Method for preparing collagen matrix multifunctional spontaneous power heart valve repair material

Abstract

The application discloses a preparation method of a collagen matrix multifunctional self-power generation heart valve repair material, and specifically comprises the following steps: firstly, stirring and mixing an ammonium sulfate solution, gelatin, acrylic acid and acrylamide, then stirring and adding N, N-methylene bisacrylamide, and then adopting electrostatic spinning to form a film to obtain a power generation layer; secondly, preparing a bonding layer by using sodium carboxymethyl cellulose and a pyrrole monomer; and finally, using a decellularized pig dermal matrix as a support, sequentially bonding the power generation layer and the bonding layer with a chitosan solution, and the heart valve repair material is prepared. The heart valve repair material prepared by the application has the function of self-power generation, can self-generate a micro-current to implement electric stimulation on a heart valve wound site, further promotes wound healing, and can also reduce the risk of infection and pain of a patient.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of polymer material preparation, and particularly relates to a preparation method of a collagen matrix multifunctional self-powered heart valve repair material. BACKGROUND

[0002] Heart valve disease is a common disease in the world. Biological valve replacement and mechanical valve replacement are effective methods for treating valve disease, but both have their own defects: biological valves lack vitality after chemical treatment, and have a service life of about 10 to 15 years, and finally fail and functionally impaired due to tissue degeneration and calcification, etc., which brings the burden and risk of reoperation to patients; mechanical valves have a long service life, but need to be anticoagulated for life, which can be accompanied by complications such as bleeding and embolism, and even potential dangers such as disability and death. Therefore, valve repair is the most ideal treatment method.

[0003] Normal heart valves include special cells and extracellular matrix (ECM) and can respond to local biomechanical changes and reshape in time. The research of a successful tissue engineering valve repair material must be combined with its components, not only to adapt to the change of shape, but also to adapt to the requirements of pressure, shear force, compliance and durability. Collagen has a unique three-dimensional structure and is highly compatible with the human body, and is widely used in the field of medical materials such as heart valve repair and wound dressing. However, traditional collagen materials still have many problems such as fast in-vivo degradation rate and insufficient biomechanical properties, which to some extent also limit their medical applications. SUMMARY

[0004] The purpose of the present application is to provide a preparation method of a collagen matrix multifunctional self-powered heart valve repair material, which has strong adhesion and high mechanical properties.

[0005] The technical solution adopted by the present application is that the preparation method of the collagen matrix multifunctional self-powered heart valve repair material is implemented according to the following steps:

[0006] Step 1, preparing a power generation layer of the multifunctional self-powered heart valve repair material;

[0007] Step 2, preparing a bonding layer of the multifunctional self-powered heart valve repair material;

[0008] Step 3, using the decellularized pig dermal matrix as a scaffold, sequentially bonding the power generation layer obtained from step 1 and the bonding layer obtained from step 2 with chitosan solution, to obtain the collagen matrix multifunctional self-powered heart valve repair material.

[0009] The present application is also characterized in that,

[0010] In step 1, specifically:

[0011] In 0.1-1 moL / L of ammonium sulfate solution, 1-10 parts by mass of gelatin, 1-10 parts by mass of acrylic acid and 1-10 parts by mass of acrylamide are added, stirred for 30 min, then 0.003-0.03 parts by mass of N,N-methylene bisacrylamide is added and stirred for 15 min, and then electrospinning technology is used to spin into a film to prepare the power generation layer of the multifunctional self-powered heart valve repair material.

[0012] The electrospinning conditions are: the voltage is 10-15 kV, the push speed of the syringe is 0.5-1.0 mL / h, the distance from the spinneret to the collection end is 15-20 cm, and the relative running speed of the needle relative to the parallel direction of the collection end is 1-4 cm / s.

[0013] In step 2, the specific steps are:

[0014] Step 2.1: 0.1-1 parts by mass of carboxymethyl cellulose sodium is dissolved in 20-200 parts by mass of anhydrous ferric chloride solution, stirred until it is completely dissolved, 4-40 parts by mass of acrylamide is added while stirring, and copolymerization occurs, thus obtaining a mixed solution a;

[0015] Step 2.2: 0.05-0.5 parts by mass of pyrrole monomer is stirred in deionized water until it is uniformly stirred, 1-10 parts by mass of gelatin is continuously added and stirred until the gelatin is completely dissolved, then 0.003-0.03 parts by mass of N,N-methylene bisacrylamide is added, and after continuous stirring for 30 min, 0.05-0.5 parts by mass of ammonium persulfate is added and stirred for 15 min, thus obtaining a mixed solution b;

[0016] Step 2.3: The mixed solution a obtained in step 2.1 and the mixed solution b obtained in step 2.2 are stirred and mixed uniformly, and after ultrasonic treatment to remove bubbles, the solution is poured into a mold, and then dried in a vacuum drying oven to obtain the adhesive layer of the multifunctional self-powered heart valve repair material.

[0017] In step 2.3, the ultrasonic treatment frequency is 20-100 Hz, and the time is 30 s; the temperature of vacuum drying is 50-100℃, and the time is 5-10 h.

[0018] In step 3, the mass fraction of the chitosan solution is 0.5-5%.

[0019] The beneficial effects of the present application are:

[0020] (1) the decellularized pig dermal matrix adopted in the application removes epidermis and cells with immunogenicity, retains a relatively complete dermal matrix, the material has no immunogenicity, good histocompatibility, and the material is closer to a natural tissue structure and can promote in-situ tissue regeneration more;

[0021] (2) the heart valve repair material prepared in the application has strong adhesion and high mechanical properties, and is suitable for implanting into a heart valve repair;

[0022] (3) the heart valve repair material prepared in the application has low immunogenicity, and no anti-immune drug needs to be taken after being implanted into the body;

[0023] (4) the heart valve repair material prepared in the application has a self-generating function, can generate a micro-current to implement electric stimulation on a heart valve wound site, further promotes wound healing, and also can reduce the risk of infection and pain of a patient. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 is a model diagram of the collagen matrix multifunctional self-generating heart valve repair material of the application;

[0025] Figure 2 is a scanning electron microscope diagram of the support layer in the collagen matrix multifunctional self-generating heart valve repair material of the application. DETAILED DESCRIPTION

[0026] The application will be described in detail below in combination with the drawings and specific embodiments.

[0027] The preparation method of the collagen matrix multifunctional self-generating heart valve repair material of the application is specifically implemented according to the following steps:

[0028] Step 1, preparing a self-generating layer of the multifunctional self-generating heart valve repair material;

[0029] Specifically, 1-10 parts by mass of gelatin, 1-10 parts by mass of acrylic acid (MMA) and 1-10 parts by mass of acrylamide (AM) are added to 0.1-1 moL / L of an ammonium sulfate (APS) solution, magnetically stirred for 30 min, 0.003-0.03 parts by mass of N, N-methylenebisacrylamide (MBA) is then added and magnetically stirred for 15 min, and then electrospinning technology is used to spin into a film to prepare the power generation layer of the multifunctional self-generating heart valve repair material;

[0030] The electrospinning conditions are as follows: the voltage is 10-15 kV, the push speed of the syringe is 0.5-1.0 mL / h, the distance from the spinning head to the collection end is 15-20 cm, and the relative running speed of the needle relative to the parallel direction of the collection end is 1-4 cm / s;

[0031] Step 2, preparation of the adhesive layer of the multifunctional self-generating heart valve repair material, the specific steps are as follows:

[0032] Step 2.1: 0.1-1 parts by mass of sodium carboxymethyl cellulose (CMC-Na) is dissolved in 20-200 parts by mass of anhydrous ferric chloride solution, and magnetic stirring is carried out until it is completely dissolved, 4-40 parts by mass of acrylamide (AAM) is added while stirring to copolymerize, the copolymerization reaction time is 0.5-1h, and thus a mixed solution a is obtained;

[0033] The concentration of the anhydrous ferric chloride solution is 0.01-0.1 mol / L;

[0034] Step 2.2: 0.05-0.5 parts by mass of pyrrole monomer is uniformly stirred in deionized water by magnetic stirring, 1-10 parts by mass of gelatin is continuously added and stirred until the gelatin is completely dissolved, then 0.003-0.03 parts by mass of N,N-methylene bisacrylamide (MBA) is added, and the stirring is continued for 30 min, then 0.05-0.5 parts by mass of ammonium persulfate (APS) is added and stirred for another 15 min, and thus a mixed solution b is obtained;

[0035] Step 2.3: the mixed solution a obtained in step 2.1 and the mixed solution b obtained in step 2.2 are uniformly mixed by magnetic stirring, and then the solution is poured into a mold after ultrasonic treatment to remove bubbles, and then dried in a vacuum drying oven to obtain the adhesive layer of the multifunctional self-generating heart valve repair material;

[0036] The ultrasonic treatment frequency is 20-100 Hz, and the time is 30 s; the temperature of vacuum drying is 50-100 DEG C, and the time is 5-10 h.

[0037] Step 3, using decellularized pig dermal matrix (PADM) as a scaffold, the power generation layer obtained in step 1 and the adhesive layer obtained in step 2 are sequentially adhered with chitosan (CS) solution, and thus the collagen matrix multifunctional self-generating heart valve repair material is obtained.

[0038] The mass fraction of the chitosan solution is 0.5-5%;

[0039] The model diagram of the collagen matrix multifunctional self-generating heart valve repair material of the application is shown in Figure 1 It can be seen that the material is composed of a support layer, a power generation layer and an adhesive layer from bottom to top;

[0040] The scanning electron microscope image of the support layer in the collagen matrix multifunctional self-generating heart valve repair material of the application is shown in Figure 2 It can be seen that the collagen fiber surface of the support layer has a certain rough structure.

[0041] The application adopts tissue engineering technology to completely remove epidermis of pig skin by a unique processing method, and thoroughly and clearly removes all cell components in dermis, and on this basis, retains the spatial structure of dermis, to prepare acellular porcine dermal matrix (PADM).

[0042] Example 1

[0043] The preparation method of the collagen matrix multifunctional self-powered heart valve repair material is specifically implemented according to the following steps:

[0044] Step 1, preparing the self-powered layer of the multifunctional self-powered heart valve repair material: 1g of gelatin, 1g of acrylic acid and 1g of acrylamide are added into 10mL of 0.1moL / L ammonium sulfate solution, and magnetic stirring is performed for 30 minutes, then 0.003g of N,N-methylene bisacrylamide is added and magnetic stirring is performed for 15 minutes, and then electrospinning is performed to form a film, and the electrospinning conditions are that the voltage is 10kV, the injection speed of the syringe is 0.5mL / h, the distance from the spinneret to the collection end is 15cm, and the relative running speed of the needle relative to the parallel direction of the collection end is 1cm / s, to prepare the self-powered layer of the multifunctional self-powered heart valve repair material;

[0045] Step 2, preparing the adhesive layer of the multifunctional self-powered heart valve repair material: 0.1g of sodium carboxymethyl cellulose is dissolved in 20mL of 0.01mol / L anhydrous ferric chloride solution, and magnetic stirring is performed until it is completely dissolved, and 4g of acrylamide is added while stirring to cause copolymerization, and thus a mixed solution a is obtained;

[0046] Poly-pyrrole is compounded by an oxidative polymerization method: 0.05g of pyrrole monomer is weighed into 20mL of ionized water, and magnetic stirring is performed to uniformly stir, and then 1g of gelatin is continuously added and stirred until the gelatin is completely dissolved, and then 0.003g of N,N-methylene bisacrylamide is added, and stirring is continuously performed for 30 minutes, and then 0.05g of ammonium persulfate is added and stirring is performed for 15 minutes, and thus a mixed solution b is obtained;

[0047] The mixed solution a and the mixed solution b obtained in step 2.2 are uniformly mixed by magnetic stirring, and after 20Hz ultrasonic debubbling for 30s, the solution is poured into a mold, and then the mold is placed in a 50℃ vacuum drying box for drying for 5h, and thus the adhesive layer of the multifunctional self-powered heart valve repair material is obtained;

[0048] Step 3: The acellular porcine dermal matrix is used as a support, and the self-powered layer and the adhesive layer are sequentially adhered with a chitosan solution (0.5wt%) in sequence.

[0049] Example 2

[0050] The preparation method of the collagen matrix multifunctional self-powered heart valve repair material is implemented according to the following steps:

[0051] Step 1, preparing the self-powered layer of the multifunctional self-powered heart valve repair material: 50 mL, 0.5 moL / L of ammonium sulfate solution is added to 5 g of gelatin, 4 g of acrylic acid and 4 g of acrylamide, and magnetic stirring is performed for 30 minutes, then 0.01 g of N, N-methylene bisacrylamide is added and magnetic stirring is performed for 15 minutes, and then electrospinning is performed to form a film, and the electrospinning conditions are a voltage of 13 kV, a push speed of the syringe of 0.6 mL / h, a distance between the spinneret and the collection end of 16 cm, and a relative running speed of the needle relative to the parallel direction of the collection end of 2 cm / s, so as to prepare the self-powered layer of the multifunctional self-powered heart valve repair material;

[0052] Step 2, preparing the adhesive layer of the multifunctional self-powered heart valve repair material: 0.5 g of sodium carboxymethyl cellulose is dissolved in 100 mL, 0.02 mol / L of anhydrous ferric chloride solution, and magnetic stirring is performed until it is completely dissolved, and then 4 g of acrylamide is added to cause copolymerization, so as to obtain a mixed solution a;

[0053] The polypyrrole is compounded by an oxidative polymerization method: 0.2 g of pyrrole monomer is weighed in 20 mL of ionized water, magnetic stirring is performed to uniformly stir, 5 g of gelatin is continuously added and stirred until the gelatin is completely dissolved, then 0.01 g of N, N-methylene bisacrylamide is added, and stirring is continuously performed for 30 minutes, then 0.1 g of ammonium persulfate is added and stirring is performed for 15 minutes, and thus a mixed solution b is obtained;

[0054] The mixed solution a and the mixed solution b obtained in step 2.2 are uniformly mixed by magnetic stirring, and after 80 Hz ultrasonic defoaming for 30 s, the solution is poured into a mold, and then the mold is placed in a 60℃ vacuum drying box for drying for 7 h, and thus the adhesive layer of the multifunctional self-powered heart valve repair material is obtained;

[0055] Step 3, using the decellularized pig dermal matrix as a scaffold, the self-powered layer and the adhesive layer are sequentially adhered with a chitosan solution, and thus the multifunctional self-powered heart valve repair material is obtained.

[0056] Example 3

[0057] The preparation method of the collagen matrix multifunctional self-powered heart valve repair material is implemented according to the following steps:

[0058] Step 1, preparing a self-generating layer of a multifunctional self-generating heart valve repair material: adding 100 mL of a 1 mol / L ammonium sulfate solution to 10 g of gelatin, 10 g of acrylic acid, and 10 g of acrylamide, and magnetically stirring for 30 minutes, then adding 0.03 g of N,N-methylenebisacrylamide and magnetically stirring for 15 minutes, and then spinning into a film using electrospinning technology. The electrospinning conditions are a voltage of 15 kV, a syringe push speed of 1 mL / h, a distance from the spinneret to the collecting end of 20 cm, and a relative running speed of the needle in a direction parallel to the collecting end of 4 cm / s, to prepare the power generation layer of the multifunctional self-generating heart valve repair material;

[0059] Step 2, preparing an adhesive layer of a multifunctional self-generating heart valve repair material: dissolving 1 g of sodium carboxymethyl cellulose in 50 mL of a 0.1 mol / L anhydrous ferric chloride solution, stirring magnetically until the solution is completely dissolved, and adding 4 g of acrylamide while stirring to copolymerize the solution, thereby obtaining a mixed solution a;

[0060] The polypyrrole was composited by oxidative polymerization: the specific steps were as follows: 0.5 g of pyrrole monomer was weighed and added to 20 mL of deionized water, and the mixture was stirred evenly by magnetic stirring. 10 g of gelatin was added and stirred continuously until the gelatin was completely dissolved, and then 0.03 g of N, N-methylenebisacrylamide was added. After stirring for 30 minutes, 0.5 g of ammonium persulfate was added and stirred for another 15 minutes to obtain a mixed solution b;

[0061] Mixed solution a and mixed solution b obtained in step 2.2 were mixed evenly by magnetic stirring, and after being debubbled by 100 Hz ultrasonic wave for 30 seconds, the solution was poured into a mold, and then dried in a vacuum drying oven at 100°C for 10 hours to obtain an adhesive layer of a multifunctional self-generating heart valve repair material;

[0062] Step 3: Using the decellularized porcine dermal matrix as a scaffold, the power generation layer and the adhesive layer are sequentially adhered with chitosan solution (5% wt).

[0063] Table 1 Performance test results of collagen matrix multifunctional self-generating heart valve repair material

[0064]

[0065] Table 1 shows the performance test results of the collagen matrix multifunctional self-generating heart valve repair material prepared by the method of the present invention. From Table 1, it can be concluded that the material has large elastic deformation and strong adhesion ability while also having a certain self-generating ability.

Claims

1. A method for preparing a collagen matrix multifunctional self-generating heart valve repair material, characterized in that: Please follow the steps below to implement: Step 1: preparing the power generation layer of the multifunctional self-generating heart valve repair material; specifically: 1-10 parts by mass of gelatin, 1-10 parts by mass of acrylic acid, and 1-10 parts by mass of acrylamide are added to a 0.1-1 mol / L ammonium sulfate solution, stirred for 30 minutes, and then 0.003-0.03 parts by mass of N,N-methylenebisacrylamide is added and stirred for 15 minutes. Then, electrospinning technology is used to form a film to prepare the power generation layer of the multifunctional self-generating heart valve repair material; Step 2: preparing an adhesive layer of a multifunctional self-generating heart valve repair material; the specific steps are: Step 2.1: dissolving 0.1-1 parts by weight of sodium carboxymethyl cellulose in 20-200 parts by weight of anhydrous ferric chloride solution, stirring until completely dissolved, and adding 4-40 parts by weight of acrylamide while stirring to copolymerize the mixture, thereby obtaining a mixed solution a; Step 2.2: Dissolve 0.05-0.5 parts by weight of pyrrole monomer in deionized water and stir until uniformly mixed. Add 1-10 parts by weight of gelatin and continue stirring until the gelatin is completely dissolved. Then, add 0.003-0.03 parts by weight of N,N-methylenebisacrylamide and continue stirring for 30 minutes. Then, add 0.05-0.5 parts by weight of ammonium persulfate and continue stirring for 15 minutes to obtain a mixed solution b. Step 2.3: The mixed solution a obtained in step 2.1 and the mixed solution b obtained in step 2.2 are stirred and mixed uniformly, and after ultrasonic treatment to remove bubbles, the solution is poured into a mold, and then placed in a vacuum drying oven to dry, thereby obtaining an adhesive layer of the multifunctional self-generating heart valve repair material; Step 3: Using the decellularized porcine dermal matrix as a scaffold, the power generation layer obtained in step 1 and the adhesive layer obtained in step 2 are sequentially bonded with chitosan solution, wherein the mass fraction of the chitosan solution is 0.5-5%; a collagen matrix multifunctional self-generating heart valve repair material is obtained; the material consists of a support layer, a power generation layer, and an adhesive layer from bottom to top.

2. The method for preparing the collagen matrix multifunctional self-generating heart valve repair material according to claim 1, characterized in that: The electrospinning conditions are as follows: voltage of 10-15 kV, syringe pushing speed of 0.5-1.0 mL / h, distance from spinneret to collecting end of 15-20 cm, and relative running speed of needle in parallel direction with respect to collecting end of 1-4 cm / s.

3. The method for preparing the collagen matrix multifunctional self-generating heart valve repair material according to claim 1, characterized in that: In the step 2.3, the ultrasonic treatment frequency is 20-100 Hz and the time is 30 s; the vacuum drying temperature is 50-100° C. and the time is 5-10 h.

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