HM45 antibody staining (40x magnification); (C) Skin cells (fibroblasts and keratinocytes) and A375SM cells stained with S100 antibody staining for A375SM cells (40x magnification); (D) Skin cells (fibroblasts and keratinocytes) and C8161 cells in DED, AE1AE3 antibody staining for keratinocytes only (40x magnification). (DED = De-Epidermised acellular Dermis; F = Fibroblasts; K = Keratinocytes).</p

Quantitation of melanoma cell invasion into the 3D skin models.

<p>The composition of cells in these models is given followed by the number of skin constructs. The maximum invasion score is indicated in brackets. The histograms show the mean +1SEM of all scores. * indicates p<0.05; ** indicates <0.01. (F = fibroblasts, K = keratinocytes).</p