(a) Temporary cover of an area after necrectomy originally evaluated as 3rd degree burn. XD was adhered immediately and stopped bleeding Ixazomib Ki instantly. (b) XD incrusted by …In vitro: keratinocytes that were grown submerged in the medium formed 1-2 cell layers on the XD matrix (Figures 5(a) and 5(c)). After the cultures were lifted to the air-liquid interface, the epidermis became stratified and formed 5�C15 cell layers in the course of one week (Figures 5(b) and 5(d)). The multilayered structure closely resembled well-differentiated epidermis in vivo composed of distinct basal, spinous, granular, and cornified layers (Figure 5).Figure 5Keratinocytes cultured and differentiated on XD. (a) Keratinocytes grown submerged formed 1-2 layers on XD. (b) Keratinocytes grown at the air-liquid interface formed the basal, spinous, and granular layers.

Keratohyalin granules (arrows) in the granular …3.3. Expression and Distribution of Keratinocyte Differentiation MarkersParaffin sections of the normal skin, recombined skin (keratinocytes cultured on XD at the air-liquid interface), and of the wounds treated with XD or Grassolind were analyzed using a panel of antibodies detecting selected markers of keratinocyte differentiation. The reactivity of the antibodies in the cultured keratinocytes and in the freshly healing skin is shown in Table 1. Immunohistochemical staining of HMW CK, involucrin, p63, and CD29 of keratinocytes cultured on XD confirmed the organization and differentiation of keratinocytes similar to that of normal epidermis (Figure 6).

HMW CK was strongly expressed in all keratinocytes cultured on XD (Figure 6(a)/(A2)). Involucrin was expressed in the granular and horny layers (Figure 6(b)/(B2)). CD29 was positive in the basal layer of keratinocytes (Figure 6(d)/(D2)). A similar differentiation pattern was observed in the newly formed epidermis after treatment with XD (Figure 6(a)/(A3), Figure 6(b)/(B3), Figure 6(d)/(D3)). The wound treated with Grassolind did not epithelize, although Carfilzomib adnexa remnants or individual keratinocytes were identified in the biopsies (Figure 6(a)/(A4), Figure 6(c)/(C4)).Figure 6Expression of differentiation markers of the keratinocytes cultured on XD at the air-liquid interface (A2, B2, C2, D2) compared to the normal skin (A1, B1, C1, D1), to freshly healing wound after necrectomy treated with XD (A3, B3, C3, D3), and to freshly …Table 1Keratinocyte differentiation in vitro and in vivo.The nuclear protein p63 was expressed in vitro in all keratinocytes of the submerged cultures and in the basal layer of the cultures grown at the air-liquid interface (Figure 6(c)/(C2)). In vivo, p63 was expressed not only in the basal layer but also in a subpopulation of suprabasal cells (Figure 6(c)/(C3)).