Nano-particle and gel for promoting dentin regeneration as well as preparation method and application of nano-particle and gel
A particle and matrigel technology, applied in the field of biomedicine, can solve problems such as the probability of tooth fracture, unsatisfactory long-term curative effect, and increase
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[0028] The invention provides a preparation method of AZD2858-CD-chitosan nanoparticles for promoting dentin regeneration, comprising the following steps: the concentration of AZD2858 in the diluent is 50-150nM;
[0029] The normal saline solution of DMSO and β-cyclodextrin is also included in the stock solution, and the volume ratio of the normal saline solution of DMSO and β-cyclodextrin is 1:(9~19); DMSO in the stock solution The volume percentage of β-cyclodextrin is 10% to 5%, and the mass percentage of β-cyclodextrin in the stock solution is 5 to 25%;
[0030] The mass ratio of the volume of the diluent to the chitosan is 400 μl: 1-3 mg.
[0031] The present invention firstly prepares AZD2858 (3-amino-6-[4-[(4-methyl-1-piperazinyl)sulfonyl]phenyl]-N-3-pyridyl-pyrazinecarboxamide) into 2.5 ~7.5mM stock solution, the solvent of described stock solution is the mixed solution that comprises the physiological saline solution of DMSO and β-cyclodextrin; The volume ratio of DM...
Embodiment 1
[0040] 1. Preparation of AZD2858-CD-chitosan nanoparticles
[0041] 1. Add 2g of β-cyclodextrin to 10ml of normal saline to prepare 20% β-cyclodextrin in normal saline solution.
[0042] 2. Weigh 2.12 mg of AZD2858, and use a mixed solution (10% DMSO + 90% physiological saline solution containing 20% β-cyclodextrin) to prepare a 2.5 mM stock solution.
[0043] 3. Take 25 μl stock solution + 975 μl 20% β-cyclodextrin physiological saline solution, and dilute to 50 nM.
[0044] 4. Weigh 1.5 mg chitosan and disperse it in 400 μl of 50 nM solution to obtain a solution containing AZD2858-CD-chitosan nanoparticles. The solution is dropped on the mica sheet, and after the solvent is evaporated to dryness, it is detected under a scanning electron microscope, and it can be seen that the AZD2858-CD-chitosan nanoparticle is a uniform particle structure ( figure 1 Middle A).
[0045] 2. Preparation of AZD2858-CD-chitosan@Matrix Gel
[0046] Add 100 μL Matrix Gel (BD Company, 10 mL) ...
Embodiment 2
[0051] Effects of AZD2858 and AZD-CD-chitosan on migration ability of apical papilla stem cells (SCAP)
[0052] SCAP was isolated and cultured in vitro, according to the cell density of 1×10 4 One per well was inoculated in the upper chamber of the transwell co-culture system, and non-specific regulatory bioactive factors amelogenin (AML), AZD2858 and AZD2858-CD-chitosan nanoparticles were added to the lower chamber of the co-culture system of each group. After 24 hours of culture, the migrating SCAP was digested, collected and counted. The results showed that AZD2858 and AZD-CD2858-chitosan nanoparticles could significantly promote the migration ability of SCAP, and there was no significant difference between the AZD2858 and AZD2858-CD-chitosan nanoparticles groups ( figure 2 ).
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