Zein and dopamine graft and application thereof

Abstract

The invention discloses a zein and dopamine graft and application thereof.The zein and dopamine graft is prepared through the following steps that S1, zein is weighed and dissolved in dimethyl sulfoxide, and a solution 1 is obtained; s2, N-hydroxysuccinimide and N, N '-ethyl carbodiimide hydrochloride are added into the solution 1, dopamine is added after the N-hydroxysuccinimide and the N, N'-ethyl carbodiimide hydrochloride are fully dissolved, dialysis and freeze-drying are performed after stirring reaction, and the zein and dopamine graft is obtained. The zein and dopamine graft prepared by the invention can be used for preparing a corresponding electrospun fibrous membrane as a scaffold material through a solution electrospinning technology. The zein and the dopamine used in the invention belong to natural source materials, so that the prepared zein and dopamine graft material has good biocompatibility, and can be used as a blood contact scaffold material due to the anti-biological adhesion effect of the zein and dopamine graft material.

Description

Technical Field

[0001] The invention belongs to the field of natural polymer material modification and biomedical application thereof, and in particular, relates to a zein and dopamine graft and its application in blood contact stent materials. Background Art

[0002] Repair and regeneration through implantation of stent materials in the body is of great significance for patients with tissue defects. However, after the stent material is implanted, if it needs to be in contact with blood for a long time, hemolysis or even thrombosis may occur, further blocking the blood vessels and causing restenosis after stent implantation. The most important factor causing restenosis after stent implantation is the damage to the vascular endothelium and intima after stent implantation, which causes an imbalance of procoagulant and anticoagulant factors in the blood vessels. Treatments for restenosis after stent implantation include balloon angioplasty, stent-in-stent therapy, rotational plaque resection, directional plaque resection, excimer laser angioplasty and vascular brachytherapy. In order to resist the thrombogenicity of foreign surfaces, antiplatelet drugs or anticoagulants are usually required for preventive treatment. However, long-term use of such drugs increases the risk of bleeding. As a blood-contact material, the most effective and simplest way to implant stents is to modify and transform the stent, that is, to modify the surface of the stent by physical, biological, or chemical methods or to evenly cover the stent with polymer materials or polymers, so that conventional stents have sufficient blood compatibility and anti-thrombotic properties, thereby reducing the incidence of restenosis after stent implantation.

[0003] Zein is a natural macromolecular protein with good biocompatibility and biodegradability. In recent years, it has attracted the attention of domestic and foreign scholars as a tissue engineering scaffold material. Dopamine is an endogenous substance in the human body. It has attracted widespread attention because it has the same catechol structure as the adhesive protein structure in mussels. It is often used to modify the surface of materials to enhance cell adhesion. However, recent studies have found that the modification of dopamine may inhibit the adhesion and proliferation of cells on the surface of materials. At present, there has been no research work on its application in the modification of blood scaffold contact materials to inhibit thrombosis. Summary of the invention

[0004] The primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide a zein and dopamine graft, the main materials of which are zein and dopamine, both of which have good biocompatibility and are safe and non-toxic.

[0005] Another object of the present invention is to provide an electrospun fiber membrane of a graft of zein and dopamine.

[0006] Another object of the present invention is to provide the above-mentioned zein and dopamine grafts and the application of the electrospun fiber membrane of the zein and dopamine grafts.

[0007] To achieve the above object, the technical solution of the present invention is:

[0008] In a first aspect, the present invention provides a zein-dopamine graft, which is prepared by the following steps:

[0009] S1, weighing zein and dissolving it in dimethyl sulfoxide to obtain solution 1;

[0010] S2. Add N-hydroxysuccinimide and N,N'-ethylcarbodiimide hydrochloride to solution 1, add dopamine after fully dissolving, stir for reaction, dialyze and freeze-dry to obtain a graft of zein and dopamine.

[0011] Furthermore, in step S1, the ratio of zein to dimethyl sulfoxide is 0.1 g to 10 g / 100 mL.

[0012] Furthermore, in step S2, the mass ratio of zein to N-hydroxysuccinimide is 100:1 to 1:1.

[0013] Furthermore, in step S2, the mass ratio of zein to N,N'-ethylcarbodiimide hydrochloride is 100:1 to 1:1.

[0014] Furthermore, in step S2, the mass ratio of zein to dopamine is 100:1 to 1:1.

[0015] Furthermore, in step S2, the reaction time is 12 to 72 hours.

[0016] In a second aspect, the present invention provides an electrospun fiber membrane of a graft of zein and dopamine, which is prepared by the following steps:

[0017] S01, weighing zein and dopamine grafted products and dissolving them in hexafluoroisopropanol to obtain an electrospinning solution;

[0018] S02. Perform solution electrospinning under preset conditions to obtain an electrospun fiber membrane of a graft of zein and dopamine.

[0019] Furthermore, in step S01, the concentration of the electrospinning solution is 20% to 60% (w / v).

[0020] Furthermore, in step S02, the preset conditions are: voltage 15-30 kV, and receiving distance 10-30 cm.

[0021] In a third aspect, the present invention provides an application of the above-mentioned zein and dopamine graft or the electrospun fiber membrane of zein and dopamine graft in blood contact stent materials.

[0022] The beneficial effects of the present invention are:

[0023] (1) The main raw material zein used in the present invention is a natural plant protein, and the modified material dopamine is an endogenous substance in the human body. Both have good biocompatibility and are safe and non-toxic.

[0024] (2) The zein and dopamine grafts prepared in the present invention can be used to prepare scaffold materials using solution electrospinning technology and applied in the field of tissue engineering.

[0025] (3) The zein and dopamine graft prepared by the present invention can achieve anti-cell adhesion effect without the need for the addition of antiplatelet drugs, has good blood compatibility, and can be used as a blood stent contact material. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The accompanying drawings are part of the present invention and are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention, but do not constitute an improper limitation of the present invention. Obviously, the drawings described below are only some embodiments. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. In the accompanying drawings:

[0027] Figure 1 It is a grafted product of zein and dopamine in d 6 -NMR spectra in DMSO;

[0028] Figure 2 It is a scanning electron microscope characterization picture of the electrospun membrane of zein and dopamine grafted with different proportions, in which A corresponds to the electrospun fiber membrane of zein, B corresponds to the electrospun fiber membrane of ZD1, C corresponds to the electrospun fiber membrane of ZD2, and D corresponds to the electrospun fiber membrane of ZD3;

[0029] Figure 3 It is the adhesion and proliferation of human umbilical vein endothelial cells on the surface of zein electrospun fiber membrane (zein) and zein and dopamine graft electrospun membrane with different dopamine contents (ZD1, ZD2, ZD3).

[0030] It should be noted that these drawings and textual descriptions are not intended to limit the conceptual scope of the present invention in any way, but are intended to illustrate the concept of the present invention for those skilled in the art by referring to specific embodiments. DETAILED DESCRIPTION

[0031] To make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the reagents and raw materials used in the present invention can be obtained commercially.

[0032] [Example 1]

[0033] Preparation of Zein-dopamine Graft (ZD1) Scaffold Material

[0034] (1) Synthesis of zein-dopamine grafted product (ZD1)

[0035] 1.0 g of zein was weighed and dissolved in 10 mL of dimethyl sulfoxide, 50 mg of N-hydroxysuccinimide and 50 mg of N,N'-ethylcarbodiimide hydrochloride were added, and after fully dissolved, 0.1 g of dopamine was added, and the mixture was stirred for reaction for 48 h and then dialyzed and freeze-dried to obtain zein and dopamine grafted product ZD1.

[0036] Figure 1 It is a grafted product of zein and dopamine in d 6 -NMR spectrum in DMSO, from which the NMR peaks of zein and dopamine structures can be clearly observed, thus confirming the successful NMR grafting of zein and dopamine.

[0037] (2) Preparation of electrospun fiber membranes of zein-dopamine grafted products

[0038] Weigh 2 g of zein and dopamine graft (ZD1), dissolve it in 5 mL of hexafluoroisopropanol to obtain an electrospinning solution with a concentration of 40% (w / v), and perform electrospinning of the solution at a voltage of 25 kV and a receiving distance of 20 cm to obtain an electrospun fiber membrane of zein and dopamine graft (ZD1).

[0039] [Example 2]

[0040] The method of Example 1 is followed, except that the amount of dopamine added in step (1) is replaced with 0.2 g to prepare a zein and dopamine graft ZD2, and an electrospun fiber membrane of the zein and dopamine graft (ZD2) is prepared by solution electrospinning technology.

[0041] [Example 3]

[0042] The method of Example 1 is followed, except that the amount of dopamine added in step (1) is replaced with 1.0 g to prepare a zein and dopamine graft ZD3, and an electrospun fiber membrane of the zein and dopamine graft (ZD3) is prepared by solution electrospinning technology.

[0043] Figure 2 This is a scanning electron microscope characterization of the electrospun membrane of zein and dopamine grafts in different proportions (A in the figure corresponds to the zein electrospun fiber membrane, B corresponds to the ZD1 electrospun fiber membrane, C corresponds to the ZD2 electrospun fiber membrane, and D corresponds to the ZD3 electrospun fiber membrane). It can be seen that samples with different dopamine addition amounts can obtain relatively uniform filamentous structures through electrospinning technology.

[0044] Figure 3 The adhesion and proliferation of human umbilical vein endothelial cells on the surface of zein electrospun fiber membrane (zein) and zein and dopamine graft electrospun fiber membrane (ZD1, ZD2, ZD3) with different dopamine contents. Specifically, zein, ZD1, ZD2, ZD3, and electrospun fiber membrane were cut into discs with a diameter of 6 mm and placed in a 96-well plate. Human umbilical vein endothelial cells were inoculated on the above-mentioned electrospun fiber membranes at a cell density of 8×103 cells / well. After culturing in an incubator at 37°C and a CO2 content of 5% for 1, 2, and 3 days, the cell survival rate was determined using a CCK-8 kit. The results show that the modification of dopamine can effectively inhibit the adhesion and proliferation of cells, and the higher the dopamine content, the lower the cell adhesion and proliferation. The anti-cell adhesion effect can be achieved without the addition of antiplatelet drugs. It has good blood compatibility and can be used as a blood stent contact material.

[0045] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0046] The above description is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modifications to equivalent embodiments of equivalent changes by using the technical content suggested above without departing from the scope of the technical solution of the present invention. The implementation scheme in the above embodiment can also be further combined or replaced. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present invention without departing from the content of the technical solution of the present invention still falls within the scope of the solution of the present invention.

Claims

1. A grafted product of zein and dopamine, characterized in that: Prepared using the following steps: S1, weighing zein and dissolving it in dimethyl sulfoxide to obtain solution 1; S2. Add N-hydroxysuccinimide and N,N'-ethylcarbodiimide hydrochloride to solution 1, add dopamine after fully dissolving, stir for reaction, dialyze and freeze-dry to obtain a graft of zein and dopamine.

2. The zein-dopamine graft according to claim 1, characterized in that: In step S1, the ratio of zein to dimethyl sulfoxide is 0.1 g to 10 g / 100 mL.

3. The zein-dopamine grafted product according to claim 1, characterized in that: In step S2, the mass ratio of zein to N-hydroxysuccinimide is 100:1 to 1:

1.

4. The zein-dopamine grafted product according to claim 1, characterized in that: In step S2, the mass ratio of zein to N,N'-ethylcarbodiimide hydrochloride is 100:1 to 1:

1.

5. The zein-dopamine grafted product according to claim 1, characterized in that: In step S2, the mass ratio of zein to dopamine is 100:1 to 1:

1.

6. The zein-dopamine grafted product according to claim 1, characterized in that: In step S2, the reaction time is 12 to 72 hours.

7. An electrospun fiber membrane of a graft of zein and dopamine, characterized in that: Prepared using the following steps: S01, weighing zein and dopamine grafted products and dissolving them in hexafluoroisopropanol to obtain an electrospinning solution; S02. Perform solution electrospinning under preset conditions to obtain an electrospun fiber membrane of a graft of zein and dopamine.

8. The electrospun fiber membrane of the grafted product of zein and dopamine according to claim 7, characterized in that: In step S01, the concentration of the electrospinning solution is 20% to 60% (w / v).

9. The electrospun fiber membrane of zein and dopamine grafts according to claim 7, characterized in that: In step S02, the preset conditions are: voltage 15-30 kV, receiving distance 10-30 cm.

10. Use of the zein and dopamine graft according to any one of claims 1 to 6 or the electrospun fiber membrane of the zein and dopamine graft according to any one of claims 7 to 9 in blood contact stent materials.

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