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01�C0.001). The early wound environment contains multiple inflammatory cytokines; in particular IL-1��, IL-6 and TNF-�� have been shown to be markedly increased in early skin wounds or injured NHEK [11, 13]. To examine whether a proinflammatory environment similar to that found at sites of skin injury might influence PEDF expression, NHEK was exposed to a range of concentrations of IL-1��, IL-6 or TNF-�� for 24?h. Treatment with IL1�� or TNF-�� at the highest concentration of 50?ng/ml resulted in a statistically significant decrease in PEDF mRNA expression (Fig.?1b, P?CHIR-99021 clinical trial result was further validated upon review of an available microarray GEO profile of IL-1�� treated keratinocytes (ID?35957232) [14]. IL-6 treatment also decreased the expression of PEDF at the two highest concentrations of 2 and 50?ng/ml, although the decrease did not reach statistically significance (Fig.?1b, P?>?0.05). Similar to the mRNA findings, levels of PEDF protein production were significantly decreased by each independent cytokine treatment at the 50?ng/ml dose (Fig.?1c, P?selleck screening library shown to inhibit migration and proliferation in the human keratinocyte cell line HaCAT [15] and in endothelial cells [1]; therefore, we wondered if PEDF would have similar effects on primary NHEK. Using a scratch wound assay on a monolayer of NHEK, cells were treated with exogenous purified recombinant human PEDF. The results revealed that treatment with PEDF at concentrations ranging from 0.2 to 500?ng/ml significantly inhibited NHEK migration in the scratch assay (Fig.?2a,b). This effect was further examined using a transwell migration assay. PEDF had a dose-dependent inhibitory effect on migration in the transwell migration assay (Fig.?2c), providing additional evidence that PEDF negatively affects keratinocyte migration. Furthermore, a similar exposure to PEDF resulted in a significant increase in the in vitro adhesion capacity of NHEK (Fig.?d). In contrast, PEDF treatment did not affect NHEK proliferation (Fig.?e). The above results suggest that the Fleroxacin early decrease in PEDF that is observed in wounds may be due to the surge of inflammatory cytokines and/or mechanical disruption of the keratinocyte layer. As the healing process progresses into the proliferative phase, the inflammatory response subsides, and PEDF returns to its uninjured level. In the resolving wound, restored levels of PEDF may inhibit further wound angiogenesis and encourage vascular regression. PEDF may modulate epithelial migration and thus support the return to skin homoeostasis. We speculate that secreted PEDF may play an important role in the dynamic reciprocity between the epidermal and dermal layers of a healing wound. Future studies regarding the in vivo role of PEDF are needed to fully understand its function in wound healing.