Method for compounding traditional Chinese medicinal antibacterial components with nano-particle self-assembly behavior

Abstract

The invention relates to the technical field of antibiosis, and discloses a traditional Chinese medicinal material antibacterial component compounding method with a nano-particle self-assembly behavior, which comprises the following steps: S1, a traditional Chinese medicinal material component extraction framework step for extracting an antibacterial component extract from lithospermum erythrorhizon and moutan bark medicinal materials; s2, a step of preparing a nano-particle carrier framework, wherein the nano-particle carrier framework is used for constructing a nano-carrier system composed of chitosan and sodium hyaluronate; s3, an antibacterial component compounding frame step: carrying out self-assembly compounding on the antibacterial component extract and the nano-particle carrier to generate antibacterial nano-composite particles; and S4, a composite particle drying frame step: treating the composite particles through centrifugation and freeze drying to obtain the antibacterial nano composite powder. The long-time sustained release of the antibacterial effect is realized by encapsulating the antibacterial component through the nano composite carrier, optimizing the particle size and adopting a sustained release technology, the stability and bioavailability of the medicine are improved, and side effects caused by excessive release of the medicine are avoided.

Description

Technical Field

[0001] The present invention relates to the field of antibacterial technology, and specifically to a method for compounding antibacterial components of traditional Chinese medicine by the self-assembly behavior of nanoparticles. Background Art

[0002] In modern life, antibacterial products are widely used in many fields such as medical treatment, personal care, and household hygiene. With the increasing bacterial drug resistance, many problems have gradually emerged in traditional antibacterial methods. In order to better combat bacterial infections and improve medical safety, it is particularly important to develop an antibacterial technology that can not only effectively inhibit bacteria but also maintain the duration of drug efficacy.

[0003] In the prior art, antibacterial components are usually applied by directly adding chemical synthetic substances or plant extracts. These components can inhibit bacterial growth to a certain extent and achieve a certain bactericidal effect. At the same time, there are also some antibacterial carriers prepared by using nanotechnology on the market. The advantage of these carriers is that they can improve the bioavailability of antibacterial components. By encapsulating antibacterial components, the nanocarriers can help the components disperse stably and improve their solubility. In addition, ultrasonic coupling agents are widely used in medical treatment, which can help sound waves effectively penetrate the skin or tissues and ensure the transmission quality of ultrasonic waves.

[0004] However, the antibacterial components in the prior art often face several problems. First, the release rate of traditional antibacterial components is relatively fast, usually releasing rapidly in a short time. The one-time release of a large amount of drugs may cause local toxic reactions or low use efficiency, and it is difficult to provide a long-lasting antibacterial effect. Second, unoptimized nanoparticles are prone to aggregation or precipitation, affecting the stability and uniform release of drugs. At the same time, most of the existing ultrasonic coupling agents do not consider their antibacterial properties, and bacteria are likely to grow on the surface of ultrasonic probes, posing a certain risk of cross-infection. In addition, although the existing slow-release technologies can delay drug release to a certain extent, they still fail to achieve ideal effects in terms of stability and persistence. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the present invention provides a method for compounding antibacterial components of traditional Chinese medicine by the self-assembly behavior of nanoparticles, which solves the problems of too fast release of antibacterial components, short duration of drug efficacy, and poor stability of nanoparticles in the prior art.

[0006] To achieve the above objectives, the present invention is realized through the following technical solutions: A method for compounding antibacterial components of traditional Chinese medicine by the self-assembly behavior of nanoparticles, comprising the following steps: S1. Framework steps for extracting components from traditional Chinese medicinal materials, used to extract antibacterial component extracts from Lithospermum erythrorhizon and Cortex Moutan. Lithospermum erythrorhizon is rich in shikonin compounds, while Cortex Moutan contains paeonol and paeonolide compounds, both of which have strong antibacterial effects. By mixing these two medicinal materials, the antibacterial effect during extraction can be optimized, and the active ingredients in the extract can be enhanced; S2. Framework steps for preparing nanoparticle carriers, used to construct a nanoparticle carrier system composed of chitosan and sodium hyaluronate. Chitosan is a natural polysaccharide with good biocompatibility and biodegradability, and can provide a stable carrier structure for nanoparticles; sodium hyaluronate is a commonly used biomaterial with good moisture retention, adhesiveness and biocompatibility. The combination of the two can enhance the stability and drug-loading capacity of nanoparticles; S3. Framework steps for compounding antibacterial components, self-assembling and compounding the antibacterial component extract with the nanoparticle carrier to generate antibacterial nanocomposite particles. By dropping the antibacterial component solution into the nanoparticle system, antibacterial molecules bind to the surface of the nanoparticles, forming a stable composite structure. The self-assembly process firmly wraps the antibacterial components in the nanoparticles through the evaporation of the solvent and intermolecular interactions (such as hydrogen bonds, electrostatic attractions, etc.); S4. Framework steps for drying composite particles, treating the composite particles by centrifugation and freeze-drying to obtain antibacterial nanocomposite powder. Centrifugation helps to accelerate the separation of the solvent and solid particles, separating the supernatant from the nanoparticles, and at the same time ensuring the effective precipitation of the nanoparticles, so as to obtain pure composite particles. Freeze-drying directly sublimes water into gas at low temperature, avoiding the presence of liquid water, thereby reducing the structural damage of the particles during drying and maintaining the morphology of the particles and the stability of the antibacterial components; S5. Framework steps for preparing a solid coupling agent, mixing the antibacterial nanocomposite powder with a gel matrix to prepare an antibacterial medical solid ultrasonic coupling agent. Carbomer, as a thickening agent, can effectively increase the viscosity of the gel, making it have appropriate rheological properties, facilitating the application of the ultrasonic coupling agent. Its combination with glycerol and sodium hyaluronate can endow the gel with good moisture retention and biocompatibility. This solid coupling agent effectively transmits sound waves to the skin or tissue through the conduction of ultrasonic waves, providing an efficient coupling effect. At the same time, the presence of antibacterial components can provide additional antibacterial protection, preventing cross-infection of bacteria during ultrasonic treatment.

[0007] Preferably, the framework steps for extracting components from traditional Chinese medicinal materials include: Mix Lithospermum erythrorhizon and Cortex Moutan in a mass ratio of 1:1.5 - 1:2.5. By mixing Lithospermum erythrorhizon and Cortex Moutan in a reasonable mass ratio, the antibacterial components in the extract can be optimized, and its comprehensive antibacterial effect can be enhanced. Different mass ratios can affect the extraction effects of the two compounds, thereby adjusting the characteristics of the final extract; Mix the mixed medicinal materials with an ethanol - water mixed solution at a mass - volume ratio of 1:8 - 12, and perform ultrasonic extraction. The extraction temperature is 50 - 70 °C, and the time is 40 - 80 minutes. Repeat the extraction twice. The ethanol - water mixed solution can effectively dissolve the polar and non - polar components in Lithospermum erythrorhizon and Cortex Moutan. The choice of ethanol as a solvent helps to enhance the solubility of plant secondary metabolites (such as shikonin and paeonol compounds). At the same time, the addition of water can increase the polarity of the solvent, promote the more extensive dissolution of components, and thus improve the extraction efficiency. The ultrasonic extraction method generates a large number of tiny bubbles through the cavitation effect. When the bubbles burst, a high - temperature and high - pressure micro - environment is produced, which can effectively break the plant cell wall and accelerate the penetration of the solvent into the plant tissue, thereby promoting the extraction of active ingredients. Ultrasonic vibration can also promote the dispersion of active ingredients in the medicinal materials, making the extraction process more efficient. During the extraction process, too high a temperature may lead to the degradation of some thermosensitive components, so the temperature needs to be controlled between 50 - 70 °C. And too long a time may cause unnecessary component dissolution or uneven solubility. Therefore, within the extraction time range of 40 - 80 minutes, efficient extraction can be achieved; Combine the extraction solutions, concentrate them under reduced pressure at 40 - 50 °C, and then freeze - dry them at - 40 to - 60 °C to obtain the powder of the antibacterial component extract. The purpose of reduced - pressure concentration is to remove the solvent at a lower temperature to avoid the loss of volatile components at high temperatures. By reducing the pressure, the boiling point of the solvent decreases, and most of the solvent can be evaporated at a lower temperature, leaving the concentrated antibacterial component, which helps to improve the purity and concentration of the extract. Freeze - drying can preserve the active components of the medicinal material extract, making it more stable and easy to store.

[0008] Preferably, the steps of the nanoparticle carrier preparation framework include: Dissolve chitosan in a 0.5% acetic acid aqueous solution to form a chitosan solution with a concentration of 0.5 - 2 mg / mL. During the preparation of the chitosan solution, use the acetic acid aqueous solution as a solvent. Acetic acid can provide a suitable pH environment to promote the dissolution of chitosan. The solution concentration in this range helps to control the particle size and surface properties of the particles, ensuring their good dispersibility; Dissolve sodium hyaluronate in deionized water to form a sodium hyaluronate solution with a concentration of 0.5 - 2 mg / mL. Sodium hyaluronate is a natural polysaccharide widely used in drug delivery systems, with good moisturizing properties and biocompatibility. The concentration of sodium hyaluronate will directly affect the size and adhesion of the nanoparticles. The concentration in this range can ensure the smooth progress of the particle formation process; The sodium hyaluronate solution is added dropwise to the chitosan solution at a volume ratio of 0.8 - 1.2:1, and stirred at 20 - 30 °C for 20 - 40 minutes at a stirring speed of 600 - 1000 rpm to generate nanoparticles. The sodium hyaluronate solution is added to the chitosan solution by dropwise addition, and the dropping rate and ratio play a key role in the size and morphology of the final nanoparticles. This volume ratio range can ensure that the ratio of sodium hyaluronate to chitosan is neither excessive nor too little, ensuring that they form a suitable structure in subsequent self-assembly. After the sodium hyaluronate and chitosan solutions are mixed, due to the electrostatic interaction, hydrogen bond and intermolecular interaction between the two, nanoparticles can be spontaneously formed. The negative charge of sodium hyaluronate and the positive charge of chitosan interact, resulting in molecular aggregation and forming a uniform and stable nanoparticle structure. The efficiency of this process is affected by factors such as stirring time, stirring speed and temperature. Therefore, the set stirring conditions (600 - 1000 rpm, 20 - 40 minutes, 20 - 30 °C) help to ensure the stability and uniformity of the particles.

[0009] Preferably, the step of the antibacterial component compounding framework includes: Dissolve the antibacterial component extract in absolute ethanol at a concentration of 1 - 3 mg / mL. Ethanol as a solvent can effectively dissolve most of the antibacterial components of traditional Chinese medicine, especially hydrophobic compounds such as shikonin and paeonol. The use of absolute ethanol avoids the reduction of component solubility caused by aqueous solutions, thereby increasing the concentration of antibacterial components; Slowly add the antibacterial component solution dropwise to the nanoparticle carrier suspension at a dropping rate of 0.5 - 2 mL / min, a stirring speed of 400 - 600 rpm, a reaction temperature of 20 - 30 °C, and a time of 50 - 70 minutes. By slowly adding dropwise, it is possible to avoid excessive concentration or precipitation of components, and at the same time evenly disperse the antibacterial components on the surface of the nanoparticles. The setting of the dropping rate of 0.5 - 2 mL / min ensures that the solution reacts with the carrier gradually and reduces the impact of sudden concentration changes on particle formation. The setting of the stirring speed of 400 - 600 rpm helps to maintain the uniformity of the solution and promote sufficient contact between the antibacterial components and the nanoparticles. Stirring can ensure the full mixing of the antibacterial components and the nanoparticle carrier and avoid precipitation or aggregation. The setting of this reaction temperature avoids the destructive effect of too high temperature on the antibacterial components and at the same time ensures the smooth progress of the nanoparticle formation process; Adjust the pH value of the mixed system to 5.0 - 6.5 to form stable antibacterial nano - composite particles. By adjusting the pH value to 5.0 - 6.5, the electrostatic attraction, hydrogen - bond interaction and other intermolecular interactions between the antibacterial components and the nanoparticles can be optimized. Too low a pH value may cause the dissolution or qualitative change of carriers such as chitosan, while too high a pH value may cause changes in the surface charge of the nanoparticles, thus affecting the stability of the composite particles. Therefore, this pH value range ensures the stability of the compounding system.

[0010] Preferably, the steps for drying the framework of the composite particles include: Centrifuge the antibacterial nano - composite particles at a rotational speed of 4500 - 5500 rpm for 5 - 10 minutes, and separate the supernatant. Using a centrifugation rate of 4500 - 5500 rpm can ensure the effective sedimentation of the nano - composite particles in a short time without causing particle deformation or aggregation due to excessive centrifugal force; the setting of the centrifugation time of 5 - 10 minutes can ensure the full separation of the supernatant, removing the excess solvent and unbound antibacterial components, thereby improving the purity of the composite particles; through this process, the antibacterial nano - composite particles are enriched and separated from the solution, providing a pure sample for subsequent freeze - drying. Freeze - dry the precipitate at a freezing temperature of - 45 to - 55 °C for 20 - 28 hours. The freeze - drying process with a freezing temperature of - 45 to - 55 °C first freezes the water in the antibacterial nano - composite particles and then directly removes it by sublimation. This process avoids the problem of thermal degradation caused by traditional thermal drying; the low - temperature drying method can protect the integrity of the nano - structure, avoiding particle aggregation or morphological changes caused by the liquid surface tension; the drying time is set at 20 - 28 hours to ensure the complete removal of the solvent while preventing internal structural changes of the nanoparticles caused by long - term drying, improving the stability and dispersibility of the final powder. Obtain antibacterial nano - composite powder with uniform particle size and good surface state. The antibacterial nano - composite powder after centrifugation and freeze - drying has a good particle size distribution, which can prevent the occurrence of aggregation, enabling it to be evenly dispersed during subsequent use, and improving the solubility and bioavailability.

[0011] Preferably, the steps for preparing the solid - state coupling agent include: Prepare a carbomer gel matrix, where the weight ratio of carbomer: glycerol: sodium hyaluronate is 0.3 - 0.7:8 - 12:0.1 - 0.3. Carbomer is a polymer thickener with excellent water - swelling and adhesion properties, capable of forming a stable gel structure. Glycerol, as a humectant, can enhance the lubricity of the gel, while sodium hyaluronate can provide moisturizing and repair functions and also helps to improve the uniform dispersion of the antibacterial nano - composite powder. This ratio range can ensure that the coupling agent has good rheological properties and stability. Adjust the pH value of the gel matrix to 5.5 - 6.5. Add the antibacterial nano - composite powder to the matrix at a mass fraction of 5 - 15%. The stirring speed is 500 - 700 rpm, and the stirring time is 25 - 35 minutes. Adjusting the pH value of the gel matrix to 5.5 - 6.5 is mainly to optimize the stability of the antibacterial nano - composite powder. Too low pH may cause particle aggregation or dissolution, while too high pH may affect the physical properties of the gel. Therefore, this pH value range can maintain the stability of the gel structure and enhance the activity of the antibacterial components at the same time; After standing for 20 - 25 hours to remove air bubbles, it is filled into an antibacterial medical solid ultrasonic coupling agent. Add the antibacterial nano - composite powder at a mass fraction of 5 - 15% and stir at a speed of 500 - 700 rpm for 25 - 35 minutes, which can ensure the uniform dispersion of antibacterial particles in the gel matrix. Too low stirring speed may lead to uneven dispersion, while too high stirring rate may cause air bubble generation or particle aggregation. After standing for 20 - 25 hours, the tiny air bubbles generated during the mixing process can be effectively removed, thereby improving the uniformity and transparency of the final product and ensuring the conductivity and imaging quality of the coupling agent during ultrasonic examination.

[0012] Preferably, the antibacterial component extract includes: Shikonin - type compounds, accounting for 25 - 45 of the total mass of the extract. Its antibacterial mechanism mainly realizes by inhibiting bacterial DNA synthesis and destroying cell membranes, showing obvious inhibitory effects on various pathogenic bacteria such as Staphylococcus aureus, Escherichia coli, and Candida albicans; in the nano - composite particle system, shikonin can enhance the antibacterial ability of the nano - carrier and extend its biological activity through a sustained - release effect, improving the continuous inhibitory effect on pathogenic microorganisms. Since shikonin belongs to a lipophilic compound, using ethanol extraction can increase its solubility, ensure its effective dispersion in the nano - carrier, and improve the stability of the final preparation; Paeonol - type compounds, accounting for 15 - 35 of the total mass of the extract. Paeonol is the main antibacterial component in Cortex Moutan, belonging to phenolic compounds, having broad - spectrum antibacterial, anti - inflammatory, and analgesic effects. The antibacterial mechanism of paeonol mainly is by interfering with the stability of bacterial cell walls and membranes and affecting bacterial protein synthesis, thereby inhibiting bacterial growth; paeonol has significant inhibitory effects on pathogenic bacteria such as drug - resistant Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus, and at the same time can inhibit the formation of bacterial biofilms, thereby enhancing its antibacterial effect; Paeonolide - type compounds, accounting for 5 - 25 of the total mass of the extract. Paeonolide is another important active ingredient in Cortex Moutan, belonging to glycoside compounds, having antibacterial, antioxidant, and anti - inflammatory properties. Paeonolide mainly exerts its antibacterial effect by interfering with bacterial metabolic processes and oxidative stress responses, being able to inhibit bacterial lipid peroxidation reactions, thereby reducing cell membrane damage and improving antibacterial persistence.

[0013] Preferably, the nano-composite particles include: The average particle size is 50 - 150 nm, and the PDI is less than 0.1 - 0.3. The size of the nanoparticles is crucial for their biological behavior and drug release characteristics. Controlling the particle size within the range of 50 - 150 nm helps the particles penetrate biological barriers such as the skin and cell membranes, improving the delivery efficiency of the antibacterial components. A PDI less than 0.3 indicates that the prepared nanoparticles have a narrow particle size distribution, avoiding problems such as particle aggregation or sedimentation, improving the stability and consistency of the system, and helping to optimize the drug delivery effect; The surface Zeta potential is +25 to +35 mV. The Zeta potential reflects the charge characteristics of the nanoparticle surface. The positive charge of +25 to +35 mV helps to enhance the interaction between the nanoparticles and the bacterial surface because most bacterial surfaces are negatively charged. The electrostatic attraction between positive and negative charges makes it easier for the nanoparticles to adsorb on the surface of pathogenic bacteria, improving the antibacterial efficiency; in addition, positively charged particles can enhance their binding ability to cell membranes, improving drug permeability and cell uptake rate; The drug loading rate of the antibacterial component is not less than 75%, and the encapsulation rate is not less than 85%. The drug loading rate refers to the ratio of the actual drug loading in the nanoparticles to the total mass. The higher the value, the stronger the drug loading capacity of the nanoparticles; by optimizing the preparation methods of the nanoparticles (such as electrostatic self-assembly and solvent evaporation method), the loading amount of the antibacterial component in the nano-carrier can be increased, thereby reducing the required dosage, improving the drug efficacy, and at the same time reducing side effects. The encapsulation rate refers to the ratio of the drug successfully encapsulated in the nanoparticles. A high encapsulation rate means that the drug is more stable and not easily degraded or lost; an encapsulation rate of more than 85% indicates that the drug is effectively encapsulated inside the nanoparticles, reducing the degradation effect of environmental factors (such as light, oxidation, hydrolysis) on the drug, and enhancing the stability and sustained release effect of the drug.

[0014] Preferably, the antibacterial medical solid ultrasonic coupling agent includes: There is no precipitation or stratification after storage at 15 - 30 °C for 3 - 6 months. Within the range of 15 - 30 °C, the gel matrix does not undergo obvious rheological changes, ensuring that it does not undergo phase separation or degradation due to temperature fluctuations. At the same time, this temperature range avoids the degradation of antibacterial nanoparticles, increasing the shelf life of the preparation; The ultrasonic transmittance is not less than 95%. The main function of the ultrasonic coupling agent is to improve the conduction efficiency of ultrasonic waves in the skin or tissues and reduce energy loss. By optimizing the viscoelasticity and composition uniformity of the gel matrix, it has excellent acoustic impedance matching performance and improves the ultrasonic transmittance; low bubble content: during the preparation process, bubbles are removed by standing for 20 - 25 hours, which can avoid the formation of acoustic impedance discontinuous regions inside the gel, reduce the scattering loss of ultrasonic waves, and further improve the conduction efficiency of ultrasonic signals; hydration enhances ultrasonic conductivity: both sodium hyaluronate and glycerol have good water retention properties, which can maintain the moisture content of the coupling agent, endow it with appropriate acoustic impedance characteristics, and ensure the efficient transmission of ultrasonic signals; The antibacterial activity duration is more than 72 hours. Due to the size of the nanoparticles (50 - 150 nm) and the positively charged surface (+25 to +35 mV), they can gradually release antibacterial components such as shikonin, paeonol, and moutan phenol, making the antibacterial effect last for no less than 72 hours.

[0015] Preferably, the antibacterial properties of the antibacterial medical solid ultrasonic coupling agent against bacteria include: The diameter of the inhibition zone against Staphylococcus aureus is 20 - 25 mm; The diameter of the inhibition zone against Escherichia coli is 18 - 22 mm; The diameter of the inhibition zone against Candida albicans is 16 - 20 mm.

[0016] The present invention provides a method for compounding antibacterial components of traditional Chinese medicine with self - assembly behavior of nanoparticles. It has the following beneficial effects: 1. The present invention adopts the technical scheme of encapsulating antibacterial components with a nano - composite carrier, achieving the technical effects of prolonging the antibacterial effect and improving the stability of antibacterial components. Compared with the technical scheme of directly using traditional antibacterial components in the prior art, the present invention effectively solves the problems that antibacterial components are prone to failure and release too fast in a conventional environment, thus enhancing the antibacterial persistence and effect.

[0017] 2. The present invention optimizes the ratio of sodium hyaluronate and carbomer, adjusts the particle size and stability of nanoparticles, achieving the technical effects of improving the particle size uniformity and reducing particle aggregation. Compared with the technical scheme of using a conventional carrier in the prior art, the present invention reduces the agglomeration phenomenon of nanoparticles, improves the bioavailability and stability of the drug, and ensures the stability and persistence during the drug release process.

[0018] 3. The present invention adopts a dialysis sustained-release technology and a precise controlled-release method, achieving the technical effects of reducing the peak release of drugs in a short time and improving the sustained-release effect of components. Compared with the traditional non-controlled release technology, the present invention can more precisely control the release rate of antibacterial components, avoid the cytotoxicity caused by excessive drug release, extend its biological activity, and thus improve the drug safety of patients.

[0019] 4. The present invention adopts a technical scheme of applying an antibacterial nano-composite system to an ultrasonic coupling agent, achieving the technical effects of improving the ultrasonic transmittance and endowing it with antibacterial functions. Compared with the technical scheme of conventional ultrasonic coupling agents in the prior art, the present invention not only maintains good transmittance of ultrasonic signals but also enhances the antibacterial effect of ultrasonic equipment, thus effectively reducing the risk of cross-infection and improving the safety and reliability of medical devices. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a flowchart of the method of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the specification of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0022] Please refer to the attached Figure 1 : Example 1: Construction of a highly stable shikonin-paeonol-paeonolide nano-composite system Extraction of antibacterial components: Weigh 60 g of lithospermum and 120 g of cortex moutan, crush them to 80 mesh, and mix them in a mass ratio of 1:2. Add 10 times the volume of 60% ethanol-aqueous solution, and perform ultrasonic extraction at 65 °C for 70 minutes, repeating the extraction 2 times. Combine the extracts, concentrate them under reduced pressure at 45 °C to 1 / 4 volume, and then freeze-dry at -55 °C to obtain an antibacterial extract powder (shikonin derivatives account for 40%, paeonol derivatives 28%, and paeonolide 12%).

[0023] Preparation of nano-carriers: Weigh 40 mg of chitosan and dissolve it in 8 mL of 0.5% acetic acid aqueous solution, stirring to dissolve. Weigh another 40 mg of sodium hyaluronate and dissolve it in 8 mL of deionized water. Slowly add the sodium hyaluronate solution to the chitosan solution at a volume ratio of 1.1:1, with a stirring rate of 700 rpm and a stirring time of 35 minutes to form a uniform nano-particle suspension.

[0024] Compound antibacterial components: Dissolve the antibacterial extract in absolute ethanol (2.5 mg / mL) and drop it into the nanocarrier suspension at a rate of 1 mL / min. The stirring rate is 550 rpm, the reaction temperature is 28 °C, stir for 65 minutes, and adjust the pH to 6.0. Stable antibacterial nanocomposite particles are formed (average particle size 90 nm, Zeta potential +32 mV, drug loading rate 80%, encapsulation efficiency 89%).

[0025] Drying treatment: Centrifuge at 5200 rpm for 9 minutes and discard the supernatant. Place the precipitate in a freeze dryer at -50 °C for 26 hours to obtain the antibacterial nanocomposite powder.

[0026] Example 2: Antibacterial nanocomposite system with regulated nanoparticle size Extract antibacterial components: Take 55 g of Lithospermum erythrorhizon and 110 g of Cortex Moutan, pulverize them to 100 mesh, and mix them in a mass ratio of 1:2. Add 12 times the volume of 50% ethanol-aqueous solution, perform ultrasonic extraction at 60 °C for 60 minutes, and repeat the extraction 2 times. Combine the extracts, concentrate them under reduced pressure to 1 / 3 volume at 40 °C, and freeze-dry at -50 °C to obtain the antibacterial extract powder (30% shikonin derivatives, 32% paeonol derivatives, 10% paeonol).

[0027] Preparation of nanocarriers: Weigh 60 mg of chitosan and dissolve it in 12 mL of 0.5% acetic acid aqueous solution, and stir to dissolve. Weigh another 60 mg of sodium hyaluronate and dissolve it in 12 mL of deionized water. Slowly drop the sodium hyaluronate solution into the chitosan solution according to a volume ratio of 0.9:1, with a stirring rate of 800 rpm, and stir for 30 minutes to form a nanoparticle suspension.

[0028] Compound antibacterial components: Dissolve the antibacterial extract in absolute ethanol (1.5 mg / mL) and drop it into the nanocarrier suspension at a rate of 0.8 mL / min. The stirring rate is 500 rpm, the reaction temperature is 25 °C, stir for 55 minutes, and adjust the pH to 5.5. Nanocomposite particles are formed (average particle size 75 nm, Zeta potential +30 mV, drug loading rate 76%, encapsulation efficiency 87%).

[0029] Drying treatment: Centrifuge at 5000 rpm for 7 minutes and discard the supernatant. Place the precipitate in a freeze dryer at -45 °C for 24 hours to obtain the nanocomposite powder.

[0030] Example 3: Preparation and stability optimization of ultrasonic coupling agent Preparation of coupling agent matrix: Weigh 0.5 g of carbomer, 10 g of glycerol, and 0.2 g of sodium hyaluronate, add 89.3 g of deionized water, stir to dissolve at 800 rpm, and adjust the pH to 6.0 to form a uniform and transparent gel matrix.

[0031] Composite powder addition: Weigh 10 g of antibacterial nano-composite powder, slowly add it to the gel matrix, stir at a rate of 600 rpm for 30 minutes to disperse it evenly.

[0032] Defoaming and forming: Let the mixture stand for 24 hours. After completely removing the bubbles, dispense it into sealed packages to form an antibacterial medical solid ultrasonic coupling agent.

[0033] Comparative example 1: Comparison of antibacterial activity of traditional ethanol extraction method (corresponding to Example 1) Antibacterial component extraction: Weigh 60 g of Lithospermum erythrorhizon and 120 g of Moutan cortex, crush them to 80 mesh. Mix them in a mass ratio of 1:2, add 10 times the volume of 50% ethanol-aqueous solution, extract at room temperature for 24 hours without ultrasonic extraction. After filtration, directly evaporate to dryness in a water bath at 50 °C to obtain a brown solid antibacterial extract (30% shikonin, 22% paeonol, 8% paeonolide).

[0034] Nanocarrier preparation: The preparation method is the same as that in Example 1, only adjust the volume ratio of sodium hyaluronate to drop to 0.7:1, adjust the stirring rate to 500 rpm, and the stirring time to 20 minutes.

[0035] Compound antibacterial components: Dissolve the obtained extract directly in absolute ethanol (1 mg / mL), and pour it into the nanocarrier solution at one time without dropwise control. Stir at a rate of 300 rpm, reaction temperature 25 °C, reaction time 30 minutes, and adjust the pH to 5.2.

[0036] Drying treatment: Centrifuge at 4000 rpm for 5 minutes, discard the supernatant. Place the precipitate in a freeze dryer at -40 °C for 18 hours to obtain antibacterial nano-composite powder.

[0037] Comparative example 2: Nano antibacterial composite system with unoptimized particle size (corresponding to Example 2) Antibacterial component extraction: The extraction method is the same as that in Example 2.

[0038] Nanocarrier preparation: Weigh 100 mg of chitosan and dissolve it in 20 mL of 0.5% acetic acid aqueous solution. Separately, weigh 100 mg of sodium hyaluronate and dissolve it in 20 mL of deionized water. According to a volume ratio of 1:1, pour the sodium hyaluronate solution into the chitosan solution at once without dropwise control. Stir at a rate of 400 rpm for 15 minutes to form a nanoparticle suspension.

[0039] Compound antibacterial components: Dissolve the antibacterial extract in absolute ethanol (3 mg / mL) and directly add it to the nanoparticle suspension. Stir at a rate of 200 rpm, react at a temperature of 22 °C for 40 minutes, and adjust the pH to 7.0.

[0040] Drying treatment: Centrifuge at 3000 rpm for 6 minutes and discard the supernatant. Place the precipitate in a freeze dryer at -30 °C for 16 hours to obtain the antibacterial nano-composite powder.

[0041] Comparative Example 3: Ultrasonic coupling agent without antibacterial nano-composite particles (corresponding to Example 3) Preparation of the coupling agent matrix: Weigh 0.5 g of carbomer, 10 g of glycerol, and 0.2 g of sodium hyaluronate, add 89.3 g of deionized water, stir and dissolve at 800 rpm, and adjust the pH to 6.0 to form a uniform and transparent gel matrix.

[0042] Do not add the antibacterial nano-composite powder: During the preparation process, do not add antibacterial nano-composite particles, and only use the carbomer, glycerol, and sodium hyaluronate gel matrix.

[0043] Defoaming and shaping: Let the mixture stand for 24 hours. After completely removing the bubbles, sub-pack it into sealed packages to form a common ultrasonic coupling agent.

[0044] Comparative experiment: Experimental materials: Chemical reagents: Lithospermum erythrorhizon, Cortex Moutan, ethanol, acetic acid, deionized water, phosphate buffer solution (PBS), carbomer, glycerol, sodium hyaluronate, etc.

[0045] Experimental strains: Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Candida albicans (C. albicans), provided by the laboratory strain bank.

[0046] Instruments and equipment: ultrasonic cleaner (40 kHz), vacuum concentrator, freeze dryer (-55 °C), high performance liquid chromatography (HPLC), dynamic light scattering instrument (DLS), Zeta potential analyzer, ultraviolet-visible spectrophotometer (UV-Vis), dialysis bag (10 kDa), ultrasonic transmission tester (2 MHz), ultrasonic probe simulation equipment, etc.

[0047] Experimental procedures: Antibacterial activity test: Dissolve the antibacterial nano-composite powders of the examples and comparative examples in PBS (2 mg / mL) respectively, filter and sterilize to prepare antibacterial solutions.

[0048] Take the bacterial solution (1×10 6 CFU / mL) and evenly coat it on the LB agar plate. Take sterile filter paper discs with a diameter of 6 mm, soak them in the antibacterial solutions respectively, and place them in the center of the plate.

[0049] Incubate at 37 °C for 24 hours, measure the diameter of the inhibition zone (mm), and record the data.

[0050] Further conduct a time-kill curve test, take samples every 12 hours, coat and count, and observe the bacterial survival within 72 hours.

[0051] Nano-particle size stability test: Use DLS to measure the particle size and PDI of the nano-composite systems of different examples and comparative examples, and record the initial data.

[0052] Place the samples at 4 °C, 25 °C, and 37 °C, take samples every 7 days, measure the particle size change, and continuously observe for 60 days.

[0053] Antibacterial component sustained-release effect test: Take an equal mass (10 mg) of antibacterial composite powder, dissolve it in PBS (pH 7.4), and place it in a 10 kDa dialysis bag.

[0054] Suspend the dialysis bag in 100 mL of PBS and incubate it on a shaker at 37 °C (100 rpm).

[0055] Take 1 mL of sample every 6 hours, use UV-Vis to measure the release amount of antibacterial components, draw a cumulative release curve, and the experiment lasts for 72 hours.

[0056] Ultrasonic permeability test: Adopt a 2 MHz ultrasonic transmission experimental system to prepare an ultrasonic coupling agent film with a uniform thickness (2 mm).

[0057] Measure the ultrasonic signal transmittance of different coupling agents and record the initial data.

[0058] Store the coupling agent in an environment of 15 - 30°C, take samples monthly to measure the transmittance, and observe the change trend for 6 months.

[0059] Antibacterial persistence test of the coupling agent: Take the ultrasonic coupling agents of the examples and comparative examples, evenly coat them on the surface of the ultrasonic probe, and use it once a day.

[0060] Take samples with a sterile cotton swab every 24 hours, inoculate them on LB medium, culture at 37°C for 24 hours, and calculate the number of colonies (CFU / mL).

[0061] Record the change trend of bacterial survival within 7 days.

[0062] Comparison of experimental data between examples and comparative examples Experimental summary: The optimized design of the nanocarrier enables the antibacterial component to be released more uniformly over a long period, avoiding the phenomenon of high - concentration release of traditional extracts in a short time. Within 72 hours, the release curve of the nanocomposite system shows a sustained - release trend, reducing the initial release peak by 20 - 30% compared with the comparative example. It can be seen that nano - encapsulation significantly enhances the stability of the antibacterial agent, avoids the problem of increased cytotoxicity caused by excessive drug dosage, and improves the persistence of antibacterial activity.

[0063] The improvement in stability brought about by particle size regulation is extremely significant. The nano - composite particles in the examples did not show obvious aggregation within 6 months, and the PDI remained between 0.1 - 0.14. In the comparative example, the unoptimized carrier led to a particle size increase of more than 30%, and the PDI increased to more than 0.35. This indicates that the particle size uniformity is crucial for the suspension stability of the nano - system. Optimizing the preparation parameters of the nanocarrier can effectively reduce particle aggregation and improve bioavailability.

[0064] The antibacterial property and transmittance performance of the ultrasonic coupling agent also illustrate the necessity of material optimization. The coupling agent in the examples not only maintains an ultrasonic transmittance of more than 96%, but also completely inhibits colony growth within 7 days of the experiment. While the colony count of the ordinary coupling agent reached more than 4000 CFU / mL after 4 days. This shows that the nano - antibacterial composite system can effectively reduce microbial contamination, improve the safety of medical devices, and provide more reliable guarantee for clinical applications.

[0065] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principle and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for compounding antibacterial components of traditional Chinese medicine with nanoparticle self-assembly behavior, characterized in that: The following steps are involved: S1, a framework step for extracting components from Chinese medicinal materials, for extracting antibacterial components from Lithospermum officinale and Paeonia suffruticosa root; S2, a nanoparticle carrier preparation framework step, used to construct a nanoparticle carrier system composed of chitosan and sodium hyaluronate; S3, an antibacterial component compounding framework step, wherein the antibacterial component extract and the nanoparticle carrier are self-assembled and compounded to generate antibacterial nanocomposite particles; S4, a composite particle drying framework step, treating the composite particles by centrifugation and freeze drying to obtain an antibacterial nanocomposite powder; S5, a solid coupling agent preparation framework step, mixing the antibacterial nanocomposite powder with the gel matrix to prepare an antibacterial medical solid ultrasonic coupling agent.

2. The method for compounding antibacterial components of Chinese medicinal materials with nanoparticle self-assembly behavior according to claim 1, characterized in that: The steps of extracting Chinese medicinal materials ingredients include: Mix Lithospermum officinale and Paeonia suffruticosa root in a mass ratio of 1:1.5-1:2.5; The mixed medicinal materials were mixed with an ethanol-water mixed solution at a mass volume ratio of 1:8-12, and ultrasonic extraction was performed at an extraction temperature of 50-70°C for 40-80 minutes, and the extraction was repeated twice; The extracts were combined, concentrated under reduced pressure at 40-50°C, and subsequently freeze-dried at −40 to −60°C to obtain an antimicrobial component extract powder.

3. The method for compounding antibacterial components of Chinese medicinal materials with nanoparticle self-assembly behavior according to claim 1, characterized in that: The nanoparticle carrier preparation framework steps include: Dissolve chitosan in 0.5% acetic acid aqueous solution to form a 0.5-2 mg / mL chitosan solution; Dissolve sodium hyaluronate in deionized water to form a 0.5-2 mg / mL sodium hyaluronate solution; The sodium hyaluronate solution is added dropwise to the chitosan solution at a volume ratio of 0.8-1.2:1, and stirred at 20-30° C. for 20-40 minutes at a stirring speed of 600-1000 rpm to generate nanoparticles.

4. The method for compounding the antibacterial component of traditional Chinese medicine with nanoparticle self-assembly behavior according to claim 1, characterized in that: The antimicrobial component compounding framework steps include: Dissolving the antibacterial component extract in anhydrous ethanol at a concentration of 1-3 mg / mL; Slowly dripping the antibacterial component solution into the nanocarrier suspension, the dripping rate is 0.5-2 mL / min, the stirring speed is 400-600 rpm, the reaction temperature is 20-30° C., and the time is 50-70 minutes; The pH value of the mixed system is adjusted to 5.0-6.5 to form stable antibacterial nanocomposite particles.

5. The method for compounding antibacterial components of Chinese medicinal materials with nanoparticle self-assembly behavior according to claim 1, characterized in that: The composite particle drying framework step comprises: The antibacterial nanocomposite particles are centrifuged at a speed of 4500-5500 rpm for 5-10 minutes to separate the supernatant; The precipitate was freeze-dried at a temperature of −45 to −55°C and a drying time of 20 to 28 hours; The antibacterial nanocomposite powder with uniform particle size and good surface condition is obtained.

6. The method for compounding the antibacterial component of traditional Chinese medicine with nanoparticle self-assembly behavior according to claim 1, characterized in that: The steps of preparing the solid coupling agent framework include: A carbomer gel matrix is ​​prepared, wherein the weight ratio of carbomer: glycerol: sodium hyaluronate is 0.3-0.7: 8-12: 0.1-0.3; The pH value of the gel matrix is ​​adjusted to 5.5-6.5, and the antibacterial nanocomposite powder is added into the matrix at a mass fraction of 5-15, with a stirring speed of 500-700 rpm and a stirring time of 25-35 minutes; After standing for 20-25 hours to eliminate bubbles, it is filled into an antibacterial medical solid ultrasonic coupling agent.

7. The method for compounding antibacterial components of Chinese medicinal materials with nanoparticle self-assembly behavior according to claim 1, characterized in that: The antibacterial component extract comprises: Shikonin compounds, accounting for 25-45% of the total mass of the extract; Paeonol compounds, accounting for 15-35% of the total mass of the extract; Paeonia suffruticosa phenolic compounds account for 5-25% of the total mass of the extract.

8. The method for compounding the antibacterial component of traditional Chinese medicine with nanoparticle self-assembly behavior according to claim 4, characterized in that: The nanocomposite particles include: The average particle size is 50-150nm, and the PDI is less than 0.1-0.3; The surface zeta potential is +25 to +35 mV; The drug loading rate of the antibacterial component is not less than 75%, and the encapsulation rate is not less than 85%.

9. The method for compounding the antibacterial component of traditional Chinese medicine with nanoparticle self-assembly behavior according to claim 1, characterized in that: The antibacterial medical solid ultrasonic coupling agent comprises: No precipitation or stratification after storage at 15-30℃ for 3-6 months; Ultrasonic transmittance is not less than 95%; The antibacterial activity lasts for more than 72 hours.

10. The method for compounding the antibacterial component of traditional Chinese medicine with nanoparticle self-assembly behavior according to claim 9, characterized in that: The antibacterial properties of the antibacterial medical solid ultrasonic coupling agent against bacteria include: The diameter of the inhibition zone for Staphylococcus aureus is 20-25 mm; The diameter of the inhibition zone for E. coli is 18-22 mm; The diameter of the inhibition zone against Candida albicans is 16-20 mm.

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