Application of α2-adrenaline receptor antagonists in the preparation of drugs to promote tendon-bone interface repair

Abstract

The invention discloses the application of α2-adrenaline receptor antagonists in the preparation of drugs for promoting tendon-bone interface repair, and belongs to the technical field of drug preparation. The application of α2-adrenaline receptor antagonists in the preparation of drugs for promoting tendon-bone interface repair, α2-adrenaline receptor antagonists, by inhibiting the expression of Adra2a at the tendon-bone interface, shields the sympathetic nerve regulation of rotator cuff injury, promotes the expression of new bone and cartilage markers at the rotator cuff tendon-bone interface, promotes rotator cuff tear repair, promotes osteogenesis and cartilage differentiation at the tendon-bone interface, promotes collagen regeneration at the rotator cuff tendon-bone interface, realizes new bone formation and cartilage band reconstruction at the tendon-bone interface, and promotes rotator cuff injury repair. It provides a new and effective rotator cuff repair and tendon-bone interface repair and reconstruction drug, which is easy to use.

Description

Technical Field

[0001] The present invention relates to the technical field of drug preparation, and more specifically to the application of an alpha 2-adrenaline receptor antagonist in the preparation of a drug for promoting the repair of the tendon-bone interface of a rotator cuff injury. Background Art

[0002] The rotator cuff includes a group of muscles surrounding the shoulder and tendons. It is a sheath-like structure composed of the tendinous parts of the subscapularis, supraspinatus, infraspinatus and teres minor muscles, which surrounds the glenohumeral joint, humeral head and joint capsule to form a nearly continuous sleeve-like structure. Rotator cuff tear is a tear that can involve tendons, muscle-tendon junctions or muscles, or avulsion from bones. Rotator cuff tear is a common shoulder joint sports injury disease, manifested as persistent shoulder pain and limited movement. The incidence of rotator cuff tear is 20.7%, which is common in elderly patients with rotator cuff degeneration. Clinically, it is mainly treated by arthroscopic rotator cuff repair, and the re-tear rate is 20% to 94%. Rotator cuff repair surgery is to suture the ruptured tendon to the bone again. The healing of the tendon-bone interface is the key to the success or failure of rotator cuff repair surgery. The tendon-bone junction is a gradual transition from tendon to cartilage to bone, which can be divided into four parts, namely tendon, non-calcified fibrocartilage, calcified fibrocartilage and bone, and the tendon is anchored in the deep layer of bone through gradual transformation. Reconstruction of the tendon-bone interface requires new bone formation and reconstruction of the cartilage band. However, the existing postoperative healing of the tendon-bone interface is mainly based on fibrosis repair, and the fibrocartilage band is difficult to regenerate. After repair, the buffering capacity of the tendon-bone interface is weakened, and it is easy to tear again. At present, research focuses on promoting osteogenic and chondrogenic differentiation through tissue engineering, that is, scaffold materials supplemented with seed cells or growth factors - structural repair of the tendon-bone interface. However, tissue engineering has always been in the exploratory stage in promoting regeneration through the spatiotemporal synergistic effect of stem cells and multiple differentiation-promoting factors.

[0003] Chronic high sympathetic tone is a hallmark of aging and obesity, leading to cardiovascular diseases such as hypertension and heart failure. Hypertensive patients are four times more likely to have giant tears than non-hypertensive people, and their risk of large tears is twice that of non-hypertensive people. Studies have shown that sympathetic tone regulates the occurrence and development of rotator cuff injuries, and the living conditions and mental states of rotator cuff injury patients vary, making them difficult to assess. With increasing age and changes in physiological state, rotator cuff injury patients have different stem cell reserves and differentiation abilities, and the microenvironment for stem cell transplantation is also different. Individual differences in rotator cuff injury patients and the complexity of rotator cuff tendon-bone interface reconstruction make rotator cuff injury repair treatment difficult to advance.

[0004] The main treatment in clinic is arthroscopic rotator cuff repair, with a re-tear rate of 20% to 94%. Traditional rotator cuff repair cannot be adjusted to the individual differences of rotator cuff injury patients, and the repaired tendon-bone interface cannot restore the gradual transition of tendon-cartilage-bone, resulting in poor mechanical strength and buffering capacity of the repaired tendon-bone complex, which is prone to re-tear. Summary of the invention

[0005] The invention discloses a medicine which is independent of a transplant and can promote osteogenic and chondrogenic differentiation at the tendon-bone interface and can promote structural repair of the tendon-bone interface of rotator cuff injuries.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] Application of α2-adrenaline receptor antagonists in the preparation of drugs for promoting tendon-bone interface repair.

[0008] The use of α2-adrenaline receptor antagonists in the rehabilitation of rotator cuff tears after rotator cuff repair surgery.

[0009] The α2-adrenaline receptor antagonist includes one or more of idazoxan, phentolamine, phenoxybenzamine, prazosin and yohimbine.

[0010] The drug is injected into the joint cavity after rotator cuff repair surgery. The dosage is 1-10 mg / kg, injected once every 1-2 days, and continuously injected for 2-4 weeks.

[0011] The α2-adrenaline receptor antagonist inhibits the expression of Adra2a at the tendon-bone interface, shields the sympathetic nerve regulation of rotator cuff injury, promotes the expression of new bone and cartilage markers at the rotator cuff tendon-bone interface, promotes the repair of rotator cuff tears, promotes osteogenesis and chondrogenesis at the tendon-bone interface, promotes collagen regeneration at the rotator cuff tendon-bone interface, achieves new bone formation and cartilage band reconstruction at the tendon-bone interface, and promotes the repair of rotator cuff injuries.

[0012] Compared with the prior art, the present invention has the following advantages and beneficial effects: it provides a new and effective tendon-bone interface repair and reconstruction drug for rotator cuff repair surgery, and the drug is easy to use. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Diagram of the modeling process for a rotator cuff tear.

[0014] Figure 2 Intra-articular injection of IDA inhibits adrenergic receptor α2a (Adra2a) and promotes the expression and distribution of new bone marker osteoin (Ostn) and cartilage marker aggrecan (Acan).

[0015] Figure 3 Intra-articular injection of IDA promotes the recovery of bilateral upper limb function after rotator cuff tear surgery.

[0016] Figure 4 Intra-articular injection for IDA improves the biomechanical strength of the tendon-bone complex in rotator cuff tear repair.

[0017] Figure 5Intra-articular injection of IDA improves bone density and bone volume at the rotator cuff tendon-bone interface. A–E) 2 weeks after surgery; F–J) 6 weeks after surgery. Figure 6 Injection into the IDA joint cavity promotes collagen regeneration at the rotator cuff tendon-bone interface. A, B) Masson staining and semi-quantitative analysis results 2 weeks after surgery. C, D) Masson staining and semi-quantitative analysis results 6 weeks after surgery.

[0018] Figure 7 Injection into the IDA joint cavity promotes cartilage regeneration at the rotator cuff tendon-bone interface. A, B) Safranin O fast green staining and semi-quantitative analysis results 2 weeks after surgery. C, D) Safranin O fast green staining and semi-quantitative analysis results 6 weeks after surgery.

[0019] Figure 8 Intra-articular injection of IDA improves the maturity and stability of the tendon-bone interface of rotator cuff tear repair. A, B) HE staining results and scores 2 weeks after surgery. C, D) HE staining results and scores 6 weeks after surgery. DETAILED DESCRIPTION

[0020] The technical solution of the present invention is further described below with reference to the accompanying drawings and through specific implementation methods.

[0021] Example 1

[0022] Studies have found that the sympathetic nerve tension of the rotator cuff increases after rotator cuff tear repair surgery, and the expression of adrenergic receptor α2a (Adra2a) at the rotator cuff tendon-bone interface increases. Therefore, the inventors tried to regulate the sympathetic nerve tension of the rotator cuff by inhibiting the expression of Adra2a, thereby achieving the purpose of promoting rotator cuff tear repair.

[0023] Idazoxan (IDA) is an α2-adrenergic receptor antagonist and an imidazoline receptor antagonist, which can competitively antagonize the central hypotensive effect of oxazoline-like active molecules. Idazoxan is used in experimental studies of depression, Alzheimer's disease, and Parkinson's disease. This example uses the α2-adrenergic receptor antagonist Idazoxan (IDA) as an example to study the relationship between the expression of Adra2a and the repair of the tendon-bone interface of the rotator cuff tear. The specific process is:

[0024] (1) Modeling of rotator cuff tear

[0025] Rats were anesthetized by intraperitoneal injection of sodium pentobarbital (40 mg / kg). After the rats were completely anesthetized, the right rotator cuff of the rats was prepared and the rats were subjected to a unilateral acute supraspinatus transection and repair model: first, the surgical area was disinfected, the right upper limb was abducted and moderately externally rotated, a longitudinal incision of about 2 cm was made with the acromion as the center of the incision, the subcutaneous fat and muscle fascia were bluntly separated, the deltoid muscle was exposed, the deltoid muscle was split to find the attachment point of the supraspinatus tendon, the supraspinatus tendon was lifted from the gap between the supraspinatus tendon and the greater tubercle of the humerus with an ophthalmic retractor, the supraspinatus attachment point was fixed with a 3-0 Prolene suture using the modified Masonallen method, the supraspinatus was transversely severed from the supraspinatus junction in the "footprint area" of the greater tubercle of the humerus with a scalpel, and the remaining cartilage layer was scraped off. Use a No. 22 needle to open two intersecting oblique bone tunnels around the "footprint area", and use a thread passing through the bone tunnel to fix the end of the supraspinatus muscle to the "footprint area" of the greater tubercle of the humerus without tension, so that it can return to its physiological state before transection. After the operation, the deltoid muscle and the incision skin are closed layer by layer, and the rats are changed to two cages after they wake up, and they can eat and move freely. The wound healing is observed every day within one week after the operation to prevent wound infection.

[0026] (2) Intra-articular injection of idazoxan

[0027] On the second day after rotator cuff tear repair surgery, intra-articular IDA injection was performed at a dose of 6 mg / kg, once every 2 days for 2 consecutive weeks.

[0028] After treatment, the mice were divided into the following three groups:

[0029] (A) Sham operation group SHAM: only sham operation of rotator cuff tear was performed, that is, after splitting the deltoid muscle, the supraspinatus muscle was found and pulled out with an ophthalmic retractor for inspection without being cut or repaired. The rest of the treatment was consistent with the above surgical procedure, and the samples were collected 2 and 6 weeks after the operation.

[0030] (B) Modeling group RC: The above-mentioned rotator cuff tear was repaired and modeling was performed without local IDA injection. The samples were collected 2 and 6 weeks after surgery.

[0031] (C) Modeling and medication group RC+IDA: The above-mentioned rotator cuff tear and repair model were performed, and local IDA injection was performed for 2 consecutive weeks, once every two days, and samples were collected 2 weeks and 6 weeks after surgery.

[0032] Two weeks after rotator cuff tear repair surgery (in the proliferative phase of rotator cuff tear repair, i.e. the early stage after surgery, which usually occurs 7 to 21 days after surgery in rats; cell regeneration and tissue healing at the trauma site begin, which is crucial for the healing of rotator cuff tears), the expression of Adra2a, the expression distribution of new bone markers and cartilage markers at the tendon-bone interface were detected to determine the effect of IDA on osteogenic and chondrogenic differentiation.

[0033] Figure 2 The figure shows the expression and distribution comparison of new bone marker osteoin (Ostn) and cartilage marker aggrecan (Acan) at the rotator cuff tendon-bone interface in different treatment groups 2 weeks after surgery. It was found that the expression and distribution of Adra2a at the tendon-bone junction increased 2 weeks after rotator cuff tear repair, while IDA reduced the expression and distribution of Adra2a at the tendon-bone junction. 2 weeks after surgery, the expression of Ostn and Acan at the tendon-bone junction decreased, while the expression and distribution of Ostn and Acan in the drug-treated group increased, indicating that IDA promotes osteogenic and chondrogenic differentiation at the rotator cuff tendon-bone interface.

[0034] At 2 weeks and 6 weeks after the repair surgery (during the remodeling period of rotator cuff tear repair, scar tissue begins to reconstruct and form mature scar tissue, and it is a stage where the strength, stability and range of motion of the shoulder need to be gradually restored), the effects of intra-articular IDA injection on rotator cuff function recovery, tendon-bone complex mechanical strength, bone mass, collagen repair, cartilage band reconstruction, interface maturity and stability were evaluated, and the short-term and long-term effects of idazoxan injection after rotator cuff injury repair surgery were tested.

[0035] Figure 3 The figure is a comparison of the maximum grip strength of the upper limbs of rats in different treatment groups 2 weeks and 6 weeks after surgery. It was found that the grip strength of the upper limbs of rats decreased significantly 2 weeks after rotator cuff tear. Compared with the modeling group RC that only underwent surgery, the grip strength of the upper limbs of rats in the modeling medication group RC+IDA recovered to a certain extent. Even if the medication was not continued 2 to 6 weeks after surgery, the rotator cuff function of rats in the modeling medication group RC+IDA was still stronger than that of the modeling RC group. The results show that IDA promotes the recovery of rotator cuff function after rotator cuff tear repair surgery.

[0036] Figure 4 The figure is a comparison of the biomechanical strength of the tendon-bone complex of rotator cuff tear repair in different treatment groups 6 weeks after surgery. It was found that the maximum failure load (Maximum failure load) and stiffness (Stiffness) of the RC group were significantly lower than those of the RC+IDA group and the SHAM group; the maximum failure load and stiffness of the RC+IDA group were not significantly different from those of the SHAM group. From the results, it can be seen that inhibiting Adra2a during the proliferation period of rotator cuff tear promotes the recovery of the new biomechanical properties of the rotator cuff tendon-bone complex during the rotator cuff tear repair and remodeling period, and reaches a level close to that before injury. The results show that IDA can improve the biomechanical strength of the tendon-bone complex after repair.

[0037] Figure 5The figure shows the comparison of bone density and bone mass at the rotator cuff tendon-bone interface of rats in different treatment groups 2 weeks and 6 weeks after surgery. 2 weeks after surgery, the bone density (BMD), bone volume fraction (BV / TV), and trabecular thickness (TB.TH) of the SHAM group were significantly higher than those of the RC group, and there was no significant difference in the number of trabecular bones (TB.N) and trabecular separation (TB.SP); at the same time, the TB.TH and TB.SP of the SHAM group were significantly higher than those of the RC+IDA group, while the TB.N of the SHAM group was lower than that of the RC+IDA group. There was no significant difference between the SHAM group and the RC+IDA group in terms of BMD and BV / TV. The BV / TV and TB.N of the RC+IDA group were higher than those of the RC group, and the TB.SP was lower than that of the RC group, while there was no significant difference in BMD and TB.TH between the two groups. At 6 weeks after surgery, the BMD and BV / TV of the SHAM group were higher than those of the RC group, and there were no significant differences in TB.TH, TB.N and TB.SP; at the same time, there were no significant differences in BMD, BV / TV, TB.TH, TB.N and TB.SP between the SHAM group and the RC+IDA group; the BV / TV and TB.N of the RC+IDA group were higher than those of the RC group, and the TB.SP was lower than that of the RC group, while there were no significant differences in BMD and TB.TH between the two groups. The results showed that IDA improved the bone mass at the rotator cuff tendon-bone junction of rats in both the early and late stages of rotator cuff healing, thereby promoting rotator cuff bone healing.

[0038] Figure 6 Comparison of collagen regeneration at the rotator cuff tendon-bone interface in different treatment groups 2 weeks and 6 weeks after surgery. At 2 weeks after surgery (Figure A), new bone (blue) and mature bone (red) were generated at the rotator cuff tendon-bone junction in each group. Among them, the RC+IDA group had a large amount of mature bone at the tendon-bone junction, which was significantly more than the other two groups; blue new bone was found on the tendon side of the three groups, indicating that there was obvious bone ingrowth in the early stage of rotator cuff tear repair. Semi-quantitative analysis showed (Figure B) that the amount of new bone in the rotator cuff tendon-bone complex in the RC group was significantly less than that in the SHAM group and the RC+IDA group, suggesting that IDA promotes bone regeneration in the early stage of rotator cuff tear repair.

[0039] Six weeks after surgery (Figure C), new bone and mature bone were found at the rotator cuff tendon-bone junction in each group, with a large amount of new bone on the tendon side in the RC group and the RC+IDA group. Semi-quantitative results showed (Figure D) that the new bone in the rotator cuff tendon-bone complex in the RC+IDA group was more than that in the SHAM group and the RC group. The results showed that IDA promoted bone regeneration in the late stage of rotator cuff tear repair.

[0040] Figure 7Comparison of cartilage regeneration at the rotator cuff tendon-bone interface in different treatment groups at 2 weeks and 6 weeks after surgery. 2 weeks after surgery (Figure A), a large area of ​​cartilage tissue (red) was present at the rotator cuff tendon-bone interface in both the RC+IDA group and the SHAM group, while only a small amount of cartilage tissue was produced in the RC group; semi-quantitative analysis showed (Figure B) that the cartilage tissue in the rotator cuff tendon-bone complex in the RC group was significantly less than that in the SHAM group and the RC+IDA group. The results showed that IDA promoted cartilage regeneration in the early stage of rotator cuff tear repair.

[0041] At 6 weeks after surgery (Figure C), cartilage tissue was present at the rotator cuff tendon-bone junction in each group, with the most in the RC+IDA group and the least in the RC group; semi-quantitative results (Figure D) also confirmed this. The results showed that IDA promoted the reconstruction of the cartilage band in the late stage of rotator cuff tear repair.

[0042] Figure 8 The following are comparisons of the maturity and stability of the rotator cuff tendon-bone interface in each group at 2 weeks and 6 weeks after surgery. At 2 weeks (Figure A) and 6 weeks (Figure C) after surgery, the number of cells at the tendon-bone interface in the RC group and the RC+IDA group increased significantly, and the regularity of cell arrangement, tissue structure and regularity, blood supply level and interface structural integrity were all worse than those in the SHAM group, with the RC group being the worst. The tendon maturity score results showed that at 2 weeks (Figure B), the tendon maturity in the RC group was significantly worse than that in the SHAM group, and there was no difference between the other groups; at 6 weeks (Figure D), the tendon maturity score in the RC group was significantly lower than that in the RC+IDA group and the SHAM group, and there was no difference between the other groups. The above HE staining results show that IDA improves the maturity and stability of the tendon-bone interface after rotator cuff tear repair.

[0043] The above results show that IDA joint cavity injection can shield the sympathetic nerve regulation of the rotator cuff injury site, promote osteogenesis and cartilage differentiation at the tendon-bone interface, and promote the repair of rotator cuff injury.

[0044] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. Application of α2-adrenaline receptor antagonist idazoxan in the preparation of drugs to promote tendon-bone interface repair.

2. The use according to claim 1, characterized in that: The medicine is an injection.

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