Tissue engineered bone-cartilage complex tissue graft and preparation method thereof
A technology of composite tissue and tissue engineering, applied in medical science, prosthesis, etc., can solve the problems of poor quality of cartilage part and poor combination of osteochondral interface, etc., and achieve the effect of increasing adhesion ability
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Embodiment 1
[0031] 1. In vitro culture of BMSCs
[0032] One-month-old SD rats were killed by dislocation of the neck, and the bilateral femurs were removed, and the soft tissues were removed, soaked in 75% alcohol for 5 minutes, washed twice with PBS, cut off one end of the femur, and the volume fraction was 10% with a syringe with a No. 5 needle. Wash the bone marrow cavity of the femur with DMEM (containing 100 U / ml penicillin, 100 μg / ml streptomycin, and 20 U / ml heparin) of fetal bovine serum, and slowly inject 4 ml of the mixed bone marrow fluid into the pre-prepared centrifuge tube containing 6 ml of lymphocyte separation medium , centrifuge at 800G for 20min, take out the mononuclear cells in the middle layer and inoculate them on 25cm 2 Culture flask, 37°C, 5% CO 2 Culture in an incubator. Change the medium for the first time after 5 days, and change the medium every 3 days thereafter. Passage after overgrown, and take the third generation of cells for experiments.
[0033] 2. ...
Embodiment 2
[0038] As another preferred embodiment, the difference between this embodiment and embodiment 1 is:
[0039] In the synthesis steps of PLGA preparation, in PBS solution 1, the concentration of bFGF was 1 μg / ml; in PBS solution 2, the concentration of TGF-β1 was 1 μg / ml; in PBS solution 3, the concentration of BMP-2 was 1 μg / ml.
[0040] The pore diameter of the osteogenic part of the scaffold material is 50 μm, and the porosity is 85%; the pore diameter of the chondrogenic part of the scaffold material is 200 μm, and the porosity is 95%.
Embodiment 3
[0042] As another preferred embodiment, the difference between this embodiment and embodiment 1 is:
[0043] In the synthesis steps of PLGA preparation, in PBS solution 1, the concentration of bFGF was 3 μg / ml; in PBS solution 2, the concentration of TGF-β1 was 3 μg / ml; in PBS solution 3, the concentration of BMP-2 was 3 μg / ml.
[0044] The pore diameter of the osteogenic part of the scaffold material is 80 μm, and the porosity is 88%; the pore diameter of the chondrogenic part of the scaffold material is 180 μm, and the porosity is 92%.
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