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Alginate hydrogel microsphere carrier containing galactosyl chitosan molecule and application thereof

A hydrogel microsphere and chitosan-based technology, applied in the direction of fixing on/in the organic carrier, can solve the problems of affecting the medical effect of cell secretion products, poor sphericity, waste of raw materials, etc., to maintain immunity The effects of isolation performance, good mechanical strength, and excellent sphericity

Active Publication Date: 2017-05-24
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first two methods all have the following problems. One is that due to the limitation of the instantaneous reaction speed of the two macromolecules, chitosan and alginic acid, the particle size of the formed microcapsules is about 1mm or even larger, and the excessively large microcapsules are likely to cause cell damage. If the cluster grows too large, the central cells will be necrotic due to the limitation of mass transfer and oxygen transfer, which will affect the cell activity and then affect the medical effect of the products secreted by the cells. The mechanical strength of the microcapsules, when transplanted into the body, the microcapsules are prone to damage and cause the body's inflammatory response; third, no matter whether alginic acid or chitosan is used as a gel bath, alginic acid and chitosan dissolved in calcium solution cannot fully react Or chitosan can't fully combine with alginic acid, both lead to the waste of raw materials
The third method is to drop alginic acid into the divalent salt solution to form an embedded hydrogel microsphere carrier with controllable size, sphericity and mechanical strength, but then add the embedded hydrogel microsphere When the carrier is immersed in the chitosan solution to further react to form a film to form a traditional AC microcapsule, chitosan will not enter the interior of the microcapsule, which forms a core-shell microcapsule with alginic acid as the core and chitosan as the shell. structure, chitosan does not contact the cells inside the microcapsules, making functional chitosan unable to play its due role

Method used

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  • Alginate hydrogel microsphere carrier containing galactosyl chitosan molecule and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1) Preparation of sodium alginate solution: 2.0 g of alginic acid was dissolved in 100 mL of physiological saline to prepare a 20 g / L sodium alginate solution, wherein the molecular weight of alginic acid was 350 kDa.

[0036] 2) Preparation of galactosyl chitosan solution: Dissolve galactosyl chitosan in physiological saline to prepare galactosyl shells with concentrations of 0, 15g / L, 20g / L, 30g / L and pH 7.0 polysaccharide solution.

[0037] 3) Preparation of gel bath solution: 11 g of anhydrous calcium chloride was dissolved in 1 L of deionized water.

[0038] 4) Take 2mL of sodium alginate solution, mix 2mL of different concentrations of galactosyl chitosan solutions obtained in step 2) respectively, and stir at room temperature for 3h. At this time, the concentration of sodium alginate is 10g / L, and the concentration of galactosyl chitosan 0, 7.5g / L, 10g / L, 15g / L in turn, using the electrostatic droplet method to prepare calcium alginate / galactosyl chitosan embedd...

Embodiment 2

[0044] 1) Preparation of sodium alginate solution: 3.0 g of alginic acid was dissolved in 100 mL of physiological saline to prepare a 30 g / L sodium alginate solution, wherein the molecular weight of alginic acid was 350 kDa.

[0045] 2) Preparation of FITC-labeled galactosyl chitosan solution: FITC-labeled galactosyl chitosan was dissolved in physiological saline to prepare a galactosyl chitosan solution with a concentration of 10 g / L and a pH of 7.0.

[0046] 3) Preparation of gel bath solution: 11 g of anhydrous calcium chloride was dissolved in 1 L of deionized water.

[0047] 4) Take 2 mL of sodium alginate solution, mix 2 mL of the galactosyl chitosan solution obtained in step 2) respectively, and stir at room temperature for 3 h. At this time, the concentration of sodium alginate is 15 g / L, and the concentration of galactosyl chitosan is 5 g / L. L, Calcium alginate / galactosyl chitosan-embedded hydrogel microsphere carrier prepared by electrostatic droplet method.

Embodiment 3

[0049] 1) Preparation of sodium alginate solution: 2.0 g of alginic acid was dissolved in 100 mL of physiological saline to prepare a 20 g / L sodium alginate solution, wherein the molecular weight of alginic acid was 350 kDa, and it was sterilized by filtration.

[0050]2) Preparation of galactosyl chitosan solution: 0.4 g of galactosyl chitosan was dissolved in 10 mL of physiological saline to prepare a 40 g / L chitosan solution, adjusted to pH 7.2, and sterilized by filtration.

[0051] 3) Preparation of gel bath solution: 11 g of anhydrous calcium chloride was dissolved in 1 L of deionized water, and sterilized by filtration.

[0052] 4) Preparation of α-polylysine solution: α-polylysine was dissolved in physiological saline to prepare a 5 g / L α-polylysine solution, which was sterilized by filtration. Among them, the molecular weight of α-polylysine is 30kDa.

[0053] 5) Take 3 mL of sodium alginate solution, mix with 1 mL of galactosyl chitosan solution, and stir at room te...

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Abstract

The invention relates to an alginate embedded type hydrogel microsphere carrier having the interior containing a galactosyl chitosan molecule and a preparation method thereof. A galactosyl group is grafted onto a chitosan molecule by an amidation reaction, and hepatocytes are cultured in microcapsules jointly formed by chitosan and alginic acid. Experiments prove that the microcapsules jointly prepared by the galactosyl-grafted chitosan and alginic acid are used for culturing primary hepatocytes of rats, and the activity and function of the hepatocytes are well maintained.

Description

technical field [0001] The invention relates to an alginate hydrogel microcarrier, in particular to an alginate hydrogel microsphere carrier containing galactosyl chitosan molecules inside and its preparation and application. Background technique [0002] Substances with biological activity are encapsulated in selective permeable membranes to form spherical microcapsules, which are called "biological microcapsules" [Chang TMS.Hemoglobin corpuscles.Research report honors physiology.Medical Library,McGill University,1957] . Since the 1990s, using biological microcapsules as a tool for immune isolation and delivery of cells, using genetically recombined cells or metabolites of primary cells to regulate physiological functions of the body and treat related diseases has become a research hotspot for biomedical workers [Basic D, Vacek I, Sun A M. Microencapsulation and transplantation of engineered cells: a new approach to somatic genetherapy. Art Cells Blood Subs Immob Biotechno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N11/04
Inventor 于炜婷娄茹云宋益哲郑会珍马小军
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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