Preparation method of quick dissociative type thermosensitive hyaluronic acid hydrogel

Abstract

The invention discloses a preparation method of quick dissociative type thermosensitive hyaluronic acid hydrogel, and belongs to the field of biomedical materials. The hyaluronic acid hydrogel is prepared through a carbodiimide cross-linking reaction based on biodegradable hyaluronic acid with excellent biocompatibility as a raw material and poly(N-isopropylacrylamide) containing disulfide bonds and amino radicals at both ends as a cross-linking agent. The hydrogel not only has the characteristic of being dissociated and converted from solid phase hydrogel to an aqueous liquor, but also the rigidity of the hydrogel can be agilely adjusted and controlled by changing temperature and degree of crosslinking and the like. The hyaluronic acid hydrogel prepared by the method provided by the invention has the advantages of available raw materials, mild reaction condition, quick dissociation, thermosensitivity, adjustable rigidity and the like, and has a potential use to three-dimensional culture, in particular to effective segregation of cell-hydrogel after culture and discuss of influence rule of cell growth by mechanical signals.

Description

technical field

[0001] The invention belongs to the field of biomedical materials, and in particular relates to a preparation method of a rapidly dissociable thermosensitive hyaluronic acid hydrogel suitable for three-dimensional cell culture. Background technique

[0002] More and more studies have shown that the growth behavior, protein expression, and cell function of cells cultured in two-dimensional monolayers based on culture plates cannot objectively reflect the real situation of cells in vivo. Scholars believe that this is due to the difference between the two-dimensional culture environment and the three-dimensional microenvironment of cells in vivo. The former ignores the influence of cell-cell and cell-matrix interactions on cell growth. Therefore, in recent years, three-dimensional cell culture technology has received extensive attention and developed rapidly. Compared with cells cultured in 2D, cells cultured in 3D are more physiologically meaningful in terms o...

Examples

Embodiment 1

[0031] 1) First, dissolve hyaluronic acid with a molecular weight of 300,000 in high-purity water to prepare a hyaluronic acid solution with a mass concentration of 4%;

[0032]2) Next, add 50% of the mass of hyaluronic acid to the solution, which is a temperature-sensitive macromolecular cross-linking poly(N-isopropylacrylamide) with a molecular weight of 20,000 and a narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups. After the joint agent is stirred and dissolved, adjust the pH value to 5 with hydrochloric acid or NaOH solution;

[0033] The temperature-sensitive macromolecular crosslinking agent of poly(N-isopropylacrylamide) of narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups is synthesized by the following steps:

[0034] a) Add chain transfer agent S,S'-bis(α,α'-methyl-α"-acetic acid) trithiocarbonate in sequence in anhydrous and oxygen-free organic solvent 1,4-dioxane, mono B...

Embodiment 2

[0040] 1) First, dissolve hyaluronic acid with a molecular weight of 50,000 in high-purity water to prepare a hyaluronic acid solution with a mass concentration of 3%;

[0041] 2) Next, add 30% of the mass of hyaluronic acid to the solution, which is a temperature-sensitive macromolecular cross-linking poly(N-isopropylacrylamide) with a molecular weight of 50,000 and a narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups. After the joint agent is stirred and dissolved, adjust the pH value to 5.5 with hydrochloric acid or NaOH solution;

[0042] The temperature-sensitive macromolecular crosslinking agent of poly(N-isopropylacrylamide) of narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups is synthesized by the following steps:

[0043] a) Add chain transfer agent S,S'-bis(α,α'-methyl-α"-acetic acid) trithiocarbonate and monomer N-isopropylpropene to tetrahydrofuran, an anhydrous and oxygen-...

Embodiment 3

[0048] 1) First, dissolve hyaluronic acid with a molecular weight of 200,000 in high-purity water to prepare a hyaluronic acid solution with a mass concentration of 4%;

[0049] 2) Next, add 60% of the mass of hyaluronic acid to the solution, which is a temperature-sensitive macromolecular cross-linking poly(N-isopropylacrylamide) with a molecular weight of 30,000 and a narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups. After the joint agent is stirred and dissolved, adjust the pH value to 4 with hydrochloric acid or NaOH solution;

[0050] The temperature-sensitive macromolecular crosslinking agent of poly(N-isopropylacrylamide) of narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups is synthesized by the following steps:

[0051] a) Add chain transfer agent S,S'-bis(α,α'-methyl-α"-acetic acid) trithiocarbonate in sequence in anhydrous and oxygen-free organic solvent 1,4-dioxane, mono ...

technical field

[0001] The invention belongs to the field of biomedical materials, and in particular relates to a preparation method of a rapidly dissociable thermosensitive hyaluronic acid hydrogel suitable for three-dimensional cell culture. Background technique

[0002] More and more studies have shown that the growth behavior, protein expression, and cell function of cells cultured in two-dimensional monolayers based on culture plates cannot objectively reflect the real situation of cells in vivo. Scholars believe that this is due to the difference between the two-dimensional culture environment and the three-dimensional microenvironment of cells in vivo. The former ignores the influence of cell-cell and cell-matrix interactions on cell growth. Therefore, in recent years, three-dimensional cell culture technology has received extensive attention and developed rapidly. Compared with cells cultured in 2D, cells cultured in 3D are more physiologically meaningful in terms o...

Examples

Embodiment 1

[0031] 1) First, dissolve hyaluronic acid with a molecular weight of 300,000 in high-purity water to prepare a hyaluronic acid solution with a mass concentration of 4%;

[0032]2) Next, add 50% of the mass of hyaluronic acid to the solution, which is a temperature-sensitive macromolecular cross-linking poly(N-isopropylacrylamide) with a molecular weight of 20,000 and a narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups. After the joint agent is stirred and dissolved, adjust the pH value to 5 with hydrochloric acid or NaOH solution;

[0033] The temperature-sensitive macromolecular crosslinking agent of poly(N-isopropylacrylamide) of narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups is synthesized by the following steps:

[0034] a) Add chain transfer agent S,S'-bis(α,α'-methyl-α"-acetic acid) trithiocarbonate in sequence in anhydrous and oxygen-free organic solvent 1,4-dioxane, mono B...

Embodiment 2

[0040] 1) First, dissolve hyaluronic acid with a molecular weight of 50,000 in high-purity water to prepare a hyaluronic acid solution with a mass concentration of 3%;

[0041] 2) Next, add 30% of the mass of hyaluronic acid to the solution, which is a temperature-sensitive macromolecular cross-linking poly(N-isopropylacrylamide) with a molecular weight of 50,000 and a narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups. After the joint agent is stirred and dissolved, adjust the pH value to 5.5 with hydrochloric acid or NaOH solution;

[0042] The temperature-sensitive macromolecular crosslinking agent of poly(N-isopropylacrylamide) of narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups is synthesized by the following steps:

[0043] a) Add chain transfer agent S,S'-bis(α,α'-methyl-α"-acetic acid) trithiocarbonate and monomer N-isopropylpropene to tetrahydrofuran, an anhydrous and oxygen-...

Embodiment 3

[0048] 1) First, dissolve hyaluronic acid with a molecular weight of 200,000 in high-purity water to prepare a hyaluronic acid solution with a mass concentration of 4%;

[0049] 2) Next, add 60% of the mass of hyaluronic acid to the solution, which is a temperature-sensitive macromolecular cross-linking poly(N-isopropylacrylamide) with a molecular weight of 30,000 and a narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups. After the joint agent is stirred and dissolved, adjust the pH value to 4 with hydrochloric acid or NaOH solution;

[0050] The temperature-sensitive macromolecular crosslinking agent of poly(N-isopropylacrylamide) of narrow molecular weight distribution containing disulfide bonds and double-terminal amino groups is synthesized by the following steps:

[0051] a) Add chain transfer agent S,S'-bis(α,α'-methyl-α"-acetic acid) trithiocarbonate in sequence in anhydrous and oxygen-free organic solvent 1,4-dioxane, mono ...

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