The Influence Of Nicotine,TNF-α,Epimedium Pubescen Flavonoid(EPF) On The Expression Of Osteopontin Of Rat Osteoblasts Cultured In Vitro

Chart One Identification of Rat Osteoblasts Cultured in vitroObjective:To culture and identify the in vitro functions of the rat osteoblasts (ROB) which wrere harvested from the cranial bone of normal subjects.Method: The newly born SD rats were sacrificed and the cranial bones of the rats were obtained cleanly and cut to blocks of 1mm× 1mm X lmm . After digested by collagenaseII for 15 minutes, the blocks were put into the culture flask. The phenotype,mineralized nodes and the expression of alkaline phosphatase were studied with alizarin red-S staining and modified Gomori's assays, respectively.Result:The ROB cells purified by modified method of tissue block functioned normally in vitro as observed in vivo. They showed the osteoblastic phenotypes such as their synthesis of alkaline phosphatase and osteocalcin and formatian of mineralized nodes.Conclusion : The study indicates that it is easy to purify plentiful ROBs with normal functions by modified method of tissue block, which can be used in the next research of bone metabolism.Key words: Rat osteoblast(ROB) cells; modified method of tissue block; identificationChart Two The Influence of Nicotine on the Expression of Osteopontin in Rat Osteoblasts Cultured in vitroObjective: To explore the influence of Nicotine on the expression of osteopontin in rat osteoblasts cultured in vitro.Method: When the passage cells confluenced more than 80%, the rat osteoblasts of the forth generation cultured in vitro were used for next step. Three groups were divided .With the intervention of Nicotine, Nicotine+ Atracurium Besilate(Tubocurarine) and sham control respectively, the passage cells were cultured continually for another 48...