Influence of polysaccharides from Viscum album L. on human dermal fibroblasts in vitro

Polysaccharides (Pz) play an important role in different biological activities, such as inflammation, fertilization, cell-cell adhesion, signal transduction, etc [1]. The purpose of this study was to isolate and characterize Pz from European mistletoe (Viscum album L.) and to evaluate their in vitro influence on human dermal fibroblast (HDF), in order to use them in dermatological therapy. Water-soluble Pz (PZE) was fractionated by ethanol precipitation (A1-A4 fractions) and A2 fraction was purified by ion-exchange chromatography on DEAE-Spherodex LS (B1-B6 fractions). For all these fractions, carbohydrate, uronic acid and the protein content were determined. The influence of carbohydrate-rich Pz fractions (A2 and B3) and of PZE on fibroblast cell behaviour has been evaluated under in vitro conditions by MTT assay [2], light microscopy and gelatin-zymography [3]. Our results indicated that B3 fraction concentrations ranging from 0.2 to 100µg/ml stimulated the fibroblast mitochondrial activity. The highest registered value, corresponding to a concentration of 100µg/ml B3, was 126.5±3.4% compared to the control (100±2.9%). Moreover, light micrographs showed that cells maintained the typical fibroblast morphology and a proliferation rate similar to that of the control. A2 fraction and PZE did not induce cytotoxic effects on HDF in the range 0.2–100µg/ml. On the other hand, the majority of cells cultivated in the presence of A2 fraction and PZE, in concentrations above 100µg/ml, presented an alteration of their morphology and a lower proliferation rate (65%). Similarly, the synthesis of matrix metalloproteinases in culture medium was influenced by the concentrations of Viscum album Pz. In conclusion, Viscum album Pz stimulated HDF proliferation in a dose-dependent manner and in terms of their purification degree.

Acknowledgements: This work was supported by Romanian Project BIOSTAR PN-06–400107.

References: [1] Deters, A.M. et al. (2005) Planta Med. 71: 33–39. [2] Mossman, T. (1983) J Immunol Methods 65: 55–61. [3] Cimpean, A. et al. (1998) J Med Biochem 2: 313–322.