Coating composition for preventing crusting of medical equipment as well as preparation method and application of coating composition

Abstract

The invention discloses a coating composition for preventing crusting of a medical instrument as well as a preparation method and application of the coating composition. The raw materials of the coating composition comprise the following components: cell membrane vesicle liquid, wherein the cell membrane vesicle liquid is selected from an aqueous solution of cell membrane vesicles; and a cross-linking agent, which is selected from the group consisting of the multi-arm-polyethylene glycol-succinimide carboxylic ester. The cell membrane vesicle liquid has the characteristic that the external leaflets of the cell membrane are fully exposed, and through covalent crosslinking between the aminated substrate and the cell membrane, the formed coating also has the characteristic that the external leaflets are fully exposed, has good stability, can tolerate physical stimuli such as fluid scouring, repeated bending, ultrasonic vibration and the like, and has a good application prospect. The stability of the cell membrane coating is obviously superior to that of a cell membrane coating prepared by a traditional mode (such as fusion and adsorption).

Description

Technical Field

[0001] The present invention relates to the technical field of functional materials, and more specifically, to a coating composition for preventing encrustation of medical devices, and a preparation method and application thereof. Background Art

[0002] Ureteral stents and catheters are commonly used indwelling devices in urology. Although they help to drain urine, crusting often occurs on the surface of these devices when used in the body for a long time. Crusting is a major problem that often plagues urology. It not only affects the function of the indwelling device, but also leads to serious complications such as hydronephrosis, urosepsis, and even death.

[0003] Crust is formed by the deposition of mineral crystals on the surface of the device due to the supersaturation of salt solution in urine. The formation of crust is a complex process, which is affected by factors such as retention time, urine composition, bacterial infection, physical properties of materials and urodynamics. It is generally believed that the biofilm formed by urinary bacterial infection can significantly promote the deposition of infected stones on the surface and cavity of indwelling devices. Therefore, the development of existing anti-crust materials mainly focuses on preventing the formation of biofilm to inhibit the formation of crust, such as antibacterial and hydrophilic coatings. Although these materials can effectively reduce the formation of biofilm, they still have a high risk of crusting due to ignoring the interaction between the material and mineral crystals. Clinical studies have found that patients without bacterial infection often suffer from repeated crusting. In fact, these non-infectious stones account for more than 80% of urinary stones and are also the main component of crusts.

[0004] Therefore, developing an appropriate coating that can effectively inhibit mineral crystallization on the material surface in the urinary system environment and thus reduce crusting on the surface of indwelling devices is an urgent problem to be solved in current research and application. Summary of the invention

[0005] Based on the above problems, the object of the present invention is to provide a coating composition for preventing encrustation of medical devices and a preparation method and application thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] In one aspect, the present invention provides a coating composition for preventing encrustation of medical devices, wherein the raw materials of the coating composition include the following components:

[0008] A cell membrane vesicle fluid selected from an aqueous solution of a cell membrane vesicle; and

[0009] The cross-linking agent is selected from multi-arm-polyethylene glycol-succinimidyl carboxylate.

[0010] Furthermore, the cross-linking agent is selected from one or more of two-arm-polyethylene glycol-succinimide carboxylate, four-arm-polyethylene glycol-succinimide succinate, four-arm-polyethylene glycol-succinimide glutarate, four-arm-polyethylene glycol-succinimide acetate and four-arm-polyethylene glycol-succinimide carbonate.

[0011] Furthermore, the cell membrane vesicles are vesicles prepared from cell membranes.

[0012] Furthermore, the outer leaflet of the cell membrane vesicles in the cell membrane vesicle fluid is exposed. The cell membrane is composed of an outer leaflet and an inner leaflet, the outer side of the cell membrane corresponds to the outer leaflet, and the inner side of the cell membrane corresponds to the inner leaflet. That is, in the cell membrane vesicles, the outer leaflet is exposed to the outside, and the inner leaflet is not exposed at all and is wrapped inside the structure.

[0013] Furthermore, the cell membrane vesicles in the cell membrane vesicle fluid are prepared from one or more of erythrocyte membranes, platelet membranes, macrophages and neutrophil membranes.

[0014] Furthermore, in the cell membrane vesicle liquid, the concentration of cell membrane vesicles is 0.1-0.5wt%.

[0015] Furthermore, the protein concentration in the cell membrane vesicle fluid is 0.2-5 mg / mL.

[0016] Furthermore, the preparation of the cell membrane vesicles comprises the following steps:

[0017] After mixing whole blood with PBS, centrifuge, aspirate the upper liquid, and wash to obtain cell pellets;

[0018] The cell pellet was resuspended in PBS, subjected to hypotonic treatment, and centrifuged to obtain the cell membrane vesicles.

[0019] Furthermore, the PBS is selected from Cytiva HyClone. The PBS does not contain calcium ions and has little effect on cell membranes.

[0020] Furthermore, the mixing volume ratio of the whole blood and PBS is 1:(6-10), the temperature of the centrifuged whole blood and PBS is 4°C, the centrifugal force is 800g, and the time is 5 minutes.

[0021] Furthermore, the temperature of the hypotonic treatment is 4°C and the time is 30-60 minutes.

[0022] Furthermore, the centrifugal force after the hypotonic treatment is 13000-14000g, and the time is 5-10min.

[0023] In the structure of the cell membrane vesicle prepared by the above method and conditions, the outer leaflet is exposed to the outside, while the inner leaflet is not exposed at all and is wrapped inside the structure.

[0024] In the technical solution of the present invention, by selecting a specific cross-linking agent to mix with the cell membrane vesicle liquid, the obtained coating composition can have good film-forming property, adhesion and anti-crusting ability when coating medical devices (especially urinary system devices). Further, the amount of the cross-linking agent added relative to the cell membrane vesicle liquid is 0.01-0.5wt%. At this time, the aforementioned effect is better. Exemplarily, the amount of the cross-linking agent added relative to the cell membrane vesicle liquid includes but is not limited to 0.05-0.1wt%, 0.05-0.08wt%, 0.05-0.06wt%, 0.06-0.1wt%, 0.06-0.08wt%, 0.08-0.1wt%, 0.05wt%, 0.06wt%, 0.08wt%, 0.1wt%, etc.

[0025] In another aspect, the present invention provides a method for preparing the coating composition as described above, comprising the steps of:

[0026] The cell membrane vesicle liquid is mixed with a cross-linking agent to obtain the coating composition.

[0027] In another aspect, the present invention provides a medical device with anti-crusting, wherein the structure of the medical device comprises:

[0028] substrate; and

[0029] A cell membrane coating is covalently cross-linked on the surface of the substrate, wherein the cell membrane coating is prepared from raw materials comprising the coating composition described above.

[0030] In this technical solution, there is no requirement for the thickness of the cell membrane coating, which can be selected according to actual conditions.

[0031] In the technical solution of the present invention, the cell membrane coating is completely coated on the surface of the substrate.

[0032] In the present technical solution, the cell membrane coating prepared by using the raw material containing the coating composition has the characteristics that the outer leaflet of the cell membrane (i.e., the outer side of the cell membrane) is fully exposed, while the inner leaflet is completely unexposed, and has excellent anti-fouling effect.

[0033] Furthermore, in the technical solution of the present invention, a coupling molecule containing an amino group is modified on the surface of the substrate. The cell membrane coating is covalently bonded to the amino groups on the surface of the substrate. Aminated modification of the substrate surface helps to stabilize the binding of the substrate and the cell membrane coating.

[0034] Furthermore, the material of the substrate is selected from one or more of polyurethane, silicone, rubber, silicon dioxide (such as glass), metal alloy, latex, polyvinyl chloride, polyethylene, polypropylene, polylactic acid, polyethylene terephthalate and polycaprolactone.

[0035] Furthermore, the surface potential of the cell membrane coating is -60 to -20 mV, and the contact angle is 30 to 60°.

[0036] In another aspect, the present invention provides a method for preparing the medical device as described above, comprising the following steps:

[0037] aminating the surface of the substrate;

[0038] The coating composition is coated on the surface-amino-modified substrate and dried to obtain a cell membrane coating covalently cross-linked on the surface of the substrate.

[0039] In this technical solution, there are no other requirements for the coating method, and conventional coating methods can be used, which will not be described in detail here.

[0040] Furthermore, the coating method is spin coating, spray coating, scraping coating, brush coating or dipping coating.

[0041] Furthermore, the spin coating has a rotation speed of 3000-4000 rpm and a time of 30-50 s.

[0042] Furthermore, the dipping method is: immersing the substrate with the surface amino treated in the coating composition at a temperature of 4-50° C. for 0.5-5 h.

[0043] Furthermore, the drying is room temperature drying.

[0044] Furthermore, the method for aminating the substrate surface comprises the following steps:

[0045] performing surface plasma treatment on the substrate;

[0046] A silane solution containing amino groups is used to achieve amination on the surface of the plasma-treated substrate through spin coating or dip coating.

[0047] Furthermore, before the substrate is subjected to plasma treatment, it is first ultrasonically cleaned and dried using anhydrous ethanol and deionized water, respectively.

[0048] Furthermore, the amino-containing silane solution is selected from a chitosan solution of 3-glyceropropyltrimethoxysilane, a mixed solution of (3-aminopropyl)trimethoxysilane in deionized water and acetic acid, a mixed solution of (3-aminopropyl)triethoxysilane in deionized water and acetic acid, a 1.0wt% polyethyleneimine solution of 3-glyceropropyltrimethoxysilane, and the like.

[0049] Furthermore, during the process of amination of the substrate surface, the spin coating speed is 2500-3500 rpm and the time is 1-1.5 min.

[0050] Furthermore, in the process of aminating the surface of the substrate, the immersion is performed at room temperature for 2-2.5 hours, and then the substrate is dried with nitrogen.

[0051] In another aspect, the present invention provides use of the coating composition as described above in the preparation of medical devices.

[0052] Furthermore, the medical device is selected from urinary system devices.

[0053] Furthermore, the medical device is selected from a ureteral stent and / or a urinary catheter.

[0054] The beneficial effects of the present invention are as follows:

[0055] The coating composition provided by the present invention is applied to medical devices (especially urinary system devices), and the cell membrane coating formed can inhibit various minerals in urine for a long time, including inorganic mineral crystals such as calcium oxalate, calcium phosphate, calcium carbonate, and calcium sulfate, and exhibits excellent biocompatibility. In addition, the cell membrane coating improves the stability of the coating through covalent cross-linking between the amino-modified substrate and the cell membrane, and can withstand physical stimulation such as fluid scouring, repeated bending, and ultrasonic vibration. Its stability is significantly better than that of the cell membrane coating prepared by traditional methods (such as fusion and adsorption).

[0056] The coating composition and the anti-encrusting medical device provided by the present invention have simple preparation methods, low-cost and easily available raw materials, mild conditions, and fully retain the structural characteristics of the exposed outer leaflet of the cell membrane. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] The specific implementation modes of the present invention are further described in detail below in conjunction with the accompanying drawings.

[0058] Figure 1 The scanning electron microscope photograph of the red blood cell membrane coating on the anti-crusting substrate prepared in Example 1 is shown.

[0059] Figure 2 Fluorescence staining photographs of the red blood cell membrane coating on the anti-crusting substrate prepared in Example 1 and Comparative Example 1 after various stimuli (ultrasonic vibration, fluid flushing, repeated bending, and 75% ethanol disinfection) are shown.

[0060] Figure 3 The water contact angle test photograph of the anti-crusting substrate prepared in Example 1 is shown.

[0061] Figure 4The DiD and Annexin V-FITC fluorescence staining photographs of the anti-crusting substrate prepared in Example 1 are shown.

[0062] Figure 5 The scanning electron microscope photograph showing the anti-fouling effect of the anti-crusting substrate prepared in Example 1.

[0063] Figure 6 The H&E staining pictures of the anti-crusting substrate prepared in Example 1 on the urinary system tissues after implantation in SD rats are shown. DETAILED DESCRIPTION

[0064] In order to more clearly illustrate the present invention, the present invention is further described below in conjunction with preferred embodiments and accompanying drawings. Similar components in the accompanying drawings are represented by the same reference numerals. It should be understood by those skilled in the art that the content specifically described below is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0065] Example 1

[0066] A method for preparing an anti-crusting device, the method comprising the following steps:

[0067] Preparation of aqueous solution of erythrocyte membrane vesicles:

[0068] 1) Add 1 mL of fresh whole blood to 10 mL of 1× PBS (Cytiva HyClone), centrifuge at 800 g for 5 min at 4°C, gently aspirate the upper yellow liquid, and repeat washing 3 times to obtain red blood cell pellet;

[0069] 2) Gently resuspend the erythrocyte sediment obtained above with 0.25×PBS (Cytiva HyClone), treat with hypotonicity at 4°C for 30 min, centrifuge at 13000 g for 10 min, repeat the resuspension-centrifugation operation 5 times to obtain erythrocyte membrane vesicles;

[0070] 3) At 4°C, add 3 mL of deionized water to the above red blood cell membrane vesicles, gently resuspend the solution, and prepare it for immediate use to obtain an aqueous solution of red blood cell membrane vesicles;

[0071] Preparation of amino-functionalized substrates:

[0072] 4) Use anhydrous ethanol and deionized water to coat the polyurethane substrate (2×2cm 2 , thickness d = 1 mm) was cleaned by ultrasonic cleaning for 30 min, dried in an oven at 50 ° C for 1 h, and treated with a plasma cleaner in high power mode for 10 min;

[0073] 5) Spin coating a pre-prepared 1.0 wt % chitosan aqueous solution containing 0.05 wt % 3-glyceropropyltrimethoxysilane on the substrate at a spin coating speed of 3000 rpm for 1 min, and drying in an oven at 60° C. for 5 h to obtain an amino-functionalized substrate;

[0074] Preparation of anti-crusting substrate:

[0075] 6) Add 0.05 wt % of four-arm-polyethylene glycol-succinimidyl succinate to the aqueous solution of the above-mentioned red blood cell membrane vesicles and mix well to obtain a mixed solution, and modify the mixed solution on the surface of the above-mentioned amino-functionalized substrate by spin coating, and the spin coating speed is 3000 rpm for 30 seconds, and dry at room temperature for 2 hours to obtain an anti-encrusting substrate with a red blood cell membrane coating formed on the surface.

[0076] The scanning electron microscope photograph of the red blood cell membrane coating on the anti-crusting substrate is shown in FIG. Figure 1 As shown in FIG. 1 , it is shown that the coating is composed of a uniform cell membrane vesicle structure. The fluorescent staining photos of the red blood cell membrane coating on the anti-crusting substrate after various stimulations (ultrasonic vibration, fluid flushing, repeated bending, 75% ethanol disinfection) are shown in FIG. Figure 2 As shown, this indicates that the cell membrane coating has good stability.

[0077] Example 2

[0078] A method for preparing an anti-crusting device, the method comprising the following steps:

[0079] Preparation of aqueous solution of erythrocyte membrane vesicles:

[0080] 1) Add 1 mL of fresh whole blood to 8 mL of 1× PBS (Cytiva HyClone), centrifuge at 800 g for 5 min at 4°C, gently aspirate the upper yellow liquid, and repeat washing 3 times to obtain red blood cell pellet;

[0081] 2) Gently resuspend the erythrocyte sediment obtained above with 0.25×PBS (Cytiva HyClone), treat with hypotonicity at 4°C for 60 min, centrifuge at 13600 g for 6 min, repeat the resuspension-centrifugation operation 5 times to obtain erythrocyte membrane vesicles;

[0082] 3) At 4°C, add 2 mL of deionized water to the above red blood cell membrane vesicles, gently resuspend the solution, and prepare it for immediate use to obtain an aqueous solution of red blood cell membrane vesicles;

[0083] Preparation of amino-functionalized substrates:

[0084] 4) Use anhydrous ethanol and deionized water to coat the polyurethane substrate (2×2cm 2, thickness d = 1 mm) was cleaned by ultrasonic cleaning for 30 min, dried in an oven at 50 ° C for 1 h, and treated with a plasma cleaner at high power mode for 10 min;

[0085] 5) The plasma-treated substrate was quickly immersed in 50 mL of a silane solution (containing 5 μL of acetic acid, 1.0 wt% (3-aminopropyl)trimethoxysilane, and 50 mL of deionized water), and then immersed at room temperature for 2 h. The surface of the substrate was then cleaned with deionized water and dried with nitrogen to obtain an amino-functionalized substrate;

[0086] Preparation of anti-crusting substrate:

[0087] 6) Add 0.06 wt % of four-arm-polyethylene glycol-succinimidyl glutarate to the aqueous solution of the above-mentioned red blood cell membrane vesicles and mix well to obtain a mixed solution, and immerse the above-mentioned amino-functionalized substrate in the mixed solution, soak it at 4°C for 30 minutes, then take it out and dry it at room temperature for 4 hours to obtain an anti-encrusting substrate with a red blood cell membrane coating formed on the surface.

[0088] Example 3

[0089] A method for preparing an anti-crusting device, the method comprising the following steps:

[0090] Preparation of aqueous solution of erythrocyte membrane vesicles:

[0091] 1) Add 1 mL of fresh whole blood to 8 mL of 1× PBS (Cytiva HyClone), centrifuge at 800 g for 5 min at 4°C, gently aspirate the upper yellow liquid, and repeat washing 3 times to obtain red blood cell pellet;

[0092] 2) Gently resuspend the erythrocyte sediment obtained above with 0.25×PBS (Cytiva HyClone), treat with hypotonicity at 4°C for 40 min, centrifuge at 13800 g for 5 min, repeat the resuspension-centrifugation operation 5 times to obtain erythrocyte membrane vesicles;

[0093] 3) At 4°C, add 2 mL of deionized water to the above red blood cell membrane vesicles, gently resuspend the solution, and then ultrasonicate at 40 Hz for 5 min to obtain an aqueous solution of red blood cell membrane vesicles;

[0094] Preparation of amino-functionalized substrates:

[0095] 4) Use anhydrous ethanol and deionized water to treat the glass substrate (2×2cm 2 , thickness d = 1 mm) was cleaned by ultrasonic cleaning for 30 min, dried in an oven at 50 ° C for 1 h, and treated with a plasma cleaner in high power mode for 8 min;

[0096] 5) The plasma-treated substrate was quickly immersed in 100 mL of a silane solution (containing 10 μL of acetic acid, 1.0 wt% (3-aminopropyl) triethoxysilane, and 100 mL of deionized water), and then immersed at room temperature for 2 h. The substrate surface was then cleaned with deionized water and dried with nitrogen to obtain an amino-functionalized substrate;

[0097] Preparation of anti-crusting substrate:

[0098] 6) Add 0.08 wt % of four-arm-polyethylene glycol-succinimidyl glutarate to the aqueous solution of the above-mentioned red blood cell membrane vesicles and mix well to obtain a mixed solution, and modify the mixed solution on the surface of the above-mentioned amino-functionalized substrate by spin coating, and the spin coating speed is 3000 rpm for 30 seconds, and dry at room temperature for 2 hours to obtain an anti-encrusting substrate with a red blood cell membrane coating formed on the surface.

[0099] Example 4

[0100] A method for preparing an anti-crusting device, the method comprising the following steps:

[0101] Preparation of aqueous solution of erythrocyte membrane vesicles:

[0102] 1) Add 1 mL of fresh whole blood to 6 mL of 1× PBS (Cytiva HyClone), centrifuge at 800 g for 5 min at 4°C, gently aspirate the upper yellow liquid, and repeat washing 3 times to obtain the red blood cell pellet;

[0103] 2) Gently resuspend the erythrocyte sediment obtained above with 0.25×PBS (Cytiva HyClone), treat with hypotonicity at 4°C for 30 min, centrifuge at 13000 g for 10 min, repeat the resuspension-centrifugation operation 5 times to obtain erythrocyte membrane vesicles;

[0104] 3) At 4°C, add 2 mL of deionized water to the above red blood cell membrane vesicles, gently resuspend the solution, and then ultrasonicate at 40 Hz for 6 min to obtain an aqueous solution of red blood cell membrane vesicles;

[0105] Preparation of amino-functionalized substrates:

[0106] 4) Use anhydrous ethanol and deionized water to treat the polyvinyl chloride substrate (2×2cm 2 , thickness d = 1 mm) was cleaned by ultrasonic cleaning for 30 min, dried in an oven at 50 ° C for 1 h, and treated with a plasma cleaner in high power mode for 12 min;

[0107] 5) Spin coating a pre-prepared 1.0 wt % polyethyleneimine solution containing 0.06 wt % 3-glyceropropyltrimethoxysilane at a speed of 3000 rpm for 1 min, and drying in an oven at 50° C. for 6 h to obtain an amino-functionalized substrate;

[0108] Preparation of anti-crusting substrate:

[0109] 6) Add 0.1 wt % of four-arm-polyethylene glycol-succinimidyl succinate to the aqueous solution of the above-mentioned red blood cell membrane vesicles and mix well to obtain a mixed solution, and modify the mixed solution on the surface of the above-mentioned amino-functionalized substrate by spin coating, the spin coating speed is 4000 rpm and maintained for 30 seconds, and dried at room temperature for 4 hours to obtain the red blood cell membrane coating.

[0110] Comparative Example 1

[0111] A method for preparing an anti-crusting device, the method comprising the following steps:

[0112] Preparation of aqueous solution of erythrocyte membrane vesicles:

[0113] 1) Add 1 mL of fresh whole blood to 10 mL of 1× PBS (Cytiva HyClone), centrifuge at 800 g for 5 min at 4°C, gently aspirate the upper yellow liquid, and repeat washing 3 times to obtain red blood cell pellet;

[0114] 2) Gently resuspend the erythrocyte sediment obtained above with 0.25×PBS (Cytiva HyClone), treat with hypotonicity at 4°C for 30 min, centrifuge at 13000 g for 10 min, repeat the resuspension-centrifugation operation 5 times to obtain erythrocyte membrane vesicles;

[0115] 3) At 4°C, add 3 mL of deionized water to the above red blood cell membrane vesicles, gently resuspend the solution, and prepare it for immediate use to obtain an aqueous solution of red blood cell membrane vesicles;

[0116] Pretreatment of substrate:

[0117] 4) Use anhydrous ethanol and deionized water to coat the polyurethane substrate (2×2cm 2 , thickness d = 1 mm) was cleaned by ultrasonic cleaning for 30 min, dried in an oven at 50 ° C for 1 h, and treated with a plasma cleaner in high power mode for 10 min;

[0118] Preparation of anti-crusting substrate:

[0119] 5) The aqueous solution of the red blood cell membrane vesicles is modified on the surface of the treated substrate by spin coating at a speed of 3000 rpm for 30 seconds, and dried at room temperature for 2 hours to obtain an anti-crusting substrate with a red blood cell membrane coating formed on the surface.

[0120] The fluorescent staining photos of the red blood cell membrane coating on the anti-crusting substrate after various stimulations (ultrasonic vibration, fluid flushing, repeated bending, 75% ethanol disinfection) are shown in the figure. Figure 2 As shown, this indicates that the cell membrane coating has poor stability.

[0121] Experimental example

[0122] 1. Water contact angle experiment in air:

[0123] At room temperature, the anti-crusting substrate coated with red blood cell membrane coating prepared in the above example and the uncoated substrate were used to test the wetting of the surface by water droplets (3 μL) in the air, and at least 5 different positions were tested for each sample.

[0124] Among them, the test results of the anti-crusting substrate in Example 1 are shown in Figure 3 .from Figure 3 It can be seen that, compared with the substrate without surface modification (before modification), the red blood cell membrane coating on the anti-encrustation substrate in the present application (after modification) is hydrophilic.

[0125] 2. Validation experiment of selective exposure of the outer leaflet of the cell membrane coating:

[0126] The anti-crusting substrate with the red blood cell membrane coating formed on the surface prepared in the above embodiment was cleaned with deionized water, placed in 5 mL of PBS solution containing 5 wt% Annexin V FITIC, and immersed in light at room temperature for 30 minutes. The selective exposure of the outer leaflet of the cell membrane coating was verified by fluorescence microscopy.

[0127] Among them, the DiD and Annexin V-FITC fluorescence staining results of the anti-crusting substrate with a red blood cell membrane coating formed on the surface in Example 1 are shown in FIG. Figure 4 From the photo, it can be seen that the cell membrane is stained red by DiD, while there is no green fluorescence after Annexin V-FITC staining, indicating that the phospholipids of the inner leaflet of the cell membrane are not exposed, indicating that the coating has the characteristic of selectively exposing the outer leaflet of the cell membrane.

[0128] The erythrocyte membrane coatings in the above-mentioned Examples 2 to 4 also have the characteristic of selective exposure of the outer leaflet of the cell membrane as shown in Example 1.

[0129] 3. Anti-mineral crusting test:

[0130] The experimental method includes the following steps:

[0131] First, a supersaturated mineral solution is prepared; for example, 150.0 mmol of sodium chloride, 5.0 mmol of sodium acetate, 1.4 mmol of calcium chloride, and 1.0 mM of sodium oxalate are added to 1 L of deionized water, and the pH of the solution is adjusted to 7.0 after mixing to obtain a supersaturated calcium oxalate mineral solution;

[0132] 1 L of 20 mM CaCl2 aqueous solution and 1 L of 20 mM NaHCO3 aqueous solution were mixed uniformly at a volume ratio of 1:1 to obtain a supersaturated calcium carbonate mineral solution;

[0133] 100mmol L -1 CaCl2 and 100mmol L -1 Na2SO4 was mixed in a volume ratio of 1:1 to obtain a supersaturated calcium sulfate mineral solution;

[0134] 20mmol L -1 CaCl2 and 20mmol L -1 KH2PO4 were mixed in a volume ratio of 1:1 and the pH was adjusted to 6.0 to obtain a supersaturated calcium phosphate mineral solution;

[0135] Then, the cell membrane coatings prepared in the examples and comparative examples were immersed in the above-mentioned supersaturated mineral solution and incubated at room temperature for 12 hours to observe the growth and coverage of mineralized crystals on the surface of the cell membrane coatings. The results are shown in Tables 1 and 2. Figure 5 shown.

[0136] In the following Table 1, the deposition coverage of different mineralized crystals is obtained by dividing the area of ​​crystals deposited on the substrate surface by the surface area of ​​the substrate.

[0137] Table 1

[0138]

[0139] The scanning electron microscope photograph of the anti-fouling effect of the anti-crusting substrate with the red blood cell membrane coating formed on the surface in Example 1 is as follows Figure 5 As shown, from Figure 5 It can be seen that the surface of the unmodified substrate control sample is covered with a large amount of mineral crystals, while the surface of the cell membrane coating-modified substrate (the anti-crusting substrate with a red blood cell membrane coating formed on the surface obtained in Example 1) is very clean, indicating that the coating has a significant anti-mineral crystal effect.

[0140] 4. In vivo implantation experiment:

[0141] The anti-crusting substrate with a red blood cell membrane coating formed on the surface prepared in the example was surgically implanted into the urinary bladder of SD rats after anesthesia. After 2 weeks of feeding, sections of the urinary system tissues (kidneys, ureters and bladder) of the SD rats were taken out for H&E staining to observe the inflammation and pathological levels of the tissues.

[0142] Example 1 The H&E staining pictures of the urinary system tissues of the anti-crusting substrate with a red blood cell membrane coating formed on the surface after implantation in SD rats are shown in the following figure. Figure 6 As shown, from Figure 6 It can be seen that the bladder tissues of the control sample (substrate without surface modification) and the sample containing cell membrane coating (the anti-crusting substrate with red blood cell membrane coating formed on the surface obtained in Example 1) are normal, and no obvious inflammatory cell infiltration is observed, indicating that the coating has excellent biocompatibility.

[0143] The erythrocyte membrane coatings in the above-mentioned Examples 2 to 4 also have excellent biocompatibility as shown in Example 1.

[0144] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not limitations on the implementation methods of the present invention. For ordinary technicians in the relevant field, other different forms of changes or modifications can be made based on the above description. It is impossible to list all the implementation methods here. All obvious changes or modifications derived from the technical solution of the present invention are still within the protection scope of the present invention.

Claims

1. A coating composition for preventing encrustation of medical devices, characterized in that: The raw materials of the coating composition include the following components: A cell membrane vesicle fluid selected from an aqueous solution of a cell membrane vesicle; and The cross-linking agent is selected from multi-arm-polyethylene glycol-succinimidyl carboxylate.

2. The coating composition according to claim 1, characterized in that The multi-arm-polyethylene glycol-succinimide carboxylate is selected from one or more of two-arm-polyethylene glycol-succinimide carboxylate, four-arm-polyethylene glycol-succinimide succinate, four-arm-polyethylene glycol-succinimide glutarate, four-arm-polyethylene glycol-succinimide acetate and four-arm-polyethylene glycol-succinimide carbonate; Preferably, the outer leaflets of the cell membrane vesicles in the cell membrane vesicle fluid are all exposed; Preferably, the cell membrane vesicles in the cell membrane vesicle fluid are prepared from one or more of erythrocyte membranes, platelet membranes, macrophage membranes and neutrophil membranes.

3. The coating composition according to claim 1, characterized in that The preparation of the cell membrane vesicles comprises the following steps: After mixing whole blood with PBS, centrifuge, aspirate the upper liquid, and wash to obtain cell pellets; The cell pellet is resuspended in PBS, subjected to hypotonic treatment, and centrifuged to obtain the cell membrane vesicles; Preferably, the PBS is selected from Cytiva HyClone; Preferably, the mixing volume ratio of whole blood to PBS is 1:(6-10), the temperature of centrifuging whole blood and PBS is 4°C, the centrifugal force is 800g, and the time is 5min; Preferably, the temperature of the hypotonic treatment is 4°C and the time is 30-60 min; Preferably, the centrifugal force after the hypotonic treatment is 13000-14000 g, and the time is 5-10 min.

4. The coating composition according to claim 1, characterized in that In the cell membrane vesicle solution, the concentration of cell membrane vesicles is 0.1-0.5wt%; and / or The protein concentration in the cell membrane vesicle fluid is 0.2-5 mg / mL; and / or The added amount of the cross-linking agent relative to the cell membrane vesicle fluid is 0.01-0.5 wt %.

5. The method for preparing the coating composition according to any one of claims 1 to 4, characterized in that: The steps include: The cell membrane vesicle liquid is mixed with a cross-linking agent to obtain the coating composition.

6. A medical device with anti-encrustation, characterized in that: The structure of the medical device includes: substrate; and A cell membrane coating covalently cross-linked on the surface of the substrate, wherein the cell membrane coating is prepared from a raw material comprising the coating composition according to any one of claims 1 to 4.

7. The medical device according to claim 6, characterized in that: The material of the substrate is selected from one or more of polyurethane, silicone, rubber, silicon dioxide, metal alloy, latex, polyvinyl chloride, polyethylene, polypropylene, polylactic acid, polyethylene terephthalate and polycaprolactone.

8. The method for preparing a medical device according to claim 6 or 7, characterized in that: The steps include: aminating the surface of the substrate; Applying the coating composition on the surface of the amino-modified substrate and drying the coating composition to obtain a cell membrane coating covalently cross-linked on the surface of the substrate; Preferably, the coating method is spin coating, spray coating, scraper coating, brush coating or dip coating; Preferably, the spin coating has a rotation speed of 3000-4000 rpm and a time of 30-50 s; Preferably, the dipping method is: immersing the surface-amino-treated substrate in the coating composition at a temperature of 4-50° C. for 0.5-5 h.

9. The preparation method according to claim 8, characterized in that: The method for aminating the substrate surface comprises the following steps: performing surface plasma treatment on the substrate; A silane solution containing amino groups is used to achieve amination on the surface of the plasma-treated substrate through spin coating or dip coating.

10. Use of the coating composition according to any one of claims 1 to 4 in the preparation of medical devices.

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