ABSTRACT
Hair Follicle Bulge region due to its availability and abundance is one of the areas which is easily accessible to Multi-potent stem cells that expresses Nestin marker [neuronal stem cells protein]. Stem cells bulge region in hair follicle stem cells has high potency to be differentiated to neuronal cells. Silibinin as an active component of Silybum marianum has anti-oxidant, anti-inflammatory, anti-carcinogenic, hepatoprotective, neurotrophic and neuroprotective effects. The aim of the present study was to evaluate the neurotrophic effects of silibinin on differentiation of hair follicle stem cells to neurons. Bulge area of whiskers in Rat was isolated and cultivated three weeks in supplemented DMEM/F12 and epidermal growth factor [EGF]. Then the cells were exposed over the concentrations of 0.05microg/ml, 0.1microg/ml, 0.4microg/ml, 0.5microg/ml, 0.7microg/ml Silibinin and Neurotrophin-3. Two weeks after culture, plated bulge cells were immunostained with Nestin and differentiated stem cells were immunostained with beta III tubulin by immunocytochemistry techniques. The results were evaluated by T-test student analysis. A Pvalue less than 0.05 was considered significant. The nestin marker was clearly demonstrated in bulge regions during the first week, but after two weeks, parallel to stem cells differentiating neuronal cells, beta III tubulin marker was expressed in neuronal cells. The toxic effects of 1microg/ml Silibinin on stem cells were also demonstrated, and it stopped the cell growth at the end of the first week. The maximum differentiation on stem cells in 0.5microg/ml Silibinin was observed to be significant [P<0.05]. Silibinin concentration increase led to reduced differentiation. Silibinin with neurotrophin 3 increased the differentiation of stem cells. Silibinin concentrations of 1microg/ml and more have toxic effects on hair follicles stem cell differentiation. Also, silibinin concentrations less than 0.1microg/ml had no effect on proliferation and differentiation hair follicle stem cells. Whereas 0.5microg/ml concentrations had significant effects on the differentiation processes of hair follicle stem cells to neuron