TW202317699A - A zwitterionic material capable of preventing bio-inert performance decay, its fabricating membrane and application - Google Patents

Abstract

The invention discloses a zwitterionic material capable of preventing bio-inert performance decay. In particular, the zwitterionic material is a polymer comprises amide group and zwitterionic group. The zwitterionic material and its fabricating membrane maintain excellent bio-inert performance after steam sterilization process.

Description

Diionic materials, thin films and applications thereof against biologically inert performance decay

The invention discloses a dual-ion type material, a thin film and an application thereof which are anti-biologically inert and decay-resistant. In particular, the anti-biologically inert dual-ion type material is a polymer material containing an amide structure and a dual-ion structure.

Biomedical materials are roughly divided into artificially synthesized materials and materials extracted from living organisms, which are used in human implants, replace parts of human organ systems, and directly contact human tissues for medical functions. Commonly used biomedical materials are polymer materials, such as polyethylene glycol (PEG), which are widely used as pharmaceutical carriers and biomedical materials because of their anti-adhesion properties of biomolecules. Although PEG has good resistance to protein adsorption, it will cause polymer chain cleavage under high temperature and high oxygen, which will cause stability problems after long-term use; secondly, the terminal hydroxyl group of PEG can be oxidized by alcohol dehydrogenase into Aldehydes, this kind of aldehydes can also react with proteins in the body and other substances with amine groups to cause human harm.

In terms of industrial demand, since many medical devices such as scalpels, artificial arms, metal stents and metal implants must go through sterilization procedures, the known polymer biomedical materials are not widely used due to their poor structural stability at high temperatures. has its limitations. Accordingly, the development of a biomedical polymer material that can be used in sterilization procedures and has stable properties is an urgent research topic and development field.

According to the background of the invention and the industrial needs described in the prior art, the first object of the present invention is to provide a dual-ionic material that is resistant to decay of biologically inert performance.

The biological inertness mentioned in the present invention means that when a material is in contact with a living body, it will not cause adverse reactions to the living body, or will not cause the material to stick to and adsorb biomolecules, then the material is said to be biologically inert. inertia. Accordingly, the anti-biologically inert performance decay-resistant dual-ionic material of the present invention is a dual-ionic material that will not lose its biological inertness under high temperature, high humidity, high pressure or a combination thereof.

Specifically, the above-mentioned diionic material for anti-biological inert performance decay is a polymer material including an amide structure and a diionic structure. The diionic structure comprises: phosphobetaine group (phosphobetaine), carboxybetaine group (carboxylbetaine), sulfobetaine group (sulfobetaine) or a combination thereof.

Specifically, the polymer material containing amide structure and double ion structure includes methacrylamide sulfobetaine hydrogel, polystyrene-methacrylamide sulfobetaine copolymer, polyethylene glycol methyl Acrylates-methacrylamide sultaine copolymer or polystyrene-polyethylene glycol methacrylate-methacrylamide sultaine copolymer.

The second object of the present invention is to provide a thin film of anti-biologically inert performance decay, which comprises a film layer and a base material, and the film layer is made of the double ionic material of anti-biologically inert performance decay as described in the first object of the present invention constituted and fixed on the surface of the substrate.

Specifically, the film layer is made of methacrylamide sulfobetaine water Gum, polystyrene-methacrylamide sulfobetaine copolymer, polyethylene glycol methacrylate-methacrylamide sulfobetaine copolymer or polystyrene-polyethylene glycol methacrylate - Composed of methacrylamide sulfobetaine copolymer.

Specifically, the substrate includes polytetrafluoroethylene, polyvinylidene fluoride, polyvinyl chloride, ethylene/vinyl acetate copolymer, polypropylene, polyethylene or rayon.

In particular, the water contact angle of the film layer is about≦60 degrees, preferably 20-60 degrees.

The methods for immobilizing the above-mentioned diionic materials with anti-biological inert performance decay on the surface of the substrate include coating procedures, chemical grafting, or surface modification of the substrate.

The third object of the present invention is to provide a method for maintaining the original biological inertness of the material surface after the sterilization process, the steps of which include making the anti-biological inert performance decay of the diionic material as described in the first object of the present invention in the A film layer of anti-biological inertness decay is formed on the surface of the material, so that the original biological inertness of the material can be maintained after the surface of the material is sterilized.

Specifically, the anti-biologically inert performance decay-resistant dual-ionic material includes methacrylamide sulfobetaine hydrogel, polystyrene-methacrylamide sulfobetaine copolymer, polyethylene glycol methacrylate - methacrylamide sultaine copolymer or polystyrene-polyethylene glycol methacrylate-methacrylamide sultaine copolymer.

Specifically, the water contact angle of the anti-biologically inert decay-resistant film layer is about ≦60 degrees, preferably 20-60 degrees.

Specifically, the sterilization procedure is autoclaving or steam sterilization.

In particular, the anti-biologically inert decay-resistant diionic film layer is fixed on the surface of the material by physical adsorption or chemical bonding.

In summary, the technical characteristics and effects of the present invention include: (1) designing and providing a polymer material comprising an amide structure and a double ion structure, thereby making the polymer material have the effect of resisting the decay of biologically inert performance, Can not lose original biological inertness under the operation environment of high temperature, high humidity or high pressure, the schematic diagram of technical characteristic of the present invention is as shown in Figure 1, wherein PSBMA is as the experimental control group that the present invention has unpredictable effect; (2) To design and provide a thin film with anti-biological inert performance decay, which includes a film layer composed of the above-mentioned polymer materials containing amide structure and double ion structure, which can be widely used in the biomedical industry; (3) provide a material that allows The surface maintains the original biological inertness of the material after the sterilization process, thereby improving the stability of the biomedical material and achieving the purpose of being able to be reused after the moist heat sterilization process.

[Fig. 1] A schematic diagram of the technical effect of the anti-biologically inert performance decay-resistant dual-ion material of the present invention.

[Fig. 2] A schematic diagram of the synthesis of the PS-r-PEGMA-r-PSBAA copolymer of the present invention.

[Fig. 3] Histogram of Escherichia coli coverage density before and after the sterilization procedure of the PSBAA hydrogel of the present invention.

[Fig. 4] Histogram of the relative adsorption rate of fibrin of the PSBAA hydrogel of the present invention before and after the sterilization procedure.

[Fig. 5] Microscopic images of the PSBAA hydrogel of the present invention before and after the sterilization procedure.

[Figure 6] Histogram of blood cell coverage density of PSBAA hydrogel of the present invention before and after sterilization procedures.

[Fig. 7] Schematic diagram of the structural changes of the molecular structures of SBAA and SBMA of the present invention after steam sterilization.

[FIG. 8] Mass spectrometry diagrams of the molecular structures of SBAA and SBMA of the present invention before and after steam sterilization.

[FIG. 9] The mass spectrograms of the PS-r-PEGMA-r-PSBAA copolymer and PS-r-PEGMA-r-PSBMA copolymer of the present invention before and after the sterilization procedure.

[Fig. 10] Histogram of blood cell coverage density before and after the sterilization procedure of the PSBAA hydrogel-foil film of the present invention.

[Figure 11] Microscope image of the surface structure of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention before and after the sterilization process (a); Histogram of coating density (b); Histogram of porosity (c ) and the average pore flow size bar graph (d).

[Fig. 12] The water contact angle histogram (a) and the hydration capacity histogram (b) of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention before and after the sterilization procedure.

[Fig. 13] Histogram of bacterial attachment density of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention before and after the sterilization procedure.

[FIG. 14] Histogram of blood cell attachment density of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention before and after the sterilization procedure.

[Fig. 15] Histogram of the relative adsorption rate of fibrin of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention before and after the sterilization procedure.

The aforementioned and other technical contents, features and effects of the present invention will be clearly presented in the following detailed description of a preferred embodiment with reference to the drawings. In order to provide a thorough understanding of the present invention, detailed steps and components thereof will be set forth in the following description. Obviously, the practice of the invention is not limited to specific details familiar to those skilled in the art. On the other hand, well-known components or steps are not described in detail in order to avoid unnecessarily limiting the invention. The preferred embodiments of the present invention will be described in detail as follows, but in addition to these detailed descriptions, the present invention can also be widely implemented in other embodiments, and the scope of the present invention is not limited, which is subject to the scope of the patents that follow .

The first embodiment of the present invention is to provide a dual ionic material with anti-biological inert performance decay, which is a polymer material including an amide structure and a diionic structure. The diionic structure comprises: phosphobetaine group (phosphobetaine), carboxybetaine group (carboxylbetaine), sulfobetaine group (sulfobetaine) or a combination thereof.

In a specific embodiment, the polymer material containing amide structure and diionic structure includes methacrylamide sulfobetaine hydrogel, polystyrene-methacrylamide sulfobetaine copolymer, polyethylene glycol Alcohol methacrylate-methacrylamide sultaine copolymer or polystyrene-polyethylene glycol methacrylate-methacrylamide sultaine copolymer.

In a specific embodiment, the reaction composition of the methacrylamide sulfobetaine hydrogel comprises methacrylamide sulfobetaine and a crosslinking agent, and the methacrylamide sulfobetaine Calculated based on the molar sum of the enamide sulfobetaine and the crosslinking agent, the molar percentage of the crosslinking agent is approximately equal to or greater than 25%.

In a specific embodiment, polystyrene, polyethylene glycol methacrylate and methacrylamide of the polystyrene-polyethylene glycol methacrylate-methacrylamide sulfobetaine copolymer The composition molar ratio of sultaine is about 10-40%: 10-40%: 10-30%, preferably about 30-40%: 30-40%: 20-30%.

The second embodiment of the present invention is to provide a thin film of anti-biological inert performance decay, which includes a film layer and a substrate, the film layer is made of the double ion anti-biological inert performance decay as described in the first embodiment of the present invention The type material is formed and fixed on the surface of the substrate.

In a specific embodiment, the film layer is made of methacrylamide sulfobetaine hydrogel, polystyrene-methacrylamide sulfobeet copolymer, polyethylene glycol methacrylate-methacrylic Composed of amide sulfobetaine copolymer or polystyrene-polyethylene glycol methacrylate-methacrylamide sulfobetaine copolymer.

In a specific embodiment, the substrate comprises polytetrafluoroethylene, polyvinylidene fluoride, polyvinyl chloride, ethylene/vinyl acetate copolymer, polypropylene, polyethylene or rayon.

In a specific embodiment, the water contact angle of the film layer is about ≦60 degrees, preferably 20-60 degrees.

In one embodiment, the porosity of the anti-biologically inert decay-resistant film is about ≦50%, preferably 30-40%.

In one embodiment, the film of the anti-biologically inert performance decay The average pore size is about≦0.15μm, preferably 0.05~0.15μm.

In a specific embodiment, the coating density of the biionic material with anti-biological inert performance decay is about ≦8 mg/cm ² , preferably 6~8 mg/cm ² .

In a specific embodiment, the method for immobilizing the anti-biologically inert decay-resistant diionic material on the surface of the substrate includes methods such as coating process, chemical grafting, or surface modification of the substrate.

The third embodiment of the present invention is to provide a method for maintaining the original biological inertness of the material surface after the sterilization process, the steps of which include making the anti-biological inert effect decay of the double ion type as described in the first embodiment of the present invention The material forms a film layer on the surface of the material that resists the decay of the bioinert performance, whereby the surface of the material maintains the original bioinertness of the material after the sterilization procedure.

In a specific embodiment, the anti-biologically inert performance decay-resistant dual ionic material contains methacrylamide sulfobetaine hydrogel, polystyrene-methacrylamide sulfobetaine copolymer, polyethylene glycol Acrylate-methacrylamide sulfobetaine copolymer or polystyrene-polyethylene glycol methacrylate-methacrylamide sulfobetaine copolymer.

In a specific embodiment, the water contact angle of the anti-biologically inert decay-resistant film layer is about ≦60 degrees, preferably 20-60 degrees.

In a specific embodiment, the coating density of the biionic material with anti-biological inert performance decay is about ≦8 mg/cm ² , preferably 6-8 mg/cm ² .

In a specific embodiment, the sterilization procedure is an autoclaving procedure.

In another specific embodiment, the above-mentioned anti-biological inert effectiveness decay double The film layer of ions is fixed on the surface of the material by physical adsorption or chemical bonding.

The following examples and experimental examples are experiments carried out according to the content described in the above-mentioned content of the invention and the embodiments, and are used as effect verification and description of the present invention accordingly.

Example 1: Synthesis and analysis of methacrylamide sulfobetaine hydrogel (PSBAA)

The monomer [3-(methacrylamino)propyl]dimethyl(3-thiopropyl)ammonium hydroxide (SBAA) was dissolved in deionized water, and then mixed with different proportions (3, 10, 25mol% ) cross-linking agent (methylenebisacrylamide; NMAB) solution (0.2wt%) was uniformly mixed and stirred for 30 minutes. , add a catalyst (tetramethylethylenediamine; TEMED), use a syringe to inject the liquid into the water gel mold, carry out free radical cross-linking polymerization at room temperature for one hour to form a water gel, and use a mold with a diameter of 10 mm Cut the hydrogel, immerse in a mixture of deionized water and methanol, and store in a refrigerator at 4°C. The hydrogels were placed in deionized water, and the deionized water was replaced six times to ensure that methanol was replaced and unreacted monomers and oligomers were removed. Immerse the above water gel into the deionized water carrier, take 4 μl of diiodomethane and drop it vertically on the surface of the water gel, and use the surface contact angle analyzer to measure and record the real-time contact angle angle, which is about 110°~120° .

Example 2: Synthesis and analysis of polystyrene-polyethylene glycol methacrylate-methacrylamide sulfobetaine copolymer (PS-r-PEGMA-r-PSBAA)

Reaction equation as shown in Figure 2, monomer styrene, polyethylene glycol methacrylate and methacrylamide sulfobetaine (SBAA) or [2-(methacryl) Base) ethyl] dimethyl-(3-sulfonic acid propyl) ammonium hydroxide (SBMA) was added to the mixed solution of toluene and methanol (1:1) at a molar ratio of 35:43.3:21.7 and stirred until completely dissolved , the solid content is controlled at about 30wt.%, and then the AIBN initiator is added, the amount of the AIBN is 1/150 of the sum of the moles of the above-mentioned monomers, and the temperature is raised to 65°C for free radical polymerization to obtain the target copolymer (PS-r-PEGMA-r-PSBAA) or (PS-r-PEGMA-r-PSBMA), after the reaction, the temperature is lowered to wash the crude product of the copolymer with n-hexane, and then freeze-dried to obtain the final product (PS -r-PEGMA-r-PSBAA) or (PS-r-PEGMA-r-PSBMA). By H-NMR analysis, wherein PS-r-PEGMA-r-PSBAA has characteristic peaks at chemical shifts of about 7.29, 3.73 and 3.25ppm; PS-r-PEGMA-r-PSBMA has characteristic peaks at chemical shifts of about 7.17, 3.59 and The position of 3.17 ppm has a characteristic peak. The copolymer composition analysis calculated by NMR and the molecular weight measured by GPC are shown in Table 1.

Table 1

Example 3: Preparation of thin film with anti-biological inert performance decay

Use the water glue or copolymer made in Example 1 or Example 2 and the base material such as rayon or vinylidene fluoride (PVDF) by spin-coating or dipping Coating method (dip-coating) made PSBAA-foil film, PS-r-PEGMA-r-PSBAA-PVDF film and PS-r-PEGMA-r-PSBMA-PVDF film respectively.

Analysis of Thin Film Properties

The thin films of various anti-biological inert performance decay disclosed by the present invention analyze its surface structure characteristics with a scanning electron microscope (SEM); measure the contact angle of its film layer with a surface contact angle analyzer; flow porometer) to analyze the average pore flow size; analyze the weight change of the film before and after immersion in the solvent, and calculate the porosity with formula (1), where ε represents the porosity, and W _W represents the weight of the film after it is immersed in the solvent. Weight, W _D represents the dry weight of the film, ρ _P represents the specific gravity of the substrate, such as PVDF is 1.78g/cm ³ , and ρ _b represents the specific gravity of the solvent, this example is n-butanol, the specific gravity is 0.81g/cm ³ .

Formula 1)

Bio-inert potency analysis

The relevant biological inert performance analysis of various anti-biologically inert performance decay-resistant diionic materials, films and the experimental control group of the present invention includes bacteria/Escherichia coli attachment test, protein/fibrin adsorption test, human whole body The general experimental procedures for blood cell attachment test and cell viability test are as follows.

Bacteria Attachment Test

Place the sample to be analyzed in deionized water, and replace the deionized water six times to ensure that the methanol is replaced and remove unreacted monomers and oligomers; before testing, the sample is soaked in PBS buffer solution for 24 hours , and then washed three times with PBS buffer solution before the experiment. Then add 1ml of the cultured bacteria solution to the sample to be tested, put it in an oven at 37°C with a rotation speed of 100rpm, and carry out bacterial attachment for 3 hours and 24 hours. After the attachment time is over, remove the bacteria solution, wash with PBS buffer 6 times to remove residual bacteria, and add glutaraldehyde (volume 1ml, concentration 2.5%) to fix the attached bacteria.

Protein adsorption test

Place the sample in a 24-well TCPS dish filled with deionized water, and replace the deionized water six times to ensure that methanol is replaced and remove unreacted monomers and oligomers; then soak the sample in PBS buffer After soaking for 24 hours, the samples were washed three times with PBS buffer. Bovine serum albumin (BSA) and human fibrinogen (FN) were prepared at 1 mg/ml and placed in an oven at 37°C for natural dissolution. Add 1ml of the pre-tested target protein-human fibrinogen (FN) to the sample, and place the sample in an oven at 37°C for 30 minutes. After 30 minutes, remove the protein solution and wash it with PBS buffer solution to remove excess protein. Add 1ml of 1mg/ml BSA to the samples respectively, and place the samples in an oven at 37°C for 30 minutes; the purpose is to fill the defect of the target protein that is not adsorbed on the material, remove the protein solution after 30 minutes, and then wash with PBS buffer Sample three times. Add 1ml of 1st Antibody specific to the target protein to each sample, then place the sample in a 37°C oven for 30 minutes, and wash it three times with PBS buffer. Add each sample to 1ml BSA (1mg/ml), the sample was placed in a 37°C oven for 30 minutes, and then washed three times with PBS buffer. Add 1ml of 2ndAntibody to each sample, which is specific to 1st Antibody. The samples are placed in an oven at 37°C for 30 minutes, and then washed three times with PBS buffer. Transfer the sample to a brand-new 24-well TCPS dish, and add 0.5ml of chromogenic agent (TMB), wait for 6 minutes for the chromogenic agent to act; then add 0.5ml of sulfuric acid with a concentration of 1M to each sample to stop reaction. Pipette 0.2ml of the solution in each sample (including the position of the TCPS hole) into a 96-well plate, run a microplate reader (Microplate reader), and set the wavelength to 450nm, and analyze and check the reading value by the instrument. After calculation, the relative adsorption percentage of protein can be calculated.

Human Whole Blood Cell Attachment Test

The samples were placed in deionized water, and the deionized water was replaced six times to ensure that the methanol was replaced and the unreacted monomers and oligomers were removed; then the samples were soaked in PBS buffer for 24 hours, and the Wash three times with PBS buffer. After removing the PBS solution, add 1ml of whole blood to the samples, and place them in a 37°C oven for 30 minutes. After the attachment time is over, remove the blood samples, wash them with PBS buffer for 6 times to remove residual blood samples, and add Glutaraldehyde (volume 1ml, concentration 2.5%) fixed the attached blood cells. The image analysis was carried out with a conjugate microscope and the calculation analysis program was carried out with software (Image J).

Cell Viability Test

The samples to be tested were placed in deionized water, and the deionized water was replaced six times to ensure that the methanol was replaced and the unreacted samples were removed. After cleaning, they were placed in a freeze dryer for lyophilization. Add the cell solution (L929) at a concentration of (10000 cells/ml) after culture to a 96-well TCPS well plate, and place the cells in a 37°C culture oven for 24 hours. Refer to ISO10993-5 to prepare toxicity test samples, and extract materials with cell culture fluid; the positive control group is zinc-containing compounds (ZDEC, 0.1g/ml); the negative control group is high-density polyethylene (HDPE, 0.2g/ml); Weigh 0.6g of the dried sample and prepare it with an extraction concentration of 0.2g/ml. The extraction environment conditions are set at 37°C, 150rpm, for 24 hours. After 24 hours, the cell liquid in the well plate was removed, and the extracted sample and standard extract were added, and then placed in a 37°C culture oven to culture the cells for 24 hours. After cell culture for 24 hours (cell coverage, bright field observation is about 80%), remove the cell culture medium in the culture plate, add positive, negative and test sample extracts respectively, and continue to culture cells for 24 hours. After culturing L929 cells with the sample extract for 24 hours, add 51 μl of XTT reagent to each well, wait for the reaction for 3 hours, pipette 100 μl from each well into a new 96-well plate, and collect at a wavelength of 450 nm on a micro-disc analyzer The readings are calculated relative to cell viability .

General steps for sterilization procedures

The sterilizing procedure conditions of the various diionic materials and thin films of anti-biological inert performance decay disclosed by the present invention and the experimental control group of the present invention are to put the sample to be tested into a wet sterilizer for sterilization, and the sterilizing time is 20 minutes, the temperature was 121°C.

Bioinert performance analysis of PSBAA hydrogel before and after sterilization procedures

The biological inert performance analysis of the PSBAA hydrogel of the present invention before and after the sterilization process includes E. coli attachment test, protein adsorption test, human whole blood blood cell Attachment test and cell viability test. The experimental control group was PSBMA hydrogel.

The results of the Escherichia coli attachment test of the PSBAA hydrogel of the present invention show that when the crosslinking agent content is 3 (SBAA3), the hydrogel made by 10mol% (SBAA25) will have more bacteria attached to it; but when crosslinking The hydrocolloid (SBAA25) made with a content of 25 mol% additives can maintain good anti-bacterial adhesion properties after being sterilized by moist heat in deionized water and PBS buffer. Therefore, the PSBAA hydrogel containing acrylamine structure can resist the environment of moist heat sterilization and maintain the original property of anti-biomolecular adhesion. The English letters D and P at the end of the test sample code indicate that the test environment is deionized water and PBS buffer respectively. The specific experimental results are shown in Figure 3. Obviously, the PSBAA hydrogel made of 25mol% cross-linking agent has a good anti-biologically inert performance decay effect, and still maintains good biologically inert performance after the sterilization procedure. Attach E. coli. In contrast, the amount of E. coli attached to the PSBMA water gel as the experimental control group increased significantly after the sterilization process, and the microscope image also clearly shows that its surface is covered with bright spots, which means that the PSBMA water gel does not have the E. coli of the present invention. Anti-Bioinert Potency Decay Effects described above.

The protein attachment test results of the PSBAA hydrogel of the present invention show that the protein adsorption capacity of the SBAA3D and SBAA3P samples increased by 10% to 20% after the sterilization treatment. But when the cross-linking agent reaches 25mol%, it can be seen that the sterilized SBAA25D and SBAA25P still maintain their original anti-adhesion properties. The specific experimental results are shown in Figure 4. Obviously, when the PSBAA hydrogel made of 25mol% cross-linking agent has a good anti-biological inert performance decay effect, it will not adsorb fibrin after the sterilization process, and still maintain good bioinert performance. In contrast, as the experimental control group, PSBMA The amount of fibrin adsorption of the hydrogel increased significantly after the sterilization process, which proves that the PSBMA hydrogel does not have the anti-biological inert performance decay effect described in the present invention.

The human whole blood cell adhesion test results of the PSBAA hydrogel of the present invention show that SBAA25D and SBAA25P still maintain their anti-adhesion properties after sterilization. The specific experimental results are shown in Figure 5 and Figure 6. Obviously, the PSBAA hydrogel made of 25mol% cross-linking agent has a good anti-biological inert performance decay effect, and still has a coverage density of blood cells close to 0 after the sterilization procedure, which means that it will not adsorb blood cells and still retain Very good bioinert performance. In contrast, the covered density of blood cells of the PSBMA hydrogel, which was used as the experimental control group, increased significantly after the sterilization procedure. Accordingly, it is proved again that the PSBAA hydrogel has the anti-biological inert efficacy decay effect described in the present invention, but the PSBMA hydrogel of the control group does not have the anti-biological inert efficacy decay effect.

The cell viability test result of PSBAA hydrogel of the present invention

In this experiment, the XTT test was used to calculate the cell survival rate. The lower the cell survival rate, the stronger the toxicity of the material. The cells used are L929 fibroblasts, HDPE is the positive control group, and ZDEC is the sub-control group. The purpose is to observe whether the sterilized double ionic material hydrogel still maintains its biocompatibility. The results show that the present invention contains acrylamide The cell survival density of PSBAA hydrogels (SBAA25D and SBAA25P) after sterilization is still higher than 95%, indicating that they have cytocompatibility and maintain the original biological inertness.

Extraction Analysis of PSBAA Hydrogel Before and After Sterilization Procedure

This experiment mainly utilizes liquid phase mass spectrometry (LC-MS) to carry out the extract analysis after the sterilization procedure to the PSBAA hydrogel containing acrylamine structure of the present invention, its The purpose is to verify that the SBAA containing the acrylamine structure is resistant to hydrolysis and has thermal stability, and the PSBMA hydrogel without the acrylamine structure is used as the control group of this experiment. The analysis results of the PSBAA hydrogel extract after sterilization are shown in Table 2, and the analysis results of the PSBMA hydrogel extract after sterilization are shown in Table 3.

Table 2

table 3

According to Table 2, Table 3, Figure 7 and Figure 8, under steam sterilization conditions, the hydrogel with SBMA structure has a detection signal of molecular weight 211 after the sterilization process, but the hydrogel with SBAA structure The extract after the procedure will not produce a detection signal with a molecular weight of 211. According to the analysis results of molecular bond breakage shown in Figure 7, Accordingly, it is proved that the PSBAA hydrogel containing acrylamine structure of the present invention will not cause bond breakage after steam sterilization, and has heat and humidity stability. According to Figure 9, it is the LC analysis of the PS-r-PEGMA-r-PSBAA copolymer and PS-r-PEGMA-r-PSBMA copolymer of the present invention before and after the sterilization procedure. Obviously, PS-r-PEGMA - The LC spectrum of the r-PSBAA copolymer did not change before and after the sterilization procedure, which indicated that the structure of the PS-r-PEGMA-r-PSBAA copolymer was stable. In contrast, a new peak was detected in the LC spectrum of PS-r-PEGMA-r-PSBMA copolymer after the sterilization process, and it was confirmed by comparison that this peak was the peak signal generated by SBMA bond breaking, which indicated that PS -The structure of r-PEGMA-r-PSBMA copolymer is unstable under the condition of steam sterilization and will lead to cracking, which proves that the block structure containing PSBAA has hygrothermal stability.

Bio-inert performance analysis of PSBAA-foil films before and after sterilization procedures

In this experiment, PSBAA hydrogel was used as the film material, and it was coated on the rayon to make the PSBAA-yellow film, and the PSBMA-yellow film was used as the experimental control group. The bio-inert performance analysis before and after the sterilization process showed that the PSBAA-foil film of the present invention has no change in the attached density of blood cells, and it still maintains the state of not sticking to the blood cells after the sterilization process. However, the PSBMA-foil film showed a significant increase in the attachment density of blood cells after the sterilization process, and the experimental results are shown in Figure 10, which proves that the film made of the PSBAA hydrogel of the present invention as a film material has anti-biological properties. Technical efficacy of inert potency decay.

Structural Analysis of PS-r-PEGMA-r-PSBAA-PVDF Films Before and After Sterilization Procedures

Structural analysis of PS-r-PEGMA-r-PSBAA-PVDF film before and after the sterilization procedure, including electron microscope surface analysis, coating density analysis, porosity, average pore flow size, water contact angle and hydration ability analysis, after the English letters The number indicates the concentration of the copolymer used in the coating process to make the film, for example, SBAA-1 indicates that the concentration of PS-r-PEGMA-r-PSBAA is 1mg/ml, and so on. The experimental results are shown in Figure 11 and Figure 12, showing the PS-r-PEGMA-r-PSBAA-PVDF film structure of the present invention after the surface analysis, coating density, porosity, average pore flow size, water Neither the contact angle nor the hydration capacity changed significantly, thus proving that the structure is stable and not cleaved under the moist heat sterilization procedure.

Bioinert performance analysis of PS-r-PEGMA-r-PSBAA-PVDF films before and after sterilization procedures

The biological inert performance analysis of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention before and after the steam sterilization process includes the Escherichia coli attachment test, the human whole blood blood cell attachment test and the protein adsorption test. The experimental control group was PS-r-PEGMA-r-PSBAA-PVDF film.

The results of the bacterial attachment test of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention showed that the PS-r-PEGMA-r-PSBAA coating concentration of 1mg/ml, 5mg/ml and 10mg/ml was prepared respectively The resulting PS-r-PEGMA-r-PSBAA-PVDF films (SBAA-1, SBAA-5 and SBAA-10) had no significant increase in the bacterial attachment rate before and after the steam sterilization procedure, and there were no bright spots in the microscope images, This means that the PS-r-PEGMA-r-PSBAA-PVDF thin film of the present invention The membranes still have good bioinert performance after steam sterilization procedures. However, as a control group, the bacterial attachment rate of the PS-r-PEGMA-r-PSBMA-PVDF film after the steam sterilization process increased significantly, and the microscope image also showed bright spots. The specific experimental results are shown in Figure 13 . Accordingly, the film with amide structure and double-ion structure coating design of the present invention does have the technical effect of anti-biological inert performance decay, and is suitable for application in the biomedical industry, especially in the field of surgical instruments that need to undergo sterilization procedures.

The human whole blood blood cell attachment test result of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention shows that PS-r -The blood cell attachment rate of PEGMA-r-PSBAA-PVDF film (SBAA-10) did not increase significantly before and after the steam sterilization process, and there were no bright spots in the microscope image, which proved that the PS-r-PEGMA-r of the present invention -PSBAA-PVDF film still has good bioinert performance after steam sterilization procedure. However, as a control group, the PS-r-PEGMA-r-PSBMA-PVDF film has a significantly increased blood cell attachment rate after steam sterilization, and there are also bright spots in the microscope image. The specific experimental results are shown in Figure 14 Show. Accordingly, it is proved once again that the film with amide structure and dual-ion structure coating design of the present invention does have the technical effect of anti-biological inert performance decay, and is suitable for application in the biomedical industry, especially for blood transfusions that need to undergo sterilization procedures. material field.

The results of the fibrin adsorption test of the PS-r-PEGMA-r-PSBAA-PVDF film of the present invention showed that the films prepared with PS-r-PEGMA-r-PSBAA coating concentrations of 1mg/ml, 5mg/ml and 10mg/ml respectively into PS-r-PEGMA-r-PSBAA-PVDF films (SBAA-1, SBAA-5 and SBAA-10) had no significant increase in the relative adsorption rate of fibrin before and after the steam sterilization procedure, which indicated that the PS-r- The PEGMA-r-PSBAA-PVDF film still had good bioinert performance after steam sterilization procedure. However, the relative adsorption rate of fibrin of the PS-r-PEGMA-r-PSBMA-PVDF film as a control group was greatly increased after steam sterilization. The specific experimental results are shown in Figure 15. Accordingly, the film with amide structure and double-ion structure coating design of the present invention does have the technical effect of anti-biological inert performance decay, and is suitable for application in the biotechnology industry, especially in the field of protein vaccine production that needs to undergo sterilization procedures.

Although the present invention has been described above with specific examples, the scope of the present invention is not limited thereto. Those skilled in the art understand that various modifications or changes can be made without departing from the intent and scope of the present invention. In addition, the abstract and the title are only used to assist the search of patent documents, and are not used to limit the scope of rights of the present invention.

Claims (12)

A diionic material with anti-biological inert performance decay is a polymer material containing an amide structure and a diionic structure, and the polymer material containing an amide structure and a diionic structure includes methacrylamide sulfobeet Alkaline glue, polystyrene-methacrylamide sulfobetaine copolymer, polyethylene glycol methacrylate-methacrylamide sulfobetaine copolymer or polystyrene-polyethylene glycol methyl Acrylates-methacrylamide sulfobetaine copolymer. As the dual ionic material for anti-biological inert performance decay as described in claim 1, the reaction composition of the methacrylamide sulfobetaine hydrogel includes methacrylamide sulfobetaine and a cross-linking agent, with the Calculated by the molar sum of methacrylamide sulfobetaine and the crosslinking agent, the molar percentage of the crosslinking agent is approximately equal to or greater than 25%. According to claim 2, the biionic material with anti-biological inert performance decay, the crosslinking agent comprises methylenebisacrylamide. As the dual ionic material of anti-biological inert performance decay as described in claim 1, the polystyrene, polyethylene The molar ratio of diol methacrylate and methacrylamide sulfobetaine is about 10~40%: 10~40%: 10~30%. A thin film of anti-biological inert performance decay, comprising a film layer and a base material, the film layer is made of the anti-biological inert performance decay as described in claim 1~4. The sub-form material is formed and fixed on the surface of the substrate, and the water contact angle with the film layer is about ≦60 degrees. The anti-biologically inert performance decay film as described in claim 5, the base material comprises polytetrafluoroethylene, polyvinylidene fluoride, polypropylene, polyvinyl chloride, ethylene/vinyl acetate copolymer, polyethylene or rayon . The anti-biological inert performance decay film as claimed in claim 5, the porosity of the anti-biological inert performance decay film is about ≦50%. The anti-biological inert performance decay film as claimed in claim 5, the average pore flow size of the anti-biological inert performance decay film is about ≦0.15 μm. For the anti-biologically inert performance decay thin film as claimed in item 5, the coating density of the anti-biological inert performance decay of the film layer is about ≦8 mg/cm ² . A method for maintaining the original biological inertness of the material surface after a sterilization procedure, comprising forming an anti-biological inertness on the surface of the material with the diionic material having anti-biological inertness decay as described in claims 1 to 4 A film layer that decays, whereby the surface of the material maintains the original biological inertness of the material after a sterilization process, which is an autoclave heat and humidity sterilization process. According to the method for maintaining the original biological inertness of the material surface after the sterilization process as described in Claim 10, the coating density of the double-ionic material of the anti-biological inertness performance decay film layer is ≦8mg/cm ² . Maintaining the surface of the material after a sterilization procedure as described in claim 10 The material is originally biologically inert, and the water contact angle of the anti-biologically inert performance decay film layer is about ≦60 degrees.

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