In vitro effects of different formulations of bovine collagen on cultured human skin

CONTEXT AND OBJECTIVE: The technique of obtaining human skin with dermis and epidermis reconstructed from cells isolated from patients can enable autologous skin grafting on patients with few donor sites. It also enables in vitro trials on chemicals and drugs. The objective of this work was to demonstrate a method for obtaining human skin composed of associated dermis and epidermis, reconstructed in vitro. DESIGN AND SETTING: Experimental laboratory study, in the Skin Cell Culture Laboratory of Faculdade de Ciências Médicas, Universidade Estadual de Campinas. METHODS: Cells from human fibroblast cultures are injected into bovine collagen type I matrix and kept immersed in specific culturing medium for fibroblasts. This enables human dermis reconstruction in vitro. On this, by culturing human keratinocytes and melanocytes, differentiated epidermis is formed, leading to the creation of human skin composed of associated dermis and epidermis, reconstructed in vitro. RESULTS: We showed that human skin composed of associated dermis and epidermis can be successfully reconstructed in vitro. It is histologically formed in the same way as human skin in vivo. Collagen tissue can be identified in the dermis, with cells and extracellular matrix organized in parallel to multilayer epidermis. CONCLUSIONS: It is possible to obtain completely differentiated human skin composed of associated dermis and epidermis, reconstructed in vitro, from injection of human fibroblasts into bovine collagen type I matrix and culturing of human keratinocytes and melanocytes on this matrix.

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P256 electroperturbation of the human skin barrier in vitro: effects on stratum corneum resistance, capacitance, lipid ordering and ultrastructure

European Journal of Pharmaceutical Sciences ◽

10.1016/0928-0987(94)90429-4 ◽

1994 ◽

Vol 2 (1-2) ◽

pp. 182

Author(s):

W.H.M. Craane-van Hinsberg ◽

F. Spies ◽

G.S. Gooris ◽

J.A. Bouwstra ◽

J.C. Verhoef ◽

...

Keyword(s):

Stratum Corneum ◽

Human Skin ◽

Skin Barrier ◽

In Vitro Effects

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THE EFFECT OF INSULIN AND DIABETES ON GLUCOSE METABOLISM IN HUMAN SKIN

Canadian Journal of Biochemistry ◽

10.1139/o66-098 ◽

1966 ◽

Vol 44 (6) ◽

pp. 801-808 ◽

Cited By ~ 6

Author(s):

A. Kahlenberg ◽

N. Kalant

Keyword(s):

Glucose Uptake ◽

Glucose Transport ◽

Human Skin ◽

Glucose Utilization ◽

Bicarbonate Buffer ◽

Glucose Content ◽

Basal Rate ◽

In Vitro Effects ◽

Intracellular Glucose

The in vitro effects of insulin on glucose transport and dissimilation were studied in human skin. Slices of skin obtained post-mortem were incubated in phosphate and bicarbonate media with a glucose concentration of 5.6 mM. In both buffers, insulin increased the glucose uptake in non-diabetic skin without affecting the intracellular glucose content. Similar results were obtained for diabetic skin incubated in a phosphate buffer.In bicarbonate buffer, the basal rate of glucose utilization of skin from diabetics was lower than normal; this may have been the result of a defect in the phosphofructokinase reaction. Diabetic skin also had an increased responsiveness to insulin in bicarbonate buffer.Non-diabetic skin incubated in phosphate buffer had a lower basal rate of glucose uptake and a greater responsiveness to insulin than the same tissue incubated in bicarbonate buffer.It is concluded that (a) skin from diabetic humans incubated in bicarbonate buffer had a decreased basal rate of glucose utilization and increased responsiveness to insulin; (b) insulin stimulated both glucose transport and dissimilation in diabetic and non-diabetic human skin.

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Desmosomal distribution in the epidermis of a non-contracting human skin equivalent

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124884 ◽

1992 ◽

Vol 50 (1) ◽

pp. 896-897

Author(s):

L.X. Oakford ◽

S.D. Dimitrijevich ◽

R. Gracy

Keyword(s):

Human Skin ◽

Basal Layer ◽

Skin Equivalent ◽

Tissue Component ◽

Intact Skin ◽

Similar Sequence ◽

Sequence Of Events ◽

Suprabasal Keratinocytes ◽

Human Skin Equivalent

In intact skin the epidermal layer is a dynamic tissue component which is maintained by a basal layer of mitotically active cells. The protective upper epidermis, the stratum corneum, is generated by differentiation of the suprabasal keratinocytes which eventually desquamate as anuclear comeocytes. A similar sequence of events is observed in vitro in the non-contracting human skin equivalent (HSE) which was developed in this lab (1). As a part of the definition process for this model of living skin we are examining its ultrastructural features. Since desmosomes are important in maintaining cell-cell interactions in stratified epithelia their distribution in HSE was examined.

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