Tissue Engineered Artificial Skin Composed of Dermis and Epidermis

Abstract

We made an artificial skin comprised of a stratified layer of keratinocytes and a dermal matrix with a type I collagen containing fibroblasts. In this work, we showed keratinocyte behavior under primary culture, gel contractions varying with concentration of collagen solution, and cell growth plots in the collagen gel. The optimum behavior of dermal equivalent could be obtained using 3.0 mg/ml collagen solution and attached gel culture. The attached gel culture had a jumping effect of growth factor on cell growth at the lag phase. To develop the artificial skin, 1x10(5) cells/cm2 of keratinocytes were cultured on the dermal equivalent at air-liquid interface. Finally, to overcome the problem that artificial skin of collagen gel was torn easily during suturing of grafting, we prepared histocompatible collagen mesh and attached the mesh to the bottom of the gel. Cultured artificial skins were successfully grafted onto rats.

Assessment of Toxic Potential of Industrial Chemicals Using a Cultured Human Bioartificial Skin Model: Production of Interleukin 1α and Hydroxyeicosatetraenoic Acids

“…The morphological features of a differentiating epidermal permeability barrier can be identified in the upper stratum layer. Fully developed artificial skin expresses the broad range of protein products typically found in differentiated human epidermis [17], and expression of the proteins as cytokeratin 19 and involucrin was seen in our artificial skin in a previous published report [14].…”

“…Although all the foregoing factors are relevant when a substance is applied onto the skin, other factors may explain why the response of cells in the artificial skin differs from that of monolayer cultures. It is known, for example, that fibroblasts organized in three-dimensional lattices differ intrinsically from cells grown in a monolayer, exhibiting more complete biosynthetic and morphological properties, comparable with those in vivo [14]. Elevating the epidermis to the air-liquid interface is crucial for the appearance of a multilayered stratum corneum with lamellar granule extrusion structures for generating lipid and keratin profiles approximating those of skin.…”

“…Normal keratinocytes were isolated from epidermis with 0.25% of trypsin solution and were routinely cultured with keratinocyte serum-free medium (K-SFM, Gibco BRL). The medium consisted of MCDB 153 medium supplemented with insulin (0.005 g/l), hydrocortisone (0.074 mg/l), triiodothyronine (0.0067 mg/l), bovine pituitary extract (50 mg/l) and epidermal growth factor (5 Ìg/l) [14].…”

Action of microparticles of heparin and alginate crosslinked gel when used as injectable artificial matrices to stabilize basic fibroblast growth factor and induce angiogenesis by controlling its release

“…In the construction of tissue, artificial matrices play an important role as a cellular "scaffold" and stabilizer for the mitogenic molecules, such as growth factors. 1 For instance, collagen, polyglycolic acid, and polylactic acid are used as matrices for the construction of skin, 2 cartilage, 3 and meniscus. 4 Heparin-binding growth factors (HBGFs), such as basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF), have powerful mitogenic effects on certain types of cells, while modulating cell proliferation and differentiation.…”

Construction of a Transplantable Tissue-Engineered Artificial Peritoneum

“…Cell culture techniques that involve collagen gel have become increasingly popular over the last 10 years be-cause of their successful application to many different cellular systems and pathological processes [23,24]. The application of these techniques in the current context has enabled us to develop an in vitro and transplantable human artificial peritoneum that closely resembles the physical and biochemical properties of in vivo tissue.…”