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Gelatin-chitosan-hyaluronic acid-heparan sulfate composite three-dimensional stent and preparation method thereof

A technology of hyaluronic acid and heparan sulfate, which is applied in the field of biomedicine to achieve the effect of uniform cell distribution and good proliferation and growth status

Inactive Publication Date: 2010-12-22
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] So far, the literature has reported scaffolds constructed using chitosan-gelatin, heparan sulfate-collagen, collagen-hyaluronic acid, etc. Gelatin-chitosan-hyaluronic acid-heparan sulfate composite three-dimensional scaffold

Method used

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  • Gelatin-chitosan-hyaluronic acid-heparan sulfate composite three-dimensional stent and preparation method thereof
  • Gelatin-chitosan-hyaluronic acid-heparan sulfate composite three-dimensional stent and preparation method thereof
  • Gelatin-chitosan-hyaluronic acid-heparan sulfate composite three-dimensional stent and preparation method thereof

Examples

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example 1

[0033] In a 60℃ water bath, dissolve gelatin, hyaluronic acid, and heparin sulfate in water to make 1.4% gelatin (W / V), 0.028% hyaluronic acid (W / V), and heparin sulfate 0.028% (W / V) mixed solution; dissolve chitosan powder in 2% (V / V) acetic acid to make 1.4% (W / V) chitosan solution, centrifuge, remove slag, and degas; The two solutions are quickly mixed evenly at a ratio of 5:5. The mixed solution was injected into a 96-well plate at 200 μL / well, pre-frozen at -80°C for 4 hours, and freeze-dried for 24 hours. After the stent is formed, the stent is cross-linked at room temperature with a cross-linking agent containing 50 mmol / L 2-morpholine ethanesulfonic acid, 50 mmol / L carbodiimide, and 50 mmol / L N-hydroxysuccinimide in 40% ethanol for 6 hours. Remove the cross-linking agent, and use pH 7.4, 0.1mol / L Na 2 HPO 4 Incubate the buffer for 2h at room temperature to neutralize residual acetic acid; wash 4 times with 40% (V / V) ethanol, 30min / time, rinse with double distilled wat...

example 2

[0035] In a 60℃ water bath, dissolve gelatin, hyaluronic acid and heparin sulfate in water to make 1.8% gelatin (W / V), 0.036% hyaluronic acid (W / V), and heparin sulfate 0.036% (W / V) mixed solution; dissolve the chitosan powder in 2% (V / V) acetic acid to make a 1.8% (W / V) chitosan solution, centrifuge, remove slag, and degas; The two solutions are quickly mixed evenly at a ratio of 5:5. The mixed solution was injected into a 96-well plate at 200 μL / well, pre-frozen at -80°C for 4 hours, and freeze-dried for 24 hours. After the stent is formed, the stent is cross-linked at room temperature with a cross-linking agent containing 50 mmol / L 2-morpholine ethanesulfonic acid, 50 mmol / L carbodiimide, and 50 mmol / L N-hydroxysuccinimide in 40% ethanol for 6 hours. Remove the cross-linking agent, and use pH 7.4, 0.1mol / L Na 2 HPO 4 Incubate the buffer for 2h at room temperature to neutralize residual acetic acid; wash 4 times with 40% (V / V) ethanol, 30min / time, rinse with double distille...

example 3

[0037] In a 60℃ water bath, dissolve gelatin, hyaluronic acid, and heparin sulfate in water to prepare 2% gelatin (W / V), 0.04% hyaluronic acid (W / V), and heparin sulfate 0.04% (W / V) mixed solution; dissolve the chitosan powder in 2% (V / V) acetic acid to make a 2% (W / V) chitosan solution, centrifuge, remove slag, and degas; The two solutions are quickly mixed evenly at a ratio of 5:5. The mixed solution was injected into a 96-well plate at 200 μL / well, pre-frozen at -80°C for 4 hours, and freeze-dried for 24 hours. After the stent is formed, the stent is cross-linked at room temperature with a cross-linking agent containing 50 mmol / L 2-morpholine ethanesulfonic acid, 50 mmol / L carbodiimide, and 50 mmol / L N-hydroxysuccinimide in 40% ethanol for 6 hours. Remove the cross-linking agent, and use pH 7.4, 0.1mol / L Na 2 HPO 4 Incubate the buffer for 2h at room temperature to neutralize residual acetic acid; wash 4 times with 40% (V / V) ethanol, 30min / time, rinse with double distilled ...

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Abstract

The invention relates to a gelatin-chitosan-hyaluronic acid-heparan sulfate composite three-dimensional stent and a preparation method thereof. The gelatin-chitosan-hyaluronic acid-heparan sulfate porous three-dimensional composite stent is prepared by mixing gelatin, chitosan, hyaluronic acid and heparan sulfate in different ratios to obtain mixtures at different concentrations and freeze-dryingthe mixtures. The pore diameter, porosity, degradation, water absorption and the like of the stent can be regulated and controlled through each component content and freeze drying condition and can meet the basic requirement of a stent material. The stent can be used for the study of the growth, propagation and differentiation of NS / PCs. Through the hyaluronic acid, cells are distributed in the stent uniformly, and the heparan sulfate can bond growth factors, so that the growth propagation condition of the cells is similar to in-vivo condition; and the composite stent contains more functionalgroups such as amino groups, carboxyl groups, hydroxyl groups, sulfo groups, amide groups and the like, and is suitable for the conglutination, growth, propagation and the like of the cells.

Description

Technical field [0001] The invention belongs to the field of biomedical technology, and relates to a tissue engineering three-dimensional scaffold material and its preparation, in particular to the construction of a class suitable for neural stem / progenitor cells (NS / PCs) adhesion, growth, proliferation and differentiation The gelatin-chitosan-hyaluronic acid-heparin sulfate composite three-dimensional scaffold and its preparation method. Background technique [0002] Tissue engineering includes three elements: seed cells, three-dimensional scaffolds and growth factors, among which the preparation of three-dimensional scaffold materials is very important. Biological materials include inorganic materials, non-degradable and degradable macromolecule materials, and proteins. Inorganic materials and organic non-degradable polymer materials are not suitable for nerve tissue engineering due to their high hardness, poor elasticity, and slow degradation; the degradation products of orga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/22A61L27/20A61L27/50
Inventor 关水林小敏刘天庆崔占峰葛丹马学虎
Owner DALIAN UNIV OF TECH
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