Fig 3 - uploaded by Alberto Calligaro
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3. Three-dimensional epithelial construct. (A) The epidermal construct is macroscopically visible as a threedimensional opalescent sheet. (B) Immunostaining of the construct revealed that epithelial cells were positive for several proliferation and differentiation markers. For example, all basal cells displayed a cytoplasmic immunoreactivity for laminin (brown staining, scale bar: 10 mm). (C) Epidermal constructs can be observed using laser scanning confocal microscopy. Cells were stained with antibodies against proteins expressed at different stages of keratinocyte differentiation . The image was obtained by acquiring fluorescence signals at 0.3 mm intervals (section thickness 10 mm). Example of this analysis is shown in (C), where basal cells show a cytoplasmic immunostaining for laminin (red fluorescence). Nuclear DNA was stained with Hoechst 33258 (blue fluorescence). x–y cross-section shows the cytoplasmic expression of laminin in the cells of the lower layer alone. An orthogonal projection of two cross-sectional cuts x–z and y–z is also shown (upper side of the projections is the lower basal layer of the epidermal construct).
Source publication
Biological risk management is required in modern tissue engineering. Particular attention should be paid to the culture medium and the scaffold used. In this perspective, it is important to define minimal culture conditions which allow proper growth and differentiation of epithelial cells in vitro. We propose a simple experimental system which perm...
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... Eyelid samples from healthy subjects collected after surgical excision were irradiated (UVA1 355 nm) for 2 min, 3 min, and 4 min for a total transmitted energies, respectively, of 50, 75, and 100 J/cm 2 and maintained with control samples 100% UVA1 protected (0 J/cm 2 ) for 1 hour at 37 ∘ C in a culture medium optimized for skin [6]. ...
Fluorescent or metal halide lamps are widely used in therapeutic applications in dermatological diseases, with broadband or narrow band emission UVA/UVA1 (320–400 nm) obtained with suitable passive filters. Recently, it has been possible for us to use a new machine provided with solid state source emitting pulsed monochromatic UVA1 355 nm. In order to evaluate the effects of this emission on immunocells of the skin, human skin samples were irradiated with monochromatic 355 nm UVA1 with different energetic fluences and after irradiation Langerhans cells were labeled with CD1a antibodies. The immunohistochemical identification of these cells permitted evaluating their modifications in terms of density into the skin. Obtained results are promising for therapeutical applications, also considering that a monochromatic radiation minimizes thermic load and DNA damage in the skin tissues.
