A Study On The Influence Of FN And Fluid Shear Stress On The Expression Of COX-2 In HPDLF

ObjectiveThe design of experiment is to utilize the fluid shear stress to simulate occlude wound in vitro. Giving different density of FN to HPDLF and measure the amount of COX -2 mRNA of HPDLF on different time. By this, to move forward a single step to utilize the influence of the FN to HPDLF in oral, especially when it was in the occlude wound.Materials and Methods1. Tissue resource:To collect young healthy permanent teeth which were plucked for the straightening out purpose from the dentofacial surgery in hospital for stomatology of China Medicial University. Primary culture the HPDLF with the method of tissue mas.2. Primary culture the HPDLF and AppreciationPrimary culture the HPDLF with the method of tissue mas. With the i detection of the mmuocytochemistry, it shows that anti - filamin is masculine while anti - ceratin is negative. This evid that this kind of cell is come from me-soblastema. We take the 3 generation cell as the study target3. Sample dealThere are two groups in this study , A and B. Group A were cultivated with the fluid shear stress given by a special swing bed. Group B were cultivated quietly. We put different density of FN (0ug/ml 20ug/ml,40ug/ml,60ug/ml) to Group A snd B. Without seroculture for 6 and 12 hours . The speed ot the swingbed is 35r/min.4. Semiquantitative RT - PCR< 1 > To extract con - RNA <2> RT-PCR< 3 > Electrophoetic analysis of the production of RT - PCR< 4 > Statistics analysisResults1. The Results of primary culture the HPDLF and AppreciationAfter 24 hours, 80% tissue mas adherenced. After 7-10 days,there are cells come out from the suround of the tissue mas, After about 14 days, the cell overgrow the culture flask. With the i detection of the mmuocytochemistry, it shows that anti - filamin is masculine while anti - ceratin is negative. This evid that this kind of cell is come from mesoblastema.2. The Results of RT - PCRElectrophoretic analysis of the production of RT - PCR and densitometric scan. Then we use t - test to analysis all the optical density values. The result is t = 13. 45, P < 0. 01. The express of COX - 2 mRNA shows after 6 hours in group A, and the amount of the express grows downwards with the increase of the amount of FN . The express of COX -2 mRNA shows after 12 hours in group A is higher than the amount of the express shows after 6 hours. The result of the e-lectrophoresis for group B is negative. The electrophoresis screenage optical density of (3 - actin amplification fragment is uniform.Conclusion1. The fluid shear stress, as a wound factor simulated in vitro, can revoke the inflammatory reaction of HPDLF as other factors causing phlegmasia. And this kind of inflammatory reaction will strengthen with increase of the length of stressing time.2. Human plasma FN can grow downwards the express amount of COX - 2mRNA of HPDLF, and different density of FN shows different growing downwards the express amount of COX - 2 mRNA of HPDLF. There is dose and time dependent between the two factors.