Extensive oral mucosa loss from a variety of conditions is associated with significant functional morbidity and mortality. Although it is known that keratinocytes are a rich source of wound healing promoting factors such as transforming growth factor-β1(TGF-β1), it is not clear whether differentiated keratinocytes in a multi-layer form release this multi-functional growth factor. This study examined the hypothesis that keratinocytes in mono- and multi-layer forms expressed different levels of TGF-β1. When NHOK reached confluency in serum free medium(KBM), in test medium containing 1.2 mM Ca++ KBM NHOK were allowed to form multi-layers and differentiate. The purpose of this study were to investigate the mRNA level of TGF-β1, FGF-2, and TIMP-1 by RT-PCR analysis and also to evaluate the expression of TGF-β1 and involucrin in keratinocytes at different times of the onset of differentiation. The numbers and sizes of these nodules were increased as the process of keratinocyte differentiation proceed. Cultured NHOK in preconfluency under KBM medium expressed a significantly higher level of TGF-β1 relative to those grown in multi-layer forms, while the level of TGF-β1 mRNA gradually reduced to its lowest level at 7 days of growing cells in test medium. Cultured NHOK in preconfluency of KBM medium expressed a lower level of FGF-2 and TIMP-1 relative to those grown in multi-layer forms, while the level of FGF-2 and TIMP-1 mRNA showed the highest level at 3 days at gradually reduced to its lowest level at 7 days of growing cells in test medium. As a differentiation marker for keratinocytes at different time points, the highest level of involucrin mRNA expression was found at the later stage of cell differentiation. It suggested that the expression of TGF-β1 mRNA be consistent with the expression of FGF-2 and TIMP-1 mRNA in NHOK grown in high calcium medium during the terminal differentiation. But differentiated NHOK expressing higher involucrin mRNA could show constant espression of TGF-β1, FGF-2 and TIMP-1.

Hematopoietic stem cells (HSCs) can self-renew and can differentiate to a variety of specialized blood cells). The proliferation and homing of HSCs are strictly regulated both in the system level and local level. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a hematopoietic growth factor, potent species-specific stimulator of granulocyte-macrophage, eosinophil, megakaryocyte and erythroid progenitors. In clinical purpose GM-CSF has been used as hematopoietic growth factor. It can promote the mobilization of HSC from bone marrow to peripheral blood. The number of HSCs mobilized into blood can be modulated by the kinds of cytolines. However, the information for the cytokine which promote mobilization is limited. Basic fibroblast growth factor (bFGF or FGF-2) induces the change of niche and affects the maintenance and differentiation of HSCs. FGF-2 positively regulates hematopoiesis, by acting on stroma cells, on early and committed hematopoietic progenitors, and possibly on some mature blood cells. In this study, we investigated the effect of FGF-2 on HSCs mobilization and proliferation compare to GM-CSF. GM-CSF and FGF-2 were injected for 2 or 5 days into peritoneum of CD-1 mice (6～8 wks old) and sampling the bone marrow and peripheral blood. The bone marrow cells and peripheral blood were analyzed using FACS. In GM-CSF group, the number of HSCs was significantly increased by 2 days of injection but was significantly decreased in 5 days of injection. On the other hand the number of HSCs was significantly increased by the administration of FGF2 both in 2 days and 5 days. GM-CSF and FGF-2 are all increased the number of HSC both in bone marrow and peripheral blood. From these results, it is revealed that chronic administration of GM-CSF does not cause of the increase the number of HSCs. On the other hand, FGF2 can stimulate the proliferation of HSC without inhibition by the treatment period. It is suggested that GM-CSF and FGF2 may use different mechanisms to stimulate the HSCs proliferation. IP: 220.149.***.