KAOLIN-BASED HEMOSTATIC MATERIAL WITH ANTI-INFLAMMATORY FUNCTION AND PREPARATION METHOD AND USE THEREOF

Abstract

The present disclosure relates to the technical field of hemostatic materials, and particularly relates to a kaolin-based hemostatic material with an anti-inflammatory function and a preparation method and use thereof. The kaolin-based hemostatic material with the anti-inflammatory function in the present disclosure includes CeO.sub.2 nanoparticle-loaded kaolin. The preparation method includes: mixing kaolin, cerium chloride, sodium carbonate, and sodium chloride in a mass ratio, and ball-milling to produce a precursor; and roasting the precursor to produce the kaolin-based hemostatic material with the anti-inflammatory function. In the present disclosure, with low-cost and abundantly-available kaolinite and the conventional CeO.sub.2 as raw materials, a CeO.sub.2-loaded kaolin-based nanomaterial is prepared through mechanochemical synthesis. The prepared CeO.sub.2/K nanomaterial has synergistic anti-inflammatory and hemostatic activities.

Claims

1. A preparation method of a kaolin-based hemostatic material with the anti-inflammatory function, comprising: mixing kaolin, cerium chloride, sodium carbonate, and sodium chloride in a mass ratio, and ball-milling to produce a precursor; and roasting the precursor to produce the kaolin-based hemostatic material with the anti-inflammatory function; wherein the mass ratio of the kaolin, the cerium chloride, the sodium carbonate, and the sodium chloride is 1:(0.2-2.35):1.25:1.15; wherein the roasting is conducted at 200 C. to 800 C. for 0.5 h to 4 h; and wherein the ball-milling is conducted for 0.5 h to 4 h at a rotational speed of 400 rpm to 560 rpm with a ball-to-material ratio of 130:(7.2-10.7).

2. The preparation method according to claim 1, wherein a heating rate for the roasting is 4 C./min to 6 C./min.

3. The preparation method according to claim 2, wherein a container for the ball-milling is a 100 mL zirconia jar; media adopted for the ball-milling are zirconia balls with diameters of 10 mm, 8 mm, and 5 mm that are in a mass ratio of 2:3:5; and the ball-milling is conducted with a planetary ball mill according to a ball-milling strategy with a 30 min forward rotation and a 30 min reverse rotation in each cycle and a 5 min pause between every two cycles.

4. The preparation method according to claim 2, wherein before the ball-milling, the kaolin, the cerium chloride, the sodium carbonate, and the sodium chloride each are dried at 180 C. for 13 h.

5. The preparation method according to claim 2, wherein after being roasted, the precursor is subjected to centrifugal washing 2 times to 8 times, and then oven-dried at 45 C.

6. A kaolin-based hemostatic material with the anti-inflammatory function produced with the preparation method according to claim 1.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0019] FIG. 1 is a schematic diagram of a synthetic process of CeO.sub.2/K;

[0020] FIG. 2 shows X-ray diffraction patterns of Kaol and CeO.sub.2/K;

[0021] FIG. 3 shows a scanning electron microscopy (SEM) image of Kaol;

[0022] FIG. 4 shows an SEM image of CeO.sub.2/K;

[0023] FIG. 5 shows blood clotting indexes (BCIs) of CeO.sub.2/K at 200 C. and 800 C.;

[0024] FIG. 6 shows BCIs of Kaol and CeO.sub.2/K;

[0025] FIG. 7 shows in vitro nanozyme activities of Kaol and CeO.sub.2/K; and

[0026] FIG. 8 shows a change in a tumor necrosis factor- (TNF-) level in lipopolysaccharide (LPS)-induced macrophages after a CeO.sub.2/K action.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0027] The technical solutions of the present disclosure are described in further detail below with reference to the specific embodiments and accompanying drawings, but the present disclosure is not limited thereto.

[0028] A preparation method of a kaolin-based hemostatic material with an anti-inflammatory function is as follows:

[0029] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite are first weighed. The Na.sub.2CO.sub.3, the NaCl, and the anhydrous CeCl.sub.3 each are dried overnight at a high temperature. Ball-milling is then conducted for a specified period of time in a planetary ball mill at a rotational speed of 560 rpm with a specified ball-to-material ratio to allow a mechanochemical action to produce a precursor for the kaolinite-based nanomaterial. The precursor is then placed in a muffle furnace and roasted at a specified temperature for a specified period of time. A roasting product is cleaned with deionized water under stirring, then dehydrated, and finally oven-dried at 45 C. for 24 h to produce a final product, which is the CeO.sub.2-loaded kaolin-based nanomaterial (CeO.sub.2/K).

Example 1

[0030] In this example, CeO.sub.2/K was prepared by the above method with different material ratios (Kaol:CeCl.sub.3:Na.sub.2CO.sub.3:NaCl=1:(0.235-2.35):1.25:1.15).

[0031] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 0.47 g, 1.41 g, 2.342 g, 3.279 g, and 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite were first weighed. The Na.sub.2CO.sub.3, NaCl, and anhydrous CeCl.sub.3 each were dried overnight at a high temperature. The raw materials were ball-milled for 4 h in a planetary ball mill at a rotational speed of 560 rpm with a ball-to-material ratio of 130:10.7 to allow mechanochemical synthesis. A ball-milled material produced after the ball-milling was placed in a muffle furnace, heated at a heating rate of 5 C./min to 400 C., and then roasted at 400 C. for 1 h. A powder material produced after the roasting was transferred to a 50 mL centrifuge tube, subjected to centrifugal washing 6 times at 8,000 r/min, kept at 45 C. for 720 min, and dried to produce CeO.sub.2/K of different material ratios.

Example 2

[0032] In this example, CeO.sub.2/K was prepared by the above method with different ball-milling times (0.5 h to 4 h).

[0033] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite were first weighed. The Na.sub.2CO.sub.3, NaCl, and anhydrous CeCl.sub.3 each were dried overnight at a high temperature. The raw materials were ball-milled for 0.5 h, 1 h, 2 h, 3 h, or 4 h in a planetary ball mill at a rotational speed of 560 rpm with a ball-to-material ratio of 130:10.7 to allow mechanochemical synthesis. A ball-milled material produced after the ball-milling was placed in a muffle furnace, heated at a heating rate of 5 C./min to 400 C., and then roasted at 400 C. for 1 h. A powder material produced after the roasting was transferred to a 50 mL centrifuge tube, subjected to centrifugal washing 6 times at 8,000 r/min, kept at 45 C. for 720 min, and dried to produce CeO.sub.2/K.

Example 3

[0034] In this example, CeO.sub.2/K was prepared by the above method with different ball-to-material ratios (130:7.2, 130:8, 130:8.8, 130:10.7, and 130:12.6).

[0035] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite were first weighed. The Na.sub.2CO.sub.3, NaCl, and anhydrous CeCl.sub.3 each were dried overnight at a high temperature. The raw materials were ball-milled for 4 h in a planetary ball mill at a rotational speed of 560 rpm with a ball-to-material ratio of 130:7.2, 130:8, 130:8.8, 130:10.7, or 130:12.6. A ball-milled material produced after the ball-milling was placed in a muffle furnace, heated at a heating rate of 5 C./min to 400 C., and then roasted at 400 C. for 1 h. A powder produced after the roasting was transferred to a 50 mL centrifuge tube, subjected to centrifugal washing 6 times at 8,000 r/min, kept at 45 C. for 720 min, and dried to produce CeO.sub.2/K.

Example 4

[0036] In this example, CeO.sub.2/K was prepared by the above method with different roasting temperatures (200 C. to 800 C.).

[0037] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite were first weighed. The Na.sub.2CO.sub.3, NaCl, and anhydrous CeCl.sub.3 each were dried overnight at a high temperature. The raw materials were ball-milled for 4 h in a planetary ball mill at a rotational speed of 560 rpm with a ball-to-material ratio of 130:10.7. A ball-milled material produced after the ball-milling was placed in a muffle furnace, heated at a heating rate of 5 C./min to 200 C., 300 C., 400 C., 500 C., or 800 C., and then roasted for 1 h at 200 C., 300 C., 400 C., 500 C., or 800 C. A powder produced after the roasting was transferred to a 50 mL centrifuge tube, subjected to centrifugal washing 6 times at 8,000 r/min, kept at 45 C. for 720 min, and dried to produce CeO.sub.2/K.

Example 5

[0038] In this example, CeO.sub.2/K was prepared by the above method with different roasting times (0.5 h to 4 h).

[0039] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite were first weighed. The Na.sub.2CO.sub.3, NaCl, and anhydrous CeCl.sub.3 each were dried overnight at a high temperature. The raw materials were ball-milled for 4 h in a planetary ball mill at a rotational speed of 560 rpm with a ball-to-material ratio of 130:10.7. A ball-milled material produced after the ball-milling was placed in a muffle furnace, heated at a heating rate of 5 C./min to 400 C., and then roasted at 400 C. for 0.5 h, 1 h, 2 h, 3 h, or 4 h. A powder produced after the roasting was transferred to a 50 mL centrifuge tube, subjected to centrifugal washing 6 times at 8,000 r/min, kept at 45 C. for 720 min, and dried to produce CeO.sub.2/K.

Example 6

[0040] In this example, CeO.sub.2/K was prepared by the above method with different numbers of centrifugal washing times (2 times, 4 times, 6 times, and 8 times) to remove the unsuccessfully-synthesized impurities.

[0041] 2.5 g of Na.sub.2CO.sub.3, 2.3 g of NaCl, 4.7 g of anhydrous CeCl.sub.3, and 2 g of kaolinite were first weighed. The Na.sub.2CO.sub.3, NaCl, and anhydrous CeCl.sub.3 each were dried overnight at a high temperature. The raw materials were ball-milled for 4 h in a planetary ball mill at a rotational speed of 560 rpm with a ball-to-material ratio of 130:10.7. A ball-milled material produced after the ball-milling was placed in a muffle furnace, heated at a heating rate of 5 C./min to 400 C., and then roasted for 1 h at 400 C. A powder produced after the roasting was transferred to a 50 mL centrifuge tube, subjected to centrifugal washing 2 times, 4 times, 6 times, or 8 times at 8,000 r/min, kept at 45 C. for 720 min, and dried to produce CeO.sub.2/K.

[0042] FIG. 1 is a schematic diagram of a synthetic process of CeO.sub.2/K. It can be seen from this figure that the synthetic process of CeO.sub.2/K is simple and eco-friendly.

[0043] The CeO.sub.2/K prepared in Examples 1 to 6 was subjected to X-ray diffraction analysis. According to results, with either the increase of a ball-milling time or the decrease of a ball-to-material ratio, CeO.sub.2 crystal grains are continuously refined. This is because the refinement of a powder in a ball-milling tank is accelerated with the increase of a ball-milling time and the decrease of a ball-to-material ratio to constantly reduce the crystal grains of a product. A change in the roasting temperature and the roasting time also causes a change in a crystal form of kaolinite. An increase in the roasting temperature and the roasting time improves the integrity of crystal grains of CeO.sub.2, and increases the grain size. In addition, the number of dehydration times affects the residue of NaCl as a diluent. As the number of dehydration times increases, a characteristic absorption peak of NaCl is gradually reduced and completely disappears after 6 times of dehydration.

[0044] FIG. 2 shows an X-ray diffraction pattern of CeO.sub.2/K synthesized after 6 times of dehydration in Example 6. As shown in FIG. 2, diffraction peaks observed at 2 values of 28.55, 33.08, 47.48, 56.34, 59.08, 69.41, 76.70, and 79.07 correspond to cubic fluorite-type CeO.sub.2 with a spatial group Fm3m (PDF #01-075-8371), and diffraction peaks at 2 values of 12.38, 19.85, 20.33, 21.25, 23.14, and 24.90 correspond to Kaol (PDF #01-080-0886). It can be known that the synthesized product is a kaolinite/CeO.sub.2 composite. In the CeO.sub.2/K composite, a crystal lattice of kaolinite is not destructed, and kaolinite retains its original crystalline structure.

[0045] FIG. 3 shows an SEM image of Kaol. A microscopic morphology and structure of Kaol were analyzed by SEM. Kaol mainly presents a two-dimensional lamellar structure with a smooth surface and a distinct boundary.

[0046] FIG. 4 shows an SEM image of CeO.sub.2/K synthesized after 6 times of dehydration in Example 6. It can be clearly observed that CeO.sub.2 is successfully anchored on the surface of kaolinite, CeO.sub.2 particles are aggregates of small spherical particles, and kaolinite is completely covered by CeO.sub.2 particles.

[0047] A coagulation test was carried out for the CeO.sub.2/K prepared in Examples 1 to 6. It can be seen from results that, the longer the ball-milling time, the poorer the coagulation effect after kaolinite is transformed into an amorphous state. However, the ball-to-material ratio and the number of dehydration times have little influence on the coagulation effect. The roasting temperature and the roasting time also affect the coagulation effect of the material. When the roasting temperature is 800 C. and the roasting time is 1 h, the coagulation effect of the material significantly decreases. With the increase of the roasting temperature, the coagulation effect deteriorates. However, a too-low roasting temperature will affect the generation of CeO.sub.2 particles. Therefore, the ball-milling time, roasting temperature, and roasting time can be comprehensively optimized by considering both the synergistic anti-inflammatory and hemostatic effects and the actual product requirements.

[0048] FIG. 5 shows BCIs of CeO.sub.2/K materials prepared after roasting at 200 C. and 800 C. Effective hemostasis is the key to a hemostatic material. The lower the BCI, the stronger the hemostatic ability. In the coagulation test for CeO.sub.2/K, BCI of the material produced at 200 C. decreases from 95.570.85% to 44.770.36%, and BCI of the material produced at 800 C. increases to 81.231.96%. These results confirm that the roasting temperature actually affects the coagulation effect of the material. The higher the roasting temperature, the higher the BCI. It indicates that a high temperature worsens the hemostatic performance of the material. Therefore, a reasonable roasting temperature should be selected.

[0049] FIG. 6 shows BCIs of Kaol and CeO.sub.2/K prepared after 6 times of dehydration in Example 5. In the coagulation test for CeO.sub.2/K, the BCI decreases from 105.540.9% to 69.611.16%. It should be noted that CeO.sub.2 cannot effectively stop bleeding, and even will worsen a hemostatic effect instead. However, the loading of CeO.sub.2 with kaolinite can improve the hemostatic performance of the material.

[0050] FIG. 7 shows in vitro nanozyme activities of Kaol and CeO.sub.2/K prepared after 6 times of dehydration in Example 5. The clearance of reactive oxygen species (ROS) is a key factor in the suppression of inflammatory diseases by anti-inflammatory materials. The reduction of excessive ROS can prevent the further progression of diseases or promote the regeneration of damaged inflammatory tissues. Catalase (CAT) can scavenge superoxide anions and decompose hydrogen peroxide to produce molecular oxygen, thereby slowing down the occurrence of inflammation. According to the results of in vitro CAT nanoenzyme activities of Kaol and CeO.sub.2/K, Kaol does not possess a CAT activity. However, after CeO.sub.2 is loaded, the CAT activity increases, and the oxygen production capacity is further enhanced. It can be known that CeO.sub.2/K has an in vitro CAT activity.

[0051] FIG. 8 shows a change in a TNF-a level in LPS-induced macrophages after a CeO.sub.2/K action. In order to further understand an anti-inflammatory mechanism of CeO.sub.2/K, a level of the related cytokine (TNF-) in an inflammatory tissue was quantitatively analyzed by enzyme-linked immunosorbent assay (ELISA). According to results, CeO.sub.2/K can effectively down-regulate the expression of the pro-inflammatory cytokine TNF- to suppress the inflammatory death of macrophages in an inflammation-induced model, thereby protecting the cells from oxidative damage induced by H.sub.2O.sub.2.

[0052] What is not mentioned above can be acquired in the prior art.

[0053] Although some specific embodiments of the present disclosure have been described in detail by way of examples, those skilled in the art will appreciate that the above examples are provided for illustration only and not for limiting the scope of the present disclosure. A person skilled in the art can make various modifications or supplements to the specific embodiments described or replace them in a similar manner, but it may not depart from the direction of the present disclosure or the scope defined by the appended claims. Those skilled in the art should understand that any modification, equivalent replacement, and improvement made to the above embodiments according to the technical essence of the present disclosure shall be included in the protection scope of the present disclosure.