Effect of Cyanobacteria (Nostoc species) Extracts on Osteogenesis Activities

This study focused on osteogenesis activities from Cyanobacteria (Nostoc species), including cytotoxicity, cell proliferation, cell differentiation and cell mineralization with osteoblast cell (MC3T3-E1). The four Nostoc species; N. parmelioides, N punctiforme, N. muscorum and N. paludosum were made axenic and cultured for a month. The ethanolic crude extracts were obtained by maceration extraction; the yield was in the range of 7.00 - 10.00 % (w/w). By using the TLC technique, it was observed that in all of the samples, there were phytochemical bands corresponding with β-carotene and chlorophyll a which were used as standard compounds. As a result, all crude extracts had no toxic effect on osteoblast cells after being incubated for 24 and 48 h with various concentrations (1 - 1,000 µg/mL), although they did not significantly promote osteoblast proliferation. In addition, it was found that 3 crude extracts (N. parmelioides, N punctiforme, and N. paludosum) also promoted osteoblast differentiation significantly after being induced for 9 days. Similarly, there was no difference between the relative cellular mineralization among treatment groups after being induced for 25 days. The potential ethanolic extracts on osteoblast cell differentiation were further partitioned by liquid-liquid partition chromatography (hexane, dichloromethane and aqueous fractions), and were analyzed for osteoblast differentiation with alkaline phosphatase (ALP) activity. All of hexane and dichloromethane fractions from N. parmelioides (181.72±6.75 % and 195.11±3.27 %), N punctiforme (194.34±2.36 % and 168.69±1.08 %), and N. paludosum (191.54±7.66 % and 167.12±9.46 %) had a significantly higher effect of ALP activity than their aqueous fractions. In conclusion, our data suggested that the nonpolar potential major compound, which might correspond with carotenoids (β-carotene, xanthophyll and chlorophyll a) from Nostoc species, exhibited positive effects on osteoblast differentiation by increasing the alkaline phosphatase enzyme.