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Subcritical carbon dioxide sintering method for co-loading porous microsphere scaffolds

A porous microsphere, carbon dioxide technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of unfavorable protein drug activity, difficult removal of organic solvents, poor cell compatibility, etc., and achieves favorable mass transfer. Effective with cell proliferation, volatile, and simple steps

Active Publication Date: 2016-01-20
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high temperature and organic solvents are not only detrimental to the activity of protein drugs, but also the residual organic solvents are difficult to remove, resulting in poor cell compatibility.
Although people try other methods to prepare PLGA microsphere scaffolds, such as Wang Yingjun et al. (CN103212116A) disclose a method for preparing PLGA porous microsphere scaffolds by normal temperature and low pressure drying; The method of microsphere scaffold, but it is impossible to co-load cells under the premise of ensuring the surface morphology of microspheres, or provide more adhesion sites for cells in the case of co-loading cells, so that cells can attach to a single microsphere unit. Internal growth, avoiding the problems of blocked regeneration of blood vessels and apoptosis due to blocked mass transfer

Method used

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  • Subcritical carbon dioxide sintering method for co-loading porous microsphere scaffolds

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Experimental program
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Effect test

Embodiment 1

[0041] The specific implementation steps are as follows:

[0042] a) At room temperature, add PLGA to the organic solvent dichloromethane, and obtain a homogeneous oil phase by stirring, wherein the concentration of PLGA is 6.25% (w / v), the molecular weight of PLGA is 40kDa, LA (lactic acid) and GA in PLGA (glycolic acid) in a mol ratio of 50 / 50;

[0043] b) Ammonium bicarbonate is added to distilled water, and a uniform aqueous phase is obtained by stirring, wherein the concentration of ammonium bicarbonate is 10% (w / v);

[0044] c) Disperse 2.5ml of the water phase obtained in step b into 15ml of the oil phase obtained in step a, and form a water-in-oil single emulsion through homogenization, wherein the volume ratio of the water phase to the oil phase is 1 / 6 ; The homogenization conditions are: stirring rate 8000rpm, stirring time 4min;

[0045] d) At a stirring rate of 200 rpm, disperse the single emulsion obtained in step c into a 0.5% (w / v) PVA (polyvinyl alcohol) aque...

Embodiment 2

[0050] The specific implementation steps are as follows:

[0051] a) At room temperature, add PLGA to the organic solvent dichloromethane, and obtain a homogeneous oil phase by stirring, wherein the concentration of PLGA is 6.25% (w / v), the molecular weight of PLGA is 40kDa, and the molar ratio of LA to GA in PLGA is 50 / 50;

[0052] b) Ammonium bicarbonate is added to distilled water, and a uniform aqueous phase is obtained by stirring, wherein the concentration of ammonium bicarbonate is 10% (w / v);

[0053] c) Disperse 2.5ml of the water phase obtained in step b into 12.5ml of the oil phase obtained in step a, and form a water-in-oil single emulsion through homogenization, wherein the volume ratio of the water phase to the oil phase is 1 / 5. The conditions for homogenization are: stirring rate 8000rpm, stirring time 4min;

[0054] d) at a stirring rate of 200rpm, disperse the single emulsion obtained in step c into a 0.5% (w / v) PVA aqueous solution having a volume of 400ml ...

Embodiment 3

[0059] The specific implementation steps are as follows:

[0060] a) At room temperature, add PLGA to the organic solvent dichloromethane, and obtain a homogeneous oil phase by stirring, wherein the concentration of PLGA is 6.25% (w / v), the molecular weight of PLGA is 40kDa, and the molar ratio of LA to GA in PLGA is 50 / 50;

[0061] b) Ammonium bicarbonate is added to distilled water, and a uniform aqueous phase is obtained by stirring, wherein the concentration of ammonium bicarbonate is 10% (w / v);

[0062] c) Disperse 2.5 ml of the water phase obtained in step b into 8 ml of the oil phase obtained in step a, and form a water-in-oil single emulsion through homogenization, wherein the volume ratio of the water phase to the oil phase is 1 / 3.2 ; The homogenization conditions are: stirring rate 5000rpm, stirring time 3min;

[0063] d) at a stirring rate of 200rpm, disperse the single emulsion obtained in step c into a 0.1% (w / v) PVA aqueous solution having a volume of 400ml to ...

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Abstract

The invention discloses a method for sintering a co-loading cell porous microsphere scaffold through subcritical carbon dioxide. An ammonium bicarbonate aqueous solution serves as an aqueous phase, an organic solution of PLGA serves as an oil phase, the aqueous phase is dispersed in the oil phase, and water-in-oil single emulsion is formed through homogenization; the single emulsion is dispersed in a PVA aqueous solution to obtain water-in-oil-in-water multiple emulsion; stir is continued to make dichloromethane volatilize so as to obtain cured porous microspheres; the cured porous microspheres are collected, washed and freeze-dried; a mold is filled with the free-dried porous microspheres and a certain amount of cells together, and under the condition of subcritical carbon dioxide, the porous microsphere scaffold is obtained through sintering. According to the method for sintering the co-loading cell porous microsphere scaffold through subcritical carbon dioxide, under the mild conditions, the co-loading cell porous microsphere scaffold is obtained through a one-step method, and activity of the cells is ensured while the porous form of the surface of the microspheres is ensured.

Description

technical field [0001] The invention relates to a preparation method of a three-dimensional tissue engineering scaffold, in particular to a method for subcritical carbon dioxide sintering and co-loading porous microsphere scaffolds. Background technique [0002] At present, the more mature tissue construction idea at home and abroad is the top-down (Top-down) method, that is, the bionic scaffold is prepared first, and then the seed cells are inoculated on the scaffold material, and the perfusion technology, adding growth factors, mechanical stimulation, etc. means of organizing. The main preparation methods include: solvent casting method, phase separation / freeze drying method, fiber bonding method, three-dimensional printing, melting molding method and the like. The main problems of this construction method are: high residual organic solvent, easy inactivation of biologically active factors, and poor cell compatibility. More importantly, it is impossible to precisely cont...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/56A61L27/38A61L27/18
Inventor 陈爱政王士斌马腾刘源岗吴文果
Owner HUAQIAO UNIVERSITY
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