CN110438455A - The preparation method of polylactic acid base Ciprofloxacin antibacterial film - Google Patents

Abstract

The invention discloses a preparation method of polylactic acid-based ciprofloxacin antibacterial film. In the method, ciprofloxacin powder and polylactic acid are evenly mixed, placed in the reaction chamber, the distance between the target and the clean substrate is adjusted to 20-30 cm, vacuumized, coated, the working current is set at 7-9A, and the working voltage is At 0.8-1.8kV, the antibacterial film of polylactate-based ciprofloxacin was prepared by low-power electron beam deposition technology. The antibacterial film prepared by the invention has strong binding force with the substrate, good compactness, controllable film thickness, good antibacterial property, can be used for artificial prosthesis transplantation, and eliminates local inflammatory reaction.

Description

Preparation method of polylactic acid-based ciprofloxacin antibacterial film

technical field

The invention belongs to the technical field of preparation of antibacterial films, and relates to a preparation method of polylactic acid-based ciprofloxacin antibacterial films using low-power electron beam deposition technology.

Background technique

Polylactic acid (PLA) has good thermal stability, no pollution in the production process, good biocompatibility and degradability, and has certain antibacterial properties. It is widely used, such as packaging materials, injection molding, biomedicine, etc. It is an ideal green polymer material.

Ciprofloxacin has a wide range of antibacterial effects, and has good antibacterial effects on Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa.

The principle of low-power electron beam deposition technology is that a beam of electrons is accelerated after passing through an electric field of 5-10kV, and finally gathers on the surface of the material to be evaporated. When the electron beam hits the surface of the material to be evaporated, the electrons will quickly lose their energy. , transfer energy to the material to be evaporated to melt and evaporate. Electron beam evaporation is the main method to prepare thin films with high melting point and high purity, and it has a wide range of power adjustment and is easy to use. Compared with the PLA-based composite film prepared by magnetron sputtering and electron beam deposition technology, charge accumulation will be formed during the magnetron sputtering coating process, which will lead to an increase in potential. When the potential rises to a certain level, it will cause glow discharge. Stop, and then produce the target "poisoning" phenomenon, this phenomenon makes the plasma unstable, so that the PLA-based antibacterial film obtained by sputtering is uneven and has many defects ([1] Li Guobin, et al. Preparation of white carbon black Technology research progress [J]. Huagong Science and Technology, 2014, 22 (5): 57-60; [2] Wang Hongbo, et al. Antibacterial performance of PLA-based nanostructure silver film [J]. Textile Journal, 2008, 29 (6 ):52-55.). Nguyen et al. used coaxial electrospinning technology to prepare polylactic acid/chitosan composite nanofiber materials. Antibacterial experiments against Escherichia coli showed that polylactic acid/chitosan nanocomposite fiber materials could completely inhibit bacterial growth in the first 12 hours. But after that, the bacteria began to grow gradually, and the antibacterial persistence was poor (Li Shunjiang et al. Research progress of polylactic acid-based antibacterial materials [J]. Applied Chemical Industry, 2014,43(5):916-918.).

Contents of the invention

The object of the present invention is to provide a kind of preparation method of polylactide-based ciprofloxacin antibacterial film with strong binding force between film and substrate and good compactness, and the method can deposit high-purity polymer film at room temperature.

The technical scheme that realizes the object of the present invention is as follows:

The preparation method of polylactic acid base ciprofloxacin antibacterial film, concrete steps are as follows:

Mix ciprofloxacin powder and polylactic acid evenly, place in the reaction chamber, adjust the distance between the target and the clean substrate to 20-30cm, vacuumize, coat, set the working current at 7-9A, and the working voltage at 0.8- 1.8kV, polylactate-based ciprofloxacin antibacterial film was prepared by low-power electron beam deposition technology.

Preferably, the substrate is selected from titanium sheet or silicon sheet.

Preferably, the mass ratio of polylactic acid to ciprofloxacin is 1:1.

Preferably, the vacuum degree reaches 6×10 ^-3 ~ 8×10 ^-3 Pa during the vacuuming.

Compared with the prior art, the present invention has the following advantages:

(1) Electron beam evaporation has higher energy density, can evaporate materials with high melting point, and the evaporation speed is fast, and the obtained film has high purity;

(2) The kinetic energy of the electron beam evaporated particles is large, and the antibacterial film prepared has strong bonding force with the substrate and good compactness;

(3) When the electron beam is evaporated, the film thickness can be measured during the deposition process according to the built-in film thickness meter, which is convenient for controlling the film thickness;

(4) The prepared film material has good antibacterial property, can be used for implantation of artificial prosthesis, and eliminates local inflammatory reaction.

Description of drawings

Fig. 1 is when substrate is Ti sheet, composite film (PLA: ciprofloxacin=1:1) is to the antibacterial effect figure of Staphylococcus aureus;

Fig. 2 is when substrate is Ti sheet, composite film (PLA: ciprofloxacin=1:1) is to the antibacterial effect figure of Pseudomonas aeruginosa;

Fig. 3 is when substrate is Si sheet, composite film (PLA: ciprofloxacin=1:1) is to the antibacterial effect figure of Pseudomonas aeruginosa;

Fig. 4 is respectively the infrared spectrogram of pure PLA, pure ciprofloxacin and composite membrane material (PLA: ciprofloxacin=1:1);

Fig. 5 is the sustained-release antibacterial effect figure of electron beam deposition composite film (PLA: ciprofloxacin=1:1) 24h, 48h and 72h;

Fig. 6 is the antibacterial effect figure of PLA base spraying ciprofloxacin composite film (PLA: ciprofloxacin=1:1) to Staphylococcus aureus;

Fig. 7 is the slow-release effect graph of PLA-based spraying ciprofloxacin composite film (PLA:ciprofloxacin=1:1) for 12h and 24h.

Detailed ways

The present invention will be described in further detail below in conjunction with the embodiments and accompanying drawings.

Electron beam deposition is the use of high-energy electron beams to bombard the target. The ionization of chemical bonds will occur in the bombarded target area, and its temperature will also rise rapidly, causing the target components to evaporate, so that the target will be converted into A plasma state composed of electrons, ions, atoms, and molecules, which is ejected to form an afterglow and deposited on the substrate.

Example 1

Take two slices and cut them into 1*1cm Ti slices and Si slices, put them into ethanol for ultrasonic cleaning for 15 minutes, pour off the ethanol and rinse with deionized water, repeat the above cleaning steps 3 times, and clean the Ti slices and Si slices The beakers of the slices were sealed with plastic wrap and placed in a muffle furnace for drying. Place the dried Ti sheet and Si sheet in the electron beam deposition chamber, fix the substrate with clips, and then deposit the PLA-based ciprofloxacin antibacterial film. The specific steps are as follows:

To clean the reaction chamber of contaminants:

Put the mass ratio (PLA: ciprofloxacin = 1:1) target into the reaction chamber of electron beam deposition, vacuumize with mechanical pump and molecular pump respectively, so that the vacuum degree reaches 6×10 ^-3 ~8× 10 ^-3 Pa.

(1) Deposition of PLA-based composite films

Turn on the power supply of the deposited film, adjust the working current to 7-9A, and control the working voltage to 0.8-1.8kV. When the vacuum degree is displayed at 5×10 ^-2 to 6×10 ^-2 Pa, it can be seen through the observation window that the The material in the plasma state between the target and the substrate is transferred from the target to the substrate, and whether the deposition is completed can be judged by the change of the film thickness on the film thickness meter. When the film thickness no longer changes, it means that the deposition is over. The current and voltage are slowly returned to zero, wait for 10 minutes, and after the temperature of the vacuum chamber is lowered, turn off the power of the instrument, and the coating is completed.

(2) Culture of Staphylococcus aureus

Weigh 0.5g of yeast extract powder, 1.0g of peptone, and 1.0g of NaCl, dissolve them in a beaker with deionized water, add a little NaOH solution dropwise, until the pH value is adjusted to 7, then drain with a glass rod, and pour the solution into 100mL In the Erlenmeyer flask, constant volume, shake well, take three washed small test tubes, take 5.00mL solution in each small test tube with a pipette gun, then seal the test tube mouth with tin foil, and then put the small test tube Sterilize in a high pressure steam sterilizer.

After the sterilization is completed, put the test tube, inoculation loop, and lighter into the ultra-clean workbench and irradiate it with an ultraviolet lamp for 20 minutes. After the irradiation is completed, turn on the fan to blow for 2 minutes. Then put on disposable rubber gloves and take out the strain of Staphylococcus aureus. Light the alcohol lamp, wipe your hands and the ultra-clean workbench with alcohol cotton, put the inoculation loop on the outer flame of the alcohol lamp and burn it until it burns red. Burn the test tube mouth of Staphylococcus aureus on an alcohol lamp, then extend the inoculation loop into the test tube containing Staphylococcus aureus, take an appropriate amount of bacteria and inoculate it into the liquid medium, then wrap it in tin foil, and use Tighten the rubber band tightly, repeat the above steps three times until all three test tubes are inoculated, turn off the alcohol lamp, close the ultra-clean workbench, put the inoculated Staphylococcus aureus in a shaker at 30°C, and incubate for 12 hours.

(3) Study on antibacterial performance of PLA-based ciprofloxacin composite film

Weigh 0.5g of yeast extract powder, 1.0g of peptone, 1.0g of NaCl, and 1.6g of agar powder, dissolve them in a beaker with deionized water, add a little NaOH solution dropwise until the pH value is adjusted to 7, and then drain with a glass rod, Pour the solution into a 100mL Erlenmeyer flask, constant volume, shake well, then seal the mouth of the Erlenmeyer flask with a parafilm, and put the Petri dish wrapped with newspaper, tweezers, pipette tip and liquid medium together Sterilize in a high pressure steam sterilizer.

After the sterilization is completed, put the petri dish, pipette tip, pipette gun and lighter into the ultra-clean workbench, irradiate with ultraviolet light for 20 minutes, blow with a fan for 2 minutes, and choose one of the three cultured Staphylococcus aureus test tubes The best effect is poured into the Erlenmeyer flask, fully shaken, and shaken; the prepared PLA-based ciprofloxacin composite film is cut into small squares or rounded with a puncher, and the Si sheet is used as the base. Cut into small squares, and use a puncher to make a circle with the Ti sheet as the base. Put the composite membrane material with the coating side up, and place it in the center of the culture dish, and pipette 10.0mL of the culture dish. Transfer the culture medium to the Petri dish, pay attention to avoid air bubbles as much as possible during the pipetting process, and the pipetting process should be rapid to prevent the medium from solidifying. Finally, the inoculated medium was placed in an incubator at 30°C for 24 hours, and the picture after taking it out was shown in Figure 1. From the antibacterial zone in Figure 1, it can be seen that the PLA-based ciprofloxacin composite film has good antibacterial effect on Staphylococcus aureus Effect.

Example 2

The membrane material is the PLA-based ciprofloxacin composite membrane material in Example 1, and the Staphylococcus aureus in Example 1 is replaced by Pseudomonas aeruginosa, and other steps are the same as in Example 1, and the PLA-based ciprofloxacin The antibacterial effect of the star composite film is shown in Figure 2 and Figure 3. When the substrate is Si sheet, when the strain is Staphylococcus aureus, the diameter of the antibacterial ring is 5.2cm, and when the strain is Pseudomonas aeruginosa, the diameter of the antibacterial ring is 4.6cm, so the PLA-based ciprofloxacin composite film has a better antibacterial effect on Staphylococcus aureus.

It can be known from Figure 4 that the PLA-based ciprofloxacin composite film corresponds to the transmittance at 1083.8cm ^-1 and 1743.3cm ^-1 on the PLA at the wavenumbers of 1083.7cm ^-1 and 1730.7cm ^-1 respectively, and the PLA-based ciprofloxacin The star composite film at the wave number of 1457.4cm ^-1 and 1616.8cm ^-1 corresponds to the transmittance of ciprofloxacin at 1453.1cm ^-1 and 1618.1cm ^-1 respectively, indicating that PLA and ciprofloxacin do exist in the PLA-based ciprofloxacin composite film The antibiotic ciprofloxacin.

Comparative example 1

Take three PLA-based ciprofloxacin composite films soaked in normal saline for 24h, 48h, and 72h, then take them out, and then do antibacterial experiments to observe the slow-release effect of PLA-based composite films. It can be seen from Figure 5 that after soaking for 24h, 48h After 72 hours, the diameters of the inhibition zone were 4.0cm, 1.5cm and 0.7cm. As the soaking time prolongs, the antibacterial effect of the composite membrane becomes worse, and the inhibition zone becomes smaller, but it has an antibacterial effect, indicating that the antibiotics in the membrane material have been releasing, and there is still release after 72 hours, so the membrane material has been released. Enough to meet the needs of transplantation.

Spray a saturated solution of ciprofloxacin on the deposited PLA film, the quality of ciprofloxacin is the same as the amount used during deposition, the antibacterial effect of the film material on Staphylococcus aureus is shown in Figure 6, it can be seen from the figure that PLA spray The membrane material of ciprofloxacin has an antibacterial effect on Staphylococcus aureus, but the antibacterial ring is not uniform. The reason is that there are many human factors in the spraying process, and it is difficult to control the content of antibiotics to be the same everywhere.

Take two pieces of composite films sprayed with PLA and ciprofloxacin, soak them in normal saline for 12 hours and 24 hours, take them out, and then do antibacterial experiments to observe the slow-release effect of the composite films. It can be seen from Figure 7 that the bacteriostatic zone after soaking for 12 hours is 0.8cm, there is no antibacterial zone after soaking for 24 hours, but the membrane material prepared by electron beam deposition still has an inhibitory zone after soaking for 72 hours, indicating that the membrane material prepared by spraying ciprofloxacin on PLA is far inferior to E-beam evaporated films have good adhesion to substrates. The membrane material prepared by spraying ciprofloxacin on PLA is easy to fall off when soaked in physiological saline, and the sustained release effect is poor, so it cannot be used for clinical transplantation. In summary, the PLA-based ciprofloxacin composite film prepared by electron beam deposition technology has high binding force and good stability.

Claims (4)

1. the preparation method of polylactic acid base Ciprofloxacin antibacterial film, which is characterized in that specific step is as follows:

Ciprofloxacin powder is uniformly mixed with polylactic acid, is placed in reaction chamber, adjustment target is at a distance from clean substrate 20~30cm is vacuumized, plated film, operating current is arranged in 7~9A, operating voltage is in 0.8~1.8kV, using low power electronics Polylactic acid base Ciprofloxacin antibacterial film is prepared in beam deposition technique.

2. preparation method according to claim 1, which is characterized in that the substrate is selected from titanium sheet or silicon wafer.

3. preparation method according to claim 1, which is characterized in that the mass ratio of the polylactic acid and Ciprofloxacin is 1:1。

4. preparation method according to claim 1, which is characterized in that described when vacuumizing, vacuum degree reaches 6 × 10^-3 ~8 × 10^-3Pa。