Mesoporous bioactive glass-based coaxial electrospun bioactive fiber membranes, methods of making and uses thereof

Abstract

The application belongs to the technical field of biomedical materials, and discloses a mesoporous bioactive glass-based coaxial electrospinning bioactive fiber membrane, a preparation method and application thereof. The fiber membrane is a composite fiber membrane composed of core-shell structure composite fibers prepared by coaxial electrospinning. The inner core of the core-shell structure composite fiber is composed of polycaprolactone and mesoporous bioactive glass, and the outer shell is composed of polycaprolactone, silk fibroin and nerve growth factor. The surface of the mesoporous bioactive glass particles is modified with amino groups. The mesoporous bioactive glass particles contain the following metal elements in mass fraction: calcium 64-92 parts, magnesium 5-20 parts, strontium 1-8 parts, sodium 1-3 parts, and potassium 1-5 parts. The fiber membrane can significantly promote the proliferation, differentiation, mineralization and osteogenic gene expression of bone marrow mesenchymal stem cells, and activate key signal pathways such as Runx2, and can efficiently guide the generation of high-quality new bone in a mouse calvarial critical size defect model.

Description

Technical Field

[0001] This invention belongs to the field of biomedical materials technology, and particularly relates to a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane and its preparation method, application and formulation screening method. Background Technology

[0002] Bone defects of critical size, usually caused by severe trauma, tumor resection, or congenital malformations, have always been a major challenge in clinical orthopedics and regenerative medicine because they exceed the physiological limits of bone tissue's self-repair. Currently, autologous bone grafting is considered the gold standard for bone defect repair, but its sources are limited and can cause secondary damage to the donor site and potential complications. While allogeneic bone grafting solves the source problem, it still faces issues such as immune rejection, the risk of disease transmission, and inconsistent bone integration capabilities, often leading to poor prognoses such as delayed healing, scar tissue formation, or nonunion.

[0003] Against this backdrop, bone tissue engineering, as a promising alternative strategy, aims to mimic the microenvironment of natural bone healing and guide and accelerate bone regeneration by constructing a ternary composite system of biomaterials, cells, and active factors. Among the three key elements of bone tissue engineering, bioscaffold materials are the core foundation. Bioactive ceramics, particularly since the development of bioactive glass by Hench et al. in 1971, have received extensive and in-depth research due to their excellent osteoconductivity, biocompatibility, and chemical composition similar to the bone mineral phase (hydroxyapatite). Mesoporous bioactive glass goes a step further, exhibiting unique advantages in drug / factor loading and controlled release due to its high specific surface area and ordered nanoscale pore structure, making it a cutting-edge research hotspot in bone repair materials.

[0004] Natural bone tissue is a highly complex composite material in terms of composition and structure. Its inorganic phase accounts for approximately 70% of its dry weight, with carbon-doped hydroxyapatite being the predominant crystalline form, accounting for over 95%. Notably, the remaining approximately 5% of the mineral composition contains several key functional ions, such as sodium, potassium, magnesium, and strontium. Studies have shown that these trace elements are not insignificant; they play crucial regulatory roles in bone metabolism: sodium and potassium ions participate in maintaining cell membrane potential and osmotic pressure balance; magnesium ions are cofactors for various osteogenic enzymes, and their deficiency is closely related to osteoporosis; strontium ions have been shown to bidirectionally regulate osteoclast and osteoblast activity, significantly enhancing bone strength. Furthermore, citric acid is abundant in bone minerals; it may influence crystal nucleation, growth, and dissolution through chelation with calcium ions on the surface of hydroxyapatite, and participate in cellular energy metabolism. Therefore, the design of next-generation biomimetic bone repair materials should not be limited to mimicking the main apatite phase, but should also emphasize the synergistic introduction of these functional components to more accurately replicate the biochemical microenvironment of natural bone.

[0005] However, integrating multiple functional components into a single material system presents a significant challenge—determining the optimal ratio of each component. Traditional trial-and-error experimental strategies, when faced with complex systems containing six or more components, result in an exponential increase in the number of experimental combinations, leading to lengthy development cycles, high costs, and low efficiency.

[0006] In recent years, machine learning, as a powerful data-driven tool, has demonstrated revolutionary potential in the field of materials science. It can efficiently predict material properties and guide optimized design by uncovering hidden patterns in high-dimensional data, thereby significantly accelerating the research and development of new materials. Combining machine learning with high-throughput experiments provides a novel approach to solving the challenge of optimizing the proportions of multi-component biomaterials. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the deficiencies and defects mentioned in the background art above, and to provide a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane and its preparation method, application and ratio screening method.

[0008] To solve the above-mentioned technical problems, the technical solution proposed by this invention is as follows:

[0009] In a first aspect, the present invention provides a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, which is a composite fiber membrane composed of core-shell structured composite fiber filaments prepared by coaxial electrospinning; the core of the core-shell structured composite fiber filament is composed of polycaprolactone and mesoporous bioactive glass, and the outer shell is composed of polycaprolactone, silk fibroin and nerve growth factor; the surface of the mesoporous bioactive glass particles is modified with amino groups; the mesoporous bioactive glass particles contain the following metal elements in parts by mass: calcium 64-92 parts, magnesium 5-20 parts, strontium 1-8 parts, sodium 1-3 parts, and potassium 1-5 parts.

[0010] In the aforementioned mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, the mass ratio of polycaprolactone to mesoporous bioactive glass particles in the core of the core-shell composite fiber is 8-12:1; and the mass ratio of polycaprolactone, silk fibroin, and nerve growth factor in the shell of the core-shell composite fiber is 24000-30000:600-750:1.

[0011] Furthermore, the thickness of the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane is 15μm-40μm; the fiber diameter of the core-shell structure composite fiber is 80-120nm, wherein the diameter ratio of the core to the shell is 1:1.8-2.4.

[0012] Furthermore, the mesoporous bioactive glass particles contain the following metal elements in parts by weight: 88 parts calcium, 8 parts magnesium, 1 part sodium, 2 parts potassium, and 1 part strontium.

[0013] Secondly, the present invention provides a method for preparing a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, comprising the following steps: (1) Weigh out the corresponding masses of nitrates of five metal elements: calcium, magnesium, sodium, potassium and strontium, and prepare mesoporous bioactive glass using the sol-gel method; (2) The surface of the mesoporous bioactive glass is modified with an amino group using a silane coupling agent; (3) The surface-modified mesoporous bioactive glass particles and polycaprolactone are dispersed in a solvent to form a core spinning solution. Polycaprolactone, silk fibroin and nerve growth factor solution are dissolved in a solvent to form a shell spinning solution. Then, coaxial electrospinning is performed to form a composite fiber membrane composed of core-shell structure composite fiber filaments, which is the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane.

[0014] Further, in step (1), the sol-gel method described above is as follows: the template agent and the pore-expanding agent are dissolved in an acidic solution with a pH of 0.8-1.5, and then nitrates corresponding to five metal elements, namely calcium, magnesium, sodium, potassium and strontium, are added to control the total number of cation moles to be 0.015-0.017 mol. Then, tetraethyl orthosilicate and triethyl phosphate are added and mixed and stirred to form a sol. After adjusting the pH to 10.0-11.0, the mixture is heated at 90-95℃ for 64-72 hours. The resulting precipitate is washed, dried, and then calcined at 500-600℃ for 6-8 hours to obtain the mesoporous bioactive glass particles. The template agent is a poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer, and the pore-expanding agent is 1,3,5-trimethylbenzene.

[0015] Furthermore, in step (2), the silane coupling agent is (3-aminopropyl)triethoxysilane, and the amino modification method is as follows: the mesoporous bioactive glass particles obtained in step (1) are dispersed in an organic solvent solution of (3-aminopropyl)triethoxysilane, stirred at room temperature, and then separated, washed, and dried to obtain surface-modified mesoporous bioactive glass particles.

[0016] Furthermore, in step (3), the solvent is hexafluoroisopropanol; the mass ratio of the surface-modified mesoporous bioactive glass particles to polycaprolactone is 1:8-12, the mass ratio of polycaprolactone, silk fibroin and nerve growth factor is 24000-30000:600-750:1, and the concentration of the nerve growth factor solution is 0.0001-0.0002 μg / mL.

[0017] Furthermore, in step (3), the parameters of the coaxial electrospinning are as follows: the flow rate of the outer shell is 0.6-1 mL / h, the flow rate of the inner core is 0.6-1 mL / h, the voltage is 18-22 kV, the receiving distance is 20-30 cm, the ambient temperature is 20-27℃, and the ambient humidity is 40-50%.

[0018] Thirdly, the present invention provides the application of a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane in the preparation of products for repairing bone defects, wherein the product for promoting bone defect repair is a drug or medical device.

[0019] Furthermore, the product described above is a product that repairs bone defects by activating the Runx2 signaling pathway.

[0020] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane of the present invention can significantly promote the proliferation, differentiation, mineralization and osteogenic gene expression of bone marrow mesenchymal stem cells, and activate key signaling pathways such as Runx2. In a mouse model of critical size defects in the skull, it can efficiently guide the formation of high-quality new bone, and the repair effect is significantly better than that of traditional materials.

[0021] 2. This invention prepares a mesoporous bioactive glass-based coaxial electrospun fiber membrane by using the optimal ratio of materials. It combines the advantages of loading and slow-release of mesoporous materials, the protective and controlled release ability of coaxial spinning for active factors, and the three-dimensional biomimetic structure of the fiber scaffold, thereby further enhancing its osteogenic properties.

[0022] 3. This invention achieves rational design and optimization of multi-component bone-mimicking mineral phase systems, breaking through the bottleneck of traditional "trial and error" methods in multi-component optimization. It can efficiently screen out the optimal solution from a vast chemical ratio space. The obtained material accurately simulates the key functional components of natural bone (Ca, P, Mg, Sr, Na, K), and each component works synergistically under optimal ratios, with Sr... 2+ With Mg 2+ As a signaling ion to activate the osteogenic pathway, Na + With K + Maintaining basic cellular functions, they work together to create a biomimetic microenvironment conducive to bone regeneration, thereby achieving multi-level, proactive regulation of the bone regeneration process. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0024] Figure 1 This is a typical nitrogen adsorption-desorption curve of the mesoporous bioactive glass prepared in Example 1 of the present invention.

[0025] Figure 2 This is a fluorescence staining image of the bioactive fiber membrane prepared in Example 1 of the present invention.

[0026] Figure 3 This is a scanning electron microscope image of the bioactive fiber membrane prepared in Example 1 of the present invention.

[0027] Figure 4 This is a scanning electron microscope image of the bioactive fiber membrane prepared in Example 1 of the present invention mineralized in SBF solution in vitro.

[0028] Figure 5 The infrared spectrum of the bioactive fiber membrane prepared in Example 1 of this invention is shown.

[0029] Figure 6 This diagram illustrates the effect of the bioactive fiber membrane prepared in Example 1 of this invention on the proliferation of BMSCs.

[0030] Figure 7 This is a diagram showing the proliferation and differentiation results of BMSCs on the bioactive fiber membrane prepared in Example 1 of the present invention.

[0031] Figure 8 This is a diagram showing the mineralization results of BMSCs by the bioactive fiber membrane prepared in Example 1 of this invention.

[0032] Figure 9 This is a schematic diagram of the results of qPCR (real-time quantitative polymerase chain reaction) of the bioactive fiber membrane prepared in Example 1 of the present invention.

[0033] Figure 10 This is a fluorescence staining image of BMSCs cultured from the bioactive fiber membrane prepared in Example 1 of this invention.

[0034] Figure 11 This is a fluorescent staining image of BMSCs induced by the bioactive fiber membrane prepared in Example 1 of the present invention to induce neural differentiation.

[0035] Figure 12 This is a MicroCT (micro-computed tomography) image showing the effect of the bioactive fiber membrane prepared in Example 1 of this invention on osteogenic repair in a mouse skull defect model.

[0036] Figure 13 This is a BV / TV (bone volume fraction) analysis diagram showing the effect of the bioactive fiber membrane prepared in Example 1 of the present invention on osteogenic repair in a mouse skull defect model.

[0037] Figure 14 The image shows the HE (hematoxylin-eosin) staining results of the effect of the bioactive fiber membrane prepared in Example 1 of this invention on osteogenic repair in a mouse skull defect model.

[0038] Figure 15 This is a scanning electron microscope image of the bioactive fiber membrane prepared in Example 2 of the present invention.

[0039] Figure 16 This is a scanning electron microscope image of the bioactive fiber membrane prepared in Example 2 of the present invention mineralized in SBF solution in vitro.

[0040] Figure 17 This is a MicroCT (micro-computed tomography) image showing the effect of the bioactive fiber membrane prepared in Example 2 of this invention on osteogenic repair in a mouse skull defect model.

[0041] Figure 18 This is a scanning electron microscope image of the bioactive fiber membrane prepared in Example 3 of the present invention.

[0042] Figure 19 This is a scanning electron microscope image of the bioactive fiber membrane prepared in Example 3 of the present invention mineralized in SBF solution in vitro.

[0043] Figure 20 This is a MicroCT (micro-computed tomography) image showing the effect of the bioactive fiber membrane prepared in Example 3 of this invention on osteogenic repair in a mouse skull defect model.

[0044] In the above figures, PCL is a bioactive fiber membrane composed of polycaprolactone; PCL@SF / NGF is a bioactive fiber membrane composed of polycaprolactone, silk fibroin, and nerve growth factor; MBG@SF / NGF is a bioactive fiber membrane composed of mesoporous bioactive glass, silk fibroin, and nerve growth factor; MBG / GAG(1-5)@SF / NGF is a bioactive fiber membrane composed of mesoporous bioactive glass, multi-component glycosaminoglycans in different proportions, silk fibroin, and nerve growth factor; the bone repair material is the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane of the present invention. Detailed Implementation

[0045] To facilitate understanding of the present invention, the present invention will be described more fully and in detail below with reference to preferred embodiments in the specification, but the scope of protection of the present invention is not limited to the following specific embodiments.

[0046] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention.

[0047] Unless otherwise specified, all raw materials, reagents, instruments and equipment used in this invention can be purchased from the market or prepared by existing methods.

[0048] This invention first selects five metallic elements—calcium, magnesium, sodium, potassium, and strontium—as the basic components for constructing bone-mimicking bioceramics based on the biochemical composition of bone minerals. Subsequently, this invention constructs a dataset through limited high-throughput experiments and uses machine learning models to predict and screen the optimal ratios with high osteogenic potential. Finally, this invention prepares mesoporous bioactive glass-based coaxial electrospun bioactive fiber membranes using the optimal ratio materials, and systematically verifies their high efficiency in promoting bone regeneration through in vitro cell experiments and in vivo animal models, and preliminarily explores their potential molecular mechanisms. This invention not only provides a high-performance candidate material for repairing critical-sized bone defects, but also establishes an efficient and reliable new paradigm for the design and development of complex multi-component biomaterials.

[0049] The sources of some of the reagents and equipment used in this invention are as follows: Polycaprolactone (PCL), purchased from Beijing Bailingwei Technology Co., Ltd., China; Hexafluoroisopropanol (HFIP), purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., China; Silk fibroin (SF), purchased from Suzhou Yongqinquan Intelligent Equipment Co., Ltd., China; Mesoporous bioactive glass (MBG), the specific components and preparation method are described in the examples; Nerve growth factor (NGF), purchased from PeproTech Inc., USA; Fully automated high-throughput pipetting workstation, Jiliang, Shanghai Zuofei, China; Mouse bone marrow mesenchymal stem cells (BMSCs) were purchased from Abiowell, China.

[0050] All materials and instruments used in the following examples are commercially available.

[0051] Example 1 This invention discloses a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, which is a composite fiber membrane composed of core-shell structured composite fibers prepared by coaxial electrospinning. The core of the core-shell structured composite fiber is composed of polycaprolactone and mesoporous bioactive glass in a mass ratio of 10:1, and the outer shell is composed of polycaprolactone, silk fibroin, and nerve growth factor in a mass ratio of 28000:700:1. The surface of the mesoporous bioactive glass particles is modified with amino groups. The mesoporous bioactive glass particles contain the following metal elements in parts by mass: calcium 88 parts, magnesium 8 parts, strontium 1 part, sodium 2 parts, and potassium 1 part. The thickness of the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane is 20 μm; the fiber diameter of the core-shell structured composite fiber is 100 nm, wherein the diameter ratio of the core to the outer shell is 1:2.

[0052] The method for preparing mesoporous bioactive glass-based coaxial electrospun bioactive fiber membranes by electrospinning technology specifically includes the following steps: (1) Weigh the corresponding mass of nitrate for each metal element and prepare mesoporous bioactive glass using the sol-gel method as follows: control the total molar number of cations to be 0.0159 mol, and calculate the mass of nitrate corresponding to each metal according to the ratio; dissolve 4.8 polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer (P123) in 300 mL of sterile water, adjust the pH to 1.0 with concentrated hydrochloric acid, add 6 g of TMB (1,3,5-trimethylbenzene) pore-expanding agent, and water bath. The mixture was stirred at room temperature for 4 hours. The corresponding nitrates were added according to the calculated component mass ratios, followed by 10 mL of tetraethyl orthosilicate (TEOS) and 2,6-triethyl phosphate (TEP). The mixture was stirred in a water bath (37℃, 24 hours). The pH was adjusted to 10.0 with concentrated ammonia and then transferred to a reaction vessel. The mixture was allowed to settle in an oven (90℃, 72 hours). The product was removed and centrifuged (10,000 rpm, 10 minutes). After washing three times, the precipitate was transferred to a crucible and dried in an oven (100℃, 10 hours). The precipitate was then calcined in a muffle furnace (550℃, 8 hours). The resulting powder was ignited in an oven (140℃, 8 hours), and the product was collected after natural cooling. (2) The surface of the mesoporous bioactive glass was modified with silane coupling agent to obtain MBG-NH2. The method is as follows: Take 0.1g of powder, add 10mL of 0.1M APTES chloroform solution, stir in a water bath (room temperature, 24h), filter, wash with deionized water and dichloromethane in sequence, and air dry at room temperature. (3) The surface-modified mesoporous bioactive glass particles and polycaprolactone (PCL, Mw=106000-188000) were weighed at a mass ratio of 1:10 and dispersed in hexafluoroisopropanol to form a core spinning solution. Polycaprolactone, silk fibroin (SF) and nerve growth factor solution (NGF, concentration of 0.0001μg / mL) were weighed at a mass ratio of 28000:700:1 and dissolved in hexafluoroisopropanol to form a shell spinning solution. Then, coaxial electrospinning was performed with the following parameters: the flow rate of the shell was 0.75mL / h, the flow rate of the core was 0.75mL / h, the voltage was 20kV, the receiving distance was 25cm, the ambient temperature was 25℃, and the ambient humidity was 44%. A composite fiber membrane composed of core-shell structure composite fiber filaments was formed, which is the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane.

[0053] The screening method for the formulation of mesoporous bioactive glass is as follows: (1) Calcium, magnesium, sodium, potassium and strontium were selected as the metal elements for the bioceramic scaffold of the bone matrix.

[0054] (2) High-throughput experiments and dataset construction Formula design: Using the uniform design method, 170 different molar percentage formulas were generated under the constraint of constant total molar amount of components.

[0055] Sample preparation: Using a fully automated high-throughput pipetting workstation (JiLiang, Shanghai Zuofei, China), each component salt was completely dissolved in dilute hydrochloric acid (concentration: 1.5M), then mixed according to the designed ratio and transferred to a 96-well plate (material: polystyrene). The well plates were sealed with a breathable sealing membrane, and micropores were punched in each well to ensure solvent evaporation. The plates were then placed in an oven and dried at (37) °C for 24 h to form a material coating.

[0056] Cell culture: Mouse bone marrow mesenchymal stem cells (BMSCs, source: Abiowell, China) were cultured at 5 × 10⁶ cells per well. 3 Cells were seeded at a density of 1,000 cells in 96-well plates after the above treatment and cultured in osteogenic induction medium (component: DMEM containing 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, China) and 1% penicillin-streptomycin (Gibco Life Sciences, USA)).

[0057] Performance indicator testing: Cell proliferation (day 7): CCK-8 assay was used. 10 μL of CCK-8 solution (Dojindo, Japan) was added to each well, and the cells were incubated at 37°C for 4 h. The absorbance (OD) was then measured at 450 nm using a microplate reader (Thermo, USA).

[0058] Early osteogenic differentiation (day 7): Alkaline phosphatase activity was measured. The culture medium was discarded, and after washing with PBS, freshly prepared ALP substrate working solution (0.1M Na₂CO₃, 0.1M NaHCO₃, 22mM MgCl₂, 0.1% Triton X-10, 0.8mM PNPP) was added to each well, and the mixture was incubated at 37°C for 30 min. After the reaction, the OD value was measured at 405 nm using a microplate reader.

[0059] Data label generation: To comprehensively evaluate the osteogenic induction efficiency of the materials (while also considering the impact on cell proliferation and differentiation), the osteoinduction index for each group was calculated as (ALP OD value) / (CCK-8 OD value). Based on the distribution of values, samples with an osteoinduction index ≥ 0.83 were labeled as "high osteogenic performance" (label 1), and the rest were labeled as "low osteogenic performance" (label 0), thus constructing a binary classification dataset.

[0060] (3) Machine learning modeling and prediction 1) Constructing a training dataset: In a full-scale space containing five metallic elements—calcium, magnesium, sodium, potassium, and strontium—a uniform design method was used to generate several different molar percentage ratios under the constraint of a constant total molar amount. For each ratio of material samples, the material samples were co-cultured with bone marrow mesenchymal stem cells, and the osteogenic performance index corresponding to each sample was measured. Based on the osteogenic performance index, each sample was assigned a classification label characterizing its osteogenic performance, thereby constructing a training dataset containing sample ratio data and corresponding classification labels. 2) Machine learning model training: Data preprocessing: The 170 datasets were randomly divided into training and test sets in a 7:3 ratio, using stratified sampling to maintain class proportions. Z-score standardization was applied to the features (molar percentage) of each component. To address class imbalance in the training set, the SMOTE oversampling algorithm (nearest neighbor number k=10) was applied to the training set during the cross-validation loop, while the test set remained unchanged to avoid data leakage.

[0061] Model Training and Selection: Using the Scikit-learn library, eight classic machine learning algorithms were trained on the training set: Logistic Regression, Support Vector Machine (SVM, kernel function: radial basis function), Decision Tree, Random Forest, Gradient Boosting Tree, K-Nearest Neighbors (KNN), Naive Bayes, and Multilayer Perceptron (MLP). Model performance was evaluated using 5-fold cross-validation, with key evaluation metrics including accuracy, precision, recall, and F1 score. The model with the best overall performance on the test set was selected as the final prediction model.

[0062] 3) Candidate formulation prediction and screening: Under the constraint that the total molar percentage is 100%, 10,000 virtual formulations are uniformly generated in the entire space using Latin hypercube sampling. These formulations are then predicted using a pre-trained optimal model to screen out all candidate formulations classified as "high osteogenic performance".

[0063] To verify the technical effect of the mesoporous bioactive glass-based coaxial electrospun bioactive fiber of the present invention, the following experiments were conducted: 1. Two-round screening based on mineralization capacity This invention compared the performance of eight different machine learning algorithms (Logistic Regression, SVM, Decision Tree, Random Forest, Gradient Boosting, KNN, Naive Bayes, and MLP) in predicting the ratio and osteogenic properties of multi-component biomaterials. In terms of accuracy, all algorithms except logistic regression and naive bayes performed well. In terms of precision, SVM and random forest performed better. Regarding recall, SVM, decision tree, random forest, gradient boosting, and KNN all achieved 100%. Considering all factors, SVM and random forest achieved the highest F1 scores.

[0064] Based on the SVM model that performed optimally on the test set, this invention predicted the osteogenic performance of 10,000 virtual formulations generated in a five-component search space through uniform design. Of these, 126 formulations were classified as having "high osteogenic performance" by the model, accounting for 1.26% of the total. This fully reflects the scarcity of the "optimal region" in the multidimensional component space and highlights the necessity of machine learning-guided targeted selection.

[0065] To conduct a second round of refined screening from these 126 candidate formulations, all candidate materials were synthesized (method: the same as the sample preparation method in Example 1) and co-cultured with BMSCs for osteogenic induction. After 14 days of culture, alizarin red staining was performed (PBS washing, 70% ethanol fixation, 40 mM alizarin red solution at pH 4.2). For quantitative comparison, the bound dye was dissolved in 10% hexadecylpyridine chloride solution, and the OD value was measured at 595 nm. Based on the quantitative results, the four formulations with the highest mineralized nodule deposition (MBG2, MBG3, MBG4, MBG5) were selected as the high-performance experimental groups. Simultaneously, from the formulations predicted by the model to have "low osteogenic performance," a group with weaker mineralization ability was randomly selected as the low-performance control group (MBG1). The specific formulations are as follows: MGB1: 85 parts calcium, 9 parts magnesium, 1 part sodium, 4 parts potassium, 1 part strontium; MGB2: 80 parts calcium, 8 parts magnesium, 3 parts sodium, 7 parts potassium, 2 parts strontium; MGB3: 86 parts calcium, 10 parts magnesium, 1 part sodium, 1 part potassium, 2 parts strontium; MGB4: 83 parts calcium, 6 parts magnesium, 1 part sodium, 6 parts potassium, 4 parts strontium; MGB5: 88 parts calcium, 8 parts magnesium, 1 part sodium, 2 parts potassium, 1 part strontium.

[0066] In-depth analysis of the screening results revealed common compositional characteristics of the high-performance formulations. This invention discovered that functional ions may exhibit a synergistic mechanism in bone formation. First, calcium is dominant. As a key component of bone minerals, calcium dominates in all high-performance material formulations, reflecting the mineral proportions in natural bone. In a second round of screening tests on 126 material formulations, this invention found that osteogenic performance declines when the calcium content is below 80%. Second, key functional ions may play a synergistic role. Compared to low osteogenic material formulations, high osteogenic material formulations show significantly higher proportions of enhancing effects from strontium and magnesium. 2+ It can activate the CaSR and Wnt signaling pathways, while simultaneously promoting osteoblast differentiation and inhibiting osteoclast activity. Mg 2+ As an important cofactor for many enzymes, sodium plays a role in cell adhesion, proliferation, and early alkaline phosphatase (ALP) activity. Sodium and potassium may play a role in balancing and regulating this process. + and K + These are important ions for maintaining cell membrane potential and intracellular osmotic pressure balance, laying the foundation for normal cellular physiological activities. Under the optimal ratio determined through model screening, the proportion of these two ions remains within a relatively stable and moderate range, avoiding potential cytotoxicity or functional inhibition caused by excessively high or low levels.

[0067] This invention employs a machine learning-driven two-round screening strategy, which not only efficiently identifies several highly promising high-performance bone-like ceramic formulations from a vast component space, but also, through reverse analysis of the screening results, preliminarily elucidates the complex role of multi-component functional ions in synergistically regulating osteogenic processes, laying a solid foundation for subsequent experimental verification and mechanism research.

[0068] 2. Preparation and characterization of bioactive fiber membranes Based on the five selected formulations (MBG1-5), MBG of the corresponding components were prepared using the sol-gel method combined with a template agent. Mesoporous bioactive glass-based coaxial electrospun bioactive fiber membranes were then prepared, following the same method as the sample preparation in Example 1. Control groups included coaxial fiber membranes containing only PCL (PCL@SF / NGF) and uniaxial PCL electrospun fiber membranes (PCL).

[0069] Material characterization: The morphology of the fibers was observed using scanning electron microscopy; the elastic modulus of the fiber membrane was tested using a universal testing machine; the fiber membrane was immersed in simulated body fluid (SBF) (37℃, 14 days), and after drying, the morphology of hydroxyapatite deposition on its surface was observed by SEM; the characteristic functional groups of the material were analyzed by Fourier transform infrared spectroscopy; AlexaFluor 594 (excitation / emission: (590 nm / 617 nm)) and fluorescein isothiocyanate-5 (FITC) (excitation / emission: 494 nm / 520 nm) were added to the shell and core spinning solutions, respectively, and the fluorescence distribution of the fibers was observed by laser confocal microscopy to confirm the core-shell structure; after BMSCs were seeded on the material surface and cultured for 3 days, the adhesion and spreading morphology of the cells on the material were observed by SEM.

[0070] This invention uses machine learning to screen components and prepares MBGs corresponding to the proportions of MBG1-5. The mesoporous structure is characterized using the Brunauer-Emmett-Teller (BET) method, and the results show a clear hysteresis loop, indicating that these MBGs were successfully prepared. Figure 1 Furthermore, this invention prepared a core-shell structured nanofiber scaffold using coaxial electrospinning, with MBG located in the core and SF / NGF located in the shell. The core-shell structure was determined by fluorescence staining: FITC stained the core green, and Alexa Fluor 594 stained the shell red; the combined image appeared yellow. Figure 2 Scanning electron microscopy (SEM) revealed that the surface morphology of these fibers exhibited an interwoven fiber structure. Figure 3 The mineralization effects of the high osteogenic component (MBG1@SF / NGF) and the control group (PCL@SF / NGF) were superior to those of the low osteogenic component, with MBG5@SF / NGF and MBG3@SF / NGF exhibiting the best performance. Figure 4 The infrared (FITR) results show a peak in -NH2, indicating that MBG has been successfully modified with an amino group (). Figure 5 These results indicate that the scaffold has good mineralization capacity in vitro.

[0071] 3. In vitro biological evaluation To comprehensively evaluate the osteogenic capacity of bioactive fibrous membranes screened by machine learning, this invention conducted a series of systematic in vitro cell experiments, covering the entire process from cell proliferation, early differentiation, late mineralization to related molecular mechanisms.

[0072] All in vitro experiments included two high-performance material groups (MBG3@SF / NGF, MBG5@SF / NGF), a low-performance control group (MBG1SF / NGF), a material control group (PCL@SF / NGF), and a blank control group (cell culture plate). Cell proliferation was assessed on days 1, 3, 5, and 7 of co-culture (CCK-8 assay); alkaline phosphatase activity was assessed on days 3, 6, and 9 of co-culture using the same method; extracellular matrix mineralization (alizarin red staining for quantification) was performed 14 days after osteogenic induction, using the same method; RNA was extracted on days 7 and 14 of co-culture and osteogenic-related gene expression was detected by qRT-PCR; total protein was extracted on day 7 of co-culture and osteogenic-related protein expression was detected by Western blotting, and quantitative analysis was performed using ImageJ software; immunofluorescence staining was performed on day 7 of co-culture, and observation and photography were conducted under a laser confocal microscope; total RNA was extracted on day 7 of co-culture for transcriptome sequencing (RNA-seq), and differentially expressed gene analysis, GO functional enrichment analysis, and KEGG pathway enrichment analysis were performed; BMSCs were co-cultured with the material in neural induction medium for 7 days and evaluated by cell morphology observation, qPCR detection of neural markers (β-III-tubulin, GFAP, Nestin) expression, and immunofluorescence staining.

[0073] (1) The material significantly promotes the proliferation and early osteogenic differentiation of BMSCs. The results of the CCK-8 experiment show that ( Figure 6 Compared with the control group (PCL@SF / NGF) and the low-performance group (MBG1@SF / NGF), both high-performance materials (MBG3@SF / NGF and MBG5@SF / NGF) significantly promoted the proliferation of mouse bone marrow mesenchymal stem cells, especially on days 5 and 7 of culture, where cell activity was significantly enhanced (p<0.01). Among them, the MBG5@SF / NGF group showed the most outstanding performance.

[0074] Alkaline phosphatase (ALP) is a key marker of early osteoblast differentiation. This invention detected ALP activity on days 3, 6, and 9 of culture. Figure 7 The results showed that the alkaline phosphatase (ALP) activity of BMSCs cultured on the high-performance material group, especially the MBG5@SF / NGF scaffold, was significantly higher than that on other scaffolds, and the expression level was even higher on day 3. This indicates that these materials can effectively accelerate the early differentiation process of BMSCs into osteoblast lineages.

[0075] (2) The material strongly induces extracellular matrix mineralization and osteogenic gene expression. Alizarin red staining was used to assess the late osteogenic differentiation capacity of BMSCs, namely the formation of extracellular matrix calcified nodules. After 14 days of osteogenic induction culture, numerous, deeply stained orange-red calcified nodules were observed in the high-performance material groups (MBG5@SF / NGF and MBG3@SF / NGF), while quantitative analysis (measured by dissolving CPC) showed... Figure 8 The mineralization levels of the MBG5@SF / NGF and MBG3@SF / NGF groups were significantly higher than those of the control group and MBG1@SF / NGF, further confirming that the high-performance material group has the strongest osteogenic efficacy.

[0076] To verify the osteogenic induction effect of the material at the molecular level, this invention detected the expression of key osteogenic genes using qRT-PCR. On day 7 of culture, in BMSCs treated with the high-performance material group, the expression of early transcription factors (Runx2, Osx) was significantly increased. Figure 9 The mRNA expression levels of extracellular matrix proteins (Col, Opn) and late differentiation markers (Ocn) were significantly upregulated. For Opn and Col, both MBG3@SF / NGF and MBG5@SF / NGF significantly upregulated their expression levels. The MBG3@SF / NGF group showed a greater advantage in inducing the expression of Ocn and Col genes, while MBG5@SF / NGF showed a greater advantage in inducing the expression of Osx, Bmp2, and Runx2 genes. Overall, MBG5@SF / NGF showed a more significant advantage in inducing the expression levels of these genes, which is highly consistent with the phenotypic results.

[0077] The material regulates the osteogenic process by activating key signaling pathways: Western blot analysis revealed a significant increase in the protein expression of Runx2, a key osteogenic transcription factor, in the high-performance materials group. Simultaneously, the phosphorylation level of its upstream regulator—extracellular signal-regulated kinase—was also significantly enhanced, suggesting potential activation of the MAPK / ERK signaling pathway.

[0078] Immunofluorescence staining allowed for direct observation that BMSCs cultured on MBG3@SF / NGF and MBG5@SF / NGF substrates exhibited well-developed and fully extended cytoskeleton (F-actin, red) and morphological spread. Simultaneously, Runx2 protein (green) displayed high-intensity fluorescence within the nucleus and co-localized with the nucleus (DAPI, blue), further confirming at the protein localization level that its osteogenic differentiation activity was effectively activated. Figure 10The Bmp2 and Runx2 proteins showed the same trend. Quantitative fluorescence intensity analysis further confirmed that the MBG3@SF / NGF and MBG5@SF / NGF materials exhibited superior performance.

[0079] The material exhibits the potential to induce neural differentiation: Given that good bone regeneration is often accompanied by the restoration of neural innervation, this invention preliminarily explored the neural differentiation induction potential of the materials. Under neural induction conditions, BMSCs co-cultured with materials from the MBG3@SF / NGF and MBG5@SF / NGF groups showed stronger proliferation ability. qPCR results showed that the expression levels of neural-related markers such as Map and Nse in the MBG3@SF / NGF and MBG5@SF / NGF groups were higher than those in other scaffolds. Western blot results also showed upregulated Nse expression in the MBG3@SF / NGF and MBG5@SF / NGF groups, indicating more active neurogenesis, and the cells exhibited longer processes and more typical neuron-like morphology. Figure 11 This indicates that the material may possess a certain degree of neural affinity, laying the foundation for the construction of neurogenic bone repair materials.

[0080] Based on the above in vitro experimental results, this invention confirms that the bioactive fiber membrane prepared by the optimal ratio (MBG3@SF / NGF and MBG5@SF / NGF) selected through machine learning can systematically promote the proliferation, early osteogenic differentiation, and late matrix mineralization of BMSCs by activating signaling pathways such as MAPK / ERK and upregulating the activity and expression of Runx2. It also exhibits multi-directional differentiation potential and has excellent application prospects in bone regeneration.

[0081] 4. In vivo bone repair experiment To ultimately verify the actual bone repair efficacy of the optimal formula selected by machine learning in vivo, this invention established a mouse skull critical size defect model (diameter 2.8 mm) and conducted an implantation experiment for 4 weeks.

[0082] Six-week-old male mice (strain: C57 / BL6) weighing 20g were used to create a skull defect model. Mice were randomly assigned to the following groups (n=6): blank control group (no defect treatment); PCL fibrous membrane group; MBG5@SF / NGF group; MBG3@SF / NGF group; MBG1@SF / NGF group. Bone repair material was implanted into the left defect site, and the right defect served as a blank control. Four weeks post-surgery, skull samples were collected. Scanning was performed using a Micro-CT scanner (SkyScan 1276, Bruker, Belgium). Three-dimensional reconstruction was performed using the accompanying software, and the fraction of newly formed bone, trabecular thickness, and trabecular separation in the defect area were quantitatively analyzed. Histological analysis: Hematoxylin-eosin staining was used to observe the morphology, cell distribution, and inflammatory response of the newly formed bone tissue. Masson's trichrome staining was used to distinguish collagen fibers (blue) from mineralized bone tissue (red) and to assess bone matrix maturity. Immunohistochemical staining was used to detect the expression of osteocalcin and osteopontin, and to assess osteoblast activity and mineralization status.

[0083] All in vitro experiments were performed at least three times independently. Data are expressed as mean ± standard deviation. Statistical analysis was performed using GraphPadPrism software. One-way ANOVA was used for comparisons among multiple groups, followed by Tukey's post-hoc test. Student's t-test was used for comparisons between two groups. P < 0.05 was considered statistically significant.

[0084] (1) Micro-CT three-dimensional reconstruction and quantitative analysis.

[0085] Four weeks after implantation, three-dimensional reconstruction using Micro-CT allowed for a direct observation of the repair progress in each group of defect areas. Figure 12 .

[0086] The blank control group showed clear defect boundaries with only a small amount of scattered new bone bridges growing from the edges to the center, while most areas were filled with fibrous connective tissue. The repair effect of the low-performance MBG1@SF / NGF group was similar to the blank control group, with limited new bone formation and ineffective bridging of the defect. The PCL control group showed slightly better repair than the blank group, but new bone formation remained slow and discontinuous. The high-performance material groups (MBG3@SF / NGF and MBG5@SF / NGF) both exhibited significantly better new bone formation capabilities than the control group. In the MBG5@SF / NGF group, the defect area was covered by a large amount of radiopaque, dense new bone tissue, the defect was essentially closed, and the new bone integrated well with the host bone. The MBG3@SF / NGF group also showed significant bone regeneration, but the completeness of the repair was slightly inferior to the MBG5@SF / NGF group.

[0087] This invention performed three-dimensional morphometric analysis on Micro-CT data. Quantitative data showed that 4 weeks after implantation, the MBG5@SF / NGF group had significantly higher bone volume fraction, trabecular thickness, trabecular number, and bone mineral density than all other groups, while its trabecular separation was significantly lower. The BV / TV value of the MBG5@SF / NGF group reached 1.5±0.06 (…). Figure 13 The value was significantly higher than that of the blank control group (0.6, p<0.001). This quantitatively demonstrates from a three-dimensional structural perspective that the MBG5@SF / NGF group material has the strongest guided bone regeneration capacity in the short term.

[0088] (2) Histological analysis reveals the maturity and integration of new bone. To assess the quality and maturity of newly formed bone at the tissue and cellular level, this invention decalcified skull samples four weeks after implantation and performed H&E staining, Masson trichrome staining, and immunohistochemical analysis.

[0089] H&E staining ( Figure 14 The results showed that the defect areas in the PCL@SF / NGF and MBG1@SF / NGF groups were mainly composed of loose fibrous tissue, with only a small amount of immature osteoid tissue at the periphery. In contrast, the MBG5@SF / NGF group had a large amount of newly formed bone tissue, regular trabecular structure, a large number of osteoblasts, and rich vascular structures, indicating an active and healthy osteogenic environment.

[0090] Masson's trichrome staining specifically stains mature collagen fibers blue. Results showed that the newly formed bone area in the MBG5@SF / NGF group exhibited a large, dense blue color, indicating abundant type I collagen deposition and good mineralization, with the newly formed bone showing high maturity at 4 weeks. In contrast, the control group showed only sparse blue areas.

[0091] Immunohistochemical analysis further confirmed active osteogenic activity at the protein level. In the newly formed bone region of the MBG5@SF / NGF group, the expression and localization of specific marker proteins Bmp2, Ocn, Nse, and CD31 were detected by immunohistochemical analysis. Their high expression indicates that functional bone repair integrating osteoblastogenesis and angiogenesis has occurred at 4 weeks.

[0092] (3) Assessment of material biocompatibility and degradability No severe inflammatory cell infiltration or significant tissue necrosis was observed at the implantation site in H&E-stained sections of all groups, indicating that all prepared biomaterials exhibited good biocompatibility within 4 weeks. Meanwhile, in sections of the MBG5@SF / NGF group, material residues were observed to be tightly intertwined with newly formed bone tissue, and the material edges had begun to show degradation morphology, suggesting that the material itself initiated a biodegradation process while guiding new bone ingrowth.

[0093] In summary, the four-week in vivo experiment, through Micro-CT three-dimensional quantitative analysis, a series of histological staining and immunohistochemical analyses, formed a strong chain of evidence, conclusively demonstrating that the bioactive fiber membrane prepared by the optimal ratio of MBG5@SF / NGF predicted by machine learning can rapidly and efficiently guide the generation of high-quality, highly mature new bone in a critical-size bone defect model, and achieve perfect integration with the host bone, exhibiting tremendous bone repair capacity and clinical translational potential in a short period of time.

[0094] Example 2 This invention discloses a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, which is a composite fiber membrane composed of core-shell structured composite fibers prepared by coaxial electrospinning. The core of the core-shell structured composite fiber is composed of polycaprolactone and mesoporous bioactive glass in a mass ratio of 8:1, and the outer shell is composed of polycaprolactone, silk fibroin, and nerve growth factor in a mass ratio of 24000:600:1. The surface of the mesoporous bioactive glass particles is modified with amino groups. The mass fractions of the metal elements in the mesoporous bioactive glass particles are: calcium 92 parts, sodium 5 parts, strontium 1 part, sodium 1 part, and potassium 1 part. The thickness of the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane is 15 μm; the fiber diameter of the core-shell structured composite fiber is 80 nm, wherein the diameter ratio of the core to the outer shell is 1:1.8.

[0095] The method for preparing mesoporous bioactive glass-based coaxial electrospun bioactive fiber membranes by electrospinning technology specifically includes the following steps: (1) Weigh the corresponding mass of nitrate for each metal element and prepare mesoporous bioactive glass using the sol-gel method as follows: control the total molar number of cations to be 0.0150 mol, and calculate the mass of nitrate corresponding to each metal according to the ratio; dissolve 4g of polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer (P123) in 300mL of sterile water, adjust the pH to 0.8 with concentrated hydrochloric acid, add 7g of TMB (1,3,5-trimethylbenzene) pore-expanding agent, and stir at room temperature for 5h in a water bath; add the corresponding nitrate according to the calculated mass of each component, and then add 12mL of tetraethyl orthosilicate (TEOS) and 2g of triethylphosphoric acid. The ester (TEP) was stirred in a water bath (40℃, 20h); the pH was adjusted to 10.5 with concentrated ammonia and then transferred to a reaction vessel. The product was allowed to stand in an oven to precipitate (92℃, 64h). The product was then centrifuged (11000rpm, 12min), washed three times, and the precipitate was transferred to a crucible and dried in an oven (105℃, 99h). The product was then calcined in a muffle furnace (500℃, 7h). The resulting powder was ignited in an oven (150℃, 7h), and the product was collected after natural cooling. (2) The surface of the mesoporous bioactive glass was modified with silane coupling agent to obtain MBG-NH2. The method is as follows: Take 0.15g of powder, add 15mL of 0.1M APTES chloroform solution, stir in a water bath (room temperature, 20h), filter, wash with deionized water and dichloromethane in sequence, and air dry at room temperature. (3) The surface-modified mesoporous bioactive glass particles and polycaprolactone (PCL, Mw=106000-188000) were weighed at a mass ratio of 1:8 and dispersed in hexafluoroisopropanol to form a core spinning solution. Polycaprolactone, silk fibroin (SF) and nerve growth factor solution (NGF, concentration of 0.00015μg / mL) were weighed at a mass ratio of 24000:600:1 and dissolved in hexafluoroisopropanol to form a shell spinning solution. Then, coaxial electrospinning was performed with the following parameters: the flow rate of the shell was 0.6mL / h, the flow rate of the core was 0.6mL / h, the voltage was 18kV, the receiving distance was 20cm, the ambient temperature was 20℃, and the ambient humidity was 40%. A composite fiber membrane composed of core-shell structure composite fiber filaments was formed, which is the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane.

[0096] The surface morphology of the bioactive fiber membrane prepared in this embodiment exhibits an interwoven fiber structure. Figure 15 ), and the mineralization effect is excellent ( Figure 16 Four weeks post-surgery, in this embodiment, the bone defect area induced by the bioactive fiber membrane was covered by a large amount of new bone tissue, the defect was basically closed, and the new bone integrated well with the host bone. Figure 17 ).

[0097] Example 3 This invention discloses a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, which is a composite fiber membrane composed of core-shell structured composite fibers prepared by coaxial electrospinning. The core of the core-shell structured composite fiber is composed of polycaprolactone and mesoporous bioactive glass in a mass ratio of 12:1, and the outer shell is composed of polycaprolactone, silk fibroin, and nerve growth factor in a mass ratio of 30000:750:1. The surface of the mesoporous bioactive glass particles is modified with amino groups. The mass fractions of the metal elements in the mesoporous bioactive glass particles are: calcium 64 parts, magnesium 20 parts, strontium 8 parts, sodium 3 parts, and potassium 5 parts. The thickness of the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane is 25 μm; the fiber diameter of the core-shell structured composite fiber is 120 nm, wherein the diameter ratio of the core to the outer shell is 1:2.4.

[0098] The method for preparing mesoporous bioactive glass-based coaxial electrospun bioactive fiber membranes by electrospinning technology specifically includes the following steps: (1) Weigh the corresponding mass of nitrate for each metal element and prepare mesoporous bioactive glass using the sol-gel method as follows: Control the total molar number of cations to be 0.017 mol, and calculate the mass of nitrate corresponding to each metal according to the ratio; Dissolve 7g of polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer (P123) in 300mL of sterile water, adjust the pH to 1.5 with concentrated hydrochloric acid, add 7-10g of TMB (1,3,5-trimethylbenzene) pore-expanding agent, and stir at room temperature in a water bath for 6h; Add the corresponding nitrate according to the calculated mass of each component, and then add 10-15g of nitrate. mL of tetraethyl orthosilicate (TEOS) and 3 g of triethyl phosphate (TEP) were stirred in a water bath (50℃, 24 h). The pH was adjusted to 11.0 with concentrated ammonia and then transferred to a reaction vessel. The mixture was allowed to stand in an oven to precipitate (95℃, 72 h). The product was then centrifuged (12000 rpm, 15 min), washed three times, and the precipitate was transferred to a crucible and dried in an oven (110℃, 10 h). The precipitate was then calcined in a muffle furnace (600℃, 8 h). The resulting powder was ignited in an oven (160℃, 8 h), and the product was collected after natural cooling. (2) The surface of the mesoporous bioactive glass was modified with silane coupling agent to obtain MBG-NH2. The method is as follows: Take 0.2g of powder, add 20mL of 0.1M APTES chloroform solution, stir in a water bath (room temperature, 24h), filter, wash with deionized water and dichloromethane in sequence, and air dry at room temperature. (3) The surface-modified mesoporous bioactive glass particles and polycaprolactone (PCL, Mw=106000-188000) were weighed at a mass ratio of 1:12 and dispersed in hexafluoroisopropanol to form a core spinning solution. Polycaprolactone, silk fibroin (SF) and nerve growth factor solution (NGF, concentration of 0.0002μg / mL) were weighed at a mass ratio of 30000:750:1 and dissolved in hexafluoroisopropanol to form a shell spinning solution. Then, coaxial electrospinning was performed with the following parameters: the flow rate of the shell was 1mL / h, the flow rate of the core was 1mL / h, the voltage was 22kV, the receiving distance was 30cm, the ambient temperature was 27℃, and the ambient humidity was 50%. A composite fiber membrane composed of core-shell structure composite fiber filaments was formed, which is the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane.

[0099] The surface morphology of the bioactive fiber membrane prepared in this embodiment exhibits an interwoven fiber structure. Figure 18 ), and the mineralization effect is excellent ( Figure 19 Four weeks post-surgery, in this embodiment, the bone defect area induced by the bioactive fiber membrane was covered by a large amount of new bone tissue, the defect was basically closed, and the new bone integrated well with the host bone. Figure 20 ).

Claims

1. A mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane, characterized in that, It is a composite fiber membrane composed of core-shell structured composite fibers prepared by coaxial electrospinning; the core of the core-shell structured composite fiber is composed of polycaprolactone and mesoporous bioactive glass, and the outer shell is composed of polycaprolactone, silk fibroin and nerve growth factor; the surface of the mesoporous bioactive glass particles is modified with amino groups; the mesoporous bioactive glass particles contain the following metal elements in parts by weight: calcium 64-92 parts, magnesium 5-20 parts, strontium 1-8 parts, sodium 1-3 parts, potassium 1-5 parts. The method for preparing the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane includes the following steps: (1) Weigh out the corresponding masses of nitrates of five metal elements: calcium, magnesium, sodium, potassium and strontium, and prepare mesoporous bioactive glass using the sol-gel method; (2) The surface of the mesoporous bioactive glass is modified with an amino group using a silane coupling agent; (3) The surface-modified mesoporous bioactive glass particles and polycaprolactone are dispersed in a solvent to form a core spinning solution. Polycaprolactone, silk fibroin and nerve growth factor solution are dissolved in a solvent to form a shell spinning solution. Then, coaxial electrospinning is performed to form a composite fiber membrane composed of core-shell structure composite fiber filaments, which is the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane.

2. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, In the core of the core-shell composite fiber, the mass ratio of polycaprolactone to mesoporous bioactive glass particles is 8-12:1; in the outer shell of the core-shell composite fiber, the mass ratio of polycaprolactone, silk fibroin, and nerve growth factor is 24000-30000:600-750:

1.

3. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, The thickness of the mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane is 15μm-40μm; the fiber diameter of the core-shell structure composite fiber is 80-120nm, wherein the diameter ratio of the core to the shell is 1:1.8-2.

4.

4. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, The mesoporous bioactive glass particles contain the following metal elements in parts by weight: 88 parts calcium, 8 parts magnesium, 1 part sodium, 2 parts potassium, and 1 part strontium.

5. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, In step (1), the sol-gel method is as follows: the template agent and the pore-expanding agent are dissolved in an acidic solution with a pH of 0.8-1.5, and then the nitrates corresponding to the five metal elements calcium, magnesium, sodium, potassium and strontium are added, controlling the total number of cation moles to be 0.015-0.017 mol. Then, tetraethyl orthosilicate and triethyl phosphate are added and mixed and stirred to form a sol. After adjusting the pH to 10.0-11.0, the mixture is heated at 90-95℃ for 64-72 hours. The resulting precipitate is washed, dried and then calcined at 500-600℃ for 6-8 hours to obtain the mesoporous bioactive glass particles. The template agent is a poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer, and the pore-expanding agent is 1,3,5-trimethylbenzene.

6. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, In step (2), the silane coupling agent is (3-aminopropyl)triethoxysilane, and the amino modification method is as follows: the mesoporous bioactive glass particles obtained in step (1) are dispersed in an organic solvent solution of (3-aminopropyl)triethoxysilane, stirred at room temperature, and then separated, washed, and dried to obtain surface-modified mesoporous bioactive glass particles.

7. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, In step (3), the solvent is hexafluoroisopropanol; the mass ratio of the surface-modified mesoporous bioactive glass particles to polycaprolactone is 1:8-12, the mass ratio of polycaprolactone, silk fibroin and nerve growth factor is 24000-30000:600-750:1, and the concentration of the nerve growth factor solution is 0.0001-0.0002 μg / mL.

8. The mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane according to claim 1, characterized in that, In step (3), the parameters of the coaxial electrospinning are as follows: the flow rate of the outer shell is 0.6-1 mL / h, the flow rate of the inner core is 0.6-1 mL / h, the voltage is 18-22 kV, the receiving distance is 20-30 cm, the ambient temperature is 20-27℃, and the ambient humidity is 40-50%.

9. The application of a mesoporous bioactive glass-based coaxial electrospun bioactive fiber membrane as described in any one of claims 1-4 in the preparation of products for repairing bone defects, characterized in that, The product used to repair bone defects is a drug or medical device.

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