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Protein-loaded bioactive calcium phosphate nanoparticles and preparation method thereof

A bioactive, nano-particle technology, applied in tissue regeneration, drug delivery, pharmaceutical formulations, etc., can solve problems such as protein burst release

Pending Publication Date: 2021-09-10
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this method, growth factors and calcium phosphate nanoparticles are only bound by adsorption, which easily leads to sudden protein release.

Method used

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  • Protein-loaded bioactive calcium phosphate nanoparticles and preparation method thereof
  • Protein-loaded bioactive calcium phosphate nanoparticles and preparation method thereof
  • Protein-loaded bioactive calcium phosphate nanoparticles and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0043] The preparation method of the protein-loaded bioactive calcium phosphate nanoparticles of the present invention comprises the following steps:

[0044] (1), dissolving the protein in a buffer solution of Tris-HCL (Tris-HCL), stirring and mixing evenly by magnetic force, so that the growth factor (or model protein) solution is balanced to obtain the first mixed solution;

[0045] (2), add calcium chloride solution in the first mixed solution, magnetic stirring and mixing, obtain the second mixed solution;

[0046] (3), add magnesium chloride solution in the second mixed solution, continue blowing and mixing, obtain the third mixed solution;

[0047] (4), add disodium hydrogen phosphate solution in the third mixed solution, mix well, and a large amount of precipitation can be seen, and the reaction system is centrifuged to remove the supernatant to obtain the precipitate;

[0048] (5) Add double distilled water to the precipitate, wash it repeatedly for 3 times, and free...

Embodiment 1

[0060] The preparation method of the protein-loaded bioactive calcium phosphate nanoparticles of the present embodiment comprises the following steps:

[0061] (1) Weigh an appropriate amount of protein, dissolve it in a Tris-HCL buffer solution with pH=8.0, stir and mix well, and prepare the first mixed solution with a concentration of 1 mg / mL.

[0062] (2), in the first mixed solution, add calcium chloride solution dropwise to final concentration 25mmol / L, continue to stir, obtain the second mixed solution;

[0063] (3), in the second mixed solution, magnesium chloride is added dropwise to a final concentration of 25mmol / L, stirred evenly, to obtain the third mixed solution;

[0064] (4) Add disodium hydrogen phosphate dropwise to the third mixed solution to a final concentration of 15mmol / L to generate a large amount of precipitate, centrifuge at 8000rpm for 10min at high speed, remove the supernatant, and obtain the precipitate;

[0065] (5) The collected precipitate was ...

Embodiment 2

[0067] The pH value of Tri-HCL in Example 1 was changed to obtain calcium phosphate nanoparticles with different protein loads. Studies have confirmed that its loading exceeds 10%, which is about 15.64%.

[0068] Such as Figure 5 Shown is the protein release profile of calcium phosphate nanoparticles synthesized in Tris-HCL buffer with different pH, the higher the pH, the higher the loading and the lower the release value. When pH=8.0, the loading capacity is 13.79%; when pH=9.0, the loading capacity is 15.64%.

[0069] Sustained-release process: Dissolve the cytochrome C-loaded nanoparticle sample in a centrifuge tube at a concentration of 1 mg / mL in PBS buffers of different pH (pH=4.1, 6.1, 7.2), and then place it at 37°C, React in a constant temperature shaker at 120-200rpm; collect the supernatant by centrifugation at a specific time point and store it in a -20°C refrigerator, add an equal amount of PBS buffer, and continue the reaction in a constant temperature shaker;...

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Abstract

The invention provides protein-loaded bioactive calcium phosphate nanoparticles and a preparation method thereof. The preparation method comprises the following steps: dissolving protein in a tris-hydroxymethyl aminomethane hydrochloride buffer solution, then dropwise adding calcium chloride, magnesium chloride and a disodium hydrogen phosphate solution, uniformly mixing, centrifuging and freeze-drying. According to the method, the calcium phosphate nanoparticles with biological activity are prepared by simply mixing protein and inorganic salt components and rapidly reacting in an open environment, the process method avoids harsh conditions such as high temperature and high pressure, other toxic components are not introduced, the operation is simple, the reaction is rapid, the activity of growth factors can be effectively protected, the advantages of effective load and release of the growth factors are realized, and a basis is provided for subsequent clinical application; and besides, the size of the bioactive calcium phosphate nanoparticles is about 40 nm, the biocompatibility is good, the activity of growth factors can be effectively reserved, and proliferation and differentiation of cells can be promoted.

Description

technical field [0001] The invention belongs to the technical field of nanomaterial synthesis, and in particular relates to a nanoparticle with biological activity synthesized by co-precipitation of inorganic ions and biologically active protein, that is, protein-loaded biologically active calcium phosphate nanoparticle and a preparation method thereof. Background technique [0002] In the past few decades, many studies have proved that a variety of growth factors have high biological activity and biological specificity, and play an important role in life activities such as tissue repair and regeneration. However, like most proteins, growth factors are mostly unstable in vivo and have a short half-life. Therefore, protein nanocarriers with biological activity can not only protect protein activity, but also prolong its half-life, effectively avoiding adverse reactions caused by repeated injection and oral administration. However, there are still two problems that limit the r...

Claims

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Application Information

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IPC IPC(8): A61L27/12A61L27/22A61L27/54
CPCA61L27/12A61L27/22A61L27/54A61L2400/12A61L2300/252A61L2300/602A61L2430/02
Inventor 王佐林雷婧诗
Owner TONGJI UNIV