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Enzyme-catalyzed biomaterial for controlled release of nitric oxide and preparation method thereof

A technology of nitric oxide and compounds, which is applied in the field of biomaterials and their preparations for the controlled release of nitric oxide by enzyme catalysis, can solve the problems of easy aggregation of NO donors and affect the effective use of NO in materials, and achieve the promotion of wound healing and good Biocompatibility, effect of promoting skin repair

Active Publication Date: 2015-09-30
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the difference in polarity, NO donors tend to aggregate, resulting in burst release
In addition, the hydrophobic matrix material restricts the migration of water molecules and affects the effective utilization of NO inside the material.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Synthesis of Chitosan Macromolecule CS-NO with Enzymatic Controlled Release of Nitric Oxide

[0013] 0.5 g of chitosan with a viscosity average molecular weight of 50,000 and a degree of deacetylation of 90% was stirred and dispersed in 50 ml of water, and pentynic acid was added and stirred evenly. The molar ratio of chitosan repeating unit to pentynic acid was 1:0.2 . If the chitosan still cannot be completely dissolved, add 1 mol / L hydrochloric acid solution until the chitosan is just completely dissolved, and add 1-(3-dimethylaminopropyl)-3-ethylcarbodioxide in the ice bath. Imine hydrochloride EDC, the molar ratio of pentynoic acid to EDC is 1:3, reacted for 24 hours, the product was dialyzed for 72 hours in a mixed solution of 5 mmol / L hydrochloric acid and 1% sodium chloride, and then at 1 mM Dialysis in mol / L hydrochloric acid for 48 hours, and finally in 4°C aqueous solution for 72 hours, changing the dialysate every 24 hours, and finally freeze-drying...

Embodiment 2

[0015] Example 2: Preparation of CS-NO material with enzyme-catalyzed and controlled release of nitric oxide

[0016] Preparation of CS-NO solution

[0017] Disperse 0.15 g of CS-NO in 10 ml of water and stir until it is completely dissolved to obtain a CS-NO solution with a concentration of 15 mg / ml.

[0018] Preparation of CS-NO film

[0019] The CS-NO was prepared into an aqueous solution with a concentration of 15%, and 150 microliters of the solution was coated on a cover glass with a diameter of 1 cm, and dried naturally. The dried CS-NO membrane is soaked in a 0.01 mol / L PBS buffer solution to obtain a CS-NO membrane.

[0020] Preparation of CS-NO porous scaffold

[0021] Dissolve chitosan with a viscosity-average molecular weight of 600,000 and a degree of deacetylation of 90% in 3% acetic acid and stir until completely dissolved. Use 1M sodium hydroxide to neutralize to a pH of about 6, and prepare CS-NO to 10 mg / Ml of the aqueous solution, mixed according to the mass ratio o...

Embodiment 3

[0024] Example 3: Local injection of CS-NO solution for treatment of lower limb ischemia in diabetic mice

[0025] Using C57 mice, the experimental animals were divided into three groups, namely saline group, CS group and CS-NO group, each with 6 animals. The lower limb ischemia model was created by ligating and dissecting the femoral artery of the lower limb of the mouse. A certain concentration of CS-NO solution was injected into the muscle around the ligation site at multiple points, the total amount was 50 microliters / mouse. Postoperative angiography was used to observe angiogenesis. The severity of ischemia and the degree of necrosis in the CS-NO group were significantly lighter than the other two groups. The samples were taken on 7, 14, 21, and 28 days. Immunofluorescence staining analyzes the distribution of small blood vessels and capillaries in muscle tissue. The release of NO effectively promotes angiogenesis, and the density of blood vessels is significantly increas...

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Abstract

The invention discloses a novel biological material with controlled release of nitrogen monoxide under catalysis of an enzyme, as well as a preparation method and application thereof. The novel biological material with the controlled release of the nitrogen monoxide under the catalysis of the enzyme is prepared by connecting a nitrogen monoxide donor compound with stable glycosylation with a natural polymer, namely chitosan through covalent bonds. The material has good processability and can be prepared into injectable solutions, thin films, porous supports, electrostatic spinning fiber films and other various products. The CS-NO and a composite material thereof can be used for treating diabetic lower limb ischemia, skin injures and myocardial infarction disease. As the nitrogen monoxide can be released in a controlled manner according to requirements, the material shows a great effect in treatment.

Description

Technical field [0001] This patent relates to a method for preparing biomaterials with enzyme-catalyzed and controllable release of nitric oxide, and also relates to medical products processed from such materials and their use in ischemic disease treatment, skin wound healing and cardiovascular implant devices Applications. Background technique [0002] Cardiovascular disease has now become one of the major diseases threatening human health. Both morbidity and mortality rank first among various diseases, and are recognized by the medical community as the "first killer" that endangers human health. According to the "China Cardiovascular Disease Report 2011" data released by the Cardiovascular Disease Prevention and Research Center of the Ministry of Health, the current number of cardiovascular disease patients in my country is about 230 million, which is equivalent to 2 out of every 10 adults. High medical expenses have become a huge economic burden. [0003] Nitric oxide (NO) is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/08C08J5/18C08J9/28D04H1/42A61K47/48A61K31/706A61P17/02A61P9/10A61L31/04A61L31/16
Inventor 赵强孔德领张计敏王淑芳
Owner NANKAI UNIV
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