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M Gröschl Department of Paediatrics, University of Erlangen-Nürnberg, Germany
Institute for Laboratory Medicine, University of Leipzig, Germany

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H-G Topf Department of Paediatrics, University of Erlangen-Nürnberg, Germany
Institute for Laboratory Medicine, University of Leipzig, Germany

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J Kratzsch Department of Paediatrics, University of Erlangen-Nürnberg, Germany
Institute for Laboratory Medicine, University of Leipzig, Germany

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J Dötsch Department of Paediatrics, University of Erlangen-Nürnberg, Germany
Institute for Laboratory Medicine, University of Leipzig, Germany

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W Rascher Department of Paediatrics, University of Erlangen-Nürnberg, Germany
Institute for Laboratory Medicine, University of Leipzig, Germany

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M Rauh Department of Paediatrics, University of Erlangen-Nürnberg, Germany
Institute for Laboratory Medicine, University of Leipzig, Germany

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We investigated the influence of salivary leptin, purified by affinity chromatography, on the proliferation of human oral keratinocytes. Furthermore we determined the time- and dose-dependency of the incubation with salivary leptin on the production of epidermal growth factor (EGF) and keratinocyte growth factor (KGF), which are growth factors relevant to keratinocyte proliferation. The analysis was performed both intra- and extracellularly. The relationship between the three cytokines in cell proliferation was studied by successive blocking with specific antibodies. The incubation of oral keratinocytes with recombinant and native leptin led to a significantly increased, dose-dependent cell proliferation (P<0.001). A further significant increase in proliferation was observed after incubating the cells with sterile filtered saliva (P<0.001). The increase in proliferation could not be observed by simultaneous incubation with salivary leptin and specific antibodies against either leptin or KGF (P<0.001). We found a significant dose-dependency between leptin incubation and production of KGF and EGF at the RNA and protein level. Both cytokines were expressed intracellularly and released into the culture medium, where they could be quantified by ELISA. Furthermore, there was a dose- and time-dependent increase in the phosphorylation of STAT-1 and STAT-3, indicating that Ob-Rb (the long form of the leptin receptor) expressed by the keratinocytes is functional. It is conceivable that the leptin-induced proliferation in keratinocytes is mediated by this signalling pathway. This is the first study to show a physiological role of salivary leptin as a growth factor for keratinocyte proliferation in the oral cavity. We could demonstrate its influence on the production of other growth factors essential for this proliferative effect. Based on the findings of our study we assume an important role for salivary leptin in wound healing within the vulnerable oral cavity.

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