Drug-loaded acupuncture needle, preparation method and application thereof

Abstract

The application provides a drug-loaded acupuncture needle and a preparation method and application thereof, and belongs to the technical field of combination of traditional Chinese medicine and western medicine. The preparation method of the drug-loaded acupuncture needle comprises the following steps: after the surface of the acupuncture needle is modified by silanization, the acupuncture needle is repeatedly soaked in sodium polystyrene sulfonate solution and polyallyl ammonium chloride solution in sequence, a high-molecular drug-loaded film is formed on the surface of the acupuncture needle, and the acupuncture needle is loaded with target drugs in a diclofenac sodium solution, so that the drug-loaded acupuncture needle is obtained. The acupuncture needle is combined with drugs, so that the same-acupoint drug administration can be realized. On one hand, the acupoint is mechanically stimulated, and the internal regulation of the body is exerted; on the other hand, the drug efficacy is exerted. The acupoint stimulation and drug treatment play the same effect and are complementary to each other. Animal experiment results show that the acupuncture needle and the diclofenac sodium in the drug-loaded acupuncture needle have a synergistic effect, and the acupuncture-needle-drug combination treatment mode has obvious effects on the treatment of rheumatoid arthritis.

Description

Technical Field

[0001] This invention belongs to the field of integrated Chinese and Western technology, and particularly relates to a drug-loaded acupuncture needle, its preparation method, and its application. Background Technology

[0002] Rheumatoid arthritis (RA) is a common autoimmune disease characterized by chronic inflammation of the synovium and joint destruction. It typically affects small joints such as the hands and wrists symmetrically, with joint swelling and pain as the main symptoms, leading to severe limb deformities. In addition to joint damage, it can also affect organs or tissues such as the heart, lungs, kidneys, and nervous system. This disease is often recurrent, chronic, and has a high disability rate, severely impacting patients' functional activities and quality of life, and increasing the economic burden on patients, their families, and society. Common RA treatments in modern medicine include nonsteroidal anti-inflammatory drugs (NSAIDs), corticosteroids, chronic antirheumatic drugs (CARDs), early biological agents, and traditional Chinese medicine. Western medicine is effective in slowing disease progression and controlling most symptoms, but it cannot cure RA and requires long-term maintenance therapy, with certain toxic side effects and limitations.

[0003] In Traditional Chinese Medicine (TCM), rheumatoid arthritis (RA) falls under the category of "Bi syndrome." External therapies in TCM have historically played a crucial and irreplaceable role in RA treatment, with records dating back to the pre-Qin period. Acupuncture, a method of treating internal diseases externally in TCM, possesses unique advantages in regulating immunity and relieving pain. However, relying solely on acupuncture has several drawbacks, such as a narrow therapeutic range and slower or weaker therapeutic effects for certain diseases. Currently, there are no reports of loading Western medications for treating RA onto the surface of acupuncture needles for the treatment of RA. Summary of the Invention

[0004] In view of this, the purpose of this invention is to provide a drug-loaded acupuncture needle, its preparation method and application. The combination of the needle and the drug in this invention has significant therapeutic effects in the treatment of rheumatoid arthritis.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a method for preparing a drug-loaded acupuncture needle, comprising: modifying the surface of the acupuncture needle with silanization, repeatedly immersing it in sodium polystyrene sulfonate solution and polyallyl ammonium chloride solution in sequence, and then loading it in diclofenac sodium solution to obtain a drug-loaded acupuncture needle.

[0007] Preferably, the silanization modification includes: immersing the acupuncture needle sequentially in a sodium hydroxide solution and an ethanol aqueous solution containing ATPES, and then curing it at 105-115°C to obtain a silanized acupuncture needle.

[0008] Preferably, the pH value of the sodium hydroxide solution is 9-11, and the soaking time of the sodium hydroxide solution is 20 seconds.

[0009] Preferably, the volume ratio of ethanol to water in the ethanol-water solution is 75-85:20-25; the concentration of ATPES in the ethanol-water solution is 250 mM / L, and the ATPES has been hydrolyzed for 12-36 h; the soaking time of the ethanol-water solution is 6-18 h.

[0010] Preferably, the curing time is 1 to 2 hours.

[0011] Preferably, the concentration of the sodium polystyrene sulfonate solution is 1-9 mg / mL, and the concentration of the polyallyl ammonium chloride solution is 1-9 mg / mL.

[0012] Preferably, the number of soaking times is 50 to 80.

[0013] Preferably, the concentration of the diclofenac sodium solution is 1-20 mg / mL, the loading is performed 1-16 times, and the loading time for each loading is 10-50 min.

[0014] The present invention also provides a drug-loaded acupuncture needle prepared by the above method.

[0015] The present invention also provides the application of the above-mentioned drug-loaded acupuncture needle in the preparation of medical devices for treating rheumatoid arthritis.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] This invention provides a method for preparing a drug-loaded acupuncture needle, comprising: modifying the surface of the acupuncture needle with silanization, repeatedly immersing it in sodium polystyrene sulfonate solution and polyallyl ammonium chloride solution, and then loading it in diclofenac sodium solution to obtain a drug-loaded acupuncture needle. This invention combines needles and drugs, achieving both mechanical stimulation of acupoints to promote internal regulation and the therapeutic effect of drugs. The acupoint stimulation and drug treatment complement each other, creating a synergistic and mutually reinforcing effect.

[0018] Animal experiments showed that the combined acupuncture and drug treatment regimen significantly reduced ankle swelling in rats compared to acupuncture or drug administration alone, indicating a synergistic effect between acupuncture needles and diclofenac sodium. This combined treatment approach is effective in treating rheumatoid arthritis. Furthermore, the drug-loaded acupuncture needles described in this application demonstrate good biocompatibility and hold promise for providing a new treatment strategy for rheumatoid arthritis. Attached Figure Description

[0019] Figure 1This is a process in which polymers with different charges are self-assembled layer by layer using electrostatic adsorption as the driving force.

[0020] Figure 2 The preparation process of acupuncture needles loaded with sodium dichlorophenate;

[0021] Figure 3 To optimize the fabrication process of DS-loaded acupuncture needles and to present the results of in vitro release experiments, the following were investigated: (a) the effect of polyelectrolyte solution concentration on the amount of DS loaded in the acupuncture needle; (b) the effect of the number of self-assembled layers on the amount of DS loaded in the acupuncture needle; (c) the effect of DS concentration on the amount of DS loaded in the acupuncture needle; (d) the effect of loading time on the amount of DS loaded in the acupuncture needle; (e) the effect of the number of DS loading cycles on the amount of DS loaded in the acupuncture needle; and (f) the results of in vitro release experiments.

[0022] Figure 4 The microstructures of acupuncture needles under different modification conditions are shown in the following figures: (a) Microstructure of the surface of a blank acupuncture needle; (b) Microstructure of the surface of the ATPES-modified acupuncture needle in Example 1; (c) Microstructure of the acupuncture needle loaded with 70 layers of PSS / PAH polymer film in Example 17; (d) Microstructure of the DS-loaded acupuncture needle in Example 17.

[0023] Figure 5 The elemental composition of the acupuncture needle surface under different modification conditions is as follows: (a) elemental composition of the blank acupuncture needle surface; (b) elemental composition of the ATPES-modified acupuncture needle surface in Example 1; (c) elemental composition of the acupuncture needle surface loaded with 70 layers of PSS / PAH polymer film in Example 17; (d) elemental composition of the acupuncture needle surface loaded with DS in Example 17.

[0024] Figure 6 Ankle joint images of rats in each group after treatment;

[0025] Figure 7 Safranin-Fix Green staining was performed on the ankle joints of rats in each group (×100x).

[0026] Figure 8 HE staining of ankle cartilage in rats of each group (×100x);

[0027] Figure 9 HE staining of the ankle joint synovium of rats in each group (×100x);

[0028] Figure 10 Immunohistochemical analysis of TNF-α, an inflammatory factor in ankle synovial tissue (×100-fold);

[0029] Figure 11 Immunohistochemical analysis of IL-6, an inflammatory factor in ankle synovial tissue (×100-fold);

[0030] Figure 12Immunohistochemical analysis of IL-1, an inflammatory factor in ankle synovial tissue (×100-fold);

[0031] Figure 13 HE staining images of the hearts of experimental animals in each group (×100x);

[0032] Figure 14 HE staining images of the livers of each group of experimental animals (×100x);

[0033] Figure 15 HE staining images of spleens from each group of experimental animals (×100 magnification);

[0034] Figure 16 HE staining of lung tissue from each group of experimental animals (×100 magnification);

[0035] Figure 17 HE staining images of the kidneys of each group of experimental animals (×100 magnification);

[0036] Figure 18 HE staining images of the back skin of each group of experimental animals. Detailed Implementation

[0037] This invention provides a method for preparing a drug-loaded acupuncture needle, comprising: modifying the surface of the acupuncture needle with silanization, repeatedly immersing it in sodium polystyrene sulfonate solution and polyallyl ammonium chloride solution in sequence, and then loading it in diclofenac sodium solution to obtain a drug-loaded acupuncture needle.

[0038] The silanization modification of the present invention includes: immersing the acupuncture needle sequentially in sodium hydroxide solution and an ethanol aqueous solution containing ATPES, and then curing it at 105-115°C to obtain a silanized acupuncture needle.

[0039] Prior to silanization modification, it is preferable to also include ultrasonic cleaning in acetone for 5 minutes and drying with nitrogen.

[0040] The sodium hydroxide solution described in this invention has a pH value of 9-11, and the soaking time in the sodium hydroxide solution is 20 seconds. The sodium hydroxide solution described in this invention provides an alkaline environment for the surface of acupuncture needles, allowing OH- to adhere. - .

[0041] The volume ratio of ethanol to water in the ethanol-water solution of this invention is preferably 75-85:20-25, more preferably 75:25; the concentration of ATPES in the ethanol-water solution is preferably 250 mM / L; the ATPES is preferably hydrolyzed for 12-36 hours, more preferably 24 hours; the soaking time of the ethanol-water solution is preferably 6-18 hours, more preferably 12 hours. The purpose of dissolving ATPES in the ethanol-water mixture in this invention is to ensure sufficient hydrolysis of ATPES.

[0042] The curing time described in this invention is preferably 1 to 2 hours, more preferably 1 hour. The curing process described in this invention makes the silane film more compact.

[0043] The silanization modification described in this invention results in a denser silanized film on the acupuncture needle, making the film structure more corrosion-resistant. This silanization modification also protects the surface of the acupuncture needle and allows the needle to carry amino groups, which is beneficial for subsequent modification.

[0044] The concentration of the sodium polystyrene sulfonate solution of the present invention is preferably 1-9 mg / mL, more preferably 5 mg / mL; the concentration of the polyallyl ammonium chloride solution is preferably 1-9 mg / mL, more preferably 5 mg / mL; and the number of soaking times is preferably 50-80 times, more preferably 70 times.

[0045] Polyallylammonium chloride (PAH) and sodium polystyrene sulfonate (PSS) are common polyelectrolytes; PAH carries a positive charge, and PSS carries a negative charge. These two polyelectrolytes can be used as drug carriers for drug delivery. Therefore, this invention uses these two polyelectrolytes as sources of positive and negative charges, respectively, for layer-by-layer self-assembly. Layer-by-layer self-assembly (LbL) is a method of alternating layer deposition that utilizes weak interactions between layers, such as electrostatic attraction, hydrogen bonding, and coordination bonds, to spontaneously form structurally complete and stable molecular aggregates or supramolecular structures.

[0046] The concentration of the diclofenac sodium solution of the present invention is preferably 1-20 mg / mL, more preferably 20 mg / mL, the loading frequency is preferably 1-16 times, more preferably 11 times, and the loading time for each loading is preferably 10-50 min, more preferably 40 min.

[0047] Diclofenac Sodium (DS) is a nonsteroidal anti-inflammatory drug (NSAID) with anti-inflammatory, analgesic, and antipyretic effects. Clinically, it is used for rheumatoid arthritis, adhesive spondylitis, non-inflammatory joint pain, arthritis, non-articular rheumatism, pain caused by non-articular inflammation, various types of neuralgia, cancer pain, post-traumatic pain, and fever caused by various inflammations.

[0048] The present invention also provides a drug-loaded acupuncture needle prepared by the above method.

[0049] This invention combines acupuncture and medicine. On the one hand, it uses mechanical stimulation of acupoints to enable the body to exert its internal regulatory function, and on the other hand, it uses medicine to exert its efficacy. Acupoint stimulation and drug treatment have the effect of complementing each other with similar or different effects.

[0050] The present invention also provides the application of the above-mentioned drug-loaded acupuncture needle in the preparation of medical devices for treating rheumatoid arthritis.

[0051] The present invention provides a drug-loaded acupuncture needle for treating rheumatoid arthritis using conventional acupuncture methods.

[0052] Animal experiments showed that the combined acupuncture and drug treatment regimen significantly reduced ankle swelling in rats compared to acupuncture or drug administration alone, indicating a synergistic effect between acupuncture needles and diclofenac sodium. This combined treatment approach is effective in treating rheumatoid arthritis. Furthermore, the drug-loaded acupuncture needles described in this application demonstrate good biocompatibility and hold promise for providing a new treatment strategy for rheumatoid arthritis.

[0053] The present invention does not have any special limitations on the source of the reagents used; conventional commercially available products in the field are acceptable.

[0054] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0055] Example 1

[0056] Acupuncture needle surface treatment

[0057] The acupuncture needles (stainless steel filiform needles) were ultrasonically cleaned in acetone for 5 minutes, dried with nitrogen, and then immersed in a sodium hydroxide solution with a pH of 10 for 20 seconds for alkaline washing. They were then rinsed with deionized water and dried with nitrogen. Subsequently, the needles were immersed in a mixed solution of alcohol and water at a ratio of 75:25 containing ATPES (hydrolyzed for 24 hours) at a concentration of 250 mM / L for 12 hours. The acupuncture needles were then cleaned with deionized water and anhydrous ethanol and dried with nitrogen. Finally, they were heated and cured in an oven at 110°C for 1 hour to obtain acupuncture needles with silanization modification on the surface.

[0058] Example 2-16

[0059] The surface treatment process of acupuncture needles was optimized using orthogonal experimental design, with copper sulfate titration time as the evaluation parameter. The experimental design scheme is shown in Table 1.

[0060] According to the orthogonal experimental design in Table 1, different parameters were set to obtain the technical solutions for the surface treatment of acupuncture needles in Examples 2-16, and the copper sulfate titration time was measured.

[0061] The copper sulfate titration time reflects the corrosion resistance of the silanized film. The longer it takes for the acupuncture needle to turn red, the denser and more corrosion-resistant the silanized film constructed using the modification method. The method for determining the copper sulfate titration time involves dropping a copper sulfate solution onto the acupuncture needle and recording the total time from the drop of solution onto the needle until it turns red.

[0062] Table 1. Orthogonal Design Factor Level Table

[0063]

[0064] Table 2. Surface treatment parameters and copper sulfate titration time results for acupuncture needles in Examples 1-16

[0065]

[0066]

[0067] As shown in Table 2, the silanized film of the acupuncture needles prepared using the method of Example 1 is denser and more corrosion-resistant. Therefore, the silanized acupuncture needles obtained in Example 1 were used for subsequent operations.

[0068] Example 17

[0069] Diclofenac Sodium-Loaded Acupuncture Needles Prepared Using Layer-by-Layer Self-Assembly

[0070] The silanized acupuncture needles prepared in Example 1 were repeatedly immersed in 5 mg / mL PSS solution and 5 mg / mL PAH solution. After each immersion in a polyelectrolyte, they were rinsed with deionized water and dried with nitrogen at room temperature. The immersion was repeated 70 times, with each immersion lasting 5 seconds, to assemble the PAH / PSS polyelectrolyte layer, thus obtaining the acupuncture needles with the assembled PAH / PSS polyelectrolyte layer.

[0071] The acupuncture needles with assembled PAH / PSS polyelectrolyte layers were repeatedly immersed in a DS solution with a concentration of 20 mg / mL. After each immersion, the needles were dried with nitrogen gas. This process was repeated 11 times, with each immersion lasting 40 minutes, to obtain acupuncture needles loaded with diclofenac sodium.

[0072] Example 18

[0073] Single-factor experiment

[0074] High-performance DS-loaded acupuncture needles were prepared through preliminary experiments. The optimal concentrations of PAH and PSS solutions were initially determined to be 5 mg / mL, the number of PAH / PSS assembly layers was 70, the concentration of DS solution was 20 mg / mL, the loading time of the acupuncture needles in the DS solution was 30 min, and the number of immersions of the acupuncture needles in the DS solution was 10.

[0075] Based on preliminary experiments, five factors—concentration of polyelectrolyte solution, number of self-assembled layers, concentration of DS, loading time, and number of DS loading cycles—were further optimized using a single-factor method. The polyelectrolyte solution concentration was set to four gradients: 1, 3, 5, 7, and 9 mg / mL (with the same concentrations of PSS and PAH solutions at each gradient); the number of self-assembled layers was set to four gradients: 50, 60, 70, and 80; the DS concentration was set to four gradients: 1, 5, 10, and 20 mg / mL; the loading time of the acupuncture needles in the DS solution was set to five gradients: 10, 20, 30, 40, and 50 min; and the number of DS loading cycles was set to five gradients: 1, 3, 5, 11, and 16. The optimal process parameters for each factor were explored sequentially using the single-factor experimental method. DS-loaded acupuncture needles were prepared according to the above experimental design. The change in DS concentration in the solution before and after drug loading was evaluated using high-performance liquid chromatography (HPLC), and the amount of DS loaded in a single acupuncture needle was calculated. Specific results are as follows: Figure 3 As shown in ae.

[0076] Depend on Figure 3 According to ae, during the preparation of acupuncture needles loaded with diclofenac sodium based on the layer-by-layer self-assembly method, the amount of DS drug loaded in the acupuncture needle is maximized when the concentration of PAH solution is 5 mg / mL, the concentration of PSS solution is 5 mg / mL, the number of PAH / PSS assembly layers is 70, the concentration of DS solution is 20 mg / mL, the soaking time of the acupuncture needle in the DS solution is 40 min, and the number of soaking times of the acupuncture needle in the DS solution is 11.

[0077] Example 19

[0078] Results of in vitro release experiments

[0079] Ten drug-loaded acupuncture needles prepared in Example 17 were immersed in 15 mL of PBS solution at 37°C. 1 mL of solution was taken at regular intervals and analyzed by high-performance liquid chromatography (HPLC). Simultaneously, 1 mL of fresh PBS solution was added to the original centrifuge tube. The release of DS was measured under two conditions: pH = 6.5 and pH = 7.4. Specific results are as follows: Figure 3 As shown in f.

[0080] As shown in 3f, the drug loaded on the acupuncture needle in Example 17 had been released by more than 90% in 40 minutes, and the release rate was relatively faster under weakly acidic conditions.

[0081] Example 20

[0082] The morphology of drug-loaded acupuncture needles at different stages of preparation was characterized using SEM. Elemental analysis was performed on the surface of acupuncture needles under different modification conditions. Specific results are as follows: Figure 4 and Figure 5 As shown.

[0083] Depend on Figure 4 It can be seen that a distinct film-like structure appeared on the surface of the acupuncture needle, indicating that the technical solution of the present invention can successfully modify the surface of the acupuncture needle.

[0084] Depend on Figure 5 The changes in the elements on the surface of the acupuncture needle at each step are clearly visible, proving the successful coating of the polymer film and the successful loading of DS, and also proving that the present invention can successfully obtain acupuncture needles loaded with DS.

[0085] Example 21

[0086] Clinical trial on the treatment and safety of acupuncture with sodium diclofenac for rheumatoid arthritis.

[0087] 1. Laboratory animals

[0088] Sixty male SPF-grade SD rats, aged 8 weeks and weighing 220±20g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0089] Husbandry conditions: Room temperature was controlled by air conditioning at 23±2℃, relative humidity 55%~68%, high-pressure bedding was used and changed twice a week. Standard maintenance feed was provided, and purified water was available for free. After a 7-day acclimatization period, the animals were used in experiments. The experiment was approved by the Ethics Committee of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, and met the requirements for animal experiments.

[0090] 2. Modeling and Grouping

[0091] The experimental animals were divided into six groups: Blank group, Model group, Joint injection group (T1), Diclofenac sodium gavage group (T2), Ordinary acupuncture group (T3), and Acupuncture-drug combination group (T4). The acupuncture needles used in the acupuncture-drug combination group were the acupuncture needles loaded with diclofenac sodium prepared in Example 17.

[0092] Ten rats were in each group. The control group rats were injected intradermally with sterile saline in the right hind paw pad, while the other rats were given Freund's complete adjuvant to prepare a rheumatoid arthritis model. The rats were injected intradermally with 0.15 mL of the saline after disinfecting the right hind paw pad with iodine.

[0093] 3. Treatment and Management

[0094] (1) Treatment began on the 3rd day after modeling. The ordinary acupuncture group and the combined acupuncture and drug group received acupuncture treatment at Zusanli and Kunlun acupoints. The blank group and the model group were fixed in the same way without additional intervention. The diclofenac sodium group was administered to rats by gavage at a dose of 4.16 mg / kg (calculated based on the adult dose of 50 mg / day). In the joint injection group, each rat was given a dose of 0.15 mg / time (since the needle insertion at Kunlun acupoint is 3 mm and at Zusanli acupoint is 7 mm, the drug load of each acupuncture needle is about 0.6 mg, and the needle length is 40 mm, the drug dosage during acupuncture administration is about 0.15 mg). Treatment was given every other day, and the treatment ended after 20 days. The circumference of the rat's foot and ankle was measured the day after each treatment. The swelling of the rat's toes was assessed by the change in this circumference. Swelling degree = (post-inflammatory foot circumference - pre-inflammatory foot circumference) / pre-inflammatory foot circumference × 100%. The degree of paw swelling in rats in each group was measured and recorded on days 2, 10, 16, and 20, as shown in Table 3.

[0095] The condition of the foot and ankle in each group after treatment is as follows: Figure 6 As shown.

[0096] The 50% withdrawal threshold (ED50) of different groups of rats was measured on days 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20, and changes in the rat ankle joint were recorded. The ED50 value was determined according to the method for determining the 2.650% mechanical withdrawal threshold in "The Effects of Tibetan Medicine *Tinospora sinensis* on the Expression of Inflammatory Cytokines in Serum and NMDA Receptors in Spinal Cord of Rats with Inflammatory Pain". The specific results are shown in Table 4.

[0097] The arthritis index of rats in different groups was assessed on days 10, 12, 14, 16, 18, and 20. The arthritis index was calculated from the onset of chronic arthritis. The scoring criteria were as follows: 0 points for no redness or swelling of joints; 1 point for slight redness and swelling of the little toe joint; 2 points for swelling of the toe joints and paws; 3 points for swelling of the toes below the ankle joint; and 4 points for swelling of all paws, including the ankle joint. The specific results are shown in Table 5.

[0098] Table 3. Swelling degree of rat paws in each group (%); n=10)

[0099]

[0100] Table 4. ED50 (g) of rats in each group; n=10)

[0101]

[0102]

[0103] Table 5 Arthritis index of rats in each group ( n=10)

[0104]

[0105] Note: In Tables 3-5, compared with the normal group (Normol), * P<0.05, ** P < 0.01 Model group comparison # P<0.05, ## P<0.01; Intra-articular injection of DS group (T1), oral administration of DS group (T2), acupuncture needle group (T3), and DS-loaded acupuncture needle group (T4).

[0106] The data in Table 3 reflect the swelling of the toes in the experimental animals. As the treatment time increased, the swelling of the experimental animals in each group was relieved, with the combined treatment group showing the most significant relief.

[0107] The data in Table 4 reflect the pain status of the experimental animals. As the treatment time increased, the pain level of the experimental animals in each group was relieved, with the combined treatment group showing the most significant pain relief.

[0108] Table 5 shows that the Arthritis Index score is also an important indicator for assessing the state of arthritis. Similar to pain relief, the arthritis score decreased significantly in the combined treatment group.

[0109] Depend on Figure 6 It can be seen that, compared with the conventional administration of diclofenac sodium, the combined treatment group can effectively relieve paw swelling in rats, and the "acupuncture-drug combination" treatment regimen also shows a significant difference in relieving paw swelling in rats compared with acupuncture treatment alone, indicating that the "acupuncture-drug combination" treatment method has a significant anti-swelling effect.

[0110] 4. Pathological index assessment

[0111] After 20 days of treatment, rats were anesthetized with 3% sodium pentobarbital at a dose of 1.5 ml / kg. Successful anesthesia induction was indicated by weakness in the limbs, deep and slow breathing, and loss of the righting reflex. All subsequent tissue collection was performed on ice. The rats were placed in a supine position with their limbs immobilized. The right knee, disinfected with iodine, was longitudinally incised. After peeling off the epidermis, a transverse incision was made at the lower edge of the patellar ligament. With the knee flexed, surrounding soft tissues were dissected. The knee synovium was removed, immediately wrapped in aluminum foil, and immersed in liquid nitrogen. Blood was collected from the abdominal aorta and placed in centrifuge tubes. The tubes were incubated at 4°C for 2 hours, centrifuged at 3000 rpm for 10 minutes, and the supernatant serum was collected and stored at -80°C for experimental analysis.

[0112] (1) Safranin-Fix Green staining, HE staining of ankle cartilage, and HE staining of ankle synovium were performed on the articular cartilage of each group of rats after treatment. The specific results are as follows: Figures 7-9 As shown.

[0113] Safranin-Fix Green staining refers to staining articular cartilage with basic safranin dye, while acidic Fast Green dye can stain subchondral bone.

[0114] Depend on Figure 7 It was found that the cartilage tissue in the model group was significantly damaged, while the treatment group was able to effectively inhibit the inflammatory response of the synovial membrane and protect the integrity of the articular cartilage. Among them, the "acupuncture and drug combination" group showed significant therapeutic and protective effects.

[0115] Depend on Figure 8 and Figure 9 It can be seen that the synovial tissue of the rats in the model group has severe hyperplasia and a large number of inflammatory cell infiltrations. However, the "acupuncture and drug combination" treatment can effectively inhibit inflammatory cell infiltration and thus alleviate synovial hyperplasia, thereby achieving the purpose of inhibiting synovial inflammation.

[0116] (2) Immunohistochemical analysis was performed on the inflammatory factors (IL-6, IL-1β, and TNF-α) in the synovial tissue of rats in each group. The specific results are as follows: Figures 10-12 As shown.

[0117] Results of immunohistochemistry Figures 10-12 It was found that the expression levels of inflammatory factors such as IL-6, IL-1β, and TNF-α in the model group were significantly increased in the synovial tissue of the knee and ankle joints, showing obvious brownish-yellow positive areas. After treatment, the expression levels of inflammatory factors in the synovial tissue decreased, especially in the "acupuncture and drug combination" group, which could significantly inhibit the expression of inflammatory factors such as IL-6, IL-1β, and TNF-α in the synovial tissue, playing a role in treating and alleviating the disease.

[0118] 5. Safety assessment

[0119] Safety performance is an important indicator for the evaluation of drugs and medical devices.

[0120] (1) H&E staining was performed on the major organs of rats in each group after treatment. The specific results are as follows: Figures 13-17 As shown.

[0121] Depend on Figure 13 and Figure 17 It can be seen that the "combined acupuncture and drug" treatment did not cause organic lesions in the heart, liver, spleen, lungs and kidneys of the experimental animals.

[0122] (2) The effects of combined acupuncture and drug therapy on skin stimulation were investigated.

[0123] Following the experimental design, every other day, the hairless skin on the back of rats was stimulated with syringes, ordinary acupuncture needles, and drug-loaded acupuncture needles for 20 days, with 10 stimulations. After the last stimulation, the back skin was harvested and stained with hematoxylin and eosin (HE). The results are as follows: Figure 18 As shown.

[0124] Depend on Figure 18 It is known that the combined treatment group did not cause skin inflammation or other possible adverse reactions.

[0125] In summary, the acupuncture needles containing diclofenac sodium in this application have good biocompatibility and are expected to provide a new treatment strategy for rheumatoid arthritis in the future.

[0126] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a drug-loaded acupuncture needle, characterized in that, include: After silanizing the surface of the acupuncture needle, it is repeatedly immersed in sodium polystyrene sulfonate solution and polyallyl ammonium chloride solution, and then loaded in diclofenac sodium solution to obtain a drug-loaded acupuncture needle; the silanization modification includes: immersing the acupuncture needle in sodium hydroxide solution and ethanol aqueous solution containing ATPES in sequence, and then curing it at 105-115°C to obtain a silanized acupuncture needle.

2. The preparation method according to claim 1, characterized in that, The pH value of the sodium hydroxide solution is 9-11, and the soaking time of the sodium hydroxide solution is 20 seconds.

3. The preparation method according to claim 1, characterized in that, The volume ratio of ethanol to water in the ethanol-water solution is 75-85:20-25; the concentration of ATPES in the ethanol-water solution is 250 mM / L, and the ATPES has been hydrolyzed for 12-36 h; the soaking time of the ethanol-water solution is 6-18 h.

4. The preparation method according to claim 1, characterized in that, The curing time is 1 to 2 hours.

5. The preparation method according to claim 1, characterized in that, The concentration of the sodium polystyrene sulfonate solution is 1–9 mg / mL, and the concentration of the polyallyl ammonium chloride solution is 1–9 mg / mL.

6. The preparation method according to claim 1, characterized in that, The soaking is performed 50 to 80 times.

7. The preparation method according to claim 1, characterized in that, The concentration of the diclofenac sodium solution is 1–20 mg / mL, the loading is performed 1–16 times, and the loading time for each loading is 10–50 min.

8. The drug-loaded acupuncture needle obtained by the preparation method according to any one of claims 1 to 7.

9. The use of the drug-loaded acupuncture needle obtained by the preparation method according to any one of claims 1 to 7 or the drug-loaded acupuncture needle according to claim 8 in the preparation of a medical device for treating rheumatoid arthritis.

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