Pentamidine fibrin glue patch for preventing tendon adhesion and preparation method thereof

Abstract

The invention belongs to the field of medical apparatus and instruments, and provides a pentamidine fibrin glue patch for preventing tendon adhesion and a preparation method thereof, the preparation method comprises the following steps: uniformly mixing pentamidine with a Fibrinophysis Inhibitor solution to obtain a Fibrinophysis Inhibitor-containing suspension, then uniformly mixing the Fibrinophysis Inhibitor-containing suspension with Sealer Protein Content powder to obtain a Fibrinophysis Inhibitor-carrying solution, adding the Fibrinophysis Inhibitor-carrying solution into the Fibrinophysis Inhibitor-carrying solution, uniformly mixing the Fibrinophysis Inhibitor-carrying solution, and uniformly mixing the Fibrinophysis Inhibitor-carrying solution with the Sealer Protein Content powder to obtain the pentamidine fibrin glue patch for preventing tendon adhesion. The preparation method comprises the following steps: uniformly mixing a calcium chloride solution with thrombin powder to obtain a Thrombin solution; mixing the Fibrinasis Inhibitor carrying solution with a Thrombin solution, and carrying out a reaction on the Fibrinasis Inhibitor carrying solution and the Thrombin solution, so as to obtain the Fibrinasis Inhibitor carrying solution and the Thrombin carrying solution; and paving the pentamidine fibrin glue into a container to form a patch, and sterilizing to obtain the pentamidine fibrin glue. The invention provides a novel pentamidine controlled release system by taking fibrin glue as a carrier. The system can effectively maintain the concentration of the medicine in a tendon repair area, meanwhile, systematic side effects are reduced, and continuity and stability of medicine delivery are guaranteed. In addition, the patch has good biocompatibility and biodegradability, and inflammation or anaphylaxis possibly caused by a traditional physical barrier is avoided.

Description

Technical Field

[0001] The invention belongs to the field of medical devices and relates to a pentamidine fibrin glue patch for preventing tendon adhesion and a preparation method thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Tendon repair often leads to serious functional impairment, among which tendon adhesion is a common clinical problem. Although a variety of drugs and surgical methods have been developed to prevent tendon adhesion, current treatment options still have limitations, especially in terms of the effectiveness and safety of drug delivery systems. Traditional methods for preventing tendon adhesion mainly include the following aspects: ① Physical barrier. Various substances such as membranes, gels or sponges are used as barriers to physically isolate the repair area and prevent excessive growth of surrounding tissues leading to adhesion. Common materials include polytetrafluoroethylene (PTFE) and hyaluronic acid. Anti-inflammatory or anti-fibrotic drugs are applied directly to the repaired tendon to reduce inflammatory response and fibrosis, thereby preventing the formation of adhesion. ② Drug treatment: Anti-inflammatory and anti-fibrotic drugs such as corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs), transforming growth factor β (TGF-β) inhibitors are used to reduce inflammatory response and fibrosis. ③ Surgical techniques: Improve surgical techniques, such as minimally invasive surgery, optimized suturing techniques, etc., to reduce tissue damage and inflammatory response, thereby reducing the risk of adhesion. ④Postoperative rehabilitation: Early activity and physical therapy can promote proper movement of the tendon and help prevent the formation of adhesions.

[0004] However, traditional methods for treating or preventing tendon adhesions have certain limitations. Some physical barrier materials may trigger local inflammatory or allergic reactions, and physical barriers may shift or curl after surgery, which will reduce their effectiveness. Simple physical isolation does not involve any drug treatment and may not be sufficient to prevent adhesions caused by biochemical factors. In addition, systemic use of anti-inflammatory drugs (such as corticosteroids or NSAIDs) may cause side effects such as gastrointestinal discomfort and liver and kidney damage. Long-term use of certain drugs may lead to increased drug tolerance or reduced therapeutic effect, and the effective concentration of drugs in the local area is difficult to maintain, which may require frequent reapplication or the use of special release systems. In terms of surgery, the success of surgery depends largely on the skills and experience of the surgeon. Any surgery may bring risks such as infection and bleeding, especially when multiple surgical repairs are required. Optimized surgical techniques may require longer recovery time or strict rehabilitation protocols after surgery. The compliance problem of postoperative rehabilitation requires patients to be highly cooperative, and patients who lack active participation may not achieve the ideal recovery effect. High-quality physical therapy requires specialized rehabilitation equipment and well-trained physical therapists, which may be difficult to obtain in resource-limited areas, and if postoperative rehabilitation measures are not performed properly or are delayed, the risk of adhesion formation may be increased.

[0005] In recent years, with the development of nanotechnology and materials science, researchers have tried to develop new drug delivery systems to improve the therapeutic effect after tendon repair. These systems are designed to provide controlled drug release, reduce local toxicity, while maintaining sufficient drug concentration to effectively prevent adhesion. The applicant's previous studies have shown that pentamidine is a drug with anti-fibrotic potential. As an antiprotozoan and antimicrobial drug, it effectively treats pneumocystis pneumonia and trypanosomiasis, etc.; recent studies have also found its potential to fight cancer (especially melanoma). In addition, Pentamidine also shows anti-inflammatory properties. In clinical applications, Pentamidine can be used in combination with a variety of biomaterials to support controlled release delivery of drugs and enhance therapeutic effects. These advantages demonstrate the versatility and potential of Pentamidine in existing medical treatments and future applications. The inventor's previous paper "Pentamidine-loaded gelatin decreases adhesion formation of flexor tendon" discloses a pentamidine-gelatin sheet that reduces the formation of flexor tendon adhesion, but its adhesion and plasticity still need to be improved. However, there is no research on using pentamidine fibrin glue patch as a drug delivery system for preventing adhesion after tendon repair. Summary of the invention

[0006] In order to solve the above problems, the present invention provides a pentamidine fibrin glue patch for preventing tendon adhesion and a preparation method thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] The first aspect of the present invention provides a pentamidine fibrin glue patch for preventing tendon adhesion, comprising:

[0009] The pentamidine and fibrinolysis inhibitor solution are mixed evenly to obtain a pentamidine suspension, and then mixed evenly with SealerProtein Concentrate powder to obtain a pentamidine-loaded solution;

[0010] The calcium salt solution and thrombin powder are mixed evenly to obtain Thrombin solution;

[0011] The pentamidine-loaded solution is mixed with the Thrombin solution to react and obtain pentamidine fibrin glue;

[0012] The pentamidine fibrin glue is spread in a container to form a patch, and then sterilized to obtain the product.

[0013] The present invention combines pentamidine with fibrin glue, a biocompatible material, to design a novel pentamidine-fibrin glue patch (Pentamidine-Fibrin Glue Patch, PFP). The patch utilizes the natural biodegradable properties of fibrin glue to form a drug sustained-release matrix, which can provide sustained drug release in the tendon repair area. In addition, the biocompatibility and biodegradability of the fibrin glue patch enable it to effectively reduce adhesion factors such as fibroblast migration without interfering with normal tendon healing.

[0014] The pentamidine fibrin glue patch is an innovative drug release system designed to prevent adhesion after tendon repair. This patch uses fibrin glue as a drug carrier, which not only provides excellent biocompatibility and biodegradability, but also ensures that it can be gradually absorbed and degraded by the body after completing its function, reducing potential inflammatory reactions. The patch has biosafety and plasticity, which is convenient for intuitive observation during surgery and adapts to the morphology and position of different tendons. Its precise drug control function avoids local or systemic side effects that may be caused by overdose. It is simple and easy to use and can be quickly attached to the surface of the tendon. By effectively controlling the formation of adhesions after tendon repair, the pentamidine fibrin glue patch not only helps to improve postoperative functional recovery, reduce the recurrence rate and the need for secondary surgery, but also reduces patients' postoperative pain by preventing adhesion formation, speeds up recovery, improves the quality of life, and thus reduces medical costs. In addition, the development of the present invention shows the application of modern bioengineering and drug release technology, improves the scientificity, safety and technical content of treatment, and brings an innovative solution to the field of tendon surgery.

[0015] In some embodiments, the concentration of pentamidine in the pentamidine-containing suspension is 25-50 μg / mL.

[0016] In some embodiments, the mass ratio of pentamidine to Sealer Protein Concentrate powder is 0.025-0.05:1 mg.

[0017] In some embodiments, the volume mass ratio of the calcium salt solution to the thrombin powder is 1 mL: 1-1.2 mg.

[0018] Preferably, the calcium salt is calcium chloride.

[0019] In some embodiments, the mass ratio of Pentamidine to Thrombin powder is 0.025-0.05:1 mg.

[0020] In some embodiments, the reaction temperature is 20-24°C and the pH is 7-7.5.

[0021] In some embodiments, the container is a culture dish with a size of 60 mm-100 mm.

[0022] In some embodiments, the sterilization is performed by ultraviolet sterilization.

[0023] In some embodiments, before use, the pentamidine fibrin glue patch is cut into a size suitable for covering the tendon area and sterilized again for use.

[0024] The second aspect of the present invention provides a pentamidine fibrin glue patch for preventing tendon adhesion prepared by the above method.

[0025] The third aspect of the present invention provides the use of the above-mentioned pentamidine fibrin glue patch in the preparation of a medical device for preventing tendon adhesion.

[0026] Beneficial effects of the present invention

[0027] (1) Improve the effectiveness and safety of drug delivery systems: Pentamidine is one of the drugs approved by the FDA and has been effectively used for a long time in the diagnosis and treatment of various clinical diseases. The pentamidine fibrin glue patch of the present invention provides a new type of pentamidine controlled release system by using fibrin glue as a carrier. This system can effectively maintain the concentration of the drug in the tendon repair area, while reducing systemic side effects, ensuring the continuity and stability of drug delivery. In addition, the patch has good biocompatibility and biodegradability, avoiding inflammation or allergic reactions that may be caused by traditional physical barriers.

[0028] (2) Promoting postoperative tendon function recovery and reducing recurrence: The use of the pentamidine fibrin glue patch of the present invention can effectively prevent the formation of adhesions after tendon surgery, thereby helping patients to better restore tendon function and reduce functional impairment and possible reoperation caused by adhesions. This is of great significance for improving the success rate of surgery and the quality of life of patients after surgery.

[0029] (3) Simplify surgical procedures and improve postoperative recovery efficiency: The patch of the present invention is simple and easy to use. It can be directly attached to the tendon surface during surgery and adapt to the morphology and position of different tendons, which simplifies the surgical procedure and reduces the operation time. At the same time, by effectively controlling the release of drugs, the need for frequent drug reapplication is avoided, which speeds up the patient's recovery process and reduces medical costs.

[0030] (4) Compared with the pentamidine-gelatin sheet, the pentamidine fibrin glue patch of the present invention has the advantages of good adhesion and high plasticity. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention, and the exemplary embodiments of the present invention and their description are used to explain the present invention and do not constitute improper limitations on the present invention.

[0032] Figure 1 The structural formula of pentamidine and the preparation and characterization of pentamidine fibrin patch;

[0033] Figure 2 To characterize the cell biology of the pentamidine fibrin patch;

[0034] Figure 3 To evaluate the in vivo performance and anti-adhesion properties of the pentamidine fibrin patch;

[0035] Figure 4 To observe the histological effect of pentamidine fibrin patch on tendon adhesion.

[0036] Figure 5 Mechanical testing of the pentamidine fibrin patch for the repair of tendon injuries. DETAILED DESCRIPTION

[0037] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0038] The present invention will be further described in detail below in conjunction with specific embodiments. It should be noted that the specific embodiments are intended to explain the present invention rather than to limit it.

[0039] Example 1 Design and Synthesis of Pentamidine Fibrin Patch

[0040] 1. Structural design and material selection: Pentamidine (Fresenius Kabi Company, structural formula see Figure 1 ) is one of the drugs approved by the FDA. It was selected because it has long-term follow-up and treatment and has biosafety. Fibrin glue was selected as a drug carrier. This material is widely used in tissue engineering due to its excellent biocompatibility and cell adhesion properties. In addition, fibrin glue can also be used as a carrier for controlled-release drugs to maintain the effective concentration of drugs in soft tissues, while reducing toxicity and increasing the sustained release of drugs.

[0041] 2. Processing of Fibrinolysis Inhibitor solution and Thrombin solution loaded with pentamidine: Use an electronic balance to weigh 25 μg of pentamidine for use. Use a pipette to draw 1 mL of Fibrinolysis Inhibitor solution (Baxter) into an EP tube and mix it thoroughly with 25 μg of pentamidine. Carefully stir and mix the mixed Fibrinolysis Inhibitor suspension containing pentamidine with 1 mg of Sealer Protein Concentrate powder for use to obtain a Fibrinolysis Inhibitor solution loaded with pentamidine. Use a pipette to draw 1 mL of Calcium Chloride solution (commercially available product) and 1 mg of Thrombin powder for use to obtain a Thrombin solution.

[0042] The pentamidine-loaded Fibrinolysis Inhibitor solution and the Thrombin solution were mixed at room temperature (24° C.) at a pH value of 7 to obtain pentamidine fibrin glue.

[0043] 3. Synthesis and processing of pentamidine fibrin patch:

[0044] The synthesized pentamidine fibrin glue was spread on a 60 mm culture dish to form a uniform patch, which was sterilized with ultraviolet light in a biosafety cabinet and cooled to room temperature to form a uniform patch. These patches need to be cut into a size suitable for covering the tendon area before the experiment and sterilized twice for use.

[0045] Example 2 In vitro biocompatibility test of pentamidine fibrin patch (PFP)

[0046] 1. Characterization of the Pentamidine Fibrin Patch: The surface structure of the patch was observed using a scanning electron microscope (SEM), the distribution and aggregation of pentamidine were confirmed by chemical analysis, and the properties, texture, transparency, thickness and surface drug distribution of the pentamidine fibrin patch were detected.

[0047] 2. Co-culture of Pentamidine Fibrin Patch with Tendon Cells: Co-culture the Pentamidine Fibrin Patch with in vitro cultured turkey tendon cells to evaluate its effects on cell proliferation and cell migration. Live / dead cell staining was used to evaluate cell survival and explore the biocompatibility of the Pentamidine Fibrin Patch.

[0048] 3. Anti-adhesion test of pentamidine fibrin patch: Using bovine serum albumin (BSA) as a negative control, the patch's ability to support cell biological functions was evaluated through a turkey tendon cell migration experiment. Cells treated with different doses of pentamidine were mixed with a well plate containing fibronectin, collagen I, collagen IV, laminin I, and fibrinogen to detect the anti-tendon cell adhesion ability of the pentamidine fibrin patch.

[0049] Example 3 In vivo biosafety and efficacy testing of pentamidine fibrin patch (PFP)

[0050] 1. Preparation of tendon regeneration and anti-adhesion model: The third (middle) finger of the right or left foot of each turkey (Heritage-Breed, 6 months old, weighing approximately 8–10 kg) was randomly selected for surgical repair. The other third finger that was not operated on was used as a reference. A z-shaped incision was made at the proximal interphalangeal joint of the palmar side of the third finger. The deep flexor tendon of the digitorum was exposed and torn 1 cm distal to the proximal vein. The tendon was repaired using a modified Pennington technique suture (4-0Fiberwire) and a circular running suture (6-0Prolene). Pentamidine fibrin patch was applied to the turkey tendon injury model, covering and surrounding the damaged tendon with the patch to simulate the clinical treatment process. After tendon repair, the skin was closed with sutures (3-0Vicryl). The turkeys were sacrificed and the samples were collected 6 weeks after surgery.

[0051] 2. Pentamidine fibrin patch anti-tendon adhesion scoring: The physiological and behavioral responses of the operated turkeys were observed regularly. After 6 weeks, all tendons were dissected and scored based on the tendon adhesion system to evaluate the biosafety of the patch and the biological response to the host.

[0052] 3. Tendon regeneration and biomechanical testing: After surgery, biomechanical testing (normalized work of flexion, tendon gliding resistance, repair strength) and histological observation are used to evaluate the repair effect of the tendon and the efficacy of the patch, and to analyze the recovery of tendon function and changes in tendon adhesion.

[0053] The structural formula of Pentamidine, the overall appearance of the Pentamidine fibrin patch and the scanning electron microscope image are shown in Figure 2. Figure 1 As shown. Figure 2 It can be seen that at different time points of tendon stem cell culture, there was no significant difference in cell activity and cell migration ability between 0μg / ml and 25μg / ml pentamidine fibrin patches; when fibrin patches with different concentrations of pentamidine (0-50μg / ml) were co-cultured with tendon stem cells, there was no obvious change in cell live or dead staining.

[0054] The in vivo experimental modeling process (turkey model), tendon suture method (Modified Pennington Technique and Running Suture) and the appearance of the tendons in different groups after animal sacrifice are shown in Figure 2. Figure 3 As shown, it can be seen that the tendon surface of the group combined with PFP was smoother after suture.

[0055] like Figure 4 As shown in the figure, after the animals were sacrificed, tendon HE staining and Masson staining showed that PEP was biosafe and biocompatible; Sirius Red staining showed that most of the collagen fibers were type I collagen, and the collagen was arranged irregularly.

[0056] like Figure 5 As shown in , in terms of biomechanics, tendon flexion work ( Figure 5 Middle A) shows that the distal interdigital ( Figure 5 Middle B) Flexion work and proximal interphalangeal ( Figure 5 The flexion work of the tendon was significantly lower than that of the simple suture group (p<0.05); the tendon sliding resistance ( Figure 5 Middle D) shows that the tendon sliding resistance of the PFP group is lower than that of the simple suture group ( Figure 5 E), but there was no significant difference between the two groups (p>0.05); the maximum failure strength of tendon ( Figure 5 F, G) and failure stiffness ( Figure 5 Middle (H) showed that there was no statistically significant difference between the two groups (p>0.05).

[0057] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A pentamidine fibrin glue patch for preventing tendon adhesion, characterized in that: include: The pentamidine and fibrinolysis inhibitor solution are mixed evenly to obtain a pentamidine suspension, and then mixed evenly with SealerProtein Concentrate powder to obtain a pentamidine-loaded solution; The calcium salt solution and thrombin powder are mixed evenly to obtain Thrombin solution; The pentamidine-loaded solution is mixed with the Thrombin solution to react and obtain pentamidine fibrin glue; The pentamidine fibrin glue is spread in a container to form a patch, and then sterilized to obtain the product.

2. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: In the pentamidine-containing Fibrinolysis Inhibitor suspension, the concentration of pentamidine is 25-50 μg / mL.

3. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: The mass ratio of the pentamidine to the Sealer Protein Concentrate powder is 0.025-0.05:1 mg.

4. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: The mass ratio of the calcium chloride solution to the thrombin powder is 1 mL: 1-1.2 mg.

5. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: The mass ratio of the pentamidine to Thrombin powder is 0.025-0.05:1 mg.

6. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: The reaction temperature is 20-24°C and the pH is 7-7.

5.

7. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: The container is a culture dish with a size of 60 mm to 100 mm; Alternatively, the sterilization is performed by ultraviolet sterilization.

8. The pentamidine fibrin glue patch for preventing tendon adhesion according to claim 1, characterized in that: Before use, the pentamidine fibrin glue patch was cut into a size suitable for covering the tendon area and sterilized again for use.

9. A pentamidine fibrin glue patch for preventing tendon adhesion prepared by the method according to any one of claims 1 to 8.

10. Use of the pentamidine fibrin glue patch according to claim 9 in the preparation of a medical device for preventing tendon adhesion.

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