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Functional chitosan porous microsphere for cartilage injury repair and preparation method of functional chitosan porous microsphere

A technology of porous microspheres and chitosan, applied in biochemical equipment and methods, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the particle size and Problems such as small pore size, inability to meet the function of cell microcarriers, and difficulty in removing porogens can achieve the effects of promoting differentiation into chondrocytes, good biodegradability, and increasing drug loading

Active Publication Date: 2019-09-20
EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In previous studies, a porogen was used in the preparation of chitosan porous microspheres, but the addition of the porogen made subsequent removal difficult, and the process was complicated and the environmental burden was heavy (CN1927892A)
Patent CN103804712A discloses a method for preparing porous and controllable chitosan microspheres. The invention uses electrostatic spraying and vacuum freeze-drying techniques to prepare chitosan porous microspheres with adjustable pore size and specific surface area. The particle size and pore size of chitosan porous microspheres are small, which cannot meet the function of cell microcarriers

Method used

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  • Functional chitosan porous microsphere for cartilage injury repair and preparation method of functional chitosan porous microsphere
  • Functional chitosan porous microsphere for cartilage injury repair and preparation method of functional chitosan porous microsphere
  • Functional chitosan porous microsphere for cartilage injury repair and preparation method of functional chitosan porous microsphere

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preparation example Construction

[0036] The second aspect of the present invention provides a method for preparing functionalized chitosan porous microspheres for repairing cartilage damage, the steps comprising:

[0037] S1. Chitosan is added to an acetic acid solution with a mass fraction of 1 to 3%, stirred and heated to dissolve, and cooled and defoamed after the solution is clarified to obtain the chitosan acetic acid solution as the water phase (W) for emulsification;

[0038] S2. Select a water-insoluble organic solvent as the oil phase (O), add the chitosan acetic acid solution obtained in step S1 dropwise to the pre-emulsified oil phase, and stir to form a W / O emulsion;

[0039] S3. Slowly add the W / O emulsion obtained in step S2 into the pre-cooled oil phase in step S2, and let it stand at a low temperature of -15 to -25°C. After the chitosan emulsion solidifies into a ball, suck out the n-heptane Alkanes solution to remove the oil phase, replace it with absolute ethanol and then stand at -15~-25°C ...

Embodiment 1

[0045] The present embodiment provides a kind of preparation method of chitosan porous microspheres for cartilage damage repair, comprising the steps:

[0046] 1. Weigh a certain amount of chitosan (molecular weight is 110kDa, degree of deacetylation is 95%) and add it to 1wt.% acetic acid solution, heat to 50°C and magnetically stir to dissolve, cool and defoam after the solution is clarified for use. The massfraction of chitosan is 2.5wt.%, as the aqueous phase (W) of emulsification usefulness;

[0047] 2. Prepare 72 mL of n-heptane solution (O) containing 4wt.% Span80, take 12 mL of the above-mentioned chitosan solution and slowly add it dropwise to the pre-emulsified n-heptane solution, and magnetically stir at 500 rpm for 1 hour to form a W / O emulsion;

[0048] 3. Pre-cool the glass plate (18cm) filled with n-heptane to -20°C, slowly and evenly drop the above W / O emulsion into the glass plate, and cool at -20°C for 24h. After the chitosan emulsion droplets solidified int...

Embodiment 2

[0052] The present embodiment provides a kind of preparation method of functionalized chitosan porous microspheres for cartilage damage repair, comprising the steps:

[0053] 1. Weigh a certain amount of chitosan (molecular weight is 110kDa, degree of deacetylation is 95%) and add it to 2wt.% acetic acid solution, heat to 40°C and magnetically stir to dissolve, cool and defoam after the solution is clarified for later use. The massfraction of chitosan is 3wt.%, as the aqueous phase (W) of emulsification usefulness;

[0054] 2. Prepare 65 mL of n-heptane solution (O) containing 3wt.% Span80, take 13 mL of the above-mentioned chitosan solution and slowly add it dropwise to the pre-emulsified n-heptane solution, and magnetically stir at 600 rpm for 0.5 h to form a W / O emulsion;

[0055] 3. Pre-cool the glass plate (15cm) filled with n-heptane to -23°C, slowly and evenly drop the above W / O emulsion into the glass plate, and cool at -23°C for 12h. After the chitosan emulsion dropl...

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Abstract

The invention discloses a functional chitosan porous microsphere for cartilage injury repair and a preparation method of the functional chitosan porous microsphere. The preparation method includes: adding a chitosan acetic acid solution to an organic solvent, stirring to form a W / O emulsion, dropwise adding the W / O emulsion into a precooled glass dish containing the same organic solvent, adding anhydrous ethanol for dewatering after emulsion droplets solidify into a ball, and finally adding residual acetic acid in a sodium hydroxide solution and the microsphere; adding glutaraldehyde to the microsphere washed to be neutral for further chemical cross-linking, removing free glutaaldehyde with glycine, and performing freeze-drying after cleaning to obtain the chitosan porous microsphere; connecting small molecular drug KGN (kartogenin) with the chitosan porous microsphere by chemical bonding to endow biological activity of differentiation of bone marrow mesenchymal stem cells to the chitosan porous microsphere. The chitosan porous microsphere prepared by the method has a three-dimensional network porous structure of a bionic structure, which provides space and conditions for the formation and growth of neonatal cartilage cells; the preparation method is simple, the structure is controllable, the spherical degree is good, and a broad application prospect in the field of cartilage tissue regeneration is achieved.

Description

technical field [0001] The invention belongs to the field of biomedical polymer materials, and relates to a functionalized chitosan porous microsphere for cartilage damage repair and a preparation method thereof. Background technique [0002] Cartilage damage is a very common clinical disease, especially in patients with osteoarthritis (Osteoarthritis, OA) caused by the aging population. Cartilage tissue is a terminally differentiated tissue, lacks blood vessels, and its nutrient supply mainly comes from joint fluid, so it is difficult to repair itself after injury. At present, there are various drawbacks in both drug therapy and surgical therapy clinically. The development of tissue engineering has brought new opportunities for the repair of cartilage damage. The traditional tissue engineering technique is to implant biomaterial scaffolds into cartilage defects, but secondary surgery is still unavoidable, which undoubtedly adds a lot of psychological and economic burden t...

Claims

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Application Information

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IPC IPC(8): A61K47/61A61K47/69A61K31/196A61P19/08C12N5/0775
CPCA61K47/61A61K47/6927A61K31/196A61P19/08C12N5/0663C12N2513/00C12N2533/72
Inventor 杨光李晓宏尹雅琪曹佳梦
Owner EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH
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