A degradable bone hemostatic material with anti-inflammatory and pro-osteogenic functions and its preparation method and application

Abstract

The application discloses a degradable bone hemostatic material with anti-inflammatory and osteogenesis promoting functions and a preparation method and application thereof, and belongs to the technical field of biomedical materials. The specific steps are as follows: a liquor of glycyrrhizic acid is prepared; mesoporous bioglass powder is uniformly dispersed in the liquor of glycyrrhizic acid, and a reaction is carried out after the pH value is adjusted to 3.3-4, so that the glycyrrhizic acid molecules are induced to self-assemble to form a composite hydrogel, the mesoporous bioglass is uniformly loaded in the glycyrrhizic acid gel network, and the composite hydrogel is obtained; and the composite hydrogel is subjected to dehydration treatment, so that the water content of the composite hydrogel is ensured to be 60-80%, and the degradable bone hemostatic material with the anti-inflammatory and osteogenesis promoting functions is obtained. The application provides a composite material based on mesoporous bioglass (MBG) and glycyrrhizic acid (GA), and through multi-aspect regulation, the plasticity, mechanical properties and hemostatic activity of the material are synergistically optimized.

Description

Technical Field

[0001] This invention belongs to the field of biomedical materials technology, and more specifically relates to a biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions, its preparation method and application. Background Technology

[0002] In orthopedic surgery involving procedures such as medullary cavity, cancellous bone, vertebral body, and joint replacement, intraoperative and postoperative bleeding control has always been a major clinical challenge. Persistent bleeding from the bone wound not only obscures the surgical field and prolongs surgical time, but can also lead to serious complications such as postoperative hematoma, infection, delayed healing, and even secondary surgery. Currently, commonly used bone hemostatic materials are mainly divided into two categories: one is physical sealing materials, represented by bone wax, which, although possessing excellent plasticity and immediate sealing effect, is a non-degradable material that can induce chronic inflammation, interfere with new bone formation, and increase the risk of infection; the other category is... While absorbable hemostatic materials, such as gelatin sponges and oxidized cellulose, possess biodegradability, they suffer from drawbacks including insufficient adhesion, low mechanical strength, and difficulty in sealing high-pressure bone marrow hemorrhage. In contrast, bioactive materials such as chitosan and silicate bioceramics, developed in recent years, exhibit regenerative potential but generally face mechanical compatibility challenges, making it difficult to simultaneously meet the dual requirements of plastic filling and mechanical sealing. In summary, existing bone hemostatic materials face three major technical bottlenecks: the "plasticity-strength-degradability" ternary contradiction, the challenge of integrating "hemostasis-anti-inflammatory-osteogenic" functions, and limited means of performance regulation. Therefore, there is an urgent need in this field to develop a novel bone hemostatic material that combines the plastic sealing capabilities of bone wax with the biocompatibility of absorbable materials, while also possessing rapid hemostatic activity, anti-inflammatory regulation, osteogenic activity, and adjustable mechanical properties to meet diverse clinical needs.

[0003] Based on this, the present invention is proposed. Summary of the Invention

[0004] The purpose of this invention is to provide a biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions, its preparation method, and its application, in order to solve the problems existing in the prior art. This invention provides a composite material based on mesoporous bioglass (MBG) and glycyrrhizic acid (GA), achieving synergistic optimization of material plasticity, mechanical properties, and hemostatic activity through multi-faceted regulation.

[0005] To achieve the above objectives, the present invention provides the following solution: One of the technical solutions of this invention provides a method for preparing a biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions. The biodegradable bone hemostatic material is a composite material formed by the self-assembly of mesoporous bioglass (MBG) and glycyrrhizic acid (GA), specifically including the following steps: Mesoporous bioglass powder was uniformly dispersed in glycyrrhizic acid solution. After adjusting the pH value to 3.3-4, the reaction was carried out to induce the self-assembly of glycyrrhizic acid molecules to form a composite hydrogel, so that the mesoporous bioglass powder was uniformly loaded in the glycyrrhizic acid gel network, and the composite hydrogel was obtained. The composite hydrogel is dehydrated to ensure that the water content of the composite hydrogel is 60-80%, thereby obtaining the biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions.

[0006] Preferably, the composite hydrogel has a mass percentage of 0.25-2% for the mesoporous bioglass powder and a glycyrrhizic acid solution concentration of 10-17.5 wt%.

[0007] Preferably, the composite hydrogel contains 1% by mass of the mesoporous bioglass powder, and the glycyrrhizic acid solution has a concentration of 15 wt%.

[0008] Preferably, the preparation steps of the glycyrrhizic acid solution are as follows: dissolve glycyrrhizic acid powder in phosphate buffer solution to obtain the glycyrrhizic acid solution.

[0009] Preferably, the pH value after pH adjustment is 3.5 to 3.8.

[0010] Preferably, the reaction temperature is 80~110℃ and the time is 10~20min.

[0011] Preferably, the dehydration temperature is 50°C and the time is 0~48h, and not 0.

[0012] The second technical solution of the present invention provides a biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions prepared by the above preparation method. In the biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions, glycyrrhizic acid provides a self-assembled gel network and exerts anti-inflammatory activity, and mesoporous bioglass releases active ions to induce osteogenic differentiation.

[0013] The third technical solution of the present invention provides the application of the above-mentioned biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions in the preparation of hemostatic materials for bone defect repair or repair materials for inducing bone regeneration.

[0014] The fourth technical solution of the present invention provides a hemostatic material for bone defect repair, comprising the above-mentioned biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions.

[0015] Fifth technical solution of the present invention: Provides a repair material for inducing bone regeneration, comprising the above-mentioned biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions.

[0016] The technical principle of this invention is as follows: Although glycyrrhizic acid has good biocompatibility, pH-responsive self-assembly properties and anti-inflammatory activity, pure GA hydrogel has weak mechanical properties and limited hemostatic activity. Although mesoporous bioglass has excellent osteoconductivity and ion release ability, its powder form is difficult to mold into a malleable hemostatic material on its own.

[0017] This invention successfully developed a low-crosslinked biodegradable bone hemostatic material based on glycyrrhizic acid. It employs a dehydration-induced physical entanglement strategy, overcoming the limitations of traditional methods that rely on adjusting chemical crosslinking density to regulate material properties. Traditional methods often result in limited control range, fluctuating performance, and difficulty in overcoming the trade-off between stiffness and toughness. For example, traditional highly crosslinked hydrogels, due to their rigid covalent network restricting chain movement, only experience network densification during dehydration, failing to generate new energy dissipation pathways, leading to a narrow toughness control window and a sharp loss of ductility during strengthening. This invention introduces mesoporous bioglass into the low-crosslinked glycyrrhizic acid hydrogel, constructing a dynamic network structure rich in movable chains with sparse chemical crosslinking points as the framework, while maintaining its basic composition. During dehydration, the reduced interchain spacing promotes the dynamic formation of numerous physical entanglements. These entanglements not only significantly enhance material stiffness as temporary crosslinking points but also effectively dissipate energy through slip and reconstruction mechanisms, allowing the material to maintain ductility while increasing strength. By precisely controlling the degree of dehydration, the physical entanglement density can be continuously adjusted, thereby smoothly regulating material properties across a broad spectrum from "flexible" to "tough," exhibiting a much higher degree of design freedom and performance optimization potential than highly cross-linked systems. The dehydration-induced entanglement mechanism proposed in this invention provides a new paradigm for the design of biodegradable bone waxes that achieve a synergistic improvement in both high strength and high toughness.

[0018] The present invention discloses the following technical effects: This invention provides a biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions. Through the synergistic effect of MBG and GA, it retains the self-assembly gelling ability and bioactivity of GA while optimizing coagulation performance through the microstructure of MBG. Simultaneously, by adjusting the dosage of MBG and GA and undergoing dehydration post-treatment, a broad spectrum of mechanical properties can be controlled from low to high modulus to meet the mechanical requirements of different surgical sites. This hemostatic material combines the advantages of rapid coagulation, high burst pressure sealing, and low secondary bleeding rate. It outperforms traditional materials in animal models, and due to its good plasticity and suitable mechanical strength, it can perfectly fill bone defects and provide stable support and closure, exhibiting excellent clinical adaptability. Furthermore, the material possesses good biocompatibility and controllable degradation, with a precisely controllable degradation rate, effectively avoiding the risk of long-term foreign body residue. It systematically solves the "plasticity-strength-degradation" ternary contradiction and the "hemostasis-anti-inflammatory-osteogenic" functional integration problem faced by existing bone hemostatic materials, demonstrating promising clinical translation prospects and application value. Attached Figure Description

[0019] Figure 1 This is a schematic diagram illustrating the preparation process of the biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions described in this invention. Figure 2 The compressive stress-strain curves are shown for the composite materials prepared in Example 1 with different MBG and GA ratios, as well as for the materials prepared in Comparative Examples 2 and 3. Figure 3 The macroscopic morphological changes of the MBG@GA system and the GA system under different moisture contents and compression. Figure 4 The infrared spectra and XRD results are shown for the composite materials with different MBG and GA ratios described in Example 1 and the material prepared in Comparative Example 1. Figure 5 The changes in compressive stress-strain curves, compressive modulus, and fracture toughness of the MBG@GA system under different moisture contents are shown. Figure 6 MBG1@GA with a water content of 60% was prepared for Example 1. 15 Demonstration of the ability to manually shape and mold composite hydrogel samples; Figure 7 The results are quantitative analysis of the maximum burst pressure of the composite materials with different MBG and GA ratios described in Example 1, as well as the materials prepared in Comparative Examples 2 and 3. Figure 8 The results of quantitative analysis of the plugging duration of the composite materials with different MBG and GA ratios described in Example 1, as well as the materials prepared in Comparative Examples 2 and 3; Figure 9MBG1@GA with a water content of 60% was prepared for Example 1. 15 In vivo degradation rate of composite hydrogel samples; Figure 10 The results of quantitative analysis of coagulation status and coagulation time of whole blood from SD rats in different material groups in 96-well plates; Figure 11 A comparison of femoral condyle defect filling material before and after surgery and the intramuscular hematoma situation three days after surgery; Figure 12 Quantitative analysis of blood loss and hemostasis success rate 3 minutes postoperatively in a rat femoral defect model; Figure 13 To verify the anti-inflammatory effect of MBG@GA by Western blotting; Figure 14 ALP and ARS staining results for MBG@GA; Detailed Implementation Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0020] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0021] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0022] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0023] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0024] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0025] Unless otherwise specified, all raw materials used in the following embodiments of the present invention are commercially available products, and the source of commercially available products does not affect the technical effect of the present invention.

[0026] Unless otherwise specified, the room temperature involved in this invention is 25±5℃.

[0027] Example 1 Step 1: Preparation of experimental materials The glycyrrhizic acid (GA) used in this embodiment is a pharmaceutical-grade raw material with a purity of ≥93%, CAS number 1405-86-3, purchased from Maclean Biotechnology Co., Ltd.; the mesoporous bioglass (MBG) used is prepared in the laboratory with a particle size range of 100~300nm and a pore size of 2~50nm; all other reagents are analytical grade, and the experimental water is phosphate buffer.

[0028] Step 2: Preparation of GA solutions of different concentrations Accurately weigh glycyrrhizic acid powder and add it to beakers containing phosphate buffer (pH 7.4). Stir magnetically (300-500 rpm) at room temperature until completely dissolved to prepare GA solutions with mass percentage concentrations of 10%, 15%, and 17.5%. After preparation, allow the solutions to stand at room temperature for later use.

[0029] Step 3: Dispersion of MBG powder Weigh out mesoporous bioglass powders of different mass ratios (1% and 2% of the total mass of the prepared MBG@GA composite hydrogel, respectively), and slowly add them to the GA solutions of different concentrations prepared above under continuous stirring (500-800 rpm). After the addition is complete, continue stirring for 30-60 minutes and supplement with ultrasonic treatment to ensure that the MBG powder is uniformly dispersed in the GA solution and to avoid particle agglomeration.

[0030] Step 4: pH-induced self-assembly to form a hydrogel Adjust the pH of the mixture in step 3 to 3.8, and heat it in a 100°C water bath for 10 minutes to allow it to fully self-assemble and form a uniform and stable MBG@GA composite hydrogel, which is denoted as MBG1@GA. 10 (This refers to the mesoporous bioglass powder accounting for 1% of the total mass of the prepared MBG@GA composite hydrogel, and the GA solution having a mass percentage concentration of 10%), MBG1@GA15 (This refers to the mesoporous bioglass powder accounting for 1% of the total mass of the prepared MBG@GA composite hydrogel, and the GA solution having a mass percentage concentration of 15%), MBG1@GA 17.5 (This refers to the mesoporous bioglass powder accounting for 1% of the total mass of the prepared MBG@GA composite hydrogel, and the GA solution having a mass percentage concentration of 17.5%), MBG2@GA 15 (This refers to the fact that the mesoporous bioglass powder accounts for 2% of the total mass of the prepared MBG@GA composite hydrogel, and the mass percentage concentration of the GA solution is 15%).

[0031] Step 5: Post-dehydration treatment to adjust mechanical properties Compression tests were conducted according to the international standard ISO 604 Plastics—Determination of compressive properties. The results were recorded for the MBG@GA composite hydrogel prepared above and the GA hydrogel of Comparative Example 2. 15 The stress-strain curves of the bone glue (BX) described in Comparative Example 3 are as follows: Figure 2 As shown. By Figure 2 It can be seen that MBG1@GA 15 The properties of the composite hydrogel are closest to those of bone glue, so the MBG1@GA prepared above was used. 15 The composite hydrogel was dehydrated by placing it in a constant temperature and humidity chamber (50℃, 40-60% relative humidity). Samples with different moisture contents were obtained by controlling the dehydration time (0-48h). A series of samples with moisture contents of 89% (undehydrated), 60%, 39%, and 10% were prepared. The samples were weighed periodically during dehydration, and the real-time moisture content was calculated based on the weight changes until the target moisture content was reached. The samples were then sealed and stored for later use. The samples with different moisture contents were used for subsequent mechanical property testing. Figure 3 The three images on the right are MBG1@GA 15 Images of composite hydrogel samples with different water contents.

[0032] Comparative Example 1 The performance was tested using the mesoporous bioglass (MBG) from Example 1 as a sample.

[0033] Comparative Example 2 The performance was tested using GA hydrogel as a sample. The specific preparation process of GA hydrogel is as follows: Accurately weigh glycyrrhizic acid powder and add it to a beaker containing phosphate buffer (pH 7.4). Stir magnetically (300-500 rpm) at room temperature until completely dissolved to prepare a 15% (w / w) GA solution. After solution preparation, heat in a 100°C water bath for 10 minutes to allow for complete self-assembly, forming a homogeneous and stable GA hydrogel, denoted as GA. 15.

[0034] Figure 3 The three images on the left are actual images of GA hydrogel samples with different water contents.

[0035] Figure 4 Characterization results of MBG@GA composite hydrogels prepared in Example 1 with different MBG and GA ratios, and of the sample obtained in Comparative Example 1.

[0036] Fourier transform infrared spectroscopy further confirmed MBG1@GA 15 Successful preparation of MBG1@GA 15 The absorption peak of glycyrrhizic acid can be observed in the spectrum. It is located at 3430 cm⁻¹. -1 Left and right (OH stretching vibration), 2943 / 2860cm -1 (Hydrogen bond association), 1734cm -1 (C=O stretching vibration), 1664cm -1 (C=C stretching vibration in the triterpenoid skeleton) and 1081 cm⁻¹ -1 It appears at the (CO vibration of polysaccharides). Simultaneously, at 776 cm⁻¹... -1 and 1081cm -1 The characteristic peaks can be attributed to the Si-O-Si bending and tensile vibrations of the mesoporous bioactive glass nanoparticles, respectively, indicating that MBGNs successfully exist in MBG1@GA 15 XRD results showed that MBG and MBG@GA composite systems with different ratios generally exhibited broad, diffuse peaks, indicating that the samples mainly maintained amorphous or low-crystallinity structural characteristics. Notably, GA-related samples all showed obvious broad peak signals around 13–16°, suggesting the presence of locally ordered structures formed by the stacking of GA molecules in the system. The intensity and shape of this broad peak changed accordingly with variations in the GA to MBG ratio.

[0037] Comparative Example 3 Commercially available bone glue (BX) was used as a sample for performance testing. The bone glue was purchased from Johnson & Johnson (China) Medical Devices Co., Ltd., model W810T.

[0038] Performance testing: 1. Mechanical property testing: A series of samples with different moisture contents prepared in Example 1 and a sample from Comparative Example 3 were subjected to compression tests according to the international standard ISO 604 Plastics—Determination of compressive properties. Their stress-strain curves, compressive modulus, and fracture toughness were recorded. The results are as follows: Figure 5 As shown.

[0039] Depend on Figure 5 It can be seen that MBG1@GA 15The mechanical evolution of MBG1@GA is significantly different. As the water content decreases, its compressive stress-strain curve continuously shifts upward and becomes steeper. Not only do peak stress and stiffness increase simultaneously, but it also retains a large deformation space in the medium water content range, indicating that the material does not immediately lose its energy dissipation capacity while being strengthened. Further quantitative analysis shows that its compressive modulus reaches its highest value at 39% water content, while its fracture toughness reaches its maximum at 60% water content. This result demonstrates that dehydration significantly affects the mechanical evolution of MBG1@GA. 15 The effect is not simply a monotonous enhancement, but rather, by adjusting the density and dynamics of physical entanglement, it triggers different dominant mechanical mechanisms in different moisture content ranges. Specifically, around 60% moisture content, the system has formed enough entanglement to improve load-bearing capacity, while still retaining some space for chain segment / fiber slippage and rearrangement, thus exhibiting better energy dissipation and the highest fracture toughness. However, when the moisture content further decreases to 39%, network densification and topological constraints continue to enhance, causing the material to exhibit the highest compressive modulus. At the same time, chain segment dynamics begin to be limited, and toughness decreases. This indicates that MBG1@GA... 15 There exists an "optimal entanglement window" induced by dehydration, within which reinforcement and toughening can be achieved synergistically, while deeper dehydration will cause the system to gradually become stiff and brittle.

[0040] MBG1@GA with a water content of 60% prepared in Example 1 was used. 15 The composite hydrogel samples underwent plasticity testing, and the results are as follows: Figure 6 As shown.

[0041] Depend on Figure 6 It can be seen that MBG1@GA with a water content of 60% 15 The composite hydrogel samples exhibit good manipulability, capable of being molded into various geometric shapes such as curved, elongated, and sheet-like forms under external force. This indicates that they possess similar operational adaptability to clinical bone wax, making them suitable for rapid conformal coverage of irregular bone defect surfaces. For bone surface sealing materials, this plasticity not only relates to ease of operation but also directly affects the integrity of adhesion to the bone cavity boundary and the subsequent sealing effect.

[0042] The MBG@GA composite hydrogel prepared in Example 1 was dehydrated using the same method as in Example 1 to obtain MBG@GA composite hydrogel samples with a water content of 60%. These samples, along with those from Comparative Examples 2 and 3, were subjected to burst pressure tests. The specific test steps were as follows: The burst pressure test was used to evaluate the sealing performance of the hydrogel under pressure. First, the bone fragment was fixed onto a self-made testing device, with a circular defect placed in the center of the device. Then, the composite hydrogel sample was evenly applied to the surface of the bone fragment, ensuring full adhesion to the edges. PBS solution was slowly and continuously injected into the device using a syringe pump, and the pressure changes within the system were recorded in real time using a pressure sensor. The pressure value at which liquid leakage or rupture first occurred in the sealed area of ​​the material was defined as the burst pressure of the sample. The results are as follows: Figure 7 As shown.

[0043] Depend on Figure 7 It can be seen that MBG1@GA with a water content of 60% 15 The average maximum burst pressure of the composite hydrogel sample reached 46.6 kPa, which was not significantly different from the 46.3 kPa of commercial bone wax (p=0.9861), indicating that it has excellent pressure resistance and sealing ability.

[0044] The MBG@GA composite hydrogel prepared in Example 1 was dehydrated using the same method as in Example 1 to obtain MBG@GA composite hydrogel samples with a water content of 60%. These samples, along with those from Comparative Examples 2 and 3, were tested for sealing performance. The specific testing steps were as follows: A self-made in vitro sealing device was used to determine the average sealing time. First, a bone fragment was fixed onto the testing device, and a circular defect was pre-set in the center of the device. Then, a certain amount of sample was applied to the surface of the defect area and gently pressed to ensure it adhered fully to the edge of the bone tissue. Afterward, PBS solution was continuously injected into the device at a constant flow rate using a syringe pump, and timing was started simultaneously. The sealing time was defined as the endpoint when liquid seepage appeared on the surface of the defect area. The results are as follows: Figure 8 As shown.

[0045] Depend on Figure 8 It can be seen that MBG1@GA with a water content of 60% 15 The sealing duration of the composite hydrogel sample also remained at a high level, similar to that of bone wax, indicating that the material can not only quickly establish an effective seal, but also maintain a stable seal under continuous liquid impact.

[0046] In summary, the biodegradable bone hemostatic material prepared by this invention, possessing anti-inflammatory and osteogenic functions, exhibits excellent plasticity, allowing it to be hand-shaped into complex geometries and tightly conform to irregular bone defect cavities. Regarding sealing performance, the material can achieve a burst pressure of up to 46.6 kPa (approximately 350 mmHg) and a sealing duration exceeding 600 seconds, comparable to the clinical gold standard bone wax. By adjusting the component ratio and degree of dehydration, its mechanical properties can be continuously controlled from "soft and tough" to "strong and tough" and then to "high rigidity." For example, at a water content of 60%, its compressive modulus, ultimate strength, and fracture toughness are not significantly different from bone wax.

[0047] 2. Degradation effect: To evaluate the in vivo degradation behavior and retention of the material at the femoral defect site, samples were collected at predetermined time points post-surgery to observe changes in material residue and distribution within the defect area. First, MBG1@GA with a water content of 60% was implanted into a rat femoral defect model. 15 Composite hydrogel samples, BX, and GS were collected. Rats were sacrificed at 1, 7, 14, and 21 days post-surgery, and the remaining material at the defect site was completely removed, weighed, and the degradation rate was calculated. The in vivo controllable degradation characteristics of the material and its dynamic changes during bone defect repair were comprehensively evaluated. The results are as follows: Figure 9 As shown.

[0048] Depend on Figure 9 It can be seen that at the site of femoral defect, the material degrades significantly over time, with MBG1@GA containing 60% water content showing the most significant degradation. 15 The distribution of the composite hydrogel sample in vivo decreased over time, and its degradation rate was faster than that of the gelatin sponge, while bone wax showed almost no degradation. This suggests that its controllable degradation characteristics can sustainably provide support and create a window of opportunity for bone repair. This degradation kinetics provides potential support for the continuous new bone formation observed in subsequent Micro-CT and histological observations, indicating that the material degradation and bone regeneration processes may have a good time-matching relationship.

[0049] 2. Hemostatic performance test: MBG1@GA with a moisture content of 60% 15 Composite hydrogel samples and control materials (BX, GA, GS (gelatin sponge), and blank group (CON)) were placed in 96-well plates, and fresh anticoagulated whole blood was added. Clotting time was recorded, and the results are as follows: Figure 10 As shown.

[0050] Depend on Figure 10 The 96-well plate experiment results show that different materials have significantly different effects on the whole blood coagulation process. Among them, MBG1@GA with a water content of 60% showed the most significant differences. 15 The composite hydrogel sample exhibited the shortest clotting time. This indicates that MBG1@GA with a water content of 60% showed the best clotting time. 15Composite hydrogel samples can significantly shorten the coagulation initiation time, thereby creating a more favorable time window for subsequent rapid hemostasis.

[0051] In vivo hemostasis: A femoral condyle defect hemorrhage model was established in SD rats, and MBG1@GA with a water content of 60% was injected. 15 The blood loss and hemostasis success rate within 3 minutes were recorded for the composite hydrogel sample and the control group materials (BX, GS (gelatin sponge), and blank group (CON)). Hematoma status was observed 1 week postoperatively.

[0052] The results are as follows Figure 11 and Figure 12 As shown.

[0053] Depend on Figure 11 and Figure 12 It can be seen that further validation of MBG1@GA with a water content of 60% in animal models of bone defects 15 In vivo hemostatic effect of composite hydrogel samples. First, a rat femoral condyle defect model was established, and the sealing status before and after filling with different materials was compared; observation of intramuscular hematoma 3 days post-surgery further showed that MBG1@GA with a water content of 60% was the most effective. 15 The composite hydrogel sample group effectively reduced postoperative local hematoma formation, suggesting that it can not only control bleeding during surgery but also reduce early local tissue damage caused by continuous postoperative oozing. In a rat femoral defect model, for blood loss, MBG1@GA with a water content of 60% showed significant improvement. 15 The composite hydrogel sample group was almost identical to the bone wax group, but significantly lower than the blank control group and the gelatin sponge group. Meanwhile, the hemostasis success rate at 3 minutes post-surgery in the rat model was significantly lower in BX, GS, and MBG1@GA samples with 60% water content. 15 The water content in the composite hydrogel samples was 100%, 10%, and 99%, respectively, further indicating that MBG1@GA has a water content of 60%. 15 The composite hydrogel sample showed significantly better early and rapid hemostasis than the traditional gelatin sponge.

[0054] 3. Anti-inflammatory effect test: The anti-inflammatory activity of the material was evaluated using bone marrow-derived macrophages. MBG1@GA, prepared in Example 1 with a water content of 60%, was used. 15 The composite hydrogel samples were placed in Transwell chambers and co-cultured with lipopolysaccharide (LPS)-stimulated macrophages for 24 h. Cellular proteins were extracted, and the expression of iNOS and CD206 pro-inflammatory proteins was detected by Western blotting (wb). The results are as follows: Figure 13 As shown.

[0055] Depend on Figure 13Western blot results showed that the levels of M1-related proteins iNOS and IL-1β were significantly elevated in the LPS group, while they were significantly elevated in the MBG1@GA group with a water content of 60%. 15 The levels of the composite hydrogel samples decreased significantly after treatment; simultaneously, the levels of M2-related proteins ARG-1 and CD206 decreased in MBG1@GA samples with a water content of 60%. 15 The water content was significantly upregulated in the composite hydrogel sample group, and overall higher than in the GA group. This indicates that MBG1@GA with a water content of 60% is significantly upregulated. 15 The composite hydrogel sample does not simply "reduce inflammation" in regulating macrophage polarization, but rather achieves bidirectional regulation of both M1 inhibition and M2 promotion, which is an important reason why it is superior to GA treatment alone.

[0056] 4. Promotes bone function evaluation: The osteogenic activity of the materials was evaluated using rat bone marrow mesenchymal stem cells (rBMSCs). MBG1@GA with a water content of 60% prepared in Example 1 was used. 15 Composite hydrogel samples were co-cultured with rBMSCs. After 7 days of culture, the activity of ALP, an early marker of osteogenic differentiation, was detected by alkaline phosphatase (ALP) staining and quantitative kit. After 14 days of culture, the formation of mineralized nodules was observed by Alizarin Red (ARS) staining and quantitative analysis was performed. The results are as follows: Figure 14 As shown.

[0057] Depend on Figure 14 It can be seen that ALP staining 7 days after osteogenic induction showed that MBG1@GA with a water content of 60% 15 The composite hydrogel sample treated with BCM exhibited the strongest early osteogenic activity. Furthermore, in ARS staining 14 days after osteogenic induction, the combined group also showed the strongest mineralization deposition, indicating that the 60% water content of MBG1@GA... 15 The composite hydrogel sample not only promotes early osteogenic initiation, but also continuously drives later mineralization maturation.

[0058] Experimental results show that the optimized formulation of the product combines excellent plasticity and sealing performance, with a burst pressure of 46.6 kPa, comparable to the clinical gold standard bone wax; its in vitro clotting time is significantly better than existing hemostatic materials. In rat, rabbit, and beagle dog bone defect models, the material achieved a 100% hemostasis success rate within 3 minutes, with mild postoperative hematoma, low secondary bleeding rate, and controllable degradation. In particular, the material exhibits good anti-inflammatory activity, regulating the local inflammatory microenvironment, and simultaneously inducing osteogenic differentiation of stem cells through the release of active ions from MBG, promoting mineralization and regeneration at the bone defect site. The material also possesses excellent plasticity, allowing it to be manually shaped into complex geometries to fit irregular bone defect cavities; by adjusting the component ratio and dehydration level, its mechanical properties can be continuously controlled within a broad spectrum from "soft and tough" to "strong and tough" to "high rigidity," meeting the mechanical requirements of different surgical sites. This material has multiple functions, including rapid hemostasis, anti-inflammation, bone promotion, mechanical adjustability, and controllable degradation. It can effectively avoid the risk of long-term foreign body residue and has good clinical suitability and application prospects.

[0059] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0060] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions, characterized in that, Includes the following steps: Mesoporous bioglass powder was uniformly dispersed in glycyrrhizic acid solution. After adjusting the pH value to 3.3-4, the reaction was carried out to induce the self-assembly of glycyrrhizic acid molecules to form a composite hydrogel, so that the mesoporous bioglass powder was uniformly loaded in the glycyrrhizic acid gel network, and the composite hydrogel was obtained. The composite hydrogel is dehydrated to ensure that the water content of the composite hydrogel is 60-80%, thereby obtaining the biodegradable bone hemostatic material with anti-inflammatory and bone-promoting functions.

2. The preparation method according to claim 1, characterized in that, The composite hydrogel contains 0.25-2% by mass of the mesoporous bioglass powder; the glycyrrhizic acid solution has a concentration of 10-17.5 wt%.

3. The preparation method according to claim 2, characterized in that, The composite hydrogel contains 1% by mass of the mesoporous bioglass powder; the glycyrrhizic acid solution has a concentration of 15 wt%.

4. The preparation method according to claim 1, characterized in that, The preparation steps of the glycyrrhizic acid solution are as follows: dissolve glycyrrhizic acid powder in phosphate buffer to obtain the glycyrrhizic acid solution.

5. The preparation method according to claim 1, characterized in that, The pH value after adjustment is 3.5~3.8; the reaction temperature is 80~110℃, and the time is 10~20min.

6. The preparation method according to claim 1, characterized in that, The dehydration treatment is carried out at a temperature of 50°C for a time of 0 to 48 hours, and is not 0.

7. The biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions prepared by the preparation method according to any one of claims 1 to 6.

8. The use of the biodegradable bone hemostatic material with anti-inflammatory and osteogenic functions as described in claim 7 in the preparation of hemostatic materials for bone defect repair or repair materials for inducing bone regeneration.

9. A hemostatic material for bone defect repair, characterized in that, The biodegradable bone hemostatic material comprising the anti-inflammatory and osteogenic functions as described in claim 7.

10. A repair material for inducing bone regeneration, characterized in that, The biodegradable bone hemostatic material comprising the anti-inflammatory and osteogenic functions as described in claim 7.

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