A cartilage tissue biomimetic matrix and preparation method thereof
A cartilage tissue and matrix technology, applied in the field of cartilage tissue bionic matrix and its preparation, can solve problems such as limited self-healing ability, lack of blood supply, and failure to achieve ideal results for cartilage defects, achieving no rejection and good compatibility Effect
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Embodiment 1
[0024] Chondrocyte culture: the cells were divided into 1×10 5 ~10 6 Cells / ml were seeded in a 10cm cell dish at 37°C, 5% CO 2 They were cultured in a carbon dioxide incubator, and the medium was changed once every 2 days. Cell morphology and growth were observed daily with an inverted microscope. After the cells were fused, they were digested with 0.25% trypsin, subcultured at a ratio of 1:4, cultured for 24-48 hours, digested and collected for later use. The medium is DMEM medium containing 15% fetal bovine serum, 1% glutamine and 1% 50U / mL penicillin and streptomycin.
[0025] After 24 hours of culture, the chondrocytes gradually adhered to the wall. The chondrocytes were elliptical or triangular in shape, with smooth surfaces, increased cell volume, increased cytoplasm, and multiplied vigorously. After 2 days, the cells gradually became dense and fusiform.
Embodiment 2
[0027] a. Polycaprolactone 20g is fully dissolved in 100ml trifluoroethanol to obtain a uniform polycaprolactone solution;
[0028] b. Preparation of chitosan / β-sodium glycerophosphate gel: 4°C-10°C, take 10ml 0.1mol / L β-sodium glycerophosphate solution and add dropwise to 10ml 2% (w / v) chitosan hydrochloric acid solution Stir and mix well, then add 10ml of 20% (w / v) polycaprolactone solution, and finally add the chondrocytes collected by digestion and mix well, the final concentration of cells is 5×10 5 ~10 6 cells / ml, mix the mixture evenly, put it into a 24-well culture plate and let it stand in a 37°C incubator until it solidifies. The configuration of 0.1mol / L β-sodium glycerophosphate solution: Dissolve 21.6g of β-sodium glycerophosphate in 100ml DMEM medium, in which DMEM medium contains 30% fetal bovine serum, 2% glutamine and 2% 50U / mL green , streptomycin, 10ng / ml TGF-β.
[0029] Observe the growth of the cultured cells in the chitosan / β-sodium glycerophosphate ge...
Embodiment 3
[0031] Select 40 clean-grade blue rabbits aged 6-12 months, with a body weight of 1.5-2.0 kg, half male and half male.
[0032] a. Polycaprolactone 25g is fully dissolved in 100ml trifluoroethanol to obtain a uniform polycaprolactone solution;
[0033] b. Preparation of chitosan / beta-glycerophosphate sodium gel: 4°C-10°C, get 10ml 0.1mol / L beta-glycerophosphate sodium solution and add dropwise to 10ml 3% chitosan hydrochloric acid solution, fully stir and mix, Then add 10ml of 25% polycaprolactone solution, and finally add the digested and collected chondrocytes and mix well, the final concentration of cells is 5×10 5 ~10 6 Cells / ml was calculated and the mixture was mixed evenly before use. The configuration of 0.1mol / L β-sodium glycerophosphate solution: Dissolve 21.6g of β-sodium glycerophosphate in 100ml DMEM medium, DMEM medium contains 30% fetal bovine serum, 2% glutamine and 2% 50U / mL green, Streptomycin, 10 ng / ml TGF-β.
[0034] Animal model preparation and experim...
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