Application of mesenchymal stem cells combined with platelets in preparation of preparation for treating ankle arthritis

Abstract

The invention discloses application of a mesenchymal stem cell combined platelet in preparation of a preparation for treating ankle arthritis, the mesenchymal stem cell combined platelet (platelet-rich plasma) is adopted to treat the ankle arthritis, the operation is simple, the side effect is small, the treatment effect is remarkable, the treatment period is short, and the application is safe and reliable. The platelet-rich plasma is rich in growth factors, can promote the regeneration of damaged tissue vessels, increase blood supply and nutrient substances, enhance the biological activity and migration ability of cartilage cells, accelerate the healing of damaged tissues around ankle joints, and effectively promote the growth and reproduction and osteogenic differentiation of bone marrow mesenchymal stem cells (BM-SCs). The method has the advantages that the cartilage differentiation of the mesenchymal stem cells at the joints is promoted, the cell repair and regeneration of the affected parts of the joints are realized, the functional recovery of the joints is promoted, and the ankle arthritis is treated more efficiently.

Description

Technical Field

[0001] This invention relates to the field of stem cell technology, and more particularly to the application of mesenchymal stem cells combined with platelets in the preparation of ankle arthritis preparations. Background Technology

[0002] The ankle joint, formed by the medial and lateral malleoli of the talus and tibia / fibula, is one of the most important weight-bearing and mobility joints in the human body, crucial for maintaining normal walking. Ankle arthritis mainly manifests as pain, swelling, and joint effusion. In severe cases, it can lead to limited joint movement. Causes include trauma, such as injuries sustained during exercise or daily life due to inadequate ankle protection, often affecting ligaments and potentially causing fractures. Untreated traumatic ankle arthritis (TAA) can result. Other causes include underlying diseases, such as gouty arthritis causing ankle synovitis or rheumatoid arthritis affecting the ankle joint. Autoimmune arthritis, such as ankylosing spondylitis or psoriatic arthritis, can also affect the ankle joint. Degenerative factors, such as age, overuse, and frequent use, also contribute. Drug treatment is common, using steroids like betamethasone to relieve pain and improve ankle function; however, steroids often have numerous side effects.

[0003] Platelets are small, anucleate cells found in the blood, existing in both resting and functional phases. Resting platelets are inactive, but stimulated by extravascular collagen, they rapidly transition from the resting phase to the functional phase, extending pseudopodia to form a network that prevents blood cells from escaping, thus achieving hemostasis. Platelets in tissues can also release large amounts of cytokines, promoting vascular endothelial proliferation and tissue repair. Normal platelets have a lifespan of approximately 1-2 weeks in the blood. Platelet-rich plasma (PRP) is obtained by aseptically centrifuging whole blood drawn from a vein and collecting the platelets. This PRP can be injected locally to treat local injuries and relieve pain. Autologous PRP can promote the growth of bone marrow mesenchymal stem cells at the normal cellular level, promote the repair of damaged tissue, and avoid immune rejection.

[0004] Mesenchymal stem cells (MCS) are important members of the stem cell family, originating from the mesoderm in early development. They are pluripotent stem cells found in various tissues (such as bone marrow, umbilical cord blood and tissue, placental tissue, and adipose tissue). MCS possess multipotent differentiation potential, are non-hematopoietic stem cell adult cells, and can differentiate into various mesenchymal cell lineages (such as osteoblasts, chondrocytes, and adipocytes) or non-mesenchymal cell lineages. They also possess unique cytokine secretion functions. MCS can migrate to the precise site of injury, differentiate into various cell lines, secrete soluble factors crucial for cell survival and proliferation, and regulate immune responses. As a type of adult pluripotent stem cell, MCS is capable of self-renewal, multipotent differentiation, has a wide range of sources, is easily isolated and cultured in vitro, and has low immunogenicity. These characteristics, including tissue repair and immunomodulation, make it potentially valuable for the treatment of ankle arthritis. Summary of the Invention

[0005] Therefore, based on the above background, the present invention applies mesenchymal stem cells combined with platelets to treat ankle arthritis, and provides the application of mesenchymal stem cells combined with platelets in the preparation of a formulation for treating ankle arthritis.

[0006] The technical solution of this invention is as follows:

[0007] Application of mesenchymal stem cells combined with platelets in the preparation of ankle arthritis preparations.

[0008] Furthermore, the mesenchymal stem cells are derived from allogeneic umbilical cord tissue, umbilical cord blood, bone marrow, placental tissue, or autologous adipose tissue.

[0009] Furthermore, the mesenchymal stem cells are derived from allogeneic umbilical cord tissue.

[0010] The present invention also provides a medicament for treating ankle arthritis, the medicament comprising an effective amount of mesenchymal stem cells and platelet-rich plasma.

[0011] Furthermore, the platelet-rich plasma is autologous platelet-rich plasma.

[0012] Furthermore, the mesenchymal stem cells are derived from allogeneic umbilical cord tissue, umbilical cord blood, bone marrow, placental tissue, or autologous adipose tissue.

[0013] Furthermore, the mesenchymal stem cells are derived from allogeneic umbilical cord tissue.

[0014] Furthermore, the drug dosage form is an injectable dosage form.

[0015] The beneficial effects of adopting the above technical solution are as follows:

[0016] This invention utilizes mesenchymal stem cells combined with platelets (platelet-rich plasma) to treat ankle arthritis. The procedure is simple, has few side effects, and offers significant therapeutic effects with a short treatment cycle. Platelet-rich plasma is rich in growth factors, which can promote angiogenesis in damaged tissues, increase blood supply and nutrients, enhance the biological activity and migration ability of chondrocytes, accelerate the healing of damaged tissues around the ankle joint, and effectively promote the growth, proliferation, and osteogenic differentiation of bone marrow mesenchymal stem cells (BM-SCs). This facilitates the differentiation of mesenchymal stem cells into chondrocytes in the joint, enabling cell repair and regeneration at the affected joint site, promoting joint function recovery, and providing a more efficient treatment for ankle arthritis. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. In the following embodiments, unless specific conditions are specified, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0018] Example 1: Clinical application experiment of allogeneic mesenchymal stem cells combined with platelets in the treatment of bone nonunion.

[0019] The experimental steps are as follows:

[0020] S1: Selection and grouping of experimental subjects

[0021] One hundred and twenty patients with ankle arthritis who were treated between January 2021 and January 2022 were selected, including 57 males and 63 females. There were 63 patients with traumatic ankle arthritis, 24 patients with gouty ankle arthritis, and 33 patients with osteoarthritis of the ankle. The patients were randomly divided into three groups of 40 patients each: drug group A, drug group B, and drug group C. There were no statistically significant differences in the general characteristics of the three groups (P>0.05).

[0022] Exclusion criteria for experimental subjects: ① Patients with abnormal coagulation function; ② Patients with severe heart, liver, or kidney failure; ③ Patients with severe joint deformities or functional impairments.

[0023] S2: Preparation of allogeneic mesenchymal stem cells

[0024] With parental consent, a fresh umbilical cord was obtained from the newborn infant. After removing the umbilical artery and vein, the remaining tissue was minced into 0.5-1.5 mm pieces. Type II collagenase (2 mg / ml) was added, and the mixture was incubated at 37°C with shaking for 3-4 hours. The mixture was then filtered through a 70 μm filter. The cell suspension was centrifuged at 4°C and 2000 rpm for 30 minutes, washed, and resuspended in complete culture medium. The cells were then cultured at 1×10⁻⁶ cells / min. 4 Cells were seeded at a density of / cm² in culture medium and cultured in a 37°C, saturated humidity, 5% CO₂ incubator. Non-adherent cells were removed after 72 hours, and the medium was changed every 3-4 days. Cells were passaged after trypsin digestion when they reached 80% confluence. The expression of CD14, CD31, CD34, CD45, CD73, CD90, CD105, and HLA-DR was detected by flow cytometry, and WJ-MSCs in vitro differentiation assays were performed to identify cell characteristics. All therapeutic applications used cells from passages 3-5.

[0025] S3: Add the mesenchymal stem cells prepared in step S2 to physiological saline to prepare a suspension.

[0026] S4: PRP preparation: Disinfect the skin, collect 10ml of the patient's own peripheral venous blood, place it in a heparinized tube, shake well, and centrifuge at 2500 rpm for 5 minutes. The liquid in the tube will separate into layers: plasma on top, red blood cells on the bottom, and a thin layer of platelets in the middle. Carefully extract the middle and top plasma layers, discarding the bottom red blood cells. Transfer to a sterile centrifuge tube for a second centrifugation at 3500 rpm for 8 minutes. Finally, extract approximately 1ml of the platelet-rich plasma from the bottom layer. This completes the preparation of PRP.

[0027] S5: Mix the MSCs prepared in S3 with the PRPs prepared in S4 to make a 5ml mixed formulation.

[0028] Group A received an intra-articular injection of 5 ml of the mesenchymal stem cell suspension prepared in step S3, administered once every three days, for three consecutive months. Group B received an intra-articular injection of 5 ml of PRP prepared in step S4, administered once every three days, for three consecutive months. Group C received an intra-articular injection of 5 ml of a mixture of mesenchymal stem cells and platelets prepared in step S5, administered once every three days, for three consecutive months.

[0029] After treatment, the treatment effect is divided into the following categories according to the "Clinical Disease Diagnosis and Efficacy Evaluation Criteria":

[0030] ① Cured: Clinical symptoms disappear, and the ankle joint is no different from normal;

[0031] ② Improvement: Clinical symptoms improve, ankle pain decreases, and function is basically restored;

[0032] ③ Ineffective: Clinical symptoms did not improve or worsened, and ankle joint function did not recover. Effective rate = (Cures + Improvements) / Total number of cases × 100%.

[0033] Group A of drugs: 0 cases showed significant effect, 14 cases showed effect, and 26 cases showed no effect, with an effective rate of 35%.

[0034] Drug Group B: 2 cases showed significant effect, 19 cases showed effect, and 19 cases showed no effect, with an effective rate of 52.5%.

[0035] Group C: 11 cases showed significant effect, 20 cases showed effect, and 9 cases showed no effect, with an effective rate of 77.5%.

[0036] The present invention and its embodiments have been described above. This description is not restrictive, and the embodiments shown above are merely one example of the present invention; the actual structure is not limited thereto. In conclusion, if those skilled in the art are inspired by this description and, without departing from the spirit of the invention, design similar structures and embodiments without creative effort, all such designs should fall within the protection scope of the present invention.

Claims

1. Application of mesenchymal stem cells combined with platelets in the preparation of ankle arthritis preparations.

2. The application according to claim 1, characterized in that, The mesenchymal stem cells are derived from allogeneic umbilical cord tissue, umbilical cord blood, bone marrow, placental tissue, or autologous adipose tissue.

3. The application according to claim 2, characterized in that, The mesenchymal stem cells were derived from allogeneic umbilical cord tissue.

4. A drug for treating ankle arthritis, characterized in that, The drug contains an effective amount of mesenchymal stem cells and platelet-rich plasma.

5. The medicament for treating ankle arthritis according to claim 4, characterized in that, The platelet-rich plasma is autologous platelet-rich plasma.

6. The medicament for treating ankle arthritis according to claim 5, characterized in that, The mesenchymal stem cells are derived from allogeneic umbilical cord tissue, umbilical cord blood, bone marrow, placental tissue, or autologous adipose tissue.

7. A medicament for treating ankle arthritis according to claim 6, characterized in that, The mesenchymal stem cells were derived from allogeneic umbilical cord tissue.

8. A medicament for treating ankle arthritis according to claim 5, characterized in that, The drug dosage form is an injectable dosage form.