Anti-osteoarthritis nanonuclear isotope gel and its preparation method and application

Abstract

The application provides an anti-osteoarthritis nanometer radionuclide gel and a preparation method and application thereof 177 Lu and adenosine monophosphate coordination self-assembly 177 Lu-labeled metal organic framework, chitosan forms a temperature-sensitive gel in the case of glycerophosphate sodium as a crosslinking agent. The temperature-sensitive gel encapsulates 177 Lu-labeled metal organic framework not only helps to prolong the retention time of radionuclide in the local joint cavity, and by utilizing the temperature response performance of the high molecular material in the temperature-sensitive gel, a semi-solid gel is formed in situ in the joint, which not only has good viscoelasticity, lubrication and buffering of the joint, but also can play a barrier effect to prevent the diffusion of inflammatory substances in the joint cavity, and the degradation components of chitosan can stimulate the production of collagen and glycosaminoglycan of chondrocytes, and play a role in protecting and repairing cartilage. Therefore, the nanometer radionuclide gel is an excellent RSO drug delivery system, and has good biocompatibility and degradability.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of nanobiomedicine, and particularly relates to an anti-osteoarthritis nanometer radionuclide gel and a preparation method and application thereof. BACKGROUND

[0002] With the in-depth exploration of the pathological mechanism of arthritis and the rapid development of nuclear medicine diagnosis and treatment technology, researchers have found that radionuclides can effectively inhibit joint synovial inflammation and show excellent efficacy in the treatment of arthritic diseases. Therefore, clinical workers have developed radioactive synovectomy (RSO), the principle of which is to inject radionuclide preparations into the joint cavity to inhibit the proliferation of inflammatory cells in the joint synovium by using the beta rays emitted by the radionuclides and to reduce the destruction of cartilage and bone. For RSO, appropriate radionuclides and carrier materials need to be selected. 90 Y、 186 Re、 169 Er、 32 P、 177 The commonly used radionuclides such as Y, Re, Er, P and Lu can emit beta rays and have appropriate penetration depth and half-life; inorganic colloid materials such as sulfur colloid, chromium phosphate and iron hydroxide can be effectively combined with radionuclides and are commonly used radionuclide carriers for clinical RSO treatment.

[0003] However, RSO still has many challenges in practical application: first, the commonly used inorganic colloid materials in clinical practice have certain toxicity and poor stability, and radionuclides are prone to leakage into the blood circulation system and lymphatic system, causing liver and kidney damage and bone marrow suppression; second, the required dose of radionuclides for RSO treatment is too large, and under the effective dose, normal joint tissues such as cartilage will be damaged to a certain extent; in addition, osteoarthritis is the result of joint cartilage damage and synovial inflammation, and RSO treatment alone can only prevent further damage to the cartilage and is difficult to repair damaged cartilage. Therefore, it is particularly important to achieve long-term retention of low-dose radionuclides in the joint cavity, reduce systemic toxicity and joint damage on the basis of inhibiting inflammation, and protect and repair damaged cartilage. SUMMARY

[0004] To solve the above technical problems, the application provides an anti-osteoarthritis nanometer radionuclide gel and a preparation method and application thereof, and the purpose is to construct a safe and effective radionuclide nanomedicine for treating osteoarthritis. The nanometer radionuclide gel system for joint cavity injection can inhibit the proliferation of inflammatory cells in the joint synovium on the one hand by using the beta rays emitted by the radionuclides, and on the other hand by using the nanometer gel system to reduce the systemic toxicity of the radionuclides and protect the normal joint tissues such as cartilage. 177Lu coordinates with adenosine monophosphate (AMP) to form a solid-state crosslinker, which is conducive to prolonging the residence time of radionuclides in the local joint cavity; on the other hand, the warm-sensitive gel formed by chitosan (CS) in glycerophosphate sodium (GP) as a crosslinking agent can form a semi-solid gel in situ in the joint cavity, which not only has good viscoelasticity, lubrication and buffering of joints, but also can play a barrier role to prevent the spread of inflammatory substances in the joint cavity, and the degradation components of CS can stimulate chondrocytes to produce collagen and glycosaminoglycan, thereby playing a role in protecting and repairing cartilage.

[0005] In order to achieve the above-mentioned purpose, the present application firstly provides a kind of anti-osteoarthritis nano radionuclide gel, including radioactive nuclide 177 Lu labeled metal organic nanoframe and warm-sensitive gel;The 177 Lu labeled metal organic frame is prepared by 177 Lu and adenosine monophosphate coordinate self-assembly, and the warm-sensitive gel is prepared by chitosan and glycerophosphate sodium.

[0006] As preferred, the mass ratio of chitosan and glycerophosphate sodium is 1:14-20.

[0007] As preferred, the particle size of the organic nanoframe is 132±3.4nm.

[0008] Based on a general inventive concept, the present application also provides a preparation method of an anti-osteoarthritis nano radionuclide gel, comprising the following steps:

[0009] S1, uniformly mix bovine serum albumin and adenosine monophosphate in ultrapure water, and slowly add 177 Lu 3+ solution continues to be stirred, and the lower layer precipitate is collected after centrifugation;

[0010] S2, disperse the precipitate obtained in step S1 into ultrapure water, add bovine serum albumin again, stir and mix uniformly, and ultrasonically redissolve under ice bath condition to obtain 177 Lu labeled metal organic nanoframe 177 (Lu / AMP@MOFs);

[0011] S3, dissolve chitosan and glycerophosphate sodium in hydrochloric acid solution and ultrapure water respectively, slowly add chitosan solution to glycerophosphate sodium solution under ice bath condition, and add the solution obtained in step S2, stir and mix uniformly to obtain nano radionuclide gel 177 (Lu / AMP@CS / GP).

[0012] As preferred, the solution of 177 Lu 3+ in step S1 is 177 LuCl3 solution, and the177 Lu 3+ The molar ratio of the solution to adenosine monophosphate is 1:5.

[0013] Preferably, the stirring time in the step S2 is 10 min, and the ice bath ultrasonic power is 100 W.

[0014] Preferably, the mass ratio of the bovine serum albumin added in the steps S1 and S2 is 1:1.

[0015] Preferably, the concentration of the chitosan solution and the glycerophosphate sodium solution in the step S3 is 2:55, and the volume ratio is 13:7.

[0016] Based on the overall inventive concept, the application further provides an application of the anti-osteoarthritis nanometer radionuclide gel in the treatment of osteoarthritis.

[0017] Preferably, the anti-osteoarthritis nanometer radionuclide gel is injected into the joint cavity.

[0018] The treatment principle of the anti-osteoarthritis nanometer radionuclide gel of the application is as follows:

[0019] The nanometer radionuclide gel system for joint cavity injection can on the one hand 177 Lu and AMP are coordinated to form a metal-organic nanoframe, which is conducive to prolonging the retention time of the radionuclide in the local joint cavity; on the other hand, the temperature-sensitive gel formed by CS under the regulation of GP as a crosslinking agent can be converted from a liquid state to a semi-solid gel in situ in the joint cavity through temperature corresponding performance, and the gel not only has good viscoelasticity, lubrication and buffering characteristics of the joint, but also can play a barrier role to prevent the diffusion of inflammatory substances in the joint cavity, and the degradation components of CS can stimulate chondrocytes to produce collagen and glycosaminoglycan, thereby playing a role in protecting and repairing cartilage. Low-dose 177 Lu is retained in the joint cavity for a long time, and β rays emitted by the radionuclide are used to inhibit the proliferation of inflammatory cells of the synovial membrane of the joint and reduce the destruction of cartilage and bone, thereby reducing the systemic toxicity on the basis of inhibiting joint inflammation and protecting and repairing damaged cartilage.

[0020] Compared with the prior art, the application has the following beneficial effects:

[0021] 1. The nano-radionuclide gel provided by the application avoids the adverse effects of fluid mobility by forming a semi-solid gel in situ in the joint, which is converted into a gel at human body temperature, facilitating better monitoring and control; the semi-solid gel formed in the joint cavity not only lubricates and buffers the joint to prevent joint adhesion, but also plays a barrier role to prevent the spread of inflammatory substances in the joint cavity. At the same time, the degradation component of the gel, glucosamine, can stimulate chondrocytes to produce collagen and glycosaminoglycan, protecting and repairing damaged cartilage.

[0022] 2. The application utilizes 177 Lu and AMP to form metal nano-organic frameworks through coordination self-assembly, which has a high radionuclide labeling rate and labeling stability, and emits beta rays to inhibit the proliferation of inflammatory cells in the synovial membrane of the joint and reduce cartilage and bone destruction; at the same time, the application utilizes CS to form a temperature-sensitive gel under the regulation of GP to encapsulate the metal nano-organic framework, which reduces the porosity of the CS / GP semi-solid gel after encapsulation, further avoiding 177 Lu leakage risk, which can avoid the toxicity caused by 177 Lu leakage, and the retention amount of 177 Lu in the joint cavity is more than 85% after 16 days of encapsulation, realizing long-term retention of radionuclides in the joint cavity, which is an excellent RSO drug delivery system that can avoid damage to normal tissues or organs, and the carrier components are non-toxic substances, and there is no significant change in the body weight of rats during the medication period, and the radionuclides are mainly concentrated in the joint cavity, and there is no obvious pathological change in other organs, which has good biocompatibility and degradability.

[0023] 3. The preparation process of the anti-osteoarthritis nano-radionuclide gel is rapid and simple, the reaction process is controllable and green, no additional excipients or organic solvents are added, and no other impurities are introduced. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings described below are only some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative labor.

[0025] Figure 1 The nano-radionuclide gel obtained by detecting Example 1 of the application 177 Lu / AMP@MOFs particle size distribution graph;

[0026] Figure 2 The appearance of Lu / AMP@MOFs after being placed for 0-24h 177 The transmission electron microscope imaging of Lu / AMP@MOFs after being placed for 0-24h

[0027] Figure 3 The influence of different volume ratios of CS / GP on the phase transition temperature of Lu / AMP@CS / GP in the experimental example 3 of the present application 177 The phase transition temperature influence change graph of Lu / AMP@CS / GP

[0028] Figure 4 The appearance of CS / GP, Lu@CS / GP and Lu / AMP@CS / GP under different temperature conditions obtained in the experimental example 4 of the present application 177 The appearance of CS / GP, Lu@CS / GP and Lu / AMP@CS / GP under different temperature conditions obtained in the experimental example 4 of the present application 177 The appearance of CS / GP, Lu@CS / GP and Lu / AMP@CS / GP under different temperature conditions obtained in the experimental example 4 of the present application

[0029] Figure 5 The scanning electron microscope imaging of CS / GP and Lu / AMP@CS / GP obtained in the experimental example 5 of the present application; A is the electron microscope imaging of CS / GP semi-solid gel; B is the electron microscope imaging of CS / GP loaded with Lu / AMP@MOFs 177 The scanning electron microscope imaging of CS / GP and Lu / AMP@CS / GP obtained in the experimental example 5 of the present application; A is the electron microscope imaging of CS / GP semi-solid gel; B is the electron microscope imaging of CS / GP loaded with Lu / AMP@MOFs 177 The scanning electron microscope imaging of CS / GP and Lu / AMP@CS / GP obtained in the experimental example 5 of the present application; A is the electron microscope imaging of CS / GP semi-solid gel; B is the electron microscope imaging of CS / GP loaded with Lu / AMP@MOFs

[0030] Figure 6 The in vitro release curve of Lu / AMP@CS / GP obtained in the experimental example 6 of the present application 177 The in vitro release curve of Lu / AMP@CS / GP obtained in the experimental example 6 of the present application

[0031] Figure 7 The SPECT-CT imaging of Lu, Lu@CS / GP and Lu / AMP@CS / GP after being injected into the joint cavity of rats obtained in the experimental example 7 of the present application 177 The SPECT-CT imaging of Lu, Lu@CS / GP and Lu / AMP@CS / GP after being injected into the joint cavity of rats obtained in the experimental example 7 of the present application 3+ The SPECT-CT imaging of Lu, Lu@CS / GP and Lu / AMP@CS / GP after being injected into the joint cavity of rats obtained in the experimental example 7 of the present application 177 The SPECT-CT imaging of Lu, Lu@CS / GP and Lu / AMP@CS / GP after being injected into the joint cavity of rats obtained in the experimental example 7 of the present application 177 The SPECT-CT imaging of Lu, Lu@CS / GP and Lu / AMP@CS / GP after being injected into the joint cavity of rats obtained in the experimental example 7 of the present application

[0032] Figure 8 The Lu retention amount in the joint cavity within 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 8 of the present application 177 The Lu retention amount in the joint cavity within 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 8 of the present application 3+ The Lu retention amount in the joint cavity within 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 8 of the present application 177 The Lu retention amount in the joint cavity within 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 8 of the present application 177 The Lu retention amount in the joint cavity within 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 8 of the present application 177 The Lu retention amount in the joint cavity within 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 8 of the present application

[0033] Figure 9 The Lu distribution amount in each tissue and organ after 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 9 of the present application 177 The Lu distribution amount in each tissue and organ after 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 9 of the present application 3+ The Lu distribution amount in each tissue and organ after 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 9 of the present application 177 The Lu distribution amount in each tissue and organ after 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 9 of the present application 177 The Lu distribution amount in each tissue and organ after 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 9 of the present application 177 The Lu distribution amount in each tissue and organ after 16 days of Lu, Lu@CS / GP and Lu / AMP@CS / GP obtained in the experimental example 9 of the present application

[0034] Figure 10 The results obtained by testing in Experimental Example 10 of the present invention are as follows: 177 Safranin O-fast green staining and TNF-α immunohistochemical staining of bone and joints of osteoarthritis rats after Lu / AMP@CS / GP treatment;

[0035] Figure 11 The results obtained by testing in Experimental Example 11 of the present invention are as follows: 177 Body weight changes in osteoarthritis rats after Lu / AMP@CS / GP treatment;

[0036] Figure 12 The results obtained by testing in Experimental Example 12 of the present invention are as follows: 177 H&E staining of major organs after Lu / AMP@CS / GP treatment. DETAILED DESCRIPTION

[0037] In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, a detailed description will be given below with reference to the accompanying drawings and specific embodiments.

[0038] The following examples are provided to illustrate the present invention but are not intended to limit the scope of the present invention. Without departing from the spirit and substance of the present invention, modifications or substitutions made to the methods, steps or conditions of the present invention are within the scope of the present invention.

[0039] Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art; unless otherwise specified, the reagents used in the examples are commercially available.

[0040] Example 1

[0041] preparation 177 Lu / AMP@CS / GP

[0042] S1. Dissolve 10 mg of bovine serum albumin (BSA) in 0.8 mL of ultrapure water, add 100 μL of adenosine monophosphate (AMP, 500 mM), mix well, and slowly add 100 μL of LuCl3 solution (100 mM, containing 2 mCi 177 Lu), continue stirring for 10 min, centrifuge (10000 rpm, 10 min), discard the supernatant, and collect the precipitate;

[0043] S2. The precipitate obtained in step S1 was dispersed into 1 mL of ultrapure water, and 10 mg of BSA was added, stirred and dissolved, and crushed with a probe ultrasonic instrument under ice bath conditions at a power of 100 W to obtain 177 Lu / AMP@MOFs.

[0044] S3. Dissolve 200 mg chitosan (CS) in 10 mL HCl (0.15 M) to prepare a 2% CS solution. Dissolve 5.5 g sodium glycerophosphate (GP) in 10 mL ultrapure water to prepare a 55% GP solution. In an ice bath, slowly add 13 mL of CS solution to 7 mL of GP solution and add 177 The Lu / AMP@MOFs nanoparticle solution is stirred and mixed to obtain the nuclide nanomedicine. 177 Lu / AMP@CS / GP.

[0045] According to the nuclide nanomedicine obtained in Example 1, the detection 177 The physicochemical properties and in vitro and in vivo pharmacodynamic characteristics of Lu / AMP@CS / GP, specifically including nanoparticle morphology, particle size, thermosensitive gel morphology and in vitro release behavior, and in vivo efficacy in treating osteoarthritis, are specifically described using the nuclide nanodrug obtained in Example 1.

[0046] Experimental Example 1

[0047] Investigation 177 Particle size of Lu / AMP@MOFs

[0048] The particle size of the nanoparticles was determined using a dynamic light scattering particle size analyzer: the sample solution was placed in a Marlven Nano ZS instrument and the particle size was detected using a dynamic light scattering method. The temperature of the measuring cell was set at 25°C and each sample was run in parallel for three times.

[0049] The results are as follows Figure 1 As shown, the prepared 177 The particle size of Lu / AMP@MOFs is 132±3.4nm.

[0050] Experimental Example 2

[0051] Investigation 177 Submicrostructure of Lu / AMP@MOFs after 0-24h storage

[0052] The morphology of the nanoparticles was observed using TEM: the sample was dropped onto a 400-mesh zinc grid covered with a carbon film, placed in a drying oven, and after natural drying, was observed under a transmission electron microscope Titan G2-F20.

[0053] The results are as follows Figure 2 As shown, the prepared 177 Lu / AMP@MOFs has a well-dispersed irregular morphology, and as the storage time increases, nanowires are formed, which shows that 177 Cross-linking process occurs inside Lu / AMP@MOFs.

[0054] Experimental Example 3

[0055] The effect of different volume ratios of CS and GP on the phase transition temperature of the gel was investigated 177 The effect of different volume ratios of CS and GP on the phase transition temperature of the gel was investigated

[0056] The phase transition temperature of the nanometer radionuclide gel was determined: the concentration of the CS solution was 2%, the concentration of the GP was 5%, and the gel solution with different volume ratios was placed in a water bath, and the temperature was raised at a rate of 5 min / °C from 25°C to 40°C. The gel was considered to have occurred if it did not flow within 10 seconds, and the phase transition temperature of the gel solution was recorded.

[0057] The results are shown in Table 1. Figure 3 As shown in Table 1, when the concentration of GP was 55% and the concentration of CS was 2%, the phase transition temperature of the gel gradually decreased with the increase of the amount of CS. When the volume ratio of 2% CS solution to 55% GP solution was 65:35, 177 The phase transition temperature of Lu / AMP@CS / GP was 35°C, which was suitable for phase transition in response to body temperature and converted into a semi-solid gel.

[0058] Example 4

[0059] The effect of different volume ratios of CS / GP, 177 Lu@CS / GP, and 177 Lu / AMP@CS / GP on the solidification state at different temperatures was investigated

[0060] The appearance of CS / GP, 177 Lu@CS / GP, and 177 Lu / AMP@CS / GP after being placed at different temperatures for 5 minutes was observed. The results are shown in Table 2. Figure 4 As shown in Table 2, the gel preparation could maintain a liquid state at room temperature, and after incubation at 37°C, CS / GP, 177 Lu@CS / GP, and 177 Lu / AMP@CS / GP all converted into solid gels.

[0061] Example 5

[0062] The electron micrographs of CS / GP and 177 Lu / AMP@CS / GP were investigated

[0063] The appearance and morphology of the nanometer radionuclide gel were observed by scanning electron microscopy. The results are shown in Table 3. Figure 5 As shown in Table 3, A is the electron micrograph of the CS / GP semi-solid gel, which presents an irregular pore structure, and B is the electron micrograph of the CS / GP loaded with 177 Lu / AMP@MOFs, and the results show that the pore size is reduced, further reducing the risk of radionuclide leakage.

[0064] Example 6

[0065] The effect of 177In vitro release curve of Lu in gel

[0066] The radioactivity of nanonuclides was measured in physiological medium by radioactivity analyzer. 177 The specific operation of Lu release is as follows: the above-mentioned nanonuclide gel solution is placed in a penicillin bottle, incubated at 37°C for 5 minutes to form a semi-solid gel, 3 mL of release medium (10 mM PBS, pH 7.4) is added, and placed in a constant temperature water bath shaker (37°C, 100 rpm). Samples are taken at specific time points, the radioactivity is measured, and the cumulative release of radionuclides is calculated.

[0067] The results are as follows Figure 6 As shown, under physiological conditions, 177 Lu was effectively retained in the semisolid gel, with only 20% leakage after 96 h.

[0068] Experimental Example 7

[0069] Investigation of radionuclides 177 Distribution of Lu in the body

[0070] Establishment of rat osteoarthritis model: Male SD rats (6-8 weeks old) were randomly divided into groups, and the SD rat osteoarthritis model was established by anterior and posterior cruciate ligament and medial meniscus resection.

[0071] Distribution of radionuclides in the body: Injection into the joint cavity 177 Lu, 177 Lu@CS / GP, 177 Lu / AMP@CS / GP, the radionuclide dosage was 100 μCi. The rats were anesthetized and fixed 1 hour, 2 days, 4 days, and 16 days after the administration, and the distribution of radionuclide in the body was observed using a SPECT imaging system.

[0072] The results are as follows Figure 7 As shown, 177 Lu / AMP@CS / GP was injected into the rat joint cavity, and the distribution of radionuclides was observed using a SPECT / CT imaging system over 16 days. It was found that the radionuclides were mainly concentrated in the joint cavity.

[0073] Experimental Example 8

[0074] Investigation of radionuclides 177 Lu retention in the joint cavity

[0075] Determination of joint cavity 177 Lu retention. The results are as follows Figure 8 As shown, free 177 Lu 3+ The amount of nucleoside in the joint cavity of the group gradually decreased, and after 16 days, the amount of nucleoside in the joint cavity was only 20%, while 177Lu@CS / GP and 177 Lu / AMP@CS / GP can maintain a good retention effect in the joint cavity, especially 177 Lu / AMP@CS / GP, 16 days later, the retention of Lu in the joint cavity was 177 The retention of Lu was more than 85%.

[0076] Experimental Example 9

[0077] Investigation of radionuclide 177 Distribution of Lu in various tissues and organs in vivo

[0078] The rats were sacrificed on the 16th day after injection of 177 Lu, 177 Lu@CS / GP, 177 Lu / AMP@CS / GP, respectively, and the joint, heart, liver, spleen, lung, kidney, stomach, bone, muscle, small intestine, large intestine, brain and blood were taken out, and the radioactivity was determined by using a gamma counter.

[0079] The results are shown in Table 1. Figure 9 177 The joint cavity of the Lu / AMP@CS / GP group had the most Lu, and only a small amount of Lu was distributed in other organs, proving the excellent retention effect of the nanonucleogel in the joint cavity. 177 The joint cavity of the Lu / AMP@CS / GP group had the most Lu, and only a small amount of Lu was distributed in other organs, proving the excellent retention effect of the nanonucleogel in the joint cavity.

[0080] Experimental Example 10

[0081] Investigation of the cartilage repair effect of 177 Lu / AMP@CS / GP

[0082] Safranin O-fast green staining: the paraffin sections were dehydrated in gradient concentration ethanol, washed, immersed in fast green staining solution for 5 min, washed with weak alkali solution, immersed in safranin staining solution for 5 min, dehydrated with anhydrous ethanol, and sealed with neutral gum, and observed under an optical microscope.

[0083] Detection of the levels of proinflammatory factors in the joint by immunohistochemical experiment: the rats were sacrificed, the joint tissue was peeled off, and decalcification treatment was performed, and the tissue was embedded and fixed, and sectioned. The paraffin sections were subjected to dewaxing treatment, washed three times, and subjected to antigen repair by using pepsin. After washing again, hydrogen peroxide (3%) and BSA solution (3%) were added to cover the surface of the tissue and incubated for 30 min. The primary antibody (IL-1β, TNF-α) diluted with PBS was added and incubated overnight, and the corresponding secondary antibody was added and incubated at room temperature. After washing three times, freshly prepared DAB color developing solution was added, and after staining the nucleus, the sections were dehydrated in anhydrous ethanol, sealed, and observed by using an optical microscope.

[0084] The results are shown in Table 1. Figure 10 ​As shown, the nanometer radionuclide gel all showed a certain cartilage repair effect at each radionuclide dose, and reduced the level of inflammatory factor TNF-α. These results verify the feasibility of the nanometer radionuclide gel for OA treatment.

[0085] Experimental Example 11

[0086] Investigation 177 Effect of Lu / AMP@CS / GP treatment on the body weight of rats

[0087] The body weight change of rats during treatment was detected. The results are shown in Figure 11 As shown, compared with the PBS group, 177 The body weight of rats in the Lu / AMP@CS / GP group did not decrease significantly, which indicated that 177 The Lu / AMP@CS / GP treatment had no obvious side effects on rats.

[0088] Experimental Example 12

[0089] Investigation 177 Effect of Lu / AMP@CS / GP treatment on the organs of rats

[0090] H&E staining: the paraffin sections were dehydrated in gradient concentration of ethanol, washed with ultrapure water, and then dyed in Eosin dyeing solution for 5 min. After hydrochloric acid ethanol differentiation, washing, and eosin staining, the sections were sealed with neutral gum and observed under an optical microscope.

[0091] The results are shown in Figure 12 As shown, there was no obvious pathological change in the main organs such as heart, liver, spleen, lung, and kidney, which confirmed that 177 The Lu / AMP@CS / GP had good biological safety.

[0092] The above is the preferred embodiment of the present application. It should be noted that, for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, which should also be considered as the protection scope of the present application.

Claims

1. An anti-osteoarthritic nanonucleus gel, characterized in that, Including radionuclides 177 Lu-labelled metal-organic nanoframes and thermo-sensitive gel; the thermo-sensitive gel is prepared from chitosan and sodium glycerophosphate; the mass ratio of the chitosan and sodium glycerophosphate is 1:14-20; The 177 The particle size of the Lu-labeled metal-organic nanoframe was 132 ± 3.4 nm; The 177 The method for preparing Lu-labeled metal-organic nanoframes comprises: S1, mixing bovine serum albumin, adenosine monophosphate in ultrapure water, stirring vigorously while slowly adding 177 Lu 3+ The solution continues to be stirred, and the lower precipitate is collected after centrifugation; S2, the precipitate obtained in step S1 is dispersed into ultrapure water, bovine serum albumin is added again, stirred and mixed uniformly, and ultrasonic redissolution is performed under ice bath condition to obtain 177 Lu-labeled metal-organic nanoframes.

2. A method of preparing an anti-osteoarthritic nanonucleus gel as claimed in claim 1, characterized in that, The method comprises the following steps: S1, mixing bovine serum albumin, adenosine monophosphate in ultrapure water, stirring vigorously while slowly adding 177 Lu 3+ The solution continues to stir, and the lower precipitate is collected after centrifugation; S2, the precipitate obtained in step S1 is dispersed into ultrapure water, bovine serum albumin is added again, stirred and mixed uniformly, and ultrasonic redissolution is carried out under ice bath condition to obtain 177 Lu-labeled metal-organic nanoframe; S3, dissolving chitosan and glycerophosphate sodium in hydrochloric acid solution and ultrapure water respectively, slowly adding chitosan solution into glycerophosphate sodium solution under ice bath condition, and adding the solution obtained in step S2, and stirring and mixing to obtain nanometer nucleoside gel.

3. The preparation method according to claim 2, characterized in that In the S1 step 177 Lu 3+ The solution is 177 LuCl3solution, the 177 Lu 3+ The molar ratio of the solution to adenosine monophosphate is 1:

5.

4. The preparation method according to claim 2, characterized in that The stirring time in step S2 is 10 min, and the ice bath ultrasonic power is 100 W.

5. The preparation method according to claim 2, characterized in that The mass ratio of bovine serum albumin added in steps S1 and S2 is 1:

1.

6. The preparation method according to claim 2, characterized in that In step S3, the concentration ratio of chitosan solution to glycerophosphate sodium solution is 2:55, and the volume ratio is 13:

7.

7. Use of the anti-osteoarthritis nanometer nucleoside gel of claim 1 or the anti-osteoarthritis nanometer nucleoside gel prepared by the method of any one of claims 2-6 in the preparation of a drug for treating osteoarthritis.

8. Use according to claim 7, characterized in that, The anti-osteoarthritis nanometer nucleoside gel is injected into joint cavity.

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