Bioactive glass as well as preparation method and application thereof

Abstract

The invention discloses bioactive glass as well as a preparation method and application thereof, and belongs to the technical field of bioactive substances. The invention provides a preparation method of bioactive glass, which comprises the following steps: dissolving cetyltrimethylammonium bromide in water, adding ethyl acetate, and stirring to form microemulsion drops to obtain ethyl acetate-cetyltrimethylammonium bromide-water microemulsion; tetraethoxysilane, triethyl phosphate, calcium nitrate and ferric nitrate are sequentially added, and Fe-BG sediment is obtained; and the Fe-BG precipitate is dried, calcined and ground, and Fe-BG powder is obtained. The invention further provides bioactive glass based on the preparation method and application of the bioactive glass. According to the bioactive glass provided by the invention, by doping iron ions, the Fe-BG material shows stronger procoagulant ability, the coagulation time is obviously shortened, the blood compatibility requirement of a biomedical material is met, and the bioactive glass is suitable for clinical hemostasis and wound repair.

Description

Technical Field

[0001] This invention relates to the field of bioactive substances technology, and in particular to a bioactive glass, its preparation method and application. Background Technology

[0002] Bioactive glass (BG) is a type of material capable of repairing, replacing, and regenerating body tissues, and forming bonds between tissues and materials. It is mainly composed of… , , CaO and Composed of basic components, it belongs to silicate glass and has a unique sub-nanopore structure, which is conducive to the penetration of biological fluids and the adhesion and proliferation of cells. Its mechanism of action is that through slow dissolution in vivo, it releases ions such as silicon, calcium, and phosphorus. These ions can promote cell adhesion, proliferation, and differentiation, and can also activate multiple intracellular signaling pathways. At the same time, its surface can induce the adsorption of extracellular matrix proteins, providing attachment sites for cells and promoting the deposition and aggregation of extracellular matrix components, thereby promoting tissue repair.

[0003] However, most existing bioactive glasses are dense structures prepared by high-temperature melting, with small specific surface areas and a lack of well-developed mesoporous channels, which limits their ability to rapidly absorb blood water through capillary effects to concentrate clotting factors. At the same time, their hemostatic mechanism relies solely on the release of calcium and silicon ions to activate the body's own endogenous coagulation cascade reaction. This biochemical process is relatively slow, and the material composition lacks active metal components that can rapidly chemically cross-link or physically aggregate with blood proteins. As a result, when faced with arterial spurting bleeding or impaired coagulation mechanisms, they cannot form stable blood clots in a short time, making it difficult to meet the stringent requirements of timeliness and reliability for emergency hemostasis.

[0004] Therefore, developing a bioactive glass material with high specific surface area, rapid physical liquid absorption and protein aggregation, improved blood clot strength, and enhanced vascularization function is a pressing technical problem to be solved in the field of hemostatic materials. Summary of the Invention

[0005] Based on the technical problems to be solved by the present invention, this invention proposes a bioactive glass, its preparation method, and its application. The invention employs doping with metal ions (such as iron) to significantly enhance the hemostatic ability of the bioactive glass. Based on the iron-doped mesoporous bioactive glass Fe-BG, and by optimizing the iron doping ratio, the application effect and coagulation efficiency of the bioactive glass in the field of hemostasis are effectively improved.

[0006] One objective of this invention is to provide a method for preparing bioactive glass, comprising: S1, dissolving hexadecyltrimethylammonium bromide in water, adding ethyl acetate, and stirring to form microemulsion droplets to obtain an ethyl acetate-hexadecyltrimethylammonium bromide-water microemulsion; S2, sequentially adding tetraethyl orthosilicate, triethyl phosphate, calcium nitrate, and ferric nitrate to the ethyl acetate-hexadecyltrimethylammonium bromide-water microemulsion to obtain an Fe-BG precipitate; S3, drying, calcining, and grinding the Fe-BG precipitate to obtain Fe-BG powder.

[0007] Further, in step S1, the mass concentration of the hexadecyltrimethylammonium bromide and water is 0.017~0.026 g / mL; the volume ratio of the ethyl acetate to water is 1:3.0~3.6; and the water is deionized water.

[0008] Further, the volume ratio of the tetraethyl orthosilicate to the triethyl phosphate is 17.0~19.0:1; the mass ratio of the calcium nitrate to the ferric nitrate is 0.50~1.50:1; the calcium nitrate is calcium nitrate tetrahydrate; and the ferric nitrate is ferric nitrate nonahydrate.

[0009] Furthermore, after the ethyl acetate-hexadecyltrimethylammonium bromide-water microemulsion is treated with ammonia, tetraethyl orthosilicate, triethyl phosphate, calcium nitrate, and ferric nitrate are added.

[0010] Further, the Fe-BG precipitate is dried, calcined, and ground to obtain Fe-BG powder, comprising: drying the Fe-BG precipitate at 50~70℃ for 22~26h, calcining it at 650~750℃ for 1.5~2.5h at a heating rate of 1.5~2.5℃ / min, and then grinding it to obtain Fe-BG powder.

[0011] The second objective of this invention is to provide a bioactive glass.

[0012] Furthermore, the bioactive glass is Fe-BG; the bioactive glass is nano-sized particles; the particle size of the nano-sized particles is 20~100nm.

[0013] Furthermore, the Fe-BG is amorphous; the Fe-BG has a mesoporous structure with an average pore size distribution of 7.32~8.20 nm.

[0014] The third objective of this invention is to provide an application of bioactive glass in the preparation of hemostatic and wound repair materials.

[0015] Furthermore, Fe-BG can improve the hemostatic efficiency of the hemostatic and wound repair materials, and has biocompatibility and structural stability.

[0016] Compared with existing technologies, this invention proposes a bioactive glass, its preparation method, and its application, which have the following beneficial effects: The bioactive glass proposed in this invention, through the incorporation of iron ions, exhibits stronger procoagulant ability and significantly shortens coagulation time. Furthermore, the amount of iron doping is concentration-dependent on the coagulation effect.

[0017] Furthermore, the Fe-BG prepared by this invention exhibits a low hemolysis rate (≤5%) while increasing the specific surface area and pore volume, meeting the blood compatibility requirements of biomedical materials. This indicates that it maintains excellent biocompatibility and is suitable for clinical hemostasis and wound repair.

[0018] Furthermore, the Fe-BG material prepared by this invention can effectively improve hemostasis efficiency and enhance the structural stability of the material while maintaining its biological activity. Attached Figure Description

[0019] Figure 1 The TEM, SEM, and EDX images of Fe-BG according to an embodiment of the present invention are shown; wherein, (a) is a TEM image; (b) is a SEM image; and (c) is an EDX image.

[0020] Figure 2 The following diagram illustrates the FT-IR, XRD, and Zeta potential maps of Fe-BG according to an embodiment of the present invention; wherein (a) is the FT-IR map; (b) is the XRD map; and (c) is the Zeta potential map.

[0021] Figure 3 The diagram shows a nitrogen adsorption / desorption isotherm and pore size distribution of Fe-BG according to an embodiment of the present invention.

[0022] Figure 4 The following diagram shows the WBCT and BCI test results of Fe-BG according to an embodiment of the present invention; wherein, (a) is the WBCT diagram; and (b) is the BCI diagram.

[0023] Figure 5 The diagram shows the TEG parameters of Fe-BG according to an embodiment of the present invention.

[0024] Figure 6 The following diagram shows the aPTT and PT test results of Fe-BG according to an embodiment of the present invention; wherein (a) is the aPTT diagram and (b) is the PT diagram.

[0025] Figure 7 The figure shows the hemolysis rate test results of Fe-BG according to an embodiment of the present invention.

[0026] Figure 8The diagram shows the WBCT test results of Fe-BG and Al-BG according to an embodiment of the present invention. Detailed Implementation

[0027] Exemplary embodiments of the present application will now be described in more detail with reference to the accompanying drawings. While exemplary embodiments of the present application are shown in the drawings, it should be understood that the present application may be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this application will be thorough and complete, and will fully convey the scope of the present application to those skilled in the art.

[0028] Unless otherwise specified, the technical means used in the following embodiments are conventional means well known to those skilled in the art, and the reagents and materials in this invention are obtained from the market or other public channels.

[0029] The Fe-BG of this invention is prepared using a modified sol-gel method, synthesized from raw materials such as tetraethyl orthosilicate, triethyl phosphate, calcium nitrate tetrahydrate, and ferric nitrate nonahydrate. By changing the iron doping concentration, Fe-BG samples with different properties are obtained. Furthermore, characterization of the material confirms that the iron-doped material has a higher surface area and pore volume, and a suitable pore size distribution, which contributes to improved bioactivity.

[0030] Example 1 This invention presents an experimental method for preparing iron-doped mesoporous bioactive glass (Fe-BG).

[0031] Mainly includes: Iron-doped mesoporous bioactive glass (Fe-BG) was prepared using an ethyl acetate-hexadecyltrimethylammonium bromide-water microemulsion as a soft template via a modified sol-gel method. First, 0.7 g of hexadecyltrimethylammonium bromide was dissolved in 33 mL of deionized water. Once the hexadecyltrimethylammonium bromide was completely dissolved, 10 mL of ethyl acetate was added. The mixture was stirred for 30 min to form microemulsion droplets, and then 5.6 mL of ammonia was added, followed by stirring for another 15 min. Tetraethyl orthosilicate, triethyl phosphate, calcium nitrate tetrahydrate, and different concentrations of ferric nitrate nonahydrate were added sequentially every 30 min (see Table 1). The resulting solution was then vigorously stirred for 4 h, and the clear solution gradually became opaque due to the formation of a white precipitate. The precipitate was collected by centrifugation and washed twice with deionized water and once with anhydrous ethanol. Subsequently, the precipitate was dried for 24 h and calcined at 700 °C for 2 h at a heating rate of 2 °C / min.

[0032] Table 1. Amounts of each raw material added in the preparation of Fe-BG Example 2 This embodiment presents TEM, SEM, and EDX analyses of iron-doped mesoporous bioactive glasses (Fe-BG) with different iron contents prepared in Example 1.

[0033] Mainly includes: Transmission electron microscopy (TEM) analysis: A small amount of sample was dispersed in ethanol, sonicated for 10 min to disperse evenly, dropped onto a copper grid, and dried. The microstructure of the sample was observed using a FEI Tecnai G2 F20 microscope with a working voltage of 200 kV.

[0034] Scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) analysis: A small amount of sample was directly adhered to conductive adhesive and sputtered with gold at 10 mA for 90 s using a Supro Instruments Minicoater ion sputtering system (China). Subsequently, the sample morphology and energy-dispersive X-ray mapping were performed using a ZEISS Sigma360 scanning electron microscope (Germany). The accelerating voltage for morphology imaging was 3 kV, and the accelerating voltage for energy-dispersive X-ray mapping was 15 kV. The EDX detector was an Oxford Xplore-30.

[0035] result: Please see Figure 1 (a) and Figure 1 (b) TEM and SEM analyses showed that bioactive glasses with different iron contents all exhibited nanoscale particle structures, with individual particle sizes mainly distributed in the range of 20–100 nm, exhibiting typical amorphous characteristics. With… With increasing doping concentration, the particle morphology gradually transforms from relatively regular spheres to irregular polyhedra, resulting in a significant increase in surface roughness and more compact particle aggregation. TEM analysis shows that all samples maintain good chemical homogeneity and a stable glassy structure; meanwhile, SEM analysis reveals that the increased surface roughness and morphological irregularity contribute to improving the material's specific surface area and bioactivity. (See also...) Figure 1 (c) The EDX spectrum shows that the distribution profiles of the five elements Si, O, Ca, P, and Fe are highly consistent. The Fe signal is clear and uniformly distributed within the particle size distribution, with no obvious local agglomeration or segregation observed. This indicates good chemical homogeneity of the material, and that iron has been effectively incorporated into the bioglass network structure. The evolution of the microstructure of this invention is as follows: The doping of bioactive glass provides a structural basis for enhancing its performance in hemostatic applications.

[0036] Example 3 This embodiment presents FT-IR, XRD, and Zeta potential analysis experiments on iron-doped mesoporous bioactive glasses (Fe-BG) with different iron contents prepared in Example 1.

[0037] Mainly includes: Fourier Transform Infrared Spectroscopy (FT-IR) Analysis: The samples were analyzed using a Thermo Nicolet 380 infrared spectrometer (USA). A portion of the sample was mixed with potassium bromide at a ratio of 1:100 and heated at 4000-4000 °C. Spectra were acquired within the wavenumber range, with a spectral resolution of 4. The number of scans was 32.

[0038] X-ray diffraction (XRD) analysis: The sample was laid flat on a glass slide and tested using a Rigaku SmartLab SE (Japan). The light source has a tube voltage of 40kV and a current of 40mA, with each step... speed from arrive Perform a scan at a speed of .

[0039] Zeta potential analysis: After dissolving the sample in deionized water, it was sonicated. The diluted solution was pipetted, and repeated pressing and aspiration were used to ensure homogeneous mixing. After several mixing operations, the sample was pipetted into the cuvette. The transparent surface of the cuvette was wiped with lens paper, and the test was performed using Zetasizer software (UK Malven Nano ZS90).

[0040] result: Please see Figure 2 (a) The FT-IR plot shows that Fe-BG has three classic BG peaks, the positions of which do not show significant changes, at 1087. 802 and 466 The characteristic peaks shown are Si-O-Si asymmetric stretching vibration, Si-O-Si symmetric stretching vibration, and Si-O bending vibration, respectively. Figure 2 (b) XRD patterns show that all bioactive glass samples exhibit typical amorphous characteristics, displaying continuous peaks without obvious sharp diffraction peaks in the 20-80° range. With... As the doping concentration increases, the diffraction intensity gradually increases, but the amorphous structure is still maintained, indicating that... Successfully incorporated into the silicate glass network as a network modifier. All samples were processed at approximately 22- and 30- The broadened scattering peaks appearing in the region are characteristic reflection peaks of the typical short-range ordered structure of silicate glass, indicating that... The introduction of this did not lead to the precipitation of a crystalline phase, and the amorphous structural characteristics required for bioactive glass were still maintained. Figure 2(c) The Zeta potential diagram shows that all bioactive glass samples exhibit negatively charged surface characteristics, with... With increasing doping concentration, the Zeta potential exhibits a monotonically increasing trend (although the absolute value gradually decreases). This change indicates that... The introduction of [something] gradually neutralizes the negative charge on the glass surface, reflecting [something]. As trivalent cations, they may accumulate on the glass surface. Based on the fact that the absolute values ​​of the Zeta potentials of all samples were greater than 20 mV, it is indicated that the particles have good dispersion stability in aqueous solution and are not prone to aggregation and precipitation. A moderate reduction in surface negative charge helps optimize the interaction between the iron-doped mesoporous bioactive glass and blood components, enhancing the binding ability with charged biomolecules while maintaining blood compatibility; this result is consistent with the procoagulant effect shown.

[0041] Example 4 This embodiment is a BET analysis experiment on iron-doped mesoporous bioactive glass (Fe-BG) with different iron contents prepared in Example 1.

[0042] Mainly includes: Specific surface area and porosity (BET) analysis: The sample was first degassed under vacuum at 200℃ for 6 hours, followed by adsorption and desorption tests using a Micromeritics ASAP 2460 microscope at 77K in liquid nitrogen. The specific surface area was calculated using the BET method, and the average pore size, pore volume, and pore size distribution were obtained by calculating the desorption isotherm using the BJH equivalent cylindrical model.

[0043] result: Please see Figure 3 Bioactive glasses with different iron contents exhibit unique pore structure characteristics and specific surface area variation patterns. All samples show typical type IV isotherms and H2 hysteresis loops, indicating that the materials have mesoporous structure characteristics, with average pore size distribution in the range of 7.32–8.20 nm. The effect of iron doping on specific surface area shows a nonlinear change, with Fe-BG1 having a specific surface area of ​​208.88 nm. Fe-BG2 increased significantly to 263.30. (+26.1%), Fe-BG3 fell back to 209.92. Fe-BG4 reached its highest value again at 267.22. (+27.9%). The pore volume change trend is basically consistent with the specific surface area, with Fe-BG4 exhibiting the largest pore volume (0.39%). This nonlinear change indicates that it originates from... As a network modifier, Fe-BG2 and Fe-BG4 exert complex regulatory effects on the network structure of silicate glass, resulting in different pore formation mechanisms at different doping concentrations. The high specific surface area and suitable mesoporous structure of Fe-BG2 and Fe-BG4 are beneficial for significantly enhancing the bioactivity and hemostatic properties of iron-doped mesoporous bioactive glasses.

[0044] Example 5 This embodiment is an in vitro coagulation evaluation experiment of iron-doped mesoporous bioactive glass (Fe-BG) with different iron contents prepared in Example 1.

[0045] Mainly includes: In this embodiment, whole blood coagulation time (WBCT) was used to quantitatively evaluate the hemostatic performance of the material by comparing the change in coagulation time before and after the addition of the hemostatic material. Fresh rabbit blood was collected in centrifuge tubes containing 3.8% sodium citrate. 20 mg of iron-doped mesoporous bioactive glass samples with different iron contents were placed in 5 mL glass tubes and incubated at 37°C for 5 min. Anticoagulated rabbit blood (1 mL) was mixed with the samples and incubated at 37°C for 3 min. Subsequently, 500 μL of 0.025M... The solution was set to trigger clotting and a timer was started. The glass tube was monitored at 15-second intervals by removing it from the water bath and inverting it, until the blood in the tube stopped flowing.

[0046] In this embodiment, the coagulation index (BCI) was used to quantitatively evaluate the procoagulant ability of the material. Fresh rabbit blood was collected in centrifuge tubes containing 3.8% sodium citrate. 50 mg of iron-doped mesoporous bioactive glass samples with different iron contents were added to beakers and incubated at 37°C for 5 min. Subsequently, 100 μL of fresh anticoagulated rabbit blood was added, followed by 20 μL of 0.2M sodium citrate. Solution. Subsequently, the mixture was incubated at 37°C for another 3 min; after adding 25 mL of deionized water, the mixture was incubated in a constant temperature shaker at 37°C and centrifuged; the absorbance of the supernatant at 545 nm was measured using a UV-Vis spectrophotometer (Techcomp UV2600, China). Each group underwent 3 tests and the average value was obtained. The whole blood coagulation index (BCI) was calculated by formula (1): BCI (%) = Abs sample group / Abs control group × 100% (1).

[0047] result: Please see Figure 4(a) WBCT images show that all iron-doped mesoporous bioactive glass samples exhibited significant procoagulant effects, with coagulation times significantly reduced compared to the blank control group (146.7±5.8 s). Among them, Fe-BG2 showed the best hemostatic performance, with a coagulation time of only 26.0±1.7 s, approximately 83% shorter than the blank group. With increasing iron content, the hemostatic effect initially increased and then decreased. This indicates that appropriate amounts of iron... Doping can significantly enhance the hemostatic properties of bioactive glass. Please refer to [link / reference]. Figure 4 (b) Based on the principle that a higher BCI value indicates a better procoagulant effect, the BCI plot shows that the procoagulant effects of the iron-doped mesoporous bioactive glass samples with different iron contents prepared in this invention are significantly better than those of the commercial control group LG (66.5±0.8%). Among them, Fe-BG2 shows the best coagulation performance, with a BCI value of approximately 39.4±0.8%, indicating that it has the strongest procoagulant ability. Furthermore, the coagulation effect of the iron-doped mesoporous bioactive glass samples shows a clear concentration dependence, namely Fe-BG2>Fe-BG3>Fe-BG4, and all are significantly better than the undoped Fe-BG1 (58.6±3.4%). The introduction of iron effectively enhances the procoagulant properties of bioactive glass. BCI and WBCT results corroborate each other, further confirming the superiority of iron-containing bioactive glass in hemostatic applications.

[0048] Example 6 This example is a TEG experiment on iron-doped mesoporous bioactive glass (Fe-BG) with different iron contents prepared in Example 1.

[0049] Mainly includes: Thromboelastography (TEG) records the entire dynamic process from coagulation initiation to thrombus formation and fibrinolysis by measuring the viscoelastic changes of blood samples under standardized conditions. 10 mg of each of iron-doped mesoporous bioactive glass samples with different iron contents were added to 1 mL of anticoagulated whole blood, gently shaken to mix thoroughly, and incubated at 37°C for 10 min. Subsequently, the whole blood solution (340 μL) and... The solution (20 μL, 0.2 mol / L) was added to the test cup and tested using a thromboelastography instrument. Unanticoagulated whole blood was used as a blank control group.

[0050] result: Please see Figure 5According to thromboelastography (TEG) analysis, iron-doped bioactive glass samples showed a significant advantage in improving coagulation function. R-value (coagulation reaction time) and K-value (coagulation formation time) are key indicators for evaluating coagulation rate. The R-values ​​(0.7–1.2 min) and K-values ​​(0.8 min) of all iron-doped mesoporous bioactive glass samples were significantly lower than those of the blank control group (R = 9.4 ± 0.5 min, K = 3.2 ± 0.1 min) and the commercial control LG (R = 14.2 ± 0.1 min, K = 3.8 ± 0.1 min), indicating a significant acceleration in the coagulation initiation and formation process. Among them, Fe-BG2 showed the best performance, with an R-value of only 0.7 ± 0.1 min, demonstrating the strongest procoagulant ability. The angle reflects coagulation dynamics; all iron-doped mesoporous bioactive glass samples angle All were significantly greater than the control group (51.1). ) and LG group (46.6) The results indicate a significantly accelerated fibrin polymerization rate. The maximum amplitude (MA) value showed no significant difference among the groups, suggesting similar final thrombus strength. The TEG results comprehensively confirmed that iron-containing bioactive glass can significantly shorten clotting time and accelerate the clotting process, with Fe-BG2 showing the best procoagulant effect, highly consistent with the results of WBCT and BCI tests.

[0051] Example 7 This embodiment is an aPTT and PT test analysis experiment on iron-doped mesoporous bioactive glass (Fe-BG) with different iron contents prepared in Example 1.

[0052] Mainly includes: Activated partial thromboplastin time (aPTT) is used to evaluate intrinsic coagulation pathways (factors XII, XI, IX, VIII), while prothrombin time (PT) is used to evaluate extrinsic coagulation pathways (factor VII). Both reflect the complete function of the coagulation system. In this example, anticoagulated whole blood was centrifuged at 3000 rpm for 15 min to obtain anemic platelet-rich plasma (PPP). 10 mg of each iron-doped mesoporous bioactive glass sample was mixed with 1 mL of PPP and incubated at 37°C for 30 min. Subsequently, 50 μL of plasma after sample removal was mixed with 50 μL of aPTT reagent and incubated at 37°C for 3 min. Then, 50 μL of preheated 0.025 mol / L... The solutions were added to the mixtures described above, and the time was immediately measured. The plasma coagulation time was recorded as aPTT. Similarly, 50 μL of the test plasma samples that had been co-incubated with the iron-doped mesoporous bioactive glass samples in the above steps were incubated at 37°C for 3 min. 100 μL of preheated PT reagent was added to each plasma sample, and the time was immediately measured. The plasma coagulation time was recorded as PT.

[0053] result: Please see Figure 6 (a) The aPTT plot showed no significant difference in aPTT time between the FeBG group and the blank group, indicating that Fe-BG could not activate the intrinsic pathway. Meanwhile, the aPTT test showed abnormal Fe-BG2 performance, with a prolonged clotting time of approximately 40.6 ± 0.8 s, significantly higher than the blank control group and other samples. This may be related to the fact that at this concentration... This relates to the specific effects of ions on intrinsic coagulation factors. Please refer to [link / reference]. Figure 6 (b) The PT plot shows that all iron-doped mesoporous bioactive glass samples significantly shortened prothrombin time, decreasing from 18.1±0.2 s in the control group to approximately 14-15 s, indicating a promoting effect on the extrinsic coagulation pathway. The above selective coagulation pathway influence pattern suggests that each iron-doped mesoporous bioactive glass exerts its hemostatic effect primarily by activating the extrinsic coagulation pathway.

[0054] Example 8 This embodiment is a blood compatibility experiment of iron-doped mesoporous bioactive glass (Fe-BG) with different iron contents prepared in Example 1.

[0055] Mainly includes: Anticoagulated whole blood was mixed with PBS at a ratio of 1:4 (v / v) and centrifuged at 250g for 15 min. The resulting red blood cells were then resuspended in PBS at a ratio of 1:9 (v / v) to prepare a red blood cell suspension. For the experimental groups, 5 mg of iron-doped mesoporous bioactive glass with different iron contents was added to 5 mL of the red blood cell suspension. A positive control was prepared by mixing red blood cells with deionized water at a ratio of 1:9 (v / v), while the negative control consisted of 5 mL of red blood cell suspension without any added sample. After incubation at 37°C for 1 h, all groups were centrifuged at 2000g for 15 min. The absorbance of the supernatant was measured at 540 nm (Techcomp UV2600), and the hemolysis rate was calculated using equation (2). Hemolysis rate (%) = ×100% (2) result: Please see Figure 7The hemolysis rate test results showed that all iron-doped mesoporous bioactive glass samples with different iron contents exhibited excellent blood compatibility, with hemolysis rates far below the safety standard of 5% (shown by the dashed line). The values ​​showed that Fe-BG1 had the highest hemolysis rate, approximately 2.42±0.08%, and the hemolysis rate decreased with increasing iron content: Fe-BG2 approximately 1.04±0.08%, Fe-BG3 approximately 0.71±0.04%, and Fe-BG4 the lowest, approximately 0.29±0.05%. Figure 7 The images show that the supernatant colors of the iron-doped mesoporous bioactive glass samples with different iron contents are similar to those of the negative control group, forming a stark contrast with the positive control group (which is deep red due to complete hemolysis). The results indicate... Doping not only did not increase the risk of hemolysis in the material, but also further reduced the damage to red blood cells by improving the surface properties of the material. All samples met the blood compatibility requirements for biomedical materials.

[0056] Comparative Example This invention also prepared aluminum-doped mesoporous bioactive glass Al-BG2 with the same content for in-depth comparison of hemostatic effects.

[0057] Please see Figure 8 WBCT images showed that Fe-BG2 (30.0±2.0s) achieved the shortest clotting time among all test groups, which was not only significantly better than the similar modified material Al-BG2 (46.7±2.9s), but also surpassed Kaolin (45.3±2.5s) in performance, proving that it is a highly competitive and efficient hemostatic material.

[0058] In summary, the Fe-BG samples prepared in this invention exhibit high surface area and pore volume with moderate pore size distribution. Furthermore, tests on coagulation time, coagulation index, and TEG of different Fe-BG samples confirmed that Fe-BG has significant advantages in hemostasis; the Fe-BG2 sample showed the best performance in promoting coagulation, significantly shortening the time compared to the blank group. Moreover, all Fe-BG samples with different properties exhibited low hemolysis rates (≤5%), meeting the blood compatibility requirements for biomedical materials, indicating their safety in clinical applications.

[0059] It should be noted that the term "comprising," or any other variation thereof, is 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 a process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0060] The above are merely embodiments of this application and are not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. A method of making a bioactive glass, characterized by, The preparation method comprises the following steps: S1, dissolving cetyltrimethylammonium bromide in water, adding ethyl acetate, and stirring to form microemulsion droplets to obtain an ethyl acetate-cetyltrimethylammonium bromide-water microemulsion; S2, adding tetraethyl orthosilicate, triethyl phosphate, calcium nitrate and ferric nitrate into the ethyl acetate-cetyltrimethylammonium bromide-water microemulsion in sequence to obtain Fe-BG precipitate; S3, drying, calcining and grinding the Fe-BG precipitate to obtain Fe-BG powder.

2. The method of claim 1, wherein the bioactive glass is prepared by a method comprising: In step S1, the mass concentration of cetyltrimethylammonium bromide and water is 0.017-0.026 g / mL; The volume ratio of ethyl acetate to water is 1:3.0-3.6; The water is deionized water.

3. The method of claim 1, wherein the bioactive glass is prepared by the method comprising: The volume ratio of tetraethyl orthosilicate to triethyl phosphate is 17.0-19.0:1; The mass ratio of calcium nitrate to ferric nitrate is 0.50-1.50:1; The calcium nitrate is calcium nitrate tetrahydrate; The ferric nitrate is ferric nitrate nonahydrate.

4. The method of claim 1, wherein the bioactive glass is prepared by the method comprising: The ethyl acetate-cetyltrimethylammonium bromide-water microemulsion is treated with ammonia water, and then tetraethyl orthosilicate, triethyl phosphate, calcium nitrate and ferric nitrate are added.

5. The method of claim 4, wherein the bioactive glass is prepared by a method comprising: The Fe-BG precipitate is dried, calcined and ground to obtain Fe-BG powder, which comprises the following steps: drying the Fe-BG precipitate at 50-70°C for 22-26 h, calcining at a heating rate of 1.5-2.5°C / min at 650-750°C for 1.5-2.5 h, and grinding to obtain Fe-BG powder.

6. A bioactive glass characterized by, The preparation method of the bioactive glass is prepared according to any one of claims 1-5.

7. The bioactive glass of claim 6, wherein, The bioactive glass is Fe-BG; The bioactive glass is a nano-sized particle; The nano-sized particle has a particle size of 20-100 nm.

8. The bioactive glass of claim 7, wherein, The Fe-BG is amorphous; The Fe-BG has a mesoporous structure, and the average pore size distribution is 7.32-8.20 nm.

9. The use of the bioactive glass prepared by the preparation method of any one of claims 1-5 or the bioactive glass of any one of claims 6-8 in the preparation of a hemostatic and wound repair material.

10. The use of a bioactive glass according to claim 9, characterized in that, The Fe-BG can improve the hemostatic efficiency of the hemostatic and wound repair material, has biocompatibility and structural stability.

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