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.2014;47(5):195-201.
doi: 10.1267/ahc.14015. Epub 2014 Sep 12.

Re-epithelialization of the Buccal Mucosa after Alkaline Chemical Injury

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Re-epithelialization of the Buccal Mucosa after Alkaline Chemical Injury

Saneyuki Takaichi et al. Acta Histochem Cytochem.2014.

Abstract

Alkaline conditions in the oral cavity may be caused by a variety of stimuli, including tobacco products, antacids, alkaline drinking water and bicarbonate toothpaste. However, the effects of an alkaline pH on the oral mucosa had not been elucidated. The purpose of this study was to investigate how basal keratinocytes are actively involved in re-epithelialization after alkaline chemical injury. We generated epithelial defects in the oral mucosa of mice by applying an alkaline chemical, and the localization of cytokeratin 13, cytokeratin 14, PCNA and p63 was investigated during the re-epithelialization process. PCNA- and p63-positive staining was seen in basal cells covering the wound surface at 1 day after the chemical injury. Cytokeratin 14-positive and PCNA-negative basal keratinocytes were localized in a few layers of the wound epithelium during epithelial outgrowth. Cytokeratin 14-positive and PCNA-positive basal keratinocytes, indicating proliferation, were localized over the entire layer of the epithelium at the wound margin. These results imply that basal keratinocytes at the wound margin migrate to the wound surface, provoke differentiation and keratinization during epithelial outgrowth and that epithelial cells are supplied from the wound margin to the epithelial outgrowth after alkaline chemical injury.

Keywords: chemical injury; oral mucosa; re-epithelialization; wound healing.

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Figures

Fig. 1.
Fig. 1.
Histological observations of chemically injured oral mucosa during epithelial outgrowth after 24 hr (A, A’), 72 hr (B, B’), 120 hr (C, C’) and 1 week (D, D’) after injury and in normal tissue (Control;E, E’). Higher magnifications of theboxed regions inA–E are shown asA’–E’: respectively. The epithelium of the buccal mucosa was identified as the stratified squamous type.E, E’: Epithelial elongation is observed from both sides of the wound margin toward the chemical injury.A, A’, B, B’: Re-epithelialization is observed in the degenerated muscle tissue.C, C’: Regenerated epithelia from both sides of the wound margin are connected.D, D’: The epithelium of the buccal mucosa has been completely regenerated and resembles the normal epithelium (E, E’). Bars=100 μm (A–E), 150 μm (A’–E’).
Fig. 2.
Fig. 2.
Immunohistochemical observations of PCNA, p63 and cytokeratins 13 and 14 in normal tissue and in epithelial outgrowth of the chemically injured oral mucosa.A, A’, B, B’: After 24 hr and 72 hr, PCNA-positive cells are observed at the leading edge of the epithelial outgrowth in the basal layer (white arrowhead).C, C’: After 120 hr, PCNA-positive cells are observed near the contact area of the epithelial outgrowth where the leading edges of epithelial cells migrate towards each other (white arrowheads).D, D’: After 1 week, PCNA-positive cells are observed in the basal layer as well as in normal epithelium.E, E’, F, F’: After 24 hr and 72 hr, p63-positive cells are observed at the leading edge of epithelial outgrowth in the basal layer as well as PCNA-positive cells (white arrowhead).G, G’: After 120 hr, p63-positive cells are observed in the center of the contact area where both leading edges of epithelial cells have migrated toward each other (white arrowheads).H, H’: After 1 week, p63-positive cells are observed in the basal layer as well as in normal epithelium.I, I’: After 24 hr, cytokeratin 14 is expressed in all layers of the epithelial outgrowth, but cytokeratin 13 is not expressed in the epithelial outgrowth at all. Cytokeratin 13 is expressed in the debris of the epithelium (asterisks).J, J’: After 72 hr, cytokeratin 14 is expressed in the leading edge of the epithelial outgrowth, and cytokeratin 13 is expressed in the suprabasal layer as it chases cytokeratin 14 (asterisks).K, K’: After 120 hr, cytokeratin 14 is expressed in the contact area where both leading edges of the epithelial epithelium migrate towards each other and cytokeratin 13 is expressed in the suprabasal layer as it chases cytokeratin 14 (asterisks).L, L’: After 1 week, cytokeratin 14 is expressed in the basal layer and cytokeratin 13 is expressed in the suprabasal layer as well as in normal epithelium.White dotted line inA–O, A’–L’ demarks the basement membrane. Bars=100 μm (A–O), 50 μm (A’–L’).
Fig. 3.
Fig. 3.
Schematic diagram of normal epithelial characteristics and keratinocyte proliferation and migration in normal (A) and wound epithelium (B, C, D).A: Immunohistochemical localizations of PCNA, p63 and cytokeratins 13 and 14, in the normal tissue are summarized.Red nuclei represent PCNA-positive cells.Blue nuclei represent p63-positive cells andyellow nuclei represent PCNA and p63-negative cells. Thepink color box shows cells cytokeratin 14-positive cells in the basal keratinocytes and thesky blue color box shows cytokeratin 13-positive cells in the suprabasal basal keratinocytes.B: At 24 hr after the wound injury, the basal keratinocytes are activated to migrate and lead the edge of the wound epithelium. However, PCNA- and p63-positive cells are only seen in the basal layer of the wound margin and in normal epithelium, and are not observed in the epithelial outgrowth of the wound epithelium.C: At 72 hr after the wound injury, as basal keratinocytes surrounding the wound margin evidence a positive reaction for PCNA and express cytokeratin 14, these cells are proliferating and contribute to the source of cells available for wound closure.D: At 120 hr after the wound injury, the majority of cells in the regenerated epithelium are positive for p63, thus indicating that the majority of the regenerated epithelium is formed by basal keratinocytes that have migrated from the surrounding epithelium. A small number of keratinocytes in the regenerated epithelium are negative for p63, which suggests that these cells may have originated from migrating basal keratinocytes or from original suprabasal keratinocytes.
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