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Successful testing/analysis of the efficacy of engineered human skin replacements
has long been restricted by the paucity of animal models that reliably mimic human
skin conditions. The nude (athymic) mouse is an established host for human tissues,
and has been extensively used for testing skin substitutes and skin equivalents directly
grafted to the mouse skin.
Evaluation of dermal–epidermal skin equivalents (‘composite-skin’) of human keratinocytes
in a collagen–glycosaminoglycan matrix (Integra artificial skin).
This model has also been employed to study human skin wound healing, by pre-establishing
human skin grafts on mice, and excising skin from the grafts using punch biopsies.
Here we aimed to further develop the model and establish a method that could have
future application in assessing the effectiveness of skin equivalents in a human skin
environment. As an exemplar we used Integra (Integra Life Sciences Corporation, Plainsboro,
NJ) a bilayer artificial skin replacement with a “dermal” layer composed of bovine
collagen gel cross-linked with shark chondroitin-6-sulphate that is well established
as a dermal replacement with multiple applications. Integra has also been used in
combination with cultured keratinocytes in wound healing models
Evaluation of dermal–epidermal skin equivalents (‘composite-skin’) of human keratinocytes
in a collagen–glycosaminoglycan matrix (Integra artificial skin).
The use of confocal laser scanning microscopy to assess the potential suitability
of 3-D scaffolds for tissue regeneration, by monitoring extra-cellular matrix deposition
and by quantifying cellular infiltration and proliferation.
In our study, therefore, Integra, with and without incorporated dermal cells was
introduced into full thickness punch wounds in pre-established human skin grafts on
nude mice (Figure 1).
Figure 1Procedure for testing a skin substitute containing living cells on a human skin graft
in nude mice. Full thickness human skin is transplanted on to the dorsum of an athymic
mouse. Grafts appear healthy and normal at the time of suture removal (after 7 days)
and up to 2–3 weeks (A) Around 4 weeks after grafting the surface of the grafts become
abnormally thickened and scabbed (B). The scab gradually sheds around 8 weeks post-grafting
leaving a human skin with more elasticity and smoother epithelium (C). 4 mm diameter
pieces of Integra, either untreated or inoculated with cultured dermal cells
are removed from the cell culture vessel (D). 3 mm diameter full thickness wounds
are created on the grafted human skin, and the Integra is carefully transferred to
fill in the skin wounds. (E). At 2 weeks post operation, healing of the Integra filled
wounds (white arrows) has taken place, and the implants have re-epithelialised and
are free from infection.
Evaluation of dermal–epidermal skin equivalents (‘composite-skin’) of human keratinocytes in a collagen–glycosaminoglycan matrix (Integra artificial skin).
The use of confocal laser scanning microscopy to assess the potential suitability of 3-D scaffolds for tissue regeneration, by monitoring extra-cellular matrix deposition and by quantifying cellular infiltration and proliferation.
