CN119455137A - Antibacterial central venous catheter and preparation method thereof - Google Patents
Antibacterial central venous catheter and preparation method thereof Download PDFInfo
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- CN119455137A CN119455137A CN202411399036.1A CN202411399036A CN119455137A CN 119455137 A CN119455137 A CN 119455137A CN 202411399036 A CN202411399036 A CN 202411399036A CN 119455137 A CN119455137 A CN 119455137A
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- Materials For Medical Uses (AREA)
Abstract
The invention belongs to the technical field of preparation of biomedical materials, and provides an antibacterial central venous catheter and a preparation method thereof. According to the invention, the pulse frequency in the silver layer deposition process is controlled to be 240-260 kHz, so that the density of silver nano particles is improved, the sputtering rate is improved, and the sputtered nano particles have higher energy, thereby improving the adhesion between the nano particles and a matrix and the density of the nano particles. Meanwhile, the working pressure is controlled to be 0.24-0.25 Pa in the silver layer deposition process, so that the collision times of sputtered silver nano particles and gas molecules are reduced, the lost energy is smaller, the diffusion capability of deposited particles and a matrix is improved, the compactness and adhesiveness of a coating are improved, the defect that the adhesive force between the silver nano coating and a central venous catheter in the prior art is weak is overcome, and the antibacterial central venous catheter with high adhesive force, compactness, stability and purity is prepared.
Description
Technical Field
The invention relates to the technical field of preparation of biomedical materials, in particular to an antibacterial central venous catheter and a preparation method thereof.
Background
A central venous catheter (CVC for short) is one of intravascular tubes, which is usually placed in a great vein, and is mainly made of Polyurethane (PU) or silica gel, and is mainly used for rapid infusion of blood volume deficiency, pressure monitoring of heart failure, infusion of vasoactive drugs for severe shock, drainage of various central peritoneal effusions and pericardial effusions, central venous pressure monitoring of cardiac great vessel surgery and other serious patients, massive and rapid venous infusion, long-term venous nutrition support treatment and the like in clinic. Therefore, the utility model has wide applicability in ICU ward, surgery, thoracic surgery, anesthesia department and the like. In the united states, over 500 ten thousand central venous catheters are used to treat cancer. However, widespread use of intravenous catheters can greatly increase the incidence of life-threatening complications such as catheter-related blood flow infections (CRBI), often resulting in prolonged patient hospitalization, increased medical costs, and increased mortality. Clinical practice has found that some patients will become infected after catheterization of central venous catheters, and previous studies have shown that prolonged catheterization increases the risk of acting as a reservoir for bacterial proliferation, increasing bacterial attachment and colonization, and microbial attachment to the inner and outer surfaces of the catheters, against the action of the host defense system. Therefore, it is necessary to develop a central venous catheter having good tissue compatibility and high-efficiency broad-spectrum antibacterial properties to effectively inhibit various typical microorganisms which are bred and stored in the inner and outer walls of the catheter.
The silver nano particles are smaller than 100nm and have 20-15000 silver atoms. The existing nano silver antibacterial agent has the advantages of good antibacterial effect, low toxicity, difficult generation of drug resistance and the like. Under certain conditions, free release of silver ions in the silver nanoparticles can induce cell death of mammalian cells or microbial cells, which means that the silver nanoparticles are broad spectrum antimicrobial agents.
The Chinese patent with publication number CN102380143A discloses a nano-silver surface modified polyurethane central venous catheter and a preparation method thereof, which discloses that the polyurethane central venous catheter is soaked in a mixed solvent of ethanol and deionized water, washed, taken out and dried, the dried polyurethane central venous catheter is soaked in silver nitrate solution, taken out and irradiated by ultraviolet light, or the dried polyurethane central venous catheter is soaked in the silver nitrate solution and irradiated by ultraviolet light, then washed and soaked by deionized water, water is continuously changed during the period, and finally dried, so that the nano-silver surface modified polyurethane central venous catheter is obtained. The silver nano coating obtained by the method has low adhesive force with the middle intravenous catheter and is easy to fall off.
Disclosure of Invention
Accordingly, the present invention is directed to an antimicrobial central venous catheter and a method for preparing the same. The silver layer obtained by the preparation method provided by the invention has strong adhesive force with the blank central venous catheter and high compactness, and in addition, the obtained bacterial central venous catheter has excellent antibacterial property and biocompatibility.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides a preparation method of an antibacterial central venous catheter, which comprises the following steps:
sequentially cleaning, polishing and drying the central venous catheter to obtain a pretreated central venous catheter;
sequentially performing plasma cleaning and silver layer deposition on the pretreatment central venous catheter by utilizing magnetron sputtering to obtain the antibacterial central venous catheter;
The magnetron sputtering parameters of silver layer deposition comprise argon as working gas, working pressure of 0.24-0.25 Pa, pulse width of pulse bias of 250-255 mu s and pulse frequency of 240-260 kHz.
Preferably, the magnetron sputtering parameters of the silver layer deposition further comprise that the distance between the pretreatment central venous catheter and the silver target is 170mm.
Preferably, the magnetron sputtering parameters of the silver layer deposition further comprise that the applied current of the silver target is 1.5×10 -2～2×10-2 A.
Preferably, the purity of the silver target is 99.99%.
Preferably, the magnetron sputtering parameters of silver layer deposition further comprise negative bias voltage of 180-200V.
Preferably, the silver layer is deposited for 90-210 s.
Preferably, the central venous catheter is made of polyurethane or silica gel.
Preferably, the cleaning mode is ultrasonic bath cleaning.
Preferably, the drying mode is nitrogen blow drying.
The invention also provides the antibacterial central venous catheter prepared by the preparation method, which comprises a central venous catheter and a silver layer deposited on the outer surface of the central venous catheter.
The invention provides a preparation method of an antibacterial central venous catheter.
The beneficial effects are that:
1. According to the invention, the pulse frequency in the silver layer deposition process is controlled to be 240-260 kHz, so that the density of silver nanoparticles is improved, the sputtering rate is improved, and the sputtered silver nanoparticles have higher energy, thereby improving the adhesion force of the silver nanoparticles and a matrix and the density of the silver nanoparticles. Meanwhile, the working pressure in the silver layer deposition process is controlled to be 0.24-0.25 Pa, so that the collision times of sputtered silver nano particles and gas molecules are reduced, the lost energy is smaller, the diffusion capacity of the deposited silver nano particles and a matrix is improved, the compactness and the adhesiveness of a coating are improved, the defect that the adhesive force of the silver nano coating and a central venous catheter in the prior art is weak is overcome, and the antibacterial central venous catheter with high adhesive force and high compactness is prepared.
2. The preparation method provided by the invention has the advantages of mild condition, simple and easy operation and short time consumption.
3. The antibacterial central venous catheter with high adhesive force and high compactness, which is prepared by the invention, has excellent biocompatibility, and the coating of the antibacterial central venous catheter has high purity and high stability.
4. The antibacterial central venous catheter with high adhesive force and high compactness prepared by the invention has lower friction coefficient and more excellent water lubricity compared with a commercial central venous catheter.
5. The antibacterial central venous catheter with high adhesive force and high compactness prepared by the invention has broad-spectrum strong antibacterial property.
Drawings
FIG. 1 is a photograph of a hollow white center venous catheter (blank CVC) of test example 1 and a CVC@Ag obtained in example 1;
FIG. 2 shows the water contact angle of a hollow white central venous catheter (blank CVC) of test example 2 and CVC@Ag obtained in example 1;
FIG. 3 is a scanning electron micrograph of CVC@Ag obtained in example 1 of test example 3 at different magnifications;
FIG. 4 is a scanning electron micrograph of a hollow white central venous catheter (blank CVC) of test example 3 at different magnification;
FIG. 5 is a spectroscopic image of a hollow white central venous catheter (blank CVC) of test example 3;
FIG. 6 is an image of the spectrum analysis of CVC@Ag obtained in example 1 of test example 3;
FIG. 7 is an atomic force microscope image of CVC@Ag obtained in example 1 of test example 4;
FIG. 8 is a representative plot of the colonisation of Staphylococcus aureus (S.aureus), methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E.Coli) on a blank CVC and CVC@Ag obtained in example 1 in test example 5;
FIG. 9 shows the antibacterial activity of Staphylococcus aureus (S.aureus), methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E.Coli) in test example 5 on blank CVC and CVC@Ag obtained in example 1;
FIG. 10 is a graph of relative activity after incubation of 4T1 cells with different catheter sets in test example 6;
FIG. 11 is an H & E staining image of skin tissue and blood samples at different catheter implantation sites for different days in test example 6;
FIG. 12 is a graph showing the characterization of immunofluorescence and immunohistochemistry of skin tissue and blood samples at different catheter implantation sites for different days in test example 6.
Detailed Description
The invention provides a preparation method of an antibacterial central venous catheter, which comprises the following steps:
sequentially cleaning, polishing and drying the central venous catheter to obtain a pretreated central venous catheter;
sequentially performing plasma cleaning and silver layer deposition on the pretreatment central venous catheter by utilizing magnetron sputtering to obtain the antibacterial central venous catheter;
The magnetron sputtering parameters of silver layer deposition comprise argon as working gas, working pressure of 0.24-0.25 Pa, pulse width of pulse bias of 250-255 mu s and pulse frequency of 240-260 kHz.
In the present invention, the raw materials used in the present invention are preferably commercially available products unless otherwise specified.
The central venous catheter is sequentially cleaned, polished and dried to obtain the pretreatment central venous catheter.
In the present invention, the material of the central venous catheter is preferably polyurethane (SU) or silicone.
In the invention, the cleaning mode is preferably ultrasonic bath cleaning, the power of the ultrasonic bath cleaning is preferably 40-50 kHz, and the time is preferably 2-3 min.
The parameters of the polishing are not particularly limited in the present invention, and polishing operations well known to those skilled in the art may be employed.
In the present invention, the drying mode is preferably nitrogen blow-drying, and the time of the nitrogen blow-drying is not particularly limited, so long as the drying can be performed.
After the pretreatment central venous catheter is obtained, plasma cleaning and silver layer deposition are sequentially carried out on the pretreatment central venous catheter by utilizing magnetron sputtering, so that the antibacterial central venous catheter is obtained.
The parameters of the plasma cleaning are not particularly limited in the present invention. In the invention, the plasma cleaning time is preferably 60-180 s.
In the invention, the magnetron sputtering parameters of silver layer deposition comprise argon as working gas, working pressure of 0.24-0.25 Pa, specifically 0.24Pa, 0.245Pa or 0.25Pa, pulse width of pulse bias of 250-255 μs, preferably 250 μs, specifically 250 μs, 251 μs, 252 μs, 253 μs, 254 μs or 255 μs, and pulse frequency of 240-260 kHz, preferably 250kHz, specifically 240kHz, 245kHz, 250kHz, 255kHz or 260kHz.
In the invention, the magnetron sputtering parameters of silver layer deposition further comprise that the distance between the pretreatment central venous catheter and the silver target is preferably 170mm, the applied current of the silver target is preferably 1.5X10 -2～2×10-2 A, particularly 1.5X10 -2A、1.6×10-2A、1.7×10-2A、1.8×10-2A、1.9×10-2 A or 2X 10 -2 A, the purity of the silver target is preferably 99.99%, and the negative bias is preferably 180-200V, particularly 180V, 185V, 190V, 195V or 200V. In the present invention, the deposition time of the silver layer is preferably 90 to 210s, and may specifically be 90s, 120s, 150s, 180s or 210s.
The invention also provides the antibacterial central venous catheter prepared by the preparation method, which comprises a central venous catheter and a silver layer deposited on the outer surface of the central venous catheter. In the invention, the thickness of the silver layer is preferably 10.5-20 mu m, the purity of the silver layer is preferably 99.99%, and the silver layer has high stability and high compactness.
The antibacterial central venous catheter and the method for preparing the same according to the present invention will be described in detail with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
The preparation method of the antibacterial central venous catheter comprises the following steps:
And (3) carrying out ultrasonic bath cleaning on the central venous catheter at 50Hz for 3min, polishing, and drying by nitrogen after finishing to obtain the pretreated central venous catheter.
The pretreated central venous catheter was plasma cleaned for 180s.
After the plasma cleaning is finished, the working pressure is regulated to be 0.25Pa, the current is applied to the silver target material to be 2 multiplied by 10 -2 A, the negative bias voltage is set to be 200V, the pulse width of the pulse bias voltage is set to be 250 mu s, the pulse frequency is set to be 250kHz, and the silver layer is deposited for 90s by pulling down a baffle plate between the workpiece frame and the silver target material, so that the antibacterial central venous catheter (CVC@Ag hereinafter) is obtained.
Comparative example 1
The difference from example 1 is that the pulse frequency was adjusted to 200kHz during the deposition of the silver layer, otherwise the same as in example 1.
Comparative example 2
The difference from example 1 is that the operating voltage was adjusted to 0.2Pa during the deposition of the silver layer, otherwise the same as in example 1.
The antibacterial central venous catheters obtained in comparative examples 1 and 2 have greatly increased colony numbers on the surfaces of CVC@Ag, and have antibacterial rates of 70% and 78% on three pathogenic bacteria of staphylococcus aureus (S.aureus), methicillin-resistant staphylococcus aureus (MRSA) and escherichia coli (E.Coli) respectively, so that the antibacterial rate of the antibacterial central venous catheter obtained in the combination example 1 is required to be further improved, and the important effect of pulse frequency and working voltage on the antibacterial property of the prepared antibacterial nano silver coating in the silver layer deposition process is shown.
Test example 1
Fig. 1 is a photograph of a blank central venous catheter (blank CVC) and CVC@Ag obtained in example 1, and as can be seen from fig. 1, the surface of the blank CVC is smooth and transparent, while the surface of the CVC@Ag is smooth, the nano silver coating is uniform, and the metallic luster of silver is displayed, so that the nano silver coating with high purity, high compactness and high stability is constructed on the surface of the blank CVC.
Test example 2
Fig. 2 shows the water contact angle of a blank central venous catheter (blank CVC) and CVC@Ag obtained in example 1, and fig. 2 shows that the water contact angle is obviously changed after the nano silver coating is coated, the water contact angle is changed from 96.4 degrees to 67.1 degrees, the nano silver coating is proved to obviously improve the hydrophilicity of the blank CVC, and the nano silver coating is a barrier for preventing bacteria adsorption.
Test example 3
The composition and morphology of the nano silver coating are analyzed by a scanning electron microscope and energy spectrum analysis.
Fig. 3 is a scanning electron micrograph of the cvc@ag obtained in example 1 at different magnifications, fig. 4 is a scanning electron micrograph of a blank central venous catheter (blank CVC) at different magnifications, and it can be seen from fig. 3 and 4 that the surface of the blank CVC is smooth and transparent, and shows uniform and dense metallic luster after the nano silver coating is coated, which indicates successful construction of the nano silver coating, and through microscopic morphology analysis, a huge difference exists between the surface of the cvc@ag and the surface of the blank CVC, and nano silver particles are uniformly adhered to the surface of the cvc@ag, which indicates successful construction of the silver nano particles, and Mapping further proves uniform distribution of silver elements.
Fig. 5 is a spectroscopic image of a blank central venous catheter (blank CVC), fig. 6 is a spectroscopic image of cvc@ag obtained in example 1, and it can be seen from fig. 5 and 6 that the silver nanoparticles are uniformly distributed.
Test example 4
The roughness of the nano silver coating was measured using an atomic force microscope.
Fig. 7 is an atomic force microscope image of cvc@ag obtained in example 1, and it can be seen from fig. 7 that the nano silver coating has a flat and smooth surface, and silver nanoparticles are uniformly distributed on the surface of the blank CVC.
Test example 5
In-vitro antibacterial experiments, three pathogenic bacteria of clinically common staphylococcus aureus (S.aureus), methicillin-resistant staphylococcus aureus (MRSA) and escherichia coli (E.Coli) are selected as experimental platforms to evaluate the in-vitro antibacterial performance of different catheters (blank CVC and CVC@Ag obtained in example 1). Soaking the catheter in the bacterial liquid, standing for sterilization for half an hour, collecting residual bacteria on the surface of the catheter by using a cotton swab, dispersing the bacteria in a culture medium again, inoculating the bacteria into a flat plate, culturing for 24 hours, and counting the colony count to calculate the bacteriostasis rate of the catheter.
FIG. 8 is a representative colonization pattern of Staphylococcus aureus (S.aureus), methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E.Coli) on the blank CVC and CVC@Ag obtained in example 1, FIG. 9 is an antibacterial rate of Staphylococcus aureus (S.aureus), methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E.Coli) on the blank CVC and CVC@Ag obtained in example 1, and it can be seen from FIGS. 8 and 9 that the colony number on the surface of CVC@Ag is greatly reduced, and the antibacterial rate on all three bacteria reaches 95%, indicating that CVC@Ag has good antibacterial performance.
Test example 6
In vivo antibacterial experiments based on the above experimental results, in vivo antibacterial experiments were performed, first, biocompatibility of catheters was evaluated, 4T1 cells were seeded into 96-well plates, and cultured in DMEM medium containing 10% fbs and 1% streptomycin and penicillin at 37 ℃ under an atmosphere of 5% co 2 to evaluate the biocompatibility of catheters. The catheters were divided into three groups, one group being a blank Control group, noted Control, one group being CVC and the other group being CVC@Ag. Catheter extracts were prepared according to the method specified in materials preparation and reference materials (GB/T16886.11-2017) section 12 of medical device biology evaluation criteria, cell activities were examined after 24 hours of co-culture of the extracts with cells, and the results are shown in FIG. 10, FIG. 10 shows a graph of relative activities of 4T1 cells after incubation with different catheter groups, and it can be seen from FIG. 10 that CVC@Ag extracts group cell activities remained at the highest level compared to the control group, indicating that CVC@Ag biocompatibility was good.
Subsequently, in vivo antibacterial experiments were performed on New Zealand white rabbits, and skin tissue and blood samples at the catheter implantation site were collected after 14 days, 28 days, 42 days, and 56 days of catheter implantation to detect the level of inflammation and the degree of bacterial infection. The results are shown in fig. 11, fig. 11 is an H & E stained image of skin tissue and blood samples at different catheter implantation sites for different days, and it can be seen from fig. 11 that acute inflammation occurred in both skin tissue samples within two weeks and the inflammation level gradually decreased with prolonged observation period, and the inflammation level of cvc@ag group was always lower than that of CVC group throughout the whole process.
The results of detecting the levels of cytokines (IL-1, IL-4, TNF-alpha and TGF-beta) related to inflammation through immunofluorescence and immunohistochemistry are shown in figure 12, and figure 12 is a graph showing the immunofluorescence and the immunohistochemical characterization of skin tissues and blood samples at different catheter implantation sites on different days, and as can be seen from figure 12, CVC@Ag relieves inflammation at the catheter implantation sites, reduces the levels of the cytokines (IL-1, IL-4, TNF-alpha and TGF-beta) related to inflammation, and the trend of the results is consistent with that of H & E staining, and the results show that CVC@Ag relieves inflammation at the catheter implantation sites and reduces the levels of the cytokines related to inflammation.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
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