Inducing re-epithelialization in skin wound through cultured oral mucosal keratinocytes

OBJECTIVES: The purpose of this study was to investigate the wound healing effect of primary cultured oral mucosal keratinocytes (OMKs) and to assess their roles in skin wounds. MATERIALS AND METHODS: OMK labeled with BromodeoxyUridine were scattered onto 1.5x1.5 cm skin defects of adult female nude mice (OMK group, n=15). For the control, culture media were placed on the wound (control group, n=15). Mice in both groups were sacrificed at three days (n=5), one week (n=5), and two weeks (n=5), and histomorphometric and immunoblot analyses with keratinocyte growth factor (KGF), interleukin (IL)-6, and IL-1alpha antibody were performed for the biopsied wound specimen. To verify the effect of the cytokine, rhIL-1alpha was applied instead of OMK transplantation, and the OMK and control groups were compared with regard to re-epithelialization. RESULTS: Histomorphometric analyses demonstrated faster re-epithelialization in the graft group than in the control group at the third day, first week, and second week. Newly forming epithelium showed maintenance of the histological character of the skin epithelium. The graft group showed superior expression of KGF, IL-6, and IL-1alpha protein, compared with the control group. Similar faster re-epithelialization was observed after treatment with rhIL-1alpha instead of OMK transplantation. CONCLUSION: We successfully confirmed that the graft of primary cultured OMKs promoted regeneration of skin defects. The mechanism of accelerated wound healing by primary cultured OMKs was attributed to inducement of cytokine expression as required for re-epithelialization.

The inductive capacity of primary cultured oral mucosal keratinocytes in skin wound healing of athymic nude mice

AIM: The aim of this study was to investigate the mechanism of promoted skin wound healing in skin defects with primary cultured oral mucosal keratinocytes. MATERIALS AND METHODS: Thirty adult female nude mice weighing 20+/-2g were used for the experiment. Primary cultured and suspended oral mucosal keratinocytes, labeled with BrdU, were scattered onto 1.5cm*1.5cmsized full thickness skin defects in the experimental group(N=15), and no grafts were placed the control group(N=15). They were sacrificed at 3 days, 1 week and 2 weeks after the treatment respectively. Histological examination of each wounds were performed to review the healing progress on measuring the length from the wound margin to regenerating epithelial front. The role of keratinocytes were assessed by double immunohistochemical staining with Anti-BrdU and Anti-cytokeratin AE1/3. RESULTS: In the experimental group the wound was completely covered with regenerating epithelia in 2 weeks, but partially regenerated in the control group. The immunohistochemical studies unexpectedly reveal that most of regenerating epithelial cells were induced from marginal epithelium of the margin, not from the scattered keratinocytes. CONCLUSION: We could successfully confirm that graft of primary cultured oral mucosal keratinocytes promotes the regeneration of skin defects.