A lubricin biomimetic adhesive-lubricating Janus hydrogel microsphere and a preparation method and application thereof

Abstract

This invention discloses a biomimetic adhesion-lubrication Janus hydrogel microsphere, its preparation method, and its applications, belonging to the field of biomedical materials technology. The hydrogel microsphere includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivatives and alginate; the adhesion region is made of dopamine derivatives and alginate. The hydrogel microsphere contains a double cross-linked network formed by divalent cations, alginate, and hyaluronic acid derivatives, and can also load therapeutic active ingredients, preferably curcumin. The adhesion region is used for adhesion to cartilage, reducing microsphere detachment, while the lubrication region is used for lubrication, reducing joint wear. Overall, it can enhance local drug retention, improve the cartilage lubrication environment, and reduce inflammatory responses.

Description

Technical Field

[0001] This invention belongs to the field of biomedical materials technology, specifically relating to a lubricant biomimetic adhesion-lubrication Janus hydrogel microsphere, its preparation method, and its application. Background Technology

[0002] Osteoarthritis is a common degenerative joint disease, mainly characterized by damage to articular cartilage, decreased lubrication, and increased inflammation. With an aging population, rising obesity rates, and increased sports injuries, the prevalence of osteoarthritis continues to rise, becoming a major cause of pain, limited mobility, and disability. Currently, non-surgical treatments for osteoarthritis mainly include exercise therapy, physical therapy, oral analgesics, and intra-articular injections. While these can alleviate symptoms to some extent, they are unlikely to effectively stop disease progression.

[0003] Healthy articular cartilage has an extremely low coefficient of friction, and its excellent lubrication properties are essential for maintaining long-term joint load-bearing and movement functions. During the progression of osteoarthritis, cartilage wear, lubrication failure, and inflammation mutually reinforce each other, leading to continuous degradation of the cartilage matrix. Therefore, developing therapeutic materials with both lubricating and anti-inflammatory properties is crucial for slowing the progression of osteoarthritis. Hydrogel microspheres, with their injectability, biocompatibility, biodegradability, and drug loading capacity, have been used for intra-articular lubrication replenishment and anti-inflammatory drug delivery in osteoarthritis. However, traditional hydrogel microspheres are prone to detachment from the cartilage surface and dispersion within the joint cavity under joint movement and synovial fluid flushing conditions, resulting in insufficient accumulation at the lesion site, reduced lubrication, and decreased drug release efficiency. In existing isotropic hydrogel materials that simultaneously incorporate adhesion and lubrication functions, the random distribution of adhesion and lubrication molecules can easily lead to mutual shielding effects, making it difficult to fully utilize both adhesion and lubrication properties simultaneously.

[0004] Lubricants on the surface of natural cartilage exhibit domain specialization, with the central region providing boundary lubrication and the two end domains mediating cartilage surface anchorage, thus achieving stable and efficient adhesion-enhanced lubrication. Janus hydrogels, with their asymmetric structure and partitioning function, hold promise for mimicking the spatial partitioning mechanism of lubricants, achieving spatial decoupling of adhesion and lubrication functions. Besides osteoarthritis, traumatic arthritis, inflammation-related cartilage damage, lubrication disorder-related cartilage damage, meniscus injury-related joint lubrication disorders, and decreased joint inflammation and lubrication function after cartilage repair or arthroscopy are often accompanied by increased cartilage surface friction, enhanced local inflammatory response, and an increased risk of cartilage matrix degeneration. Therefore, injectable hydrogel microspheres with cartilage surface adhesion, boundary lubrication, and anti-inflammatory active ingredient delivery functions have potential applications in cartilage-related joint diseases and related joint cavity protection scenarios. Summary of the Invention

[0005] To address the aforementioned shortcomings in the prior art, this invention provides a biomimetic adhesion-lubrication Janus hydrogel microsphere, its preparation method, and its application. The hydrogel microsphere includes an adhesion region and a lubrication region. The adhesion region is used to adhere to cartilage, reducing microsphere detachment, while the lubrication region is used for lubrication, reducing joint wear. Overall, it can enhance local drug retention, improve the cartilage lubrication environment, and reduce inflammatory responses.

[0006] To achieve the above objectives, the technical solution adopted by the present invention to solve its technical problem is as follows: A biomimetic adhesion-lubrication Janus hydrogel microsphere is disclosed. The hydrogel microsphere includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivatives and alginate. The adhesion region is made of dopamine derivatives and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate, and hyaluronic acid derivatives.

[0007] Furthermore, the hydrogel microspheres have a particle size of 200~800 μm, and the volume ratio of the lubrication zone to the adhesion zone is 1-20:1.

[0008] Furthermore, the hydrogel microspheres also contain anti-inflammatory active ingredients.

[0009] Furthermore, the anti-inflammatory component is curcumin, and the mass concentration of the curcumin is 0.1-0.5%.

[0010] The preparation method of the above-mentioned biomimetic adhesion-lubrication Janus hydrogel microspheres includes the following steps: (1) Hyaluronic acid derivative, alginate and photoinitiator are dissolved in aqueous solution to prepare lubrication zone precursor solution; dopamine derivative, alginate and photoinitiator are dissolved in aqueous solution to prepare adhesion zone precursor liquid; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. (3) Collect the hydrogel droplets in a crosslinking solution containing calcium chloride, and simultaneously irradiate them with light to obtain hydrogel microspheres.

[0011] Further, in step (1), the lubrication zone precursor fluid includes the following components by mass concentration: sodium alginate 0.5-3%, methacrylamide sodium alginate 0.5-2%, methacrylamide hyaluronic acid 1-4%, photoinitiator 0.1-0.5%, and the remainder is water; The adhesion zone precursor fluid contains the following components in the following mass concentrations: sodium alginate 0.5-3%, methacrylamide sodium alginate 1-4%, dopamine methacrylamide 1-5%, photoinitiator 0.1-0.5%, and the remainder is water.

[0012] Further, in step (1), the photoinitiator is lithium phenyl-2,4,6-trimethylbenzoylphosphonate.

[0013] Furthermore, in step (2), the flow rate of the precursor liquid in the lubrication zone is 0.5-2 mL / h, the flow rate of the precursor solution in the adhesion zone is 0.05-1 mL / h, and the flow rate of the inert gas is 3-7 L / min.

[0014] Furthermore, in step (2), the flow rate of the precursor liquid in the lubrication zone is 1 mL / h, the flow rate of the precursor solution in the adhesion zone is 0.25 mL / h, and the flow rate of the inert gas is 4 L / min.

[0015] Furthermore, in step (3), the mass percentage of calcium chloride in the crosslinking solution is 1-3%; the light wavelength is 400-410nm.

[0016] The above-mentioned lubricant biomimetic adhesive-lubricating Janus hydrogel microspheres are used in the preparation of intra-articular injection formulations for the prevention, relief or treatment of cartilage damage-related joint diseases.

[0017] The above-mentioned biomimetic adhesion-lubrication Janus hydrogel microspheres are used in the preparation of surface lubrication and protection materials for artificial joints, cartilage replacement materials or joint implant materials.

[0018] Furthermore, cartilage-related joint diseases include osteoarthritis, traumatic arthritis, inflammation-related cartilage damage, or lubrication disorder-related cartilage damage.

[0019] Intra-articular injections are used to improve joint lubrication disorders, cartilage surface wear, or intra-articular inflammatory responses associated with meniscus injuries.

[0020] Intra-articular injection preparations are used for lubrication and protection of cartilage surface, anti-inflammatory protection, or delay of cartilage degeneration after cartilage repair or arthroscopic surgery.

[0021] The beneficial effects of this invention are as follows: (1) The present invention prepares lubricant biomimetic Janus hydrogel microspheres through an oil-free gas shear microfluidic platform. The Janus hydrogel microspheres are double cross-linked hydrogel microspheres composed of a calcium alginate ion cross-linking network and a methacrylamide polymer photocross-linking network.

[0022] The Janus hydrogel microspheres possess spatially separated lubrication and adhesion zones. In the lubrication zone, hyaluronic acid and its derivatives provide hydration lubrication, forming a hydration lubrication layer on the cartilage surface. In the adhesion zone, dopamine-derived catechol groups enhance wet adhesion to the cartilage surface, reducing microsphere detachment and intra-articular dispersion caused by joint movement and synovial fluid erosion. This allows the microspheres to more stably accumulate on the cartilage surface, thereby improving boundary lubrication efficiency and reducing cartilage interfacial friction.

[0023] (2) The Janus hydrogel microspheres of the present invention can be loaded with therapeutic active ingredients, preferably with anti-inflammatory active ingredients, and even more preferably with curcumin. They can release anti-inflammatory active ingredients locally in the joint cavity and synergistically intervene in the progression of cartilage damage-related joint diseases with adhesion-enhanced lubrication, thus having both lubrication and anti-inflammatory therapeutic functions.

[0024] (3) The Janus hydrogel microspheres of the present invention have good injectability, biocompatibility and biodegradability. They can be administered via intra-articular injection for minimally invasive drug delivery. They are expected to become a precise, effective and convenient treatment or protection method for cartilage damage-related joint diseases. They can also be used for lubrication and protection of the surface of artificial joints, cartilage substitutes or joint implants. Attached Figure Description

[0025] Figure 1 The diagram shows the oil-free gas shear microfluidic platform (left) and the dual-channel coaxial needle system used in this invention (right). Figure 2 The synthesis and NMR characterization of AlgMA (A) and HAMA (B) used in this invention are shown. Figure 3 The images show the morphology of Lub-Cur@Ms (A) and Adh-Cur@Ms (B) prepared in Comparative Examples 1 and 2 of this invention. Figure 4 Two-dimensional fluorescence image, three-dimensional fluorescence image, fluorescence distribution curve and bright field image of Janus-Cur@Ms1 prepared in Example 2 of the present invention; Figure 5 Two-dimensional fluorescence image, three-dimensional fluorescence image, fluorescence distribution curve and bright field image of Janus-Cur@Ms4 prepared in Example 1 of the present invention; Figure 6 Two-dimensional fluorescence image, three-dimensional fluorescence image, fluorescence distribution curve and bright field image of Janus-Cur@Ms20 prepared in Example 3 of the present invention; Figure 7 Bright-field diagrams and particle size distribution diagrams of Janus-Cur@Ms prepared under different nitrogen flow rates in Examples 4-6 of this invention; Figure 8Scanning electron microscope (SEM) images of Janus-Cur@Ms4 prepared in Example 1 of this invention and Lub-Cur@Ms and Adh-Cur@Ms prepared in Comparative Examples 1 and 2. Figure 9 The images show the morphology of Janus-Cur@Ms before (A) and after (B) injection, prepared in Example 1 of this invention. Figure 10 The figures (C) and (D) show the residue of microspheres prepared in Comparative Examples 1, 2 and Examples 1, 2 and 3 of this invention before and after rinsing on the surface of isolated cartilage. Figure 11 Ultrasound images of the adhesion of Janus-Cur@Ms4 prepared in Example 1 and Lub-Cur@Ms prepared in Comparative Example 1 to healthy joints and osteoarthritis joints in rabbits. Figure 12 The diagram shows the changes in compartment orientation of Janus-Cur@Ms4 prepared in Example 1 of this invention before (A) and after (B) adhesion to the cartilage surface; Figure 13 The diagram shows the tribological test results of the microspheres prepared in Comparative Example 3, Comparative Example 1 and Example 1 of this invention (G), the friction coefficient-time curve on the cartilage surface (H), the friction coefficient statistics on the cartilage surface (I) and the friction coefficient statistics on the titanium alloy surface (J). Figure 14 This is an in-situ degradation diagram of Janus-Cur@Ms4 prepared in Example 1 of the present invention at different time points in the rabbit joint cavity; Figure 15 The in vitro degradation curve (L) and curcumin release curve (M) of Janus-Cur@Ms4 prepared in Example 1 of the present invention. Figure 16 The images show the live / dead staining of chondrocytes in Example 1, Comparative Example 4, the normal control group, and the PBS group (A) and the CCK-8 quantitative data (B). Figure 17 The images show ROS staining of chondrocytes stimulated by IL-1β in Example 1, Comparative Example 4, normal control group, and PBS group (A) and the statistical graph of average fluorescence intensity (B). Figure 18 Immunofluorescence staining images and average fluorescence intensity statistics (E) of chondrocytes ACAN (F), COL2A1 (G), SOX9 (H) and MMP13 (I) in Example 1, Comparative Example 4, normal control group and PBS group of the present invention; Figure 19 This is an RT-qPCR quantification diagram of COL2A1, ACAN, SOX9, MMP13 and ADAMTS5 in chondrocytes of Example 1, Comparative Example 4, normal control group and PBS group of the present invention; Figure 20 These are glycosaminoglycan staining images of chondrocytes in Example 1, Comparative Example 4, normal control group, and PBS group of the present invention. Figure 21 Gross observation of the tibial and femoral articular surfaces of the knee joints in rabbits from the Sham group, PBS group, Lub-Cur@Ms group, and Janus-Cur@Ms4 group (B), ICRS gross scoring heatmap (D), and total score statistical chart (E). Figure 22 Anteroposterior and lateral X-ray images of the knee joints of rabbits in the Sham group, PBS group, Lub-Cur@Ms group and Janus-Cur@Ms4 group (C) and statistical graph of joint space width (F); Figure 23 Micro-CT three-dimensional reconstruction images (G), subchondral bone micro-CT images (H), and quantitative images of osteophyte volume (I), BMD (J), BV / TV (K), Tb.Th (L), and Tb.N (M) of rabbit knee joints in the Sham group, PBS group, Lub-Cur@Ms group, and Janus-Cur@Ms4 group were generated. Figure 24 Histological staining of femoral articular cartilage in rabbits of the Sham, PBS, Lub-Cur@Ms, and Janus-Cur@Ms4 groups (A), immunohistochemical staining of COL2A1, ACAN, and MMP13 (B), and H&E staining of major organs (C) and skin tissue after Janus-Cur@Ms4 treatment (D). Detailed Implementation

[0026] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention; that is, the described embodiments are merely some embodiments of the invention, and not all embodiments.

[0027] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0028] It should be noted that relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes the element.

[0029] The features and performance of the present invention will be further described in detail below with reference to the embodiments and accompanying drawings.

[0030] The sodium alginate (AlgMA) and hyaluronic acid (HAMA) used in the following examples and comparative examples were prepared using the following method: 1 g of alginate or 1 g of hyaluronic acid was weighed and completely dissolved in 100 mL of deionized water, and stirred at 4°C. Subsequently, under vigorous stirring, 30 mL of methacrylic anhydride was added dropwise to the alginate solution to prepare AlgMA, and 10 mL of methacrylic anhydride was added dropwise to the hyaluronic acid solution to prepare HAMA. During the reaction, the pH of the reaction system was adjusted to approximately 8 using 0.5 M sodium hydroxide solution. The reaction was vigorously stirred at 4°C for 48 h. After the reaction, the mixture was dialyzed against deionized water for 7 days using a dialysis bag with a molecular weight cutoff of 12-14 kDa to remove unreacted methacrylic anhydride. Finally, AlgMA and HAMA were obtained by lyophilization. Characterization was performed using proton nuclear magnetic resonance (NMR), and the results are as follows: Figure 2 As shown, AlgMA and HAMA successfully completed the methacrylation modification.

[0031] The main reagents used in this invention include: alginate (A2033, Sigma-Aldrich); hyaluronic acid (H909937, Macklin); methacrylic anhydride (276685, Sigma-Aldrich); dopamine methacrylamide (N303773, Aladdin); calcium chloride (C299717, Aladdin); lithium phenyl-2,4,6-trimethylbenzoylphosphonate (Engineering for Life); fluorescent polystyrene nanoparticles (200 nm, Xi'an Ruixi); curcumin (C1386, Sigma-Aldrich); hyaluronidase (Engineering for Life); PBS (HyClone); DMEM / F-12 (Gibco); type II collagenase (40508ES60, Yeason); and Transwell chambers (0.4). μm pore size, Labselect; Calcein / PI live / dead staining kit C2015M, Beyotime; CCK-8 kit K009, ZETA Life; DCFH-DA kit S0033S, Beyotime; Hematoxylin-Eosin staining kit G1120, Solarbio; Modified Safranine O / FastGreen staining kit G1371, Solarbio; Toluidine Blue staining kit G2543, Solarbio; Envision Detection Kit K5007, Dako.

[0032] The oil-free gas shear microfluidic platform used in this invention mainly consists of an electronic syringe pump, a nitrogen cylinder, a customized dual-channel coaxial needle system, a light source, and a collection tank containing CaCl2 solution. The electronic syringe pump controls the flow rate of the precursor liquid, the nitrogen cylinder provides the shear gas flow, the dual-channel coaxial needle system delivers the precursor liquid to the lubrication chamber and the adhesion chamber respectively, the light source forms a photocrosslinking network, and the CaCl2 collection tank forms an ionic crosslinking network. The dual-channel coaxial needle system includes two 30G inner needles for delivering the two precursor liquids respectively; a 14G needle is inserted from the side into the outer needle for delivering nitrogen gas. The oil-free gas shear microfluidic platform and dual-channel coaxial needle system used in this invention are as follows: Figure 1 As shown.

[0033] In this invention, mass concentration is expressed as the mass of the solute divided by the volume of the solution.

[0034] Example 1 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v of sodium alginate, 1 w / v of methacrylamide sodium alginate, 2 w / v of methacrylamide hyaluronic acid, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v of sodium alginate, 2 w / v of methacrylamide sodium alginate, 3 w / v of dopamine methacrylamide, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres (Janus-Cur@Ms4). The microspheres are washed three times with PBS before use.

[0035] Example 2 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication is provided. The hydrogel microsphere includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 1:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v of sodium alginate, 1 w / v of methacrylamide sodium alginate, 2 w / v of methacrylamide hyaluronic acid, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v of sodium alginate, 2 w / v of methacrylamide sodium alginate, 3 w / v of dopamine methacrylamide, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 1 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres (Janus-Cur@Ms1).

[0036] Example 3 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication is provided. The hydrogel microsphere includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 20:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v of sodium alginate, 1 w / v of methacrylamide sodium alginate, 2 w / v of methacrylamide hyaluronic acid, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v of sodium alginate, 2 w / v of methacrylamide sodium alginate, 3 w / v of dopamine methacrylamide, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.05 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% of calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres (Janus-Cur@Ms20).

[0037] Example 4 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v of sodium alginate, 1 w / v of methacrylamide sodium alginate, 2 w / v of methacrylamide hyaluronic acid, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v of sodium alginate, 2 w / v of methacrylamide sodium alginate, 3 w / v of dopamine methacrylamide, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 7 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride, and irradiate with light at a wavelength of 405 nm to obtain hydrogel microspheres.

[0038] Example 5 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v of sodium alginate, 1 w / v of methacrylamide sodium alginate, 2 w / v of methacrylamide hyaluronic acid, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v of sodium alginate, 2 w / v of methacrylamide sodium alginate, 3 w / v of dopamine methacrylamide, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 5 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride, and irradiate with light at a wavelength of 405 nm to obtain hydrogel microspheres.

[0039] Example 6 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v of sodium alginate, 1 w / v of methacrylamide sodium alginate, 2 w / v of methacrylamide hyaluronic acid, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v of sodium alginate, 2 w / v of methacrylamide sodium alginate, 3 w / v of dopamine methacrylamide, 0.2 w / v of curcumin, and 0.3 w / v of lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 3 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride, and irradiate with light at a wavelength of 405 nm to obtain hydrogel microspheres.

[0040] Example 7 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 0.5 w / v% sodium alginate, 0.5 w / v% sodium alginate with methacrylamide, 0.5 w / v% hyaluronic acid with methacrylamide, 0.1 w / v% curcumin, and 0.1 w / v% lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 0.5 w / v% sodium alginate, 0.5 w / v% sodium alginate with methacrylamide, 0.5 w / v% dopamine methacrylamide, 0.1 w / v% curcumin, and 0.1 w / v% lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 1% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres. The microspheres are then washed three times with PBS for later use.

[0041] Example 8 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 3 w / v sodium alginate, 3 w / v sodium methacrylamide, 5 w / v hyaluronic acid, 0.5 w / v curcumin, and 0.5 w / v lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 3 w / v sodium alginate, 5 w / v sodium methacrylamide, 5 w / v dopamine methacrylamide, 0.5 w / v curcumin, and 0.5 w / v lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 3% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres. The microspheres are then washed three times with PBS for later use.

[0042] Comparative Example 1 A hydrogel microsphere comprising a lubrication region made of a hyaluronic acid derivative and alginate; Its preparation method includes the following steps: (1) Dissolve 1.5 w / v% sodium alginate, 1 w / v% sodium alginate methacrylamide, 2 w / v% hyaluronic acid methacrylamide, 0.2 w / v% curcumin and 0.3 w / v% lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; (2) The lubricating zone precursor fluid is introduced into the inner needle of the single-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets are formed. The flow rate of the lubricating zone precursor fluid is 1 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres (Lub-Cur@Ms). The microspheres are washed three times with PBS before use.

[0043] Comparative Example 2 A hydrogel microsphere comprising an adhesion region made of a dopamine derivative and an alginate; Its preparation method includes the following steps: (1) Sodium alginate 1.5 w / v%, sodium methacrylamide 2 w / v%, dopamine methacrylamide 3 w / v%, curcumin 0.2 w / v%, lithium phenyl-2,4,6-trimethylbenzoylphosphonate 0.3 w / v% were dissolved in an aqueous solution to prepare the adhesion zone precursor solution; (2) The precursor solution of the adhesion zone is introduced into the inner needle of the coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets are formed. The flow rate of the precursor solution of the adhesion zone is 0.25 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres (Adh-Cur@Ms). The microspheres are washed three times with PBS before use.

[0044] Comparative Example 3 An isotropic ionic crosslinked calcium alginate-loaded curcumin microsphere is prepared by the following steps: (1) Dissolve / disperse 1.5 w / v% sodium alginate and 0.2 w / v% curcumin in deionized water and mix thoroughly to obtain an isotropic precursor solution; (2) The precursor solution is introduced into the inner needle of the single-channel coaxial needle system, and an inert gas is delivered through the outer needle. Under the shear force of the inert gas, hydrogel droplets are formed. The flow rate of the precursor solution is 1 mL / h and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride, and perform only ionic crosslinking (without light exposure) to obtain isotropic ionic crosslinked calcium alginate-loaded curcumin microspheres (Cur@Ms). The microspheres are washed three times with PBS before use.

[0045] Comparative Example 4 A biomimetic adhesion-lubrication Janus hydrogel microsphere for lubrication includes a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivative and alginate. The adhesion region is made of dopamine derivative and alginate. The hydrogel microsphere includes a double cross-linked network formed by divalent cations, alginate and hyaluronic acid derivative. The volume ratio of the lubrication region to the adhesion region is 4:1. Its preparation method includes the following steps: (1) Dissolve 1.5 w / v% sodium alginate, 1 w / v% sodium methacrylamide, 2 w / v% hyaluronic acid methacrylamide, and 0.3 w / v% lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare a lubrication zone precursor solution; dissolve 1.5 w / v% sodium alginate, 2 w / v% sodium methacrylamide, 3 w / v% dopamine methacrylamide, and 0.3 w / v% lithium phenyl-2,4,6-trimethylbenzoylphosphonate in an aqueous solution to prepare an adhesion zone precursor solution; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. The flow rate of the lubrication zone precursor liquid is 1 mL / h, the flow rate of the adhesion zone precursor solution is 0.25 mL / h, and the flow rate of the inert gas is 4 L / min. (3) Collect hydrogel droplets in a crosslinking solution containing calcium chloride, with a mass percentage of 2% calcium chloride. Simultaneously, irradiate with light at a wavelength of 405 nm to prepare hydrogel microspheres (Janus@Ms4). The microspheres are washed three times with PBS before use.

[0046] Test case I. Methods 1. The Janus structure characterization method in this invention is as follows: 0.2% (w / v) of fluorescent polystyrene nanoparticles of different colors are added to the precursor liquid of the lubrication compartment and the precursor liquid of the adhesion compartment, respectively, and the compartment structure of the microsphere is observed by an inverted fluorescence microscope and a confocal laser scanning microscope.

[0047] 2. The cartilage adhesion performance test method in this invention is as follows: microspheres are applied to the surface of isolated rabbit tibial plateau cartilage, rinsed with a total amount of 1 mL of PBS, and images before and after rinsing are taken and the percentage of residual microspheres is calculated; the in vivo adhesion test is performed by injecting PBS containing microspheres into the knee joint of healthy rabbits or the knee joint of osteoarthritis, and performing ultrasound examination after the rabbits move freely for 2 hours.

[0048] 3. The lubrication performance testing method in this invention is as follows: Different groups of hydrogel microspheres are dispersed in PBS at a concentration of 10% (w / v), and the coefficient of friction is tested using a Universal Mechanical Tester. Isolated cartilage tissue and a titanium alloy disk are used as the lower surface of the friction pair, respectively, and a polyethylene sphere with a diameter of 8 mm (elastic modulus 1 GPa, Poisson's ratio 0.4) is used as the upper surface. The polyethylene sphere slides back and forth on the lower surface, and the coefficient of friction during the sliding process is recorded. The load is 1 N (to simulate the maximum physiological pressure of approximately 25 MPa at the knee joint), the amplitude is 8 mm, the frequency is 1 Hz, and the test time is 1800 s.

[0049] 4. The method for evaluating chondrocyte function in this invention is as follows: Primary chondrocytes were stimulated with 10 ng / mL IL-1β to simulate the inflammatory microenvironment of osteoarthritis. Microspheres were placed in the upper chamber of a Transwell, and chondrocytes were cultured in the lower chamber. The amount of microspheres added was 10% of the system liquid mass. The normal control group consisted of chondrocytes that were not stimulated with IL-1β and not treated with microspheres; the PBS group consisted of chondrocytes that were stimulated with IL-1β and then treated with PBS; the Janus@Ms4 group consisted of chondrocytes that were stimulated with IL-1β and then treated with Janus@Ms4 prepared in Comparative Example 4; and the Janus-Cur@Ms4 group consisted of chondrocytes that were stimulated with IL-1β and then treated with Janus-Cur@Ms4 prepared in Example 1. After intervention, the biocompatibility, antioxidant stress effect, and influence on chondrocyte anabolic and catabolitic metabolism of the microspheres were evaluated using live / dead cell staining, CCK-8 staining, DCFH-DA staining, immunofluorescence staining, RT-qPCR, and glycosaminoglycan staining.

[0050] 5. The method for evaluating the therapeutic effect of osteoarthritis in vivo in this invention is as follows: A right knee osteoarthritis model was established using 6-month-old male New Zealand rabbits via a modified Hulth method. Intra-articular injection intervention began one week after modeling, with each injection volume being 250 μL. The microsphere concentration in the treatment group was 100 mg / mL, injected once every two weeks for a total of four times. Samples were collected at week 8 post-operation. The Sham group was the sham-operated group; the PBS group was the group treated with intra-articular injection of PBS after osteoarthritis modeling; the Lub-Cur@Ms group was the group treated with Lub-Cur@Ms prepared in Comparative Example 1 after osteoarthritis modeling; and the Janus-Cur@Ms4 group was the group treated with Janus-Cur@Ms4 prepared in Example 1 after osteoarthritis modeling. The therapeutic effect and biosafety were evaluated using gross observation, ICRS scoring, X-ray, micro-CT, histological staining, immunohistochemistry, and H&E staining of major organs.

[0051] II. Results 1. The compartmental structure of the microspheres was observed using fluorescence microscopy and confocal laser scanning microscopy. Using Janus-Cur@Ms4 from Example 1, the results are as follows: Figure 5 As shown, Janus-Cur@Ms4 has a distinct dual-chamber structure, with the adhesion chambers distributed in a sub-hemispherical cap shape and the lubrication chambers occupying a larger volume.

[0052] 2. Janus-Cur@Ms4 and Lub-Cur@Ms and Adh-Cur@Ms prepared in Comparative Examples 1 and 2 were observed using scanning electron microscopy. The results are as follows: Figure 8 As shown. The morphology of Janus-Cur@Ms4 prepared in Example 1 was observed before and after injection, and the results are as follows. Figure 9 As shown. From Figure 8 It can be seen that Janus-Cur@Ms4 exhibits surface morphology characteristics of both lubrication and adhesion chambers. From Figure 9 It can be seen that Janus-Cur@Ms4 retains its structural integrity after injection, indicating that it has good injectability.

[0053] 3. Taking the Janus hydrogel microspheres in Examples 1-3 as examples, fluorescence and bright-field observations were performed on Janus hydrogel microspheres with different compartment ratios. The results are as follows: Figure 4-6 As shown, Janus-Cur@Ms1 has two compartments with roughly equal volumes; the adhesion compartment in Janus-Cur@Ms4 is subhemispherical; and the adhesion compartment in Janus-Cur@Ms20 is further reduced in size. Therefore, it can be seen that the volume ratio of the lubrication compartment and the adhesion compartment in Janus hydrogel microspheres can be controlled by changing the flow rates of the precursor solutions for the lubrication and adhesion compartments.

[0054] 4. Taking the Janus hydrogel microspheres in Examples 4-6 as examples, Janus-Cur@Ms prepared under different nitrogen flow rates were observed using a bright-field microscope, and the particle size distribution was statistically analyzed. The results are as follows: Figure 7 As shown, when the nitrogen flow rate is reduced from 7 L / min to 3 L / min, the particle size of Janus-Cur@Ms gradually increases, indicating that the present invention can achieve controllable preparation of Janus hydrogel microspheres by adjusting the nitrogen flow rate.

[0055] 5. The morphology of Lub-Cur@Ms prepared in Comparative Example 1 is as follows: Figure 3 As shown in Figure A, the scanning electron microscope image is as follows: Figure 8 As shown.

[0056] 6. The morphology of Adh-Cur@Ms prepared in Comparative Example 2 is as follows: Figure 3 As shown in B, the scanning electron microscope image is as follows. Figure 8 As shown. By Figure 8 It can be seen that the surface wrinkles of Lub-Cur@Ms are relatively wide and smooth, the surface wrinkles of Adh-Cur@Ms are relatively rough, and Janus-Cur@Ms4 has the surface morphology characteristics of both compartments.

[0057] 7. The cartilage adhesion ability of the hydrogel microspheres in Examples 1-3 and Comparative Examples 1-2 was evaluated, and the results are as follows: Figure 10-12 As shown.

[0058] Figure 10 The images show the residue status and residue rate statistics of Adh-Cur@Ms, Janus-Cur@Ms1, Janus-Cur@Ms4, Janus-Cur@Ms20, and Lub-Cur@Ms on the surface of isolated rabbit cartilage before and after rinsing. The results show that Adh-Cur@Ms, Janus-Cur@Ms1, and Janus-Cur@Ms4 have higher residue rates, significantly higher than Janus-Cur@Ms20 and Lub-Cur@Ms. Among them, Janus-Cur@Ms4 maintains strong cartilage adhesion ability despite a smaller adhesion compartment volume, indicating that it can effectively balance the proportion of cartilage adhesion and lubrication compartments.

[0059] Figure 11 Ultrasound images showing the adhesion of Lub-Cur@Ms and Janus-Cur@Ms4 within the joints of healthy rabbits and osteoarthritis-affected rabbits. It can be seen that the Janus-Cur@Ms4 group exhibits punctate hyperechoic signals on the articular cartilage surface, while no obvious deposition signal is observed in the Lub-Cur@Ms group.

[0060] Figure 12 The image shows that Janus-Cur@Ms4 adheres primarily to the cartilage surface, primarily in the lubrication compartment, indicating that its adhesion compartment can face the cartilage surface and achieve anchorage. This demonstrates that Janus-Cur@Ms4 can achieve intra-articular cartilage surface retention through its adhesion compartments.

[0061] 8. The lubrication ability of the microspheres prepared in Example 1, Comparative Example 1, and Comparative Example 3 was evaluated, and the results are as follows: Figure 13 As shown.

[0062] Figure 13 The diagram shows the tribological test results (G), the friction coefficient-time curves of Cur@Ms, Lub-Cur@Ms, and Janus-Cur@Ms4 on the cartilage surface (H), the statistical graph of the friction coefficient on the cartilage surface (I), and the statistical graph of the friction coefficient on the titanium alloy surface (J).

[0063] On the cartilage surface: the friction coefficient of Lub-Cur@Ms is lower than that of Cur@Ms, indicating that the lubrication chamber components have a lubricating effect; the friction coefficient of Janus-Cur@Ms4 is even lower than that of Lub-Cur@Ms, indicating that the adhesion chamber can reduce the off-target movement of microspheres during the friction process, thereby enhancing the boundary lubrication effect of the lubrication chamber.

[0064] On the titanium alloy surface: the order of friction coefficients of the three groups of microspheres is consistent with that of the cartilage surface, namely Cur@Ms > Lub-Cur@Ms > Janus-Cur@Ms4, with Janus-Cur@Ms4 still exhibiting the lowest friction coefficient on the titanium alloy surface. Therefore, the Janus hydrogel microspheres of this invention can not only reduce friction at the natural cartilage interface but also provide effective boundary lubrication on the surfaces of artificial materials such as titanium alloys.

[0065] 9. The degradation and curcumin release of Janus-Cur@Ms4 prepared in Example 1 were evaluated, and the results are as follows: Figure 14-15 As shown. Figure 14 The study showed that Janus-Cur@Ms4 gradually degraded in the rabbit joint cavity over time, and by day 14, the microspheres lost their complete spherical structure and degraded into fragments. Figure 15 The results showed that Janus-Cur@Ms4 gradually degrades over time in vitro and can slowly and continuously release curcumin. This demonstrates that Janus-Cur@Ms4 possesses both degradability and the ability to continuously release curcumin.

[0066] 10. The effects of the microspheres prepared in Example 1 and Comparative Example 4 on chondrocyte function were evaluated, and the results are as follows: Figure 16-20 As shown.

[0067] Figure 16 The images show the live / dead staining patterns and CCK-8 quantitative data of chondrocytes in the normal control group, PBS group, Janus@Ms4 group, and Janus-Cur@Ms4 group. It can be seen that no obvious cell death was observed in chondrocytes after different microsphere treatments, indicating that the microspheres have good biocompatibility. Figure 17 The above images show ROS staining patterns and average fluorescence intensity statistics of IL-1β-stimulated chondrocytes in each group. It can be seen that Janus@Ms4 did not significantly reduce ROS levels, while Janus-Cur@Ms4 could significantly reduce intracellular ROS accumulation. Figure 18 The images show the immunofluorescence staining and quantitative results of ACAN, COL2A1, SOX9, and MMP13 in chondrocytes of the above groups. Figure 19 The results of RT-qPCR quantification of related genes showed that Janus-Cur@Ms4 upregulated the expression of COL2A1, ACAN and SOX9, and downregulated the expression of MMP13 and ADAMTS5. Figure 20The image shows glycosaminoglycan staining, indicating that Janus-Cur@Ms4 can improve the reduced glycosaminoglycan production caused by inflammatory stimulation. This demonstrates that Janus-Cur@Ms4 can improve chondrocyte function under inflammatory conditions through curcumin loading.

[0068] 11. The therapeutic effects of the microspheres prepared in Example 1 and Comparative Example 1 on a rabbit knee osteoarthritis model were evaluated, and the results are as follows: Figure 21-24 As shown.

[0069] Figure 21 Gross images of the tibial and femoral articular surfaces of the knee joints in rabbits from the Sham, PBS, Lub-Cur@Ms, and Janus-Cur@Ms4 groups, along with ICRS gross scoring heatmaps and total score statistics, are shown. It can be seen that the PBS group exhibited severe cartilage erosion, subchondral bone exposure, osteophyte formation, and joint deformity; the Lub-Cur@Ms group showed less cartilage damage than the PBS group; the Janus-Cur@Ms4 group had a smoother and more intact cartilage surface, with no obvious defects or osteophyte formation, and its ICRS score was superior to both the PBS and Lub-Cur@Ms groups.

[0070] Figure 22 The images show anteroposterior and lateral X-ray images of the rabbit knee joints and statistical charts of joint space width for each group. It can be seen that Janus-Cur@Ms4 can delay joint space narrowing caused by osteoarthritis.

[0071] Figure 23 The images show the three-dimensional reconstruction images of the rabbit knee joints using micro-CT, the micro-CT images of the subchondral bone, and related quantitative maps for each group. It can be seen that the Janus-Cur@Ms4 group exhibited less osteophyte formation and improved abnormal subchondral bone remodeling.

[0072] Figure 24 The images show histological staining, immunohistochemical staining, and safety evaluation of the femoral articular cartilage in each group of rabbits. Histological staining revealed discontinuous cartilage surface and significantly reduced matrix staining in the PBS group; damage was reduced in the Lub-Cur@Ms group; and the cartilage structure and matrix staining were well preserved in the Janus-Cur@Ms4 group. Immunohistochemical results showed that Janus-Cur@Ms4 could restore COL2A1 and ACAN expression and inhibit MMP13 expression, indicating that it can promote cartilage matrix synthesis and inhibit cartilage matrix degradation. H&E staining of major organs and skin tissues showed no obvious inflammatory cell infiltration or pathological changes, indicating that Janus-Cur@Ms4 has good in vivo biocompatibility.

[0073] In summary, this invention prepares biomimetic adhesion-lubrication Janus hydrogel microspheres using an oil-free gas shear microfluidic platform. These microspheres possess independent and asymmetrically distributed lubrication and adhesion regions. The lubrication regions are provided with boundary lubrication by a HAMA-containing polysaccharide hydrogel network, while the adhesion regions are provided with cartilage wet adhesion by a dopamine-methylacrylamide-containing hydrogel network. The Janus spatial partitioning avoids mutual shielding of adhesion and lubrication functions in isotropic materials.

[0074] These Janus hydrogel microspheres are regularly shaped, controllable in size, injectable, and biodegradable, and can load and release therapeutic active ingredients; in a preferred embodiment, the therapeutic active ingredient is curcumin. Compared with simple lubricating microspheres, Janus-Cur@Ms4 can stably remain on the cartilage surface, reducing off-target dispersion caused by joint movement and synovial fluid erosion, thereby enhancing the lubrication effect; compared with unloaded Janus microspheres, Janus-Cur@Ms4 can reduce the ROS level of chondrocytes under inflammatory conditions, restore chondrocyte anabolic metabolism and inhibit catabolism; in a rabbit knee osteoarthritis model, Janus-Cur@Ms4 can alleviate cartilage damage, delay joint space narrowing, inhibit osteophyte formation and abnormal subchondral bone remodeling, and reduce cartilage matrix loss.

[0075] In summary, this invention proposes a method for preparing lubricant-inspired cartilage-adhesive Janus hydrogel microspheres using an oil-free gas shear microfluidic platform. These microspheres can be loaded with therapeutic active ingredients, preferably curcumin, and through adhesion, enhance the synergistic effect of lubrication and anti-inflammatory active ingredient release, achieving comprehensive intervention on pathological aspects such as cartilage lubrication failure, inflammatory response, and matrix degeneration in cartilage injury-related joint diseases. This method is expected to become a precise, effective, and convenient intra-articular injection carrier for intra-articular treatment or protection.

[0076] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A biomimetic adhesion-lubrication Janus hydrogel microsphere containing lubricant, characterized in that, The hydrogel microspheres include a lubrication region and an adhesion region. The lubrication region is made of hyaluronic acid derivatives and alginate. The adhesion region is made of dopamine derivatives and alginate. The hydrogel microspheres contain a double cross-linked network formed by divalent cations, alginate, and hyaluronic acid derivatives.

2. The lubricant-inspired biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 1, characterized in that, The hydrogel microspheres have a particle size of 200~800 μm, and the volume ratio of the lubrication zone to the adhesion zone is 1-20:

1.

3. The lubricant-inspired biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 1, characterized in that, The hydrogel microspheres also contain anti-inflammatory active ingredients.

4. The lubricant-inspired biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 3, characterized in that, The anti-inflammatory component is curcumin, and the mass concentration of curcumin is 0.1-0.5%.

5. The method for preparing the lubricant-inspired biomimetic adhesion-lubrication Janus hydrogel microspheres according to any one of claims 1-4, characterized in that, Includes the following steps: (1) Hyaluronic acid derivative, alginate and photoinitiator are dissolved in aqueous solution to prepare lubrication zone precursor solution; dopamine derivative, alginate and photoinitiator are dissolved in aqueous solution to prepare adhesion zone precursor liquid; (2) The lubrication zone precursor liquid and the adhesion zone precursor liquid are respectively introduced into the two inner needles of the dual-channel coaxial needle system. At the same time, inert gas is delivered through the outer needle. Under the action of the inert gas shear force, hydrogel droplets with an asymmetric dual-zone structure are formed. (3) Collect the hydrogel droplets in a crosslinking solution containing calcium chloride, and simultaneously irradiate them with light to obtain hydrogel microspheres.

6. The method for preparing the lubricant biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 5, characterized in that, In step (1), the lubrication zone precursor fluid includes the following components in mass concentration: sodium alginate 0.5-3%, methacrylamide sodium alginate 0.5-2%, methacrylamide hyaluronic acid 1-4%, photoinitiator 0.1-0.5%, and the remainder is water; The adhesion zone precursor fluid comprises the following components in the indicated mass concentrations: sodium alginate 0.5-3%, methacrylamide sodium alginate 1-4%, dopamine methacrylamide 1-5%, photoinitiator 0.1-0.5%, and the remainder being water.

7. The method for preparing the lubricant biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 6, characterized in that, In step (1), the photoinitiator is lithium phenyl-2,4,6-trimethylbenzoylphosphonate.

8. The method for preparing the lubricant biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 5, characterized in that, In step (2), the flow rate of the precursor liquid in the lubrication zone is 0.5-2 mL / h, the flow rate of the precursor solution in the adhesion zone is 0.05-1 mL / h, and the flow rate of the inert gas is 3-7 L / min.

9. The method for preparing the lubricant biomimetic adhesion-lubrication Janus hydrogel microspheres as described in claim 5, characterized in that, In step (3), the mass percentage of calcium chloride in the crosslinking solution is 1-3%; the light wavelength is 400-410nm.

10. The use of the lubricant biomimetic adhesion-lubricating Janus hydrogel microspheres according to any one of claims 1-4 in the preparation of intra-articular injection formulations for the prevention, relief or treatment of cartilage damage-related joint diseases.

11. The use of the lubricant biomimetic adhesion-lubricating Janus hydrogel microspheres according to any one of claims 1-4 as a surface lubrication and protective material for artificial joints, cartilage substitutes or joint implants.

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