The Experimental Study On Effects Of Chinese Recipe Of Replenishing Kidney And Activating Blood Circulation On Osteoblasts In Vitro

Objective: To investigate the effect of Chinese Recipe of Replenishing kidney and Activating Blood liquid Circulation(CRRKABCL) for the cell proliferation,cell differention,and mineralization of osteoblast cultivated in vitro hat belong to neonatal SD rat, in order to explore its mechanism of prevention and treatment of osteoporosis andprovidethe base of experimental theory for its more exploitation and investigation.Methods:Culture of osteoblast:To obtain osteoblasts from neonatal SD rat's skull using the method of twice enzyme digestion, conduct primary culture and subculturing, take the tertiary cells as experimental model, and then culture these cells with different concentration of the CRRKABCL together.Identification of osteoblast:Inverted phase contrast microscope is used to observe morphological change of the osteoblasts'growth; and then alkaline phosphatase staining are used by adopting ameliorative Gomori's calcium-cobalt.Assaying function of cell proliferation:MMT is used (to indicate the OD value at the point which wave-length is 570nm).(4)Assaying function of cell differentiation: Activity of akaline phosphatase is assayed by PNPP; content of osteocalcin in RPMI1640 is assayed by osteocalcin radioimmunoassay .Assaying function of cell mineralization: Alizarin bordeaux staining method is used, and mineralzation nodes are counted under optical microscope at 40 times.(indicating with mineralzation nodes count per eyeshot)Result: relatively rarefied osteoblasts can be obtained by means of cell culture in vitro using the method of twice enzyme digestion . CRRKABCL with different concentration(20 u g/ml 40 u g/ml 80 u g/ml)can increase quantity of osteoblasts cultivated in vitro,and there are dosagedependence and temporal positive correlation between auxo-action and drug concentration. (3) CRRKABCL can promote osteoblast to synthesize and secrete akaline phosphatase,and increase secrete of osteocalcin in osteoblast with increased dosage.CRRKABCL can stimulate osteoblast to enhance the amount of mineralization node count.Conclusion:CRRKABCL can promote cellproliferation of the osteoblast cultivated in vitro and increase cellproliferation rate. CRRKABCL can accelerate osteoblast to further differentiation and maturation.CRRKABCL can accelerate mineralization formation of the osteoblasts cultivated in vitro.CRRKABCL can advance bone formation of the osteoblast cultivated in vitro.