The Influence Of Different Concentration Of The Aprotinin And Tranexamic Acid On The Degradation Speed Of The Fibrin-chondrocyte Compound In Vitro.

Objective: In order to study the character of deferent kinds of FG bracket and find the best balance point of the degradation rate of the FG bracket and the chondrocyte propagate, maintaining of surface type and cytoplasm secretion, we add different concentration of the aprotinin and tranexamic acid into the FG bracket to see the influence of the concentration to the degradation rate and the chondrocyte propagate, maintaining of surface type and cytoplasm secretion. We have done some task to make the tissue engineer method come into the clinical phase Methods: 1.The chondrocytes from articular cartilage of 3 weeks old New Zealand rabbit were Isolated and monolayer cultured, then the cultured chondrocytes were seeded on the FG scaffold. The improved FG bracket of different concentration of the aprotinin ( 5000KIU/ml, 10000KIU/ml 20000KIU/ml) and tranexamic acid (10mg/ml> 20mg/ml 30mg/ml) and standard fibrin-chondrocyte bracket were divided into 9 groups by different concentration of the aprotinin and tranexamic acid, then cultured and amplified for 6 weeks in three-dimensional spatial in vitro. The behavior of chondrocytes cultured in fibrin glue was observed by histology and electron microscopy and type II collagen immunohistochemical test(S-P test). The mass and degradation speed of fibrin-chondrocyte constructs were evaluated in different periods. Results:1.The chondrocyte propagated under the electron microscopy in the standard group and the improved group 1,2,4,5. The cells divided in the improved group 7,8 after one week of culture, the cell shape is like the arborization. The chondrocyte is ameba like after 9 days of culture in the improved group 9. 2. The chondrocyte excreting extracellular matrix were observed under the electron microscopy in the standard group and the improved group 1,5. 3. From The type II collagen immunohistochemical test, the brown granules were seen surround the chondrocyte in the blank bubble after one week and two weeks of culture. The chondrocyte could excrete extracellular matrix in three-dimensional spatial. 4. After one weekand two weeks of culture, The difference of the quality residual between the standard group and all improved groups were statistically significant (P<0.001).the degradation speed of all the improved groups were lower than that of the standard group. The difference was statistically significant. The difference of the quality residual after 1 and 2 weeks of culture was statistically significant between improved groups 1 and 9. The FG bracket collapsed after 4 weeks of culture in the standard group, but only few FG brackets collapsed after 4 weeks of culture in the improved group. 5. The average double increase time of the chondrocyte is 4.7 days in the standard group and improved group 1,5. The average double increase time of the chondrocyte is 5.9 days in improved group 9. The cell growth curve of the standard group is similar to the improved group 1, 5 and the cell growth curve of the improved group 9 is significantly lower than the others. Conclusion: l.The degrading speed of FG in vitro slowed significantly by adding the aprotinin and the tranexamic acid. As the concentration grow higher, the rate of the degradation rate of FG become lower. 2. The chondrocyte propagating is influenced by different concentration of the aprotinin and the tranexamic acid. Higher concentration of the aprotinin and the tranexamic acid restrain the chondrocyte propagate, but when the concentration of the aprotinin is lower than 10000KIU/ml and the tranexamic acid concentration is 20mg/ml, the chondrocyte propagate was not influenced. 3. Higher concentration of the aprotinin (10000KIU/ml) influences the maintaining of the cell surface type and makes the chondrocyte show polarization change. 4. When the concentration of the aprotinin is 10000KIU/ml and the tranexamic acid concentration is 20mg/ml, the degrading speed of the FG bracket was the lowest, and the propagating of the chondrocyte, the maintaining of the surface type and the extra-cellular excrete were not influence...