A biomimetic glycosylated mineralized collagen/glycosylated chitosan/plga composite bone tissue engineering scaffold and its preparation method
A technology of bone tissue engineering and mineralized collagen, applied in tissue regeneration, pharmaceutical formulations, prostheses, etc., to achieve the effects of non-toxic molding performance, good molding performance, and good biocompatibility
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Embodiment 1
[0059] The preparation of glycosylated chitosan (HA / CTS, oHAs / CTS), the steps are as follows:
[0060] Weigh hyaluronic acid (HA) 450mg, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) 900mg, N-hydroxysuccinimide (NHS) 975mg , dissolved in 150mL morpholineethanesulfonic acid (MES) buffer (concentration 0.05M), stirred at 37°C for 1 hour, then added 900mg chitosan, continued to stir for 20 hours, centrifuged at 8000rpm for 10 minutes to obtain supernatant, and used a concentration of 0.1M Slowly adjust the pH of the NaOH solution to 7.1. During the adjustment process, the solution gradually became turbid. After continuing to stir for 4 hours, adjust the pH to 7.5. The solution became more turbid. Centrifuge at 8000rpm for 10 minutes to collect the precipitate, and wash the precipitate repeatedly with deionized water for 3 Centrifuge at 8000rpm after each washing, centrifugation time 10min / time; lyophilize and grind to obtain hyaluronic acid-modified chitosan p...
Embodiment 2
[0063] The preparation of glycosylated mineralized collagen (Col / HA / HAP, Col / oHAs / HAP) is as follows:
[0064] 1) Weigh 128 mg of collagen, 32 mg of hyaluronic acid (HA), and 48 mg of sodium cyanoborohydride and dissolve them in 8 mL of cross-linking reaction system. The mixed solution with a ratio of 3:2 was subjected to magnetic stirring at 37°C for 24 hours in the dark to obtain a cross-linking solution;
[0065] 2) Dilute the cross-linking solution in step 1) 6 times with acetic acid with a mass concentration of 5%, replace it 4 times in an ultrafiltration tube with a molecular weight cut-off of 30KDa, centrifuge at 4000g for 30min / time, and vacuum freeze-dry to obtain glycosylation collagen;
[0066] 3) Dissolving the glycosylated collagen in step 2) in 500 mL of 0.01M hydrochloric acid to prepare a glycosylated collagen solution with a concentration of 0.6 g / L;
[0067] 4) Slowly add 0.1M NaH to the glycosylated collagen solution obtained in step 3) 2 PO 4 Solution 4...
Embodiment 3
[0073] Glycosylated mineralized collagen / glycosylated chitosan / PLGA composite bone tissue engineering scaffold (Col / HA / HAP-HA / CTS-PLGA(5:1:4), Col / oHAs / HAP-oHAs / CTS- PLGA (5:1:4)), the steps are as follows:
[0074] 1) dissolving PLGA in 1,4-dioxane to obtain a PLGA solution with a mass concentration of 8%;
[0075] 2) the glycosylated chitosan (HA / CTS) and the glycosylated mineralized collagen (Col / HA / HAP) prepared in embodiment 1 and embodiment 2 are added in the PLGA solution, or, the embodiment The glycosylated chitosan (oHAs / CTS) and glycosylated mineralized collagen (Col / oHAs / HAP) prepared in Example 1 and Example 2 were added to the PLGA solution, stirred by magnetic force for 24 hours, and then ultrasonically treated at 400W for 20 minutes , continue magnetic stirring for 24h to obtain a suspension, wherein the mass ratio of Col / HA / HAP, HA / CTS to PLGA is 5:1:4, and the mass ratio of Col / oHAs / HAP, oHAs / CTS to PLGA is 5 :1:4;
[0076] 3) Inject the suspension obtained...
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