Advanced therapies combating acute and chronic skin wounds are likely to be brought about using our knowledge of regenerative medicine coupled with appropriately tissue-engineered skin substitutes. At the present time, there are no models of an artificial skin that completely replicate normal uninjured skin. Natural biopolymers such as collagen and fibronectin have been investigated as potential sources of biomaterial to which cells can attach. The first generation of degradable polymers used in tissue engineering were adapted from other surgical uses and have drawbacks in terms of mechanical and degradation properties. This has led to the development of synthetic degradable gels primarily as a way to deliver cells and/or molecules in situ, the so-called smart matrix technology. Tissue or organ repair is usually accompanied by fibrotic reactions that result in the production of a scar. Certain mammalian tissues, however, have a capacity for complete regeneration without scarring; good examples include embryonic or foetal skin and the ear of the MRL/MpJ mouse. Investigations of these model systems reveal that in order to achieve such complete regeneration, the inflammatory response is altered such that the extent of fibrosis and scarring is diminished. From studies on the limited examples of mammalian regeneration, it may also be possible to exploit such models to further clarify the regenerative process. The challenge is to identify the factors and cytokines expressed during regeneration and incorporate them to create a smart matrix for use in a skin equivalent. Recent advances in the use of DNA microarray and proteomic technology are likely to aid the identification of such molecules. This, coupled with recent advances in non-viral gene delivery and stem cell technologies, may also contribute to novel approaches that would generate a skin replacement whose materials technology was based not only upon intelligent design, but also upon the molecules involved in the process of regeneration.

https://royalsocietypublishing.org/doi/pdf/10.1098/rsif.2006.0179

Tissue engineering of replacement skin: the crossroads of biomaterials, wound healing, embryonic development, stem cells and regeneration

A guide to biological skin substitutes.

Purpose.To bioengineer a corneal surface replacement using ex vivo expanded, cultured corneal epithelial stem cells seeded on a matrix derived from amniotic membrane and use this bioengineered graft to manage difficult ocular surface disease.Methods.Fourteen patients with ocular surface disease unre

Successful transplantation of bioengineered tissue replacements in patients with ocular surface disease.

Biologically-based skin substitutes have developed as commercial products over the last 5 years. The first generation includes the collagen-based synthetic device, Integra, and Alloderm, which is based on devitalised and cross-linked human dermis. These are used as dermal replacements for third degree burns. Within the last year, the tissue-engineered product, Dermagraft-TC, has become available. While originally intended as a temporary covering for severe burns, Dermagraft-TC has proved to markedly improve the healing of deep second degree burns. The earliest living skin substitutes used autologous keratinocytes expanded in vitro. Two new products containing living cells, Dermagraft and Apligraf, are expected to be approved shortly for diabetic foot ulcers and venous stasis ulcers, respectively. Dermagraft is produced by growing human fibroblasts on a three-dimensional scaffold. The cells actively proliferate and lay down extracellular matrix to generate a papillary dermis-like device that shows a combination of angiogenic, growth factor and cell adhesion properties that enhance healing in diabetic foot ulcers. The production of Apligraf includes casting human fibroblasts in collagen, in order to generate a dermal equivalent on which is grown an epidermis. The structure is akin to a skin graft and is so applied. Despite Dermagraft and Apligraf being of allogeneic origin, rejection has not been an issue in clinical trials and possible contamination by pathogens has been eliminated as a concern through extensive testing. These developments represent a new concept and are expected to revolutionise wound care. They may also provide a platform for gene therapy applications.

Skin substitutes to enhance wound healing.

Long-Term Culture of Murine Epidermal Keratinocytes

Organotypic cultures of human skin were made using dermal fibroblasts seeded into a type I collagen gel overlaid with epidermal keratinocytes. Full-thickness excision of tattoos was performed on five patients, three of whom received sex-mismatched allografts. Patients were not immunosuppressed. Biopsies were obtained up to 3.5 years later. In situ hybridization of the PHY2,1 repetitive Y chromosome sequence revealed male fibroblasts and keratinocytes at 11 weeks and 2.5 years in the two female patients grafted with male cells. Structural components in the dermal substitute matured with time, and elastic fibers formed an interlacing meshwork by 18 months. Electron microscopy of the dermal-epidermal junction of an organotypic allograft revealed anchoring fibrils that had normal features at this time. Hyperemia of early grafts settled and contour correction was maintained, while repigmentation was variable. Hypertrophic scars did not occur, and graft contracture was never more than 20 percent. We conclude that this organotypic skin graft shows potential toward the goal of allogeneic skin replacement in a onestep procedure. (Plast. Reconstr. Surg. 96 : 166, 1995).

Survival of allogeneic cells in cultured organotypic skin-grafts

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Survival of allogeneic cells in cultured organotypic skin-grafts