Injectable magnesium-loaded lipid-based super-lubricating hydrogel microsphere, articular cavity injection formulation, and preparation method therefor

Abstract

The present invention provides an injectable magnesium-loaded lipid-based super-lubricating hydrogel microsphere, an articular cavity injection formulation, and a preparation method therefor. The method comprises the following steps: S1, preparing a magnesium-loaded multilamellar vesicular liposome suspension containing magnesium ions; S2, preparing an uncrosslinked magnesium-containing hydrogel; and S3, mixing the magnesium-loaded multilamellar vesicular liposome suspension with the magnesium-containing hydrogel to prepare hydrogel droplets, and carrying out crosslinking to form the magnesium-loaded lipid-based super-lubricating hydrogel microspheres. According to the present invention, the magnesium loading capacity of the lipid-based super-lubricating hydrogel microspheres can be greatly improved; on one hand, the biological function provided by the magnesium ions can be greatly prolonged, that is, the repair of cartilage by synovial liquid progenitor cells is continuously promoted; and on the other hand, the intrinsic lubricating effect of the lipid-based super-lubricating hydrogel microspheres can mitigate the further wear of newly formed cartilage; in addition, the magnesium ions can significantly promote the expression of PRG4 in chondrocytes, which not only facilitates the maintenance of chondrocyte homeostasis, but also further enhances articular lubrication.

Description

Injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres, joint cavity injection preparation and preparation method thereof Technical Field

[0001] The present invention relates to the field of medicine, in particular to injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres, a joint cavity injection preparation and a preparation method thereof. Background Art

[0002] Osteoarthritis (OA) is a chronic, progressive joint disease. The development and progression of OA involves multiple factors, including age, trauma, obesity, and genetics. The pathological changes of OA are characterized by cartilage degeneration, accompanied by synovial inflammation, subchondral bone changes, and osteophyte formation, which in turn cause joint pain, dysfunction, and deformity, seriously affecting the patient's quality of life. The disability rate of knee and hip OA is very high. However, there is currently no effective treatment for OA, and patients with advanced OA often require joint replacement surgery.

[0003] Recent studies show that knee and hip OA affects approximately 4% of the global population. In China, the prevalence of knee OA among people aged 60 and over is approximately 8.1%. In the United States, the average annual cost of OA treatment for each OA patient is approximately US$6,870, while in Europe, the average cost is between €1,330 and €10,425. As OA is a joint disease associated with old age, its prevalence is expected to rise annually with the aging population. According to the United Nations, by 2050, the proportion of people aged 60 and over will exceed 20% of the global population. Consequently, medical and socioeconomic expenditures related to OA are expected to increase significantly in the coming decades.

[0004] According to the latest clinical treatment guidelines for OA, oral or topical nonsteroidal anti-inflammatory drugs (NSAIDs) remain the first-line treatment. However, these drugs only temporarily relieve OA-induced joint pain symptoms and do not slow OA progression. Furthermore, oral NSAIDs carry the risk of gastrointestinal or cardiovascular complications, particularly in patients with underlying medical conditions. For OA patients who are refractory to oral or topical NSAIDs or who have contraindications to such treatment, intra-articular injection therapy becomes the preferred option. Currently, standard intra-articular injection preparations are hyaluronic acid (HA) and corticosteroids (CS), but their efficacy in OA remains controversial (Jones et al., 2019). HA is a relatively safe agent, but current clinical evidence has not yet confirmed its efficacy in alleviating OA (Jones et al., 2019). CS can provide short-term analgesia for OA patients, but recent clinical studies have shown that intra-articular injection of CS carries the risk of accelerating OA cartilage degeneration and disease progression (McAlindon et al., 2017). Several new intra-articular injection therapies are in clinical development: platelet-rich plasma (PRP) and stem cell (MSC) therapies have shown promise in promoting cartilage repair. However, due to the lack of internationally standardized preparations for these agents, safety is a significant concern, and clinical guidelines do not yet recommend these therapies.

[0005] In summary, current intra-articular injection preparations for osteoarthritis are mainly used to relieve symptoms such as pain. There is an urgent need for a new intra-articular injection preparation that can effectively promote cartilage regeneration in osteoarthritis and is safe and economical.

[0006] Magnesium (Mg) is one of the most abundant elements in the human body, with 80% of the body's magnesium being distributed in the musculoskeletal system. Magnesium is involved in numerous biochemical reactions in the body, including energy metabolism, protein, and nucleic acid synthesis, suggesting that magnesium is crucial for maintaining normal metabolism and health of the musculoskeletal system. Clinical studies have confirmed that decreased magnesium ion levels in human plasma are closely associated with the development and progression of osteoarthritis. Furthermore, animal studies have confirmed that a low-magnesium diet affects the health of articular cartilage. These findings suggest that magnesium supplementation is crucial for the treatment of osteoarthritis. Subsequent animal studies have further demonstrated that intra-articular injection of magnesium ions can effectively mitigate disease progression in animal models of arthritis (Yao et al., 2021, Yao et al., 2019).

[0007] Numerous stem / progenitor cells exist in various components of joints (e.g., synovium, cartilage, and synovial fluid), and these cells may be involved in the regeneration of degenerated or damaged cartilage. Among them, synovial progenitor cells (SPCs) within synovial fluid possess a strong capacity for chondrogenic differentiation (Jones et al., 2008, McGonagle et al., 2017). Under specific circumstances (e.g., joint distraction surgery), SPCs can adhere to the surface of damaged cartilage and further differentiate into chondrocytes, promoting cartilage regeneration (Baboolal et al., 2016). Recent studies have demonstrated that magnesium supplementation significantly promotes chondrogenic differentiation of synovial and bone marrow stem cells; in addition, magnesium can enhance the adhesion of stem cells by regulating integrins (Shimaya et al., 2010, Wang et al., 2017).

[0008] Hydrogel systems are a commonly used drug delivery system with excellent biocompatibility. Recent studies have reported that lipid-based superlubricating hydrogels exhibit exceptional lubricity, reducing surface friction by 80%-99.3% compared to conventional hydrogels (Lin et al., 2020). In a previously granted patent (Patent Application Publication No.: CN113995891A), the inventors further developed lipid-based superlubricating hydrogels into hydrogel microspheres, whose spherical structure acts as a "rolling bearing" to enhance lubrication (Lei et al., 2022). These hydrogels are suitable for treating osteoarthritis to reduce joint wear and are an excellent magnesium-carrying sustained-release system. Improving the magnesium ion loading capacity of lipid-based superlubricating hydrogels to prolong and increase the release time and amount of magnesium ions is a technical bottleneck that urgently needs to be overcome.

[0009] It should be noted that the information disclosed in the above background technology section is only used to understand the background of this application, and therefore may include information that does not constitute prior art known to ordinary technicians in this field. Summary of the Invention

[0010] The main purpose of the present invention is to overcome the defects of the above-mentioned background technology and provide an injectable magnesium-loaded lipid-based super-lubricating hydrogel microsphere, an articular cavity injection preparation and a preparation method thereof.

[0011] To achieve the above object, the present invention adopts the following technical solutions:

[0012] A method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres comprises the following steps:

[0013] S1. Preparation of a magnesium-loaded multilamellar vesicle liposome suspension containing magnesium ions;

[0014] S2. Preparation of uncrosslinked magnesium-containing hydrogel;

[0015] S3. The magnesium-loaded multilamellar vesicular liposome suspension and the magnesium-containing hydrogel are mixed to prepare hydrogel droplets, which are then cross-linked to form magnesium-loaded lipid-based super-lubricating hydrogel microspheres.

[0016] Further:

[0017] Step S1 comprises: adding hydrogenated soybean phosphatidylcholine (HSPC) to a magnesium ion-containing solution to prepare the magnesium-loaded multilamellar vesicle liposome suspension.

[0018] In step S1, hydrogenated soybean phosphatidylcholine (HSPC) is added to a magnesium ion-containing solution and ultrasonically treated at 25-60° C. for 10-30 minutes.

[0019] The magnesium ion-containing solution is magnesium chloride, magnesium sulfate solution or magnesium ascorbate solution.

[0020] The magnesium ion concentration in the magnesium ion-containing solution is 1-5M.

[0021] The concentration of the magnesium-loaded multilamellar vesicle liposome suspension is 40-100 mM.

[0022] The magnesium-containing hydrogel in step S2 is prepared by mixing a magnesium ion solution and any type of hydrogel, preferably a GelMA hydrogel.

[0023] In step S3, the mixing ratio of the magnesium-loaded multilamellar vesicular liposomes and the magnesium-containing supergel is 20:3 (w / w).

[0024] In step S3, the magnesium-loaded multilamellar vesicular liposomes and the magnesium-containing hydrogel are mixed and then mixed with sterile microfluidic oil to prepare pre-gel hydrogel droplets by a microfluidic emulsion method.

[0025] The invention discloses injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres, which are hydrogel microspheres prepared by the method.

[0026] A joint cavity injection preparation comprises the injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres.

[0027] The present invention has the following beneficial effects:

[0028] In the preparation method of injectable magnesium-loaded lipid-based super-lubricant hydrogel microspheres proposed in the present invention, in addition to loading magnesium ions into the hydrogel, magnesium ions are also loaded into liposomes in the form of oil-in-water. Hydrogel droplets are prepared by mixing a magnesium-loaded multilamellar vesicle liposome suspension in the form of oil-in-water with a magnesium-containing hydrogel. The magnesium-loaded lipid-based super-lubricant hydrogel microspheres obtained after cross-linking can greatly increase the magnesium loading amount of the lipid-based super-lubricant hydrogel microspheres; on the one hand, it can greatly prolong the biological function provided by magnesium ions, that is, continuously promote the repair of cartilage by synovial fluid progenitor cells; on the other hand, the lubricating effect of the lipid-based super-lubricant hydrogel microspheres themselves can reduce the re-wear of new cartilage; in addition, the research of the present invention also confirmed that magnesium ions can significantly promote the expression of chondrocyte PRG4 (synoviolin encoded by the PRG4 gene is the main component of joint lubrication), which can further maintain chondrocyte homeostasis and enhance joint lubrication.

[0029] Compared with the prior art, the present invention has the following advantages:

[0030] Magnesium is an essential nutrient for the normal human body. At the same time, oral and injectable preparations containing magnesium have been used in clinical practice for many years and are safe and reliable. Combinations of compounds with magnesium as the active ingredient can play a therapeutic role in delaying the progression of osteoarthritis and relieving joint pain by targeting multiple osteoarthritis mechanisms. (1) First, magnesium ions can promote the adhesion of stem / progenitor cells in synovial fluid to the surface of damaged cartilage and promote their differentiation into chondrocytes, thereby facilitating cartilage regeneration. (2) Second, magnesium ions can significantly promote the synthesis of cartilage extracellular matrix, especially synovial protein, thereby enhancing the lubricating function of the hydrogel microspheres and delaying the wear of the original and newly regenerated cartilage. (3) In addition, magnesium ions can also alleviate the inflammatory response in intra-articular tissues, including cartilage and synovium, and reduce the release of inflammatory factors and proteolytic enzymes, thereby effectively alleviating the damage to cartilage tissue caused by inflammation and the pain caused by inflammation. (4) On the other hand, by loading magnesium ions into hydrogels and liposomes respectively, the magnesium loading of lipid-based super-lubricating hydrogel microspheres can be greatly increased to provide the function of long-term sustained release of magnesium ions, and compared with bulk hydrogels, hydrogel microspheres can be more evenly distributed in the joint cavity. (5) Furthermore, different types of magnesium-containing hydrogels can be used to synthesize the magnesium-loaded lipid-based super-lubricating hydrogel microspheres to meet special requirements for the adhesion, hardness and other properties of the hydrogel microspheres, further broadening the application range of the hydrogel microspheres. (6) Finally, the joint cavity injection preparation with magnesium-loaded lipid-based hydrogel microspheres as the main raw material is locally injected by joint cavity puncture. It has high safety, low clinical operation technical difficulty, and is easy to use. It can be carried out in most regular medical institutions. The raw material source of the drug is wide, the preparation method is simple, and the cost is low, which can effectively reduce the economic burden on patients themselves and society.

[0031] Other beneficial effects of the embodiments of the present invention will be further described below. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIG1 is a flow chart of a method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to an embodiment of the present invention.

[0033] FIG2 shows the results of fast green staining of paraffin sections of knee joint samples of rats in the control group, magnesium chloride group, RGD group, and magnesium chloride + RGD group at 8 and 16 weeks after treatment (scale: 1000 μm).

[0034] Figure 3 shows the results of red-fast green staining and gene expression of chondrogenic markers in paraffin sections of chondrogenic cell pellets of the control group and magnesium chloride composition at week 6 of in vitro induced chondrogenic differentiation of rat synovial fluid stem / progenitor cells (scale bar: 100 μm).

[0035] FIG4 is a graph showing the gene expression results of cartilage markers in the control group and magnesium chloride composition in human articular cartilage explants cultured in vitro for the third week.

[0036] FIG5 is a flowchart of the preparation method of injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres of the present invention. DETAILED DESCRIPTION

[0037] The following is a detailed description of the embodiments of the present invention. It should be emphasized that the following description is only exemplary and is not intended to limit the scope of the present invention and its application.

[0038] As shown in FIG1 and FIG5 , an embodiment of the present invention provides a method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres, comprising the following steps:

[0039] S1. Preparation of a magnesium-loaded multilamellar vesicle liposome suspension containing magnesium ions;

[0040] S2. Preparation of uncrosslinked magnesium-containing hydrogel;

[0041] S3. The magnesium-loaded multilamellar vesicular liposome suspension and the magnesium-containing hydrogel are mixed to prepare hydrogel droplets, which are then cross-linked to form magnesium-loaded lipid-based super-lubricating hydrogel microspheres.

[0042] The present invention also provides an intra-articular injectable preparation for treating osteoarthritis, wherein the intra-articular injectable preparation mainly contains the magnesium-loaded lipid-based super-lubricating hydrogel microspheres as an active ingredient. The osteoarthritis includes primary and secondary osteoarthritis.

[0043] The embodiment of the present invention also provides an application of a joint cavity injectable preparation prepared with the magnesium-loaded lipid-based super-lubricating hydrogel microspheres as an effective ingredient for delaying articular cartilage loss and meniscus degeneration, promoting articular cartilage regeneration, enhancing joint lubrication, and relieving joint edema, pain or synovial inflammatory response.

[0044] In some specific embodiments, a method for preparing an intra-articular injection preparation containing magnesium-loaded lipid-based hydrogel microparticles as a main component is provided. The steps are as follows:

[0045] 1. Preparation of magnesium-loaded liposomes:

[0046] 1-1. Hydrogenated soybean phosphatidylcholine (HSPC) was added to a magnesium ion solution and ultrasonically treated at 30°C and 60°C for 15 minutes, respectively, to prepare a water-in-oil (W / O) suspension, i.e., a multilamellar vesicle-like liposome suspension containing magnesium ions.

[0047] 2. Preparation of Magnesium-containing GelMA Hydrogel:

[0048] 2-1. Add type II gelatin at a mass volume ratio of 10% to 0.25 M sodium carbonate-sodium bicarbonate buffer, heat to 55°C, and adjust the pH to 7.4.

[0049] 2-2. Methacrylic anhydride was added dropwise to the gelatin solution, and the mixture was stirred at 55°C for one hour, the pH was adjusted to 7.4 and the reaction was terminated.

[0050] 2-3. The obtained solution was dialyzed at 37°C for 7 days (compared to the molecular weight cut-off of deionized water of 12,000-14,000 Da), filtered through 0.22 μm, freeze-dried, and stored at -20°C in the dark.

[0051] 2-4. The freeze-dried GelMA powder was dissolved in a magnesium ion solution at a mass volume ratio of 8%, and the photoinitiator I2959 was added to the solution to prepare a magnesium-containing GelMA uncrosslinked hydrogel.

[0052] 3. Synthesis of magnesium-loaded lipid-based super-lubricating hydrogel microparticles: At 37°C, the magnesium-loaded multilamellar vesicular liposomes obtained in steps 1 and 2 above and the magnesium-containing GelMA hydrogel were mixed in a certain proportion to prepare liquid hydrogel droplets, which were then cross-linked by ultraviolet irradiation to obtain magnesium-loaded lipid-based super-lubricating hydrogel microparticles.

[0053] 4. The obtained magnesium-loaded lipid-based superlubricating hydrogel microparticles were washed with acetone and deionized water in turn and then immersed in sterile deionized water for storage.

[0054] Wherein, preferably, the magnesium ion-containing solution in step 1-1 is magnesium chloride, magnesium sulfate or magnesium ascorbate solution.

[0055] Furthermore, the magnesium-containing solution in step 1-1 is 1-5M.

[0056] Preferably, the concentration of the multilamellar vesicle liposome suspension containing magnesium chloride in step 1-1 is 40-100 mM.

[0057] Preferably, the magnesium ion-containing solution in steps 2-4 is magnesium chloride, magnesium sulfate or magnesium ascorbate solution.

[0058] Furthermore, the magnesium-containing solution in steps 2-4 is 1-5M.

[0059] Preferably, the ratio of the initiator I2959 in steps 2-4 is 0.05% (w / v).

[0060] Preferably, in step 3, the mixing ratio of the magnesium-loaded multilamellar vesicular liposomes obtained in steps 1 and 2 and the magnesium-containing GelMA hydrogel is 20:3 (w / w).

[0061] Preferably, the liquid hydrogel droplets in step 3 are prepared by mixing the magnesium-loaded multilamellar vesicular liposomes and the magnesium-containing GelMA hydrogel obtained in steps 1 and 2 in a certain proportion, and then mixing them with sterile microfluidic oil (prepared by mixing 95 wt% paraffin oil and 5 wt% Span 80), and using a microfluidic emulsion method to prepare liquid hydrogel droplets before gelation.

[0062] Preferably, the storage temperature in step 4 is 4°C.

[0063] Experimental example

[0064] Experimental Example 1: Magnesium ions promote cartilage regeneration in osteoarthritis by regulating the adhesion and chondrogenic differentiation of synovial fluid stem / progenitor cells.

[0065] Experimental methods

[0066] 1. Establishment of a rat osteoarthritis model induced by surgical resection of the anterior cruciate ligament and the anterior horn of the medial meniscus: After anesthesia, experimental rats (3-month-old) underwent a transpatellar approach to the right knee joint. The joint capsule was incised, the patellar fat pad and the anterior horn of the medial meniscus were removed, and the anterior cruciate ligament was exposed. The anterior cruciate ligament was then cut with a blade, and the surgical area was irrigated with saline. After successful modeling using an anterior drawer test, the joint capsule was sutured with absorbable sutures, and the skin was sutured with silk sutures.

[0067] 2. Grouping: Sixteen weeks after surgery, the rats were randomly divided into four groups and received intra-articular injections of normal saline (control group), 0.5M magnesium chloride solution, 2mg / ml RGD solution (cell adhesion blocker), or a mixture of 0.5M magnesium chloride and 2mg / ml RGD solution twice weekly for two weeks. Mice were sacrificed at eight and 16 weeks after injection, and right knee joint specimens were obtained for histological analysis to assess pathological changes in the knee joints following treatment.

[0068] Experimental results

[0069] 1. After the experimental group mice received the injection in the articular cavity, no redness or swelling of the corresponding knee joints, or abnormalities such as limited movement, were observed, indicating that the articular cavity injection is safe.

[0070] 2. Magnesium ions can promote cartilage regeneration in a rat osteoarthritis model. As shown in the red-fast green staining in Figure 2, at 8 and 16 weeks after treatment, the articular cartilage of the knee joints of mice in the normal saline group was severely worn, manifested as localized cartilage loss; while the articular cartilage thickness of the knee joints of rats in the magnesium chloride group increased significantly, and the cartilage matrix stained darker. After injection of RGD, cartilage degeneration was further aggravated compared with the normal saline group, manifested as cartilage loss with a wider range and depth. After injection of a mixed solution of RGD and magnesium chloride, the cartilage regeneration effect of magnesium chloride on arthritic rats disappeared. This result suggests that magnesium chloride can promote cartilage regeneration in osteoarthritis by regulating the adhesion and chondrogenic differentiation of synovial fluid stem / progenitor cells.

[0071] Experimental Example 2: Magnesium ions can promote the chondrogenic differentiation of synovial fluid stem / progenitor cells in vitro.

[0072] Experimental methods

[0073] 1. Isolation and culture of rat synovial fluid stem / progenitor cells: Sixteen weeks after establishing a rat osteoarthritis model induced by anterior cruciate ligament and medial meniscus resection surgery, 100 μl of PBS solution was injected into the rat knee joint under anesthesia using a 1 ml syringe with a 25G injection needle. The joint cavity was repeatedly aspirated and irrigated several times. The aspirated PBS solution containing the cell suspension was suspended in α-MEM medium containing 10% FBS, seeded into culture dishes, and cultured in a cell culture incubator at 37°C, 5% CO2.

[0074] 2. Preparation for chondrogenic differentiation: Add the following ingredients to high-glucose DMEM medium in proportion: 1% FBS, 1% ITS, 1% NEAA, 0.1 M vitamin C, 1 nM dexamethasone, 40 μg / ml L-proline, 1 mM sodium pyruvate, 1 ng / ml transforming growth factor β-1, and 500 ng / ml bone morphogenetic protein.

[0075] 3. Chondrogenic differentiation of rat stem / progenitor cells: 5*10 5 2-5th generation rat stem / progenitor cells were placed in a 15 ml centrifuge tube, 1*10 3 After centrifugation at rpm for 5 minutes and removal of the supernatant, the cells were resuspended in chondrogenic induction solution (control group) or 10 mM magnesium chloride (magnesium chloride group) and 1*10 3 Centrifuge at 100 rpm for 5 minutes, loosen the cap of the centrifuge tube, and culture the cell pellet in a 37°C, 5% CO2 cell culture incubator for 6 weeks. The resulting cell spheres were fixed or lysed and then subjected to histological analysis or RT-qPCR analysis to evaluate chondrogenic differentiation.

[0076] Experimental results

[0077] 1. Magnesium ions can significantly promote the chondrogenic differentiation of rat stem / progenitor cells. As shown in the red-fast green staining in Figure 3, compared with the control group, the addition of magnesium chloride significantly increased the number of vacuolated chondrocytes and red-stained chondrocyte extracellular matrix in the cell spheroids. The RT-qPCR results in Figure 3 show that the addition of magnesium chloride significantly increased the gene expression of chondrogenic markers, especially PRG4, suggesting that magnesium ions can effectively promote the chondrogenic differentiation of rat stem / progenitor cells.

[0078] Experimental Example 3: Magnesium ions can promote the expression of chondrogenic markers in human articular cartilage explants in vitro.

[0079] Experimental methods

[0080] 1. Human Articular Cartilage Explant Culture: Human lateral femoral condyle specimens were obtained from total knee replacement surgery. The cartilage tissue was minced into small pieces approximately 1 x 1 mm using a scalpel blade. After washing with PBS, explants of equal mass were placed in either DMEM / F12 medium containing 10% FBS (control group) or 10 mM magnesium chloride (MgCl group) in a 37°C, 5% CO2 incubator for 3 weeks. The resulting human articular cartilage explant tissue was lysed and analyzed by RT-qPCR to assess the expression of chondrogenic markers.

[0081] Experimental results

[0082] 1. Magnesium ions can significantly promote the gene expression of chondrogenic markers in human articular cartilage explants. As shown in the RT-qPCR results in Figure 4, compared with the control group, the addition of magnesium chloride significantly increased the gene expression of chondrogenic markers, especially PRG4, in cartilage explants, suggesting that magnesium ions can effectively promote the synthesis of human cartilage extracellular matrix, especially synoviolin.

[0083] The above description further details the present invention in conjunction with specific / preferred embodiments, and the specific implementation of the present invention should not be construed as being limited to these descriptions. Persons skilled in the art will appreciate that, without departing from the spirit of the present invention, they may make various substitutions or modifications to the described embodiments, and these substitutions or modifications should be considered to fall within the scope of protection of the present invention. Throughout this specification, reference to terms such as "one embodiment," "some embodiments," "preferred embodiments," "examples," "specific examples," or "some examples" indicates that the specific features, structures, materials, or characteristics described in conjunction with such embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Persons skilled in the art may combine and assemble the different embodiments or examples described in this specification, as well as features of different embodiments or examples, without conflicting opinions. Although the embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions, and modifications may be made herein without departing from the scope of protection of the patent application.

Claims

1. A method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres, characterized in that: The steps include: S1. Preparation of a magnesium-loaded multilamellar vesicle liposome suspension containing magnesium ions; S2. Preparation of uncrosslinked magnesium-containing hydrogel; S3. The magnesium-loaded multilamellar vesicular liposome suspension and the magnesium-containing hydrogel are mixed to prepare hydrogel droplets, which are then cross-linked to form magnesium-loaded lipid-based super-lubricating hydrogel microspheres.

2. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claim 1, wherein: Step S1 comprises: adding hydrogenated soybean phosphatidylcholine (HSPC) to a magnesium ion-containing solution to prepare the magnesium-loaded multilamellar vesicle liposome suspension.

3. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claim 2, wherein: In step S1, hydrogenated soybean phosphatidylcholine (HSPC) is added to a magnesium ion-containing solution and ultrasonically treated at 25-60° C. for 10-30 minutes.

4. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claim 2, wherein: The magnesium ion-containing solution is one of magnesium chloride, magnesium sulfate, or magnesium ascorbate solution.

5. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claims 2 to 4, characterized in that: The magnesium ion concentration in the magnesium ion-containing solution is 1-5M.

6. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claim 1, wherein: The concentration of the magnesium-loaded multilamellar vesicle liposome suspension is 40-100 mM.

7. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claim 1, wherein: The magnesium-containing hydrogel in step S2 is prepared by mixing a magnesium ion solution and any type of hydrogel, preferably a GelMA hydrogel.

8. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to claims 6 to 7, characterized in that: The magnesium ion concentration in the magnesium ion-containing solution is 1-5M.

9. The method for preparing injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres according to any one of claims 1 to 7, characterized in that: In step S3, preferably, the mixing ratio of the magnesium-loaded multilamellar vesicular liposomes and the magnesium-containing supramolecular hydrogel is 20:3 (w / w); preferably, the magnesium-loaded multilamellar vesicular liposomes and the magnesium-containing hydrogel are mixed and then mixed with sterile microfluidic oil to prepare pre-gelled hydrogel droplets by a microfluidic emulsion method.

10. An injectable magnesium-loaded lipid-based super-lubricating hydrogel microsphere, characterized in that: The hydrogel microspheres are prepared by the method according to any one of claims 1 to 8.

11. A preparation for intra-articular injection, characterized in that: The invention comprises the injectable magnesium-loaded lipid-based super-lubricating hydrogel microspheres as claimed in claim 9.

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