A clay mineral biomaterial and a preparation method and application thereof

Abstract

The application provides a clay mineral biomaterial and a preparation method and application thereof, and the preparation method of the clay mineral biomaterial comprises the following steps: mixing montmorillonite and a potassium permanganate solution to form a reaction solution; performing a hydrothermal reaction on the reaction solution; and performing washing and drying treatment on the product of the hydrothermal reaction to obtain the clay mineral biomaterial. The preparation method of the clay mineral biomaterial of the application uses montmorillonite as a carrier, in-situ grows nanoscale manganese dioxide particles on the montmorillonite carrier through a hydrothermal reaction, so that the manganese dioxide particles are well dispersed and fixed on the surface of the montmorillonite carrier and can prevent the agglomeration of nanoparticles; meanwhile, the hydrophilic and hydroxyl-rich characteristics of the surface of the montmorillonite carrier enable good interfacial bonding between the montmorillonite carrier and the manganese dioxide particles, so that the montmorillonite carrier can improve the SOD and CAT enzyme activities of manganese dioxide, so as to better eliminate active oxygen and reduce inflammation.

Description

Technical Field

[0001] This application belongs to the field of anti-inflammatory drug technology, and more specifically, relates to a clay mineral biomaterial, its preparation method and application. Background Technology

[0002] Inflammation is a natural physiological response designed to protect the body from external damage or internal threats. It is a normal reaction of the body's immune system to external stimuli, but prolonged or excessive inflammation can lead to tissue damage, immune system dysfunction, and the development of many diseases. Furthermore, inflammation treatment is closely related to reducing reactive oxygen species (ROS) levels. ROS, including free radicals and oxidants, are a key factor in the inflammatory process; they can damage cell membranes, DNA, and proteins, further exacerbating inflammation. Reducing ROS levels has become an important strategy for controlling inflammation. In the treatment of inflammatory diseases, some drugs and treatments also focus on regulating ROS levels to alleviate the inflammatory response. However, there remains a need for simple, economical, and more effective drugs to reduce ROS and achieve the goal of treating inflammation. Summary of the Invention

[0003] The purpose of this application is to provide a clay mineral biomaterial, its preparation method and application. The preparation process of the clay mineral biomaterial of this application is very simple, the cost is lower and it has a good anti-inflammatory effect.

[0004] To achieve the above objectives, a first aspect of this application provides a method for preparing clay mineral biomaterials, comprising the following steps:

[0005] Montmorillonite and potassium permanganate solution are mixed to form a reaction solution;

[0006] The reaction solution is subjected to a hydrothermal reaction;

[0007] The products of the hydrothermal reaction are washed and dried to obtain the clay mineral biomaterial.

[0008] Furthermore, the montmorillonite is montmorillonite refined through ball milling.

[0009] Furthermore, the ball milling process is as follows: ball milling is performed using a wet ball milling method, anhydrous ethanol is added during the ball milling process, the rotation speed is set to 350 r / min, and the ball milling time is 60 h.

[0010] Furthermore, the montmorillonite, after undergoing the ball milling process, is then subjected to vacuum freeze-drying.

[0011] Furthermore, in the reaction solution, the mass ratio of montmorillonite to potassium permanganate is (50-500) mg: 1.45 g.

[0012] Furthermore, the hydrothermal reaction is carried out at a temperature of 160°C for a duration of 48 hours.

[0013] Furthermore, the montmorillonite is medical-grade montmorillonite.

[0014] In a second aspect, this application provides a clay mineral biomaterial prepared by any of the above-described preparation methods, wherein the clay mineral biomaterial comprises a montmorillonite carrier and nano-sized manganese dioxide loaded on the montmorillonite carrier.

[0015] A third aspect of this application provides the application of a clay mineral biomaterial in the preparation of an anti-inflammatory drug, wherein the clay mineral biomaterial is prepared by any of the preparation methods described above, and the clay mineral biomaterial comprises a montmorillonite carrier and nano-sized manganese dioxide loaded on the montmorillonite carrier.

[0016] Compared with the prior art, this application has the following technical effects:

[0017] This application discloses a method for preparing clay mineral biomaterials using montmorillonite as a carrier. Nanoscale manganese dioxide particles are grown in situ on the montmorillonite carrier via a hydrothermal reaction, resulting in good dispersion and fixation of the manganese dioxide particles on the surface of the montmorillonite carrier, which also prevents nanoparticle aggregation. Simultaneously, the hydrophilic and hydroxyl-rich characteristics of the montmorillonite carrier surface create a good interfacial bond with the manganese dioxide particles, enabling the montmorillonite carrier to enhance the activity of SOD and CAT enzymes of manganese dioxide, thereby better eliminating reactive oxygen species and reducing inflammation.

[0018] The present application discloses a method for preparing clay mineral biomaterials using montmorillonite as raw material, which is abundant and has the advantage of low cost.

[0019] The method for preparing clay mineral biomaterials proposed in this application is simple in steps, easy to operate, and conducive to large-scale production.

[0020] In the clay mineral biomaterial prepared in this application, montmorillonite enhances the SOD and CAT enzyme activity of manganese dioxide, thereby improving the anti-inflammatory effect of manganese dioxide and giving the clay mineral biomaterial a good anti-inflammatory function. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of this application, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 XRD patterns of MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 provided in Embodiments 1-5 of this application;

[0023] Figure 2 The graphs show the SOD enzyme activity detection of MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 provided in Examples 2-5 of this application.

[0024] Figure 3 The CAT enzyme activity detection graphs for MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 provided in Examples 1-5 of this application are shown.

[0025] Figure 4 The in vitro anti-inflammatory effects of MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 provided in Examples 1-5 of this application are shown in the figure. Detailed Implementation

[0026] To make the technical problems, technical solutions, and beneficial effects of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0027] The terminology used in the embodiments of this application is for the purpose of describing particular embodiments only and is not intended to be limiting of this application. The singular forms “a,” “the,” and “the” used in the embodiments of this application and the appended claims are also intended to include the plural forms unless the context clearly indicates otherwise.

[0028] The weights of the relevant components mentioned in the embodiments of this application can refer not only to the specific content of each component, but also to the proportional relationship between the weights of the components. Therefore, any scaling up or down of the content of the relevant components according to the embodiments of this application is within the scope disclosed in the embodiments of this application. Specifically, the mass described in the embodiments of this application can be a mass unit known in the chemical industry, such as μg, mg, g, or kg.

[0029] The medical-grade montmorillonite used in this application embodiment is a medical-grade montmorillonite product from Shanghai Aladdin Biochemical Technology Co., Ltd., CAS number: 1318-93-0.

[0030] Example 1

[0031] Example 1 of this application provides a method for preparing montmorillonite, which includes the following steps: weighing an appropriate amount of medical-grade montmorillonite, and performing ball milling using a wet ball milling method; during the ball milling process, anhydrous ethanol is added, the rotation speed is set to 350 r / min, and the time is 60 h; after the ball milling is completed, the ball milling jar is removed, and the grinding balls are separated from the mixed slurry using a sieve; the resulting mixed slurry is then vacuum freeze-dried to obtain ultrafine montmorillonite, labeled as MMT. In this example, the mass ratio of medical-grade montmorillonite, zirconia balls, and anhydrous ethanol is 2:120:4, and the diameter of the zirconia balls is 2 mm.

[0032] Example 2

[0033] Example 2 of this application provides a method for preparing manganese dioxide, comprising the following steps: adding 1.45g of KMnO4 (potassium permanganate) to 35mL of H2O to form a reaction solution; placing the reaction solution in a reaction vessel and carrying out a hydrothermal reaction at 160℃ for 48h to generate manganese dioxide; washing and freeze-drying the precipitate to obtain the final product, labeled as MnO2.

[0034] Example 3

[0035] Example 3 of this application provides a method for preparing clay mineral biomaterials, comprising the following steps: adding 1.45g KMnO4 (potassium permanganate) and 50mg of montmorillonite ball-milled in Example 1 to 35mL H2O to form a reaction solution; transferring the reaction solution to a reaction vessel and carrying out a hydrothermal reaction at 160℃ for 48h to generate manganese dioxide on the montmorillonite; washing the precipitate with deionized water and freeze-drying to obtain the final product montmorillonite / manganese dioxide, labeled as MMT / MnO2-1.

[0036] Using ball-milled montmorillonite as a carrier can prolong the residence time of montmorillonite in animals, which is beneficial for the treatment of inflammation in animals by drugs containing clay mineral biomaterials.

[0037] Example 4

[0038] Example 4 of this application provides a method for preparing clay mineral biomaterials, comprising the following steps: adding 1.45g KMnO4 (potassium permanganate) and 100mg of montmorillonite ball-milled in Example 1 to 35mL H2O to form a reaction solution; transferring the reaction solution to a reaction vessel and carrying out a hydrothermal reaction at 160℃ for 48h to generate manganese dioxide on the montmorillonite; washing the precipitate with deionized water and freeze-drying to obtain the final product montmorillonite / manganese dioxide, labeled as MMT / MnO2-2.

[0039] Example 5

[0040] Example 5 of this application provides a method for preparing clay mineral biomaterials, comprising the following steps: adding 1.45g KMnO4 (potassium permanganate) and 500mg of montmorillonite ball-milled in Example 1 to 35mL H2O to form a reaction solution; transferring the reaction solution to a reaction vessel and carrying out a hydrothermal reaction at 160℃ for 48h to generate manganese dioxide on the montmorillonite; washing the precipitate with deionized water and freeze-drying it to obtain the final product montmorillonite / manganese dioxide, labeled as MMT / MnO2-3.

[0041] The XRD patterns of MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 prepared in Examples 1-5 of this application are shown below. Figure 1 As shown, Figure 1 This indicates that the substances in the composite clay mineral biomaterial montmorillonite / manganese dioxide prepared in Examples 3-5 of this application are composed of montmorillonite and manganese dioxide.

[0042] SOD-like enzyme activity detection

[0043] The SOD-like enzyme activities of MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 prepared in Examples 2-5 of this application were detected using a superoxide dismutase (SOD) activity assay kit (Solarbio, BC0170), and the detection was performed according to the steps in the kit's instructions.

[0044] Test results as follows Figure 2 As shown, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 inhibit O2 - The abilities to inhibit O2 were 29.20±1.10%, 28.26±2.57%, and 60.19±3.34%, respectively, all greater than the ability of MnO2 to inhibit O2. - The activity was 6.36±2.67%, which indicates that the SOD-like enzyme activities of MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 are greater than those of MnO2.

[0045] CAT-like enzyme activity detection

[0046] The CAT-like enzyme activities of MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 prepared in Examples 1-5 of this application were detected. The content of O2 produced by the decomposition of H2O2 was detected using a dissolved oxygen meter. The specific steps were as follows: 8 ml of water, 100 μL of 10M H2O2, and 30 μL of 1 mg / ml MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 were added to a 6-well plate, and after standing for 10 min, the dissolved oxygen meter was used for detection.

[0047] Test results as follows Figure 3 As shown, the O2 production amounts of the composite clay mineral biomaterials MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 prepared in Examples 3-5 of this application were 12.00±0.48ppm, 13.39±0.92ppm, and 15.23±0.59ppm, respectively, all greater than the O2 production amount of MnO2 (11.66±0.42ppm). The O2 production amount of MMT (5.21±0.31ppm) was close to that of the blank group (4.85±0.51ppm). These results indicate that the CAT-like enzyme activity of the composite clay mineral biomaterials prepared in Examples 3-5 of this application is greater than that of pure manganese dioxide and pure montmorillonite.

[0048] from Figure 2 , Figure 3 As can be seen, compared with MMT or MnO2, the enzyme activity of the composite clay mineral biomaterial montmorillonite / manganese dioxide prepared in Examples 3-5 of this application is improved, and the enzyme activity of MMT / MnO2-3 is the highest.

[0049] In vitro cell anti-inflammatory effect detection

[0050] The in vitro anti-inflammatory effects of MMT, MnO2, MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 prepared in Examples 1-5 of this application were tested. RAW264.7 cells were seeded in 12-well plates and cultured for 12 hours. The test material solution was added to a final concentration of 5 μg / ml, and the cells were cultured for another 12 hours. H2O2 was added to a final concentration of 1 mM, and the cells were cultured for 3 hours. The cells were washed three times with PBS, then stained with DCFH-DA dye, washed three times with PBS, and photographed using a confocal microscope.

[0051] Photo results as follows Figure 4As shown, the montmorillonite complexes MMT / MnO2-1, MMT / MnO2-2, and MMT / MnO2-3 containing manganese dioxide prepared in Examples 3-5 of this application produced fewer green-stained cells due to reactive oxygen species than the MnO2, MMT, and blank groups. Among them, MMT / MnO2-3 had the lowest reactive oxygen species, indicating that the clay mineral biomaterials prepared in Examples 3-5 of this application have better anti-inflammatory effects compared with MMT or MnO2.

[0052] Figure 4 Negative indicates the negative control group, in which no hydrogen peroxide or test material is added; Positve indicates the positive control group, in which hydrogen peroxide is added but no test material is added.

[0053] The above embodiments merely illustrate several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of this patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. Use of a clay mineral biomaterial for the preparation of an anti-inflammatory medicament, characterized in that, The clay mineral biomaterial comprises a montmorillonite carrier and nanoscale manganese dioxide loaded on the montmorillonite carrier; a preparation method of the clay mineral biomaterial comprises the following steps: montmorillonite and a potassium permanganate solution are mixed to form a reaction solution; the reaction solution is subjected to a hydrothermal reaction; the product of the hydrothermal reaction is washed and dried to obtain the clay mineral biomaterial; the montmorillonite is refined by ball milling; in the reaction solution, the mass ratio of the montmorillonite to the potassium permanganate is (50-500) mg: 1.45 g.

2. Use of a clay mineral biomaterial according to claim 1 for the preparation of an anti-inflammatory medicament, characterized in that, the ball milling process is wet ball milling, and anhydrous ethanol is added during the ball milling process, the rotation speed is set to 350 r / min, and the ball milling time is 60 h.

3. Use of a clay mineral biomaterial according to claim 2 for the preparation of an anti-inflammatory medicament, characterized in that, the montmorillonite is subjected to vacuum freeze-drying treatment after the ball milling.

4. Use of a clay mineral biomaterial according to claim 1 for the preparation of an anti-inflammatory medicament, characterized in that, the temperature of the hydrothermal reaction is 160 ℃, and the time is 48 h.

5. Use of a clay mineral biomaterial according to any one of claims 1 to 4 for the preparation of an anti-inflammatory medicament, characterized in that, the montmorillonite is a medical-grade montmorillonite.

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