Kaolin hemostatic material with anti-inflammatory function and its preparation method and application

Abstract

The present invention relates to the technical field of hemostatic materials, and in particular to a kaolin hemostatic material with anti-inflammatory function, and a preparation method and application thereof. The kaolin hemostatic material with anti-inflammatory function of the present invention comprises kaolin loaded with CeO2 nanoparticles. The preparation method comprises mixing kaolin with cerium chloride, sodium carbonate and sodium chloride in a certain mass ratio, ball milling the mixture and obtaining a precursor, and calcining the precursor to obtain the kaolin hemostatic material with anti-inflammatory function. The present invention adopts kaolinite, which is inexpensive and abundant in reserves, as a raw material, selects conventional CeO2 raw material, and prepares a kaolin-based nanomaterial loaded with CeO2 by a mechanochemical synthesis method, thereby preparing a CeO2 / K nanomaterial with synergistic anti-inflammatory and hemostatic activity.

Description

Technical Field

[0001] The present invention relates to the technical field of hemostatic materials, and in particular to a kaolin hemostatic material with anti-inflammatory function, a preparation method and application thereof. Background Art

[0002] In the treatment of diabetic foot, debridement is an essential step to remove necrotic tissue (including bone and abscesses). However, the common vascular lesions in diabetic patients necessitate anticoagulation therapy to prevent cardiovascular events, which also increases the risk of postoperative bleeding. Therefore, after debridement, effective hemostasis measures are necessary, as well as controlling the inflammatory response to reduce the risk of infection and promote wound healing.

[0003] Currently, the most commonly used hemostatic materials in clinical practice are sterile gauze. However, for diabetic patients who use anticoagulants for a long time, these traditional methods may not be sufficient to effectively control bleeding and may even prolong the patient's hospital stay. In order to improve the effectiveness of diabetic wound treatment, it is crucial to find materials with better hemostatic and anti-inflammatory properties.

[0004] Kaolin (K), a natural mineral, has shown great potential in the field of hemostasis due to its excellent physicochemical properties and biocompatibility. Studies have shown that kaolin dressings have significant hemostatic effects in trauma and percutaneous vascular interventional treatments. Patent [CN 117462732 A] focuses on improving the hemostatic effects of kaolin and addressing its clumping and pulverization issues. Patent [CN 109481731 B] uses nano-oxides to impart antibacterial properties to kaolin hemostatic materials, enabling them to achieve synergistic hemostatic and antibacterial effects. Patent [CN 116177587 A] describes a mesoporous CeO2 nanocomposite material and demonstrates its strong antioxidant capacity. However, there are no kaolin-based materials specifically designed for synergistic hemostasis and anti-inflammatory effects after debridement of diabetic foot surgery.

[0005] Furthermore, existing synthesis methods often use toxic solvents or reagents, such as NaOH and H2SO4, which may remain on the surface of the nanoparticles, leading to potential adverse biological reactions. The use of external stabilizers or end-capping agents may also reduce the biosafety of the materials. Furthermore, large-scale production processes present difficulties in controlling the consistency of nanoparticle size and shape, making it difficult to ensure product purity and consistency. Summary of the Invention

[0006] The purpose of the present invention is to address the above-mentioned deficiencies in the prior art and to provide a kaolin hemostatic material with anti-inflammatory function, a preparation method thereof, and an application thereof.

[0007] The invention discloses a kaolin hemostatic material with anti-inflammatory function, comprising kaolin loaded with CeO2 nanoparticles.

[0008] A method for preparing the kaolin hemostatic material with anti-inflammatory function as described above comprises mixing kaolin with cerium chloride, sodium carbonate and sodium chloride in a certain mass ratio, ball milling the mixture and calcining the precursor to obtain the kaolin hemostatic material with anti-inflammatory function.

[0009] Furthermore, the mass ratio of kaolin to cerium chloride, sodium carbonate and sodium chloride is 1:0.2 to 2.35:1.25:1.15.

[0010] Furthermore, in step S2, the calcination temperature is 200°C-800°C, and the calcination time is 0.5-4h.

[0011] Furthermore, in step S2, the calcination heating rate is 4-6°C / min.

[0012] Furthermore, the ball-to-material ratio of the ball mill is 130:7.2-10.7, the rotation speed is 400-560 rpm, and the time is 0.5-4 h.

[0013] Furthermore, the ball milling container is a 100 mL zircon jar; the medium is zircon balls with diameters of 10 mm, 8 mm, and 5 mm, and the ratio is 2:3:5; the ball milling uses a planetary ball mill, which synthesizes the material according to the ball milling strategy of forward rotation for 30 minutes, reverse rotation for 30 minutes, and stopping for 5 minutes before the next cycle.

[0014] Furthermore, the kaolin, cerium chloride, sodium carbonate and sodium chloride were dried before ball milling; the drying temperature was 180° C. and the drying time was 13 h.

[0015] Furthermore, the precursor is calcined, washed by centrifugation 2-8 times, and then dried at 45°C.

[0016] An application of the kaolin hemostatic material with anti-inflammatory function as described above is characterized in that it is used as a hemostatic material after debridement of diabetic foot.

[0017] The present invention adopts kaolinite, which is inexpensive and abundant in reserves, as a raw material, selects conventional CeO2 raw material, and prepares kaolin-based nanomaterials loaded with CeO2 through a mechanochemical synthesis method to prepare CeO2 / K nanomaterials with high consistency in nanoparticle size and shape and synergistic anti-inflammatory and hemostatic activities. In the present invention, kaolinite provides hemostasis and stabilizes CeO2 nanoparticles, and CeO2 has catalytic properties to achieve anti-inflammatory effects, thereby achieving synergistic anti-inflammatory and hemostatic effects. In addition, the present invention has a short experimental cycle, a simple operating process, and can also achieve green production and meet the needs of large-scale industrialization. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1Schematic diagram of the synthesis process of CeO2 / K;

[0019] Figure 2 is the X-ray diffraction pattern of Kaol and CeO2 / K;

[0020] Figure 3 is the scanning electron microscope image of Kaol;

[0021] Figure 4 This is the scanning electron microscope image of CeO2 / K;

[0022] Figure 5 BCI index diagram of CeO2 / K at 200 and 800℃;

[0023] Figure 6 BCI index diagram of Kaol and CeO2 / K;

[0024] Figure 7 Figure 2 is the in vitro nanozyme activity diagram of Kaol and CeO2 / K;

[0025] Figure 8 This is a graph showing the changes in TNF-α levels in LPS-induced macrophages after the action of CeO2 / K. DETAILED DESCRIPTION

[0026] The following are specific embodiments of the present invention and the accompanying drawings to further describe the technical solutions of the present invention, but the present invention is not limited to these embodiments.

[0027] The preparation method of the kaolin hemostatic material with anti-inflammatory function is as follows:

[0028] First, 2.5g of Na2CO3, 2.3g of NaCl, 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature. Then, maintaining a certain ball-to-material ratio, the planetary ball mill was rotated at 560rpm and milled for a period of time. The mechanochemical reaction product was a kaolinite-based nanomaterial precursor. The precursor was then placed in a muffle furnace and calcined at a certain temperature for a period of time. Finally, the product was washed, stirred with deionized water, washed, and then dehydrated. Finally, it was dried at 45°C for 24h to obtain the final product, i.e., CeO2-loaded kaolin nanomaterial (CeO2 / K).

[0029] Example 1

[0030] In this embodiment, CeO2 / K was prepared according to the above method using different material ratios (1:0.25-2.35:1.25:1.15).

[0031] First, 2.5g of Na2CO3, 2.3g of NaCl, 0.47g, 1.41g, 2.342g, 3.279g, and 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature. The raw materials were mechanochemically synthesized using a planetary ball mill at a ball-to-material ratio of 130:10.7 and a speed of 560 rpm for 4 hours. After ball milling, the mixture was placed in a muffle furnace and calcined at 400°C for 1 hour at a heating rate of 5° / min. After calcination, the powder was transferred to a 50mL centrifuge tube and washed six times by centrifugation at 8000 rpm. Finally, the mixture was kept at 45°C for 720 minutes and dried to obtain CeO2 / K with different material ratios.

[0032] Example 2

[0033] In this example, CeO2 / K was prepared according to the above method with different ball milling times (0.5 to 4 h).

[0034] First, 2.5g of Na2CO3, 2.3g of NaCl, 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature. The raw materials were mechanochemically synthesized using a planetary ball mill at a ball-to-material ratio of 130:10.7 and a rotation speed of 560 rpm for 0.5h, 1h, 2h, 3h, and 4h, respectively. After ball milling, the powder was calcined at 400°C in a muffle furnace at a heating rate of 5° / min for 1h. After calcination, the powder was transferred to a 50mL centrifuge tube and washed six times by centrifugation at 8000 rpm. Finally, the mixture was incubated at 45°C for 720min and dried to obtain CeO2 / K.

[0035] Example 3

[0036] In this embodiment, CeO2 / K was prepared according to the above method with different ball-to-material ratios (130:7.2, 8, 8.8, 10.7 and 12.6).

[0037] First, 2.5g of Na2CO3, 2.3g of NaCl, 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature and then ball-milled in a planetary ball mill at 560rpm for 4h. The ball-to-material ratios were 130:7.2, 8, 8.8, 10.7, and 12.6, respectively. After ball milling, the mixture was placed in a muffle furnace and calcined at 400°C for 1h at a heating rate of 5° / min. After calcination, the powder was transferred to a 50mL centrifuge tube and washed six times by centrifugation at 8000r / min. Finally, the mixture was kept at 45°C for 720min and dried to obtain CeO2 / K.

[0038] Example 4

[0039] In this embodiment, CeO2 / K was prepared according to the above method at different calcination temperatures (200-800°C).

[0040] First, 2.5g of Na2CO3, 2.3g of NaCl, 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature and then ball-milled in a planetary ball mill at a ball-to-material ratio of 130:10.7 and a rotation speed of 560rpm for 4 hours. After ball milling, the raw materials were placed in a muffle furnace and calcined at 200°C, 300°C, 400°C, 500°C, and 800°C at a heating rate of 5° / min for 1 hour. After calcination, the powder was transferred to a 50mL centrifuge tube and washed six times by centrifugation at 8000r / min. Finally, the mixture was kept at 45°C for 720min and dried to obtain CeO2 / K.

[0041] Example 5

[0042] In this example, CeO2 / K was prepared according to the above method with different calcination times (0.5-4h).

[0043] First, 2.5g of Na2CO3, 2.3g of NaCl, 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature and milled in a planetary ball mill at a ball-to-material ratio of 130:10.7 and a rotation speed of 560rpm for 4 hours. After ball milling, the raw materials were placed in a muffle furnace and calcined at 400°C at a heating rate of 5° / min for 0.5h, 1h, 2h, 3h, and 4h. After calcination, the powder was transferred to a 50mL centrifuge tube and washed six times by centrifugation at 8000r / min. Finally, the mixture was kept at 45°C for 720min and dried to obtain CeO2 / K.

[0044] Example 6

[0045] In this embodiment, CeO2 / K was prepared according to the above method by centrifugal dehydration for 2, 4, 6, and 8 times to remove impurities that were not successfully synthesized.

[0046] First, 2.5g of Na2CO3, 2.3g of NaCl, 4.7g of anhydrous CeCl3, and 2g of kaolinite were weighed. The NaCO3, NaCl, and anhydrous CeCl3 were dried overnight at high temperature and then ball-milled in a planetary ball mill at a ball-to-cement ratio of 130:10.7 and a rotation speed of 560 rpm for 4 hours. After ball milling, the raw materials were placed in a muffle furnace and calcined at 400°C for 1 hour at a heating rate of 5° / min. After calcination, the powders were transferred to 50mL centrifuge tubes and centrifuged at 8000 rpm for 2, 4, 6, and 8 wash cycles. Finally, the mixture was kept at 45°C for 720 minutes and dried to obtain CeO2 / K.

[0047] Figure 1 This is a schematic diagram of the synthesis process of CeO2 / K. It can be seen from the figure that the synthesis process of CeO2 / K is simple and green.

[0048] X-ray diffraction analysis of CeO2 / K prepared in Examples 1-6 revealed that the CeO2 grains became increasingly refined with increasing milling time and decreasing milling ratio. This is because the powder refinement rate in the milling jar accelerates, and the product grains continue to decrease with increasing milling time and decreasing milling ratio. Meanwhile, changes in roasting temperature and roasting time result in changes in the kaolinite crystal form, while increases in temperature and time promote a more complete CeO2 grain size. Furthermore, the number of dehydration cycles affects the residual amount of diluent NaCl. As the number of dehydration cycles increases, the characteristic absorption peak of NaCl gradually decreases until it completely disappears at 6 cycles.

[0049] Figure 2 The X-ray diffraction pattern of CeO2 / K synthesized after dehydration 6 times in Example 6 is shown in Figure 2 The observed diffraction peaks at 2θ of 28.55°, 33.08°, 47.48°, 56.34°, 59.08°, 69.41°, 76.70°, and 79.07° correspond to cubic fluorite CeO2 with space group Fm3m (PDF#01-075-8371). The diffraction peaks at 2θ of 12.38°, 19.85°, 20.33°, 21.25°, 23.14°, and 24.90° correspond to Kaol (PDF#01-080-0886), confirming that the synthesized product is a composite of kaolinite and CeO2. In the CeO2 / K composite, the kaolinite lattice is intact, and the kaolinite in the composite retains its original crystalline structure.

[0050] Figure 3 This is the scanning electron microscope image of Kaol. The micromorphology and structure of Kaol were analyzed by SEM. Kaol mainly presents a two-dimensional flake structure with a smooth surface and clear boundaries.

[0051] Figure 4 This is the scanning electron microscope image of CeO2 / K synthesized after dehydration 6 times in Example 6. It can be clearly observed that CeO2 is successfully anchored on the surface of kaolinite. The CeO2 particles are aggregates with small spherical particles, and the kaolinite is completely covered by CeO2 particles.

[0052] Coagulation experiments were conducted on the CeO2 / K prepared in Examples 1-6. It was found that the longer the ball milling time, the worse the coagulation effect after the kaolinite turned amorphous, but the ball-to-material ratio and the number of dehydration times had little effect on the coagulation effect. Calcination temperature and time also affect the material's coagulation effect. As the temperature and time reach 800°C and 1 hour, the coagulation effect of the material decreases significantly, and increasing the temperature shows a reverse coagulation effect. However, excessively low temperatures can affect CeO2 particle formation. Therefore, considering the synergistic effects of anti-inflammatory and hemostasis, as well as the actual product needs, the ball milling time, calcination temperature, and calcination time can be comprehensively optimized.

[0053] Figure 5 The following plots show the BCI index of CeO2 / K materials calcined at 200°C and 800°C, respectively. Effective hemostasis is key to hemostatic materials. The lower the blood clotting index (BCI), the stronger the hemostatic ability. In CeO2 / K coagulation experiments, the BCI index decreased from 95.57±0.85% to 44.77±0.36% at 200°C, and then increased to 81.23±1.96% at 800°C. This result confirms that temperature actually affects the material's coagulation effect. Higher temperatures increase the BCI, indicating that higher temperatures worsen the material's hemostatic properties. Therefore, a reasonable calcination temperature should be selected.

[0054] Figure 6 Figure 3 shows the BCI index of CeO2 / K prepared after six dehydration cycles in Kaol and Example 5. In the CeO2 / K coagulation experiment, the BCI index was found to decrease from 105.54±0.9% to 69.61±1.16%. It is worth noting that CeO2 is not effective in stopping bleeding and may even worsen the hemostatic effect. However, the use of kaolinite to load CeO2 improves the hemostatic performance of the material.

[0055] Figure 7The in vitro nanozyme activity diagram of Kaol and CeO2 / K prepared after dehydration 6 times in Example 5. Clearing ROS is the main factor for anti-inflammatory materials to inhibit inflammatory diseases. Reducing excessive ROS can prevent the further development of the disease or promote the regeneration of damaged inflammatory tissue. Peroxidase (CAT) can clear superoxide anions and tandem hydrogen peroxide to produce molecular oxygen and slow down the occurrence of inflammation. From the in vitro CAT nanozyme activity diagram of Kaol and CeO2 / K, it can be seen that Kaol does not have CAT activity, but the CAT enzyme activity increases after loading CeO2, and the oxygen production capacity is further improved. It proves that CeO2 / K has in vitro CAT enzyme activity.

[0056] Figure 8 This figure shows the changes in TNF-α levels in LPS-induced macrophages after CeO2 / K treatment. In order to further understand the anti-inflammatory mechanism of CeO2 / K, the levels of related cytokines (tumor necrosis factor α, TNF-α) in inflammatory tissues were quantitatively analyzed by enzyme-linked immunosorbent assay (ELISA). The results showed that CeO2 / K can effectively downregulate the proinflammatory cytokine TNF-α, inhibit the inflammatory death of macrophages in the inflammation-induced model, and protect cells from H2O2-induced oxidative damage.

[0057] Any matters not mentioned above shall be subject to the existing technology.

[0058] Although some specific embodiments of the present invention have been described in detail through examples, those skilled in the art should understand that the above examples are for illustration only and are not intended to limit the scope of the present invention. Those skilled in the art of the present invention may make various modifications or additions to the described specific embodiments or replace them in similar ways, but they will not deviate from the direction of the present invention or exceed the scope defined by the appended claims. Those skilled in the art should understand that any modifications, equivalent replacements, improvements, etc. made to the above embodiments based on the technical essence of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a kaolin hemostatic material with anti-inflammatory function, characterized in that: Kaolin is mixed with cerium chloride, sodium carbonate and sodium chloride in a certain mass ratio and then ball-milled to obtain a precursor, and the precursor is calcined to obtain a kaolin hemostatic material with anti-inflammatory function; The mass ratio of kaolin to cerium chloride, sodium carbonate and sodium chloride is 1:0.2~2.35:1.25:1.15; The calcination temperature is 200-800°C and the time is 0.5-4 h; The ball-to-material ratio of ball milling is 130:7.2-10.7, the rotation speed is 400-560 rpm, and the time is 0.5~4 h.

2. The preparation method according to claim 1, wherein: The calcination heating rate is 4-6℃ / min.

3. The preparation method according to claim 1, wherein: The ball milling container was a 100 mL zircon jar; the medium was zircon balls with diameters of 10 mm, 8 mm, and 5 mm, and the ratio was 2:3:5; a planetary ball mill was used for ball milling, and the material was synthesized according to the ball milling strategy of forward rotation for 30 min, reverse rotation for 30 min, and stopping for 5 min before the next round.

4. The preparation method according to claim 1, wherein: Before ball milling, kaolin, cerium chloride, sodium carbonate and sodium chloride were dried at 180°C for 13 h.

5. The preparation method according to claim 1, wherein: After the precursor is calcined, it is washed by centrifugation 2-8 times and then dried at 45°C.

6. A kaolin hemostatic material with anti-inflammatory function prepared by the preparation method according to any one of claims 1 to 5.

7. A use of the kaolin hemostatic material with anti-inflammatory function as claimed in claim 6, characterized in that: Used as a hemostatic material after debridement of diabetic foot.

Images