CN113511811A

CN113511811A - Multifunctional mesoporous biomaterial, preparation method and application

Info

Publication number: CN113511811A
Application number: CN202110281853.7A
Authority: CN
Inventors: 苟中入; 杨贤燕; 徐三中
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2021-03-16
Filing date: 2021-03-16
Publication date: 2021-10-19

Abstract

The invention discloses a multifunctional mesoporous biomaterial and a preparation method thereof. The amorphous glassy state particulate matter is formed by an outer wrapping mesoporous structure and an inner cabbage structure, the aperture of the outer wrapping ring-shaped mesoporous is 5-25 nm, and the gap pitch of the inner cabbage structure is 30-200 nm; the mesoporous biomaterial mainly comprises SiO₂‑B₂O₃CaO, further containing P₂O₅、CuO、MgO、ZnO、SrO、Li₂O、Na₂O and/or K₂And O. The invention uses the microstructure guiding agent as the microstructure guiding agent, adopts the sol-gel method to prepare the particle material with unique mesoporous pattern, has simple preparation process, is used for artery and vein hemostasis and the promotion healing or regeneration and repair of soft tissue and bone tissue wounds, has good histocompatibility of the material, can adjust inflammatory reaction in the wounds, and prevents and treats the infection of pathogenic bacteria.

Description

Multifunctional mesoporous biomaterial, preparation method and application

Technical Field

The invention relates to a multifunctional biomaterial with a unique porous structure for emergency wound hemostasis, antibiosis, soft tissue wound repair and bone defect regeneration repair, belonging to the field of biomaterial for surface and in-vivo soft/hard tissue wound regeneration repair and emergency treatment.

Background

Millions of people die each year in our country due to excessive blood loss, tissue necrosis/infection, nonunion of bone defects, delayed union, etc., caused by car accidents, fall injuries, crushing injuries, natural disasters, intra-or post-operative and other emergencies. Especially, in recent years, the aging of China is accelerated, various bad living habits are increasingly prevalent, and chronic inflammation, organ lesion or injury and the like are caused, so that various tissues and organs are damaged or reconstruction and repair problems are caused after excision. Therefore, the realization of rapid hemostasis, inflammation control, infection prevention and control and promotion of regeneration repair or healing of damaged tissues and organs for various human tissue injuries and wounds is the most clinically needed treatment scheme and manual intervention strategy.

The currently commonly used artificial biomaterials are mainly dressings, tourniquets, bandages, high molecular materials, inorganic non-metallic ceramics, bioactive glass, degradable alloys and a large number of non-degradable metals and alloys. Although some conventional dressings (such as chitosan, gelatin and collagen) are low in price and relatively convenient to use, can effectively cover the surface of a wound, can physically block part of traumatic bleeding and isolate bacterial infection, the dressing has limitations on the situations of wounds with high bleeding speed and high bleeding amount, and is easy to cause wound adhesion in the use of various hemostatic materials and dressings, so that the inflammatory reaction time of the wound surface is long, and the wound surface is difficult to replace. Meanwhile, the stimulation activity of the biological materials on the healing and repairing of large tissue wounds is very limited, and the tissue wound repair under the conditions of chronic pathological changes and inflammations of patients cannot be effectively prevented and controlled. Therefore, the development of multifunctional mesoporous biomaterials or methods that can rapidly, safely and effectively stop bleeding from wounds, promote wound healing and synergistically prevent and control inflammation and infection is a long-standing challenge in clinical medicine and biomedicine.

In recent years, a great deal of research proves that some inorganic ions necessary for human body metabolism can regulate inflammation, effectively inhibit bacteria and even kill bacteria, or promote vascularization and wound healing and repair. For example, it is found that copper, zinc and the like can effectively resist bacteria, magnesium ions and zinc ions have the function of regulating inflammation, silicon, boron, copper, magnesium, zinc and the like can regulate vascular endothelial cell factors and promote vascular growth, calcium, silicon, strontium, magnesium, zinc and the like can regulate the expression of bone formation related factors and the bone regeneration efficiency, improve bone density or strength and the like, and sodium ions and potassium ions can regulate the degradability of bioglass materials. Moreover, a large number of researches prove that the amorphous glassy material with the mesoporous structure has a hemostatic function and can promote the regeneration and healing of soft tissues. However, how to organically coordinate various functions, solve the problems of wound repair of various soft and hard tissues and synchronously prevent and control inflammation and infection is a difficult problem in the scientific field of biomedical material preparation. For example, the problem of asynchronous evolution of inflammatory reaction of soft and hard tissue injuries caused by various accidents, particularly the problem that the infection cannot be effectively eradicated due to the formation of a biofilm caused by bacterial accumulation in the hard tissue, is a problem which needs strict attention in tissue wound repair, and the problems of adverse effects on the physiological functions of patients caused by the use of anti-inflammatory and anti-infective medicaments and even drug resistance are caused. These problems are all the problems that the traditional biomaterial and apparatus products do not have a compromise.

According to the research of the prior art, a synergetic prevention and control multifunctional material which meets various pathological reactions, complications and side reactions in various wounds in a human body clinically on the aspects of chemical composition, physical and chemical properties and biological effects is urgently needed to be explored, the material not only has the compatibility of realizing the compatibility of the material on the cell and molecular level to the cells of the human body, but also actively controls the vascularization, and simultaneously, the microstructure of the material, especially the porous material under the condition that the pore channels are mutually communicated can synchronously play the effects of the material on the aspects of promoting the blood coagulation, promoting the wound healing, preventing and treating infection and understanding the inflammatory immunoreaction. Obviously, the conventional materials such as microporous aluminosilicate, mesoporous silicon, mesoporous bioglass, bioactive ceramics, high molecular materials and the like which rely on the conventional inorganic particles with high specific surface area cannot meet the requirements, and the materials can become a new generation of multifunctional mesoporous biomaterial only by innovative design and optimized construction on chemical composition and microstructure, so that the problems of clinical soft and hard tissue wound emergency treatment, regenerative repair and complication prevention and control are solved.

Disclosure of Invention

In order to solve the technical blank of the deficiency in the background technology, the invention provides a multifunctional mesoporous biomaterial and a preparation method thereof, which have unique porous structures, are used for promoting the healing or the regeneration and the repair of various soft and hard tissue wound surfaces, especially can be used for the efficient bacteriostasis, the infection prevention and the rapid healing and the repair of the wound surfaces with inflammation problems in the chronic pathological change environment, and fill the blank in the prior art.

The dual-channel microstructure of the invention enables the outside of the particles to have extremely high specific surface and the inside to have channels with larger size, has extremely high-efficiency procoagulant ability to traumatic hemorrhage, can adsorb active factors of tissue regenerative cells, is beneficial to cell adhesion, growth, proliferation and differentiation, and the material can be degraded, absorbed and completely biocompatible.

In addition, the composition and the microstructure design of the material are extremely beneficial to the released inorganic ion composition which is necessary for human physiology and can efficiently inhibit bacteria by adjusting the calcination temperature and the sodium/potassium ion content level of the material and regulating and controlling the mesoporous structure.

The material provided by the invention meets the multifunctional requirements of rapid hemostasis, efficient bacteriostasis, long-acting anti-infection and promotion of the multifunctional synergetic regeneration and repair of soft/hard tissues, thereby achieving the ideal multifunctional mesoporous biomaterial standard and providing a superior novel multifunctional mesoporous biomaterial for solving clinical problems.

The technical scheme adopted by the invention is as follows:

a multifunctional mesoporous biomaterial comprises:

the multifunctional mesoporous biomaterial is a unique mesoporous structure particulate matter, and has a cabbage structure positioned inside and a wrapping mesoporous structure positioned outside.

The gap pitch between adjacent cabbage sheets in the internal cabbage structure is 30-200 nm.

The pore diameter of the external wrapping mesopores is 5-25 nm.

The ultrafine particles with the mesoporous structure are primary granulated particles of ultrafine powder of 100 nm-180 mu m or secondary granulated particles of 300 mu m-3 mm.

The granularity of the primary granulated particles is 100 nm-180 mu m, and the granularity of the secondary granulated particles is 300 mu m-3 mm.

The chemical substance with the pore channel structure consisting of the outer wrapping mesopores and the inner cabbage is in an amorphous glass state.

The main component of the multifunctional mesoporous biomaterial is SiO₂-B₂O₃-CaO。

The multifunctional mesoporous biomaterial also contains P₂O₅、CuO、MgO、ZnO、SrO、Li₂O、Na₂O and/or K₂O, one or more of the following.

The mesoporous structure particulate matter comprises the following components in parts by mole:

SiO₂24 to 60 portions of

B₂O₃4 to 30 portions of

10-40 parts of CaO

P₂O₅0 to 6 portions of

0 to 10 parts of MgO

0-5 parts of CuO

0 to 10 parts of ZnO

0 to 10 parts of SrO

Li₂0 to 5 parts of O

Na₂0 to 5 parts of O

K₂0-5 parts of O.

The invention uses silicon and boron oxide SiO₂And B₂O₃Is composed of a pore channel skeleton and amorphous glassy ultrafine particles hybridized by other multi-component oxides in the skeleton structure, the ultrafine particles take a cabbage structure as an inner core and are wrapped by a bagThe wound mesoporous pore structure is approximately spherical particles formed on the outer layer, and the ultrafine particles can form micron-millimeter-sized large particles through secondary granulation.

The multifunctional mesoporous biomaterial is prepared by adopting a template-mediated sol-gel method, and is subjected to secondary granulation to prepare particles. The type of the template agent is not strictly limited, and the template agent can be used for synthesizing the multifunctional mesoporous biomaterial as long as the active agent generated by the cabbage structure-wrapping mesoporous structure is not influenced.

The invention utilizes the microstructure guiding agent to prepare the ultrafine particle powder with a unique mesoporous structure in a sol-gel way to serve as the multifunctional mesoporous biomaterial.

Secondly, a preparation method of the multifunctional mesoporous biomaterial, which comprises the following steps:

1) adding the microstructure guiding agent into deionized water, stirring for dissolving, adjusting the pH value with an acid solution, and continuously stirring until the solution is clear;

2) adding tetraethoxysilane, fully hydrolyzing, sequentially adding a phosphorus source, a boron source and soluble metal salt, and fully stirring in a closed manner to obtain sol;

3) drying the sol, calcining to obtain powder, ball milling the powder to obtain glass state superfine mesoporous powder, and granulating the glass state mesoporous powder twice to obtain particles as the multifunctional mesoporous biomaterial.

The soluble metal salt comprises soluble calcium salt and soluble magnesium salt. The soluble metal salt also comprises one or more of soluble copper salt, soluble strontium salt, soluble lithium salt, soluble potassium salt, soluble sodium salt and soluble zinc salt.

The method comprises the following specific steps:

1) adding the microstructure guiding agent into deionized water according to the mass ratio of 1 (20-30), mechanically stirring and dissolving at 10-60 ℃, adjusting the pH value of the solution to 1.0-3.0 by using an acid solution, and continuously stirring until the solution is clear;

2) adding tetraethoxysilane into the solution, fully hydrolyzing, sequentially adding a phosphorus source, a boron source and soluble metal salt, sealing, and fully stirring for 4-36 hours at the ambient temperature of 40-95 ℃ to obtain uniform sol;

3) drying the sol obtained in the step 2) at 60-160 ℃, calcining at 550-750 ℃, ball-milling to obtain glassy state ultrafine mesoporous powder, and performing secondary granulation on the glassy state mesoporous powder to obtain particles serving as the multifunctional mesoporous biomaterial.

The microstructure directing agent is a polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer, preferably one or any combination of Pluronic P123(PEO20-PPO70-PEO20), Pluronic F127(PEO106-PPO70-PEO106) and Pluronic F108(PEO133-PPO50-PEO 133).

The phosphorus source is triethyl phosphate or phosphoric acid; the boron source is boric acid.

The calcium salt is calcium nitrate or calcium acetate; the magnesium salt is magnesium nitrate or magnesium acetate; the copper salt is copper nitrate; the lithium salt is lithium nitrate or lithium carbonate; the potassium salt is potassium nitrate or potassium carbonate; the sodium salt is sodium nitrate or sodium carbonate; the zinc salt is zinc nitrate; the strontium salt is strontium nitrate.

The acid solution is nitric acid or hydrochloric acid solution.

The secondary granulation is one of viscous solution hanging drop wrapping and viscous solution spraying wrapping;

the viscous solution hanging drop wrapping specifically comprises the following steps:

4) carrying out ball milling treatment on the mesoporous powder obtained in the step 3), adding the powder subjected to ball milling treatment into an organic binder solution with the mass fraction of 1% -10% according to the solid/liquid mass ratio of 1 (2-8), uniformly stirring to form a paste, then dropwise adding the paste into the organic binder solution with opposite charges, separating out microparticles, drying, and calcining at 500-700 ℃ to obtain particles with the granularity of 300-3 mm.

The viscous solution spray wrapping specifically comprises the following steps:

4) ball-milling the mesoporous powder obtained in the step 3), adding the ball-milled powder into 1-10% of organic binder solution according to the solid/liquid mass ratio of 1 (4-20), uniformly stirring to form extremely thin slurry, spraying the thin slurry into hot air atmosphere at the temperature of 60-100 ℃ through a high-pressure sprayer to rapidly dry slurry particles, and calcining at the temperature of 500-700 ℃ to obtain particles with the granularity of 300-3 mm.

The organic binder is one of hyaluronic acid, chitosan, carboxymethyl chitosan and sodium alginate.

The molar ratio of the microstructure directing agent to the tetraethoxysilane is 100: (3000-6000); the molar ratio of the phosphorus source to the tetraethoxysilane is 100: (0-2000), wherein the molar ratio of the boron source to the tetraethoxysilane is 100: (80-3000), wherein the molar ratio of the calcium salt to the ethyl orthosilicate is 100: (60-600), wherein the molar ratio of the calcium salt to the zinc salt, the magnesium salt or the strontium salt is 100: (0-25), wherein the molar ratio of the calcium salt to the copper salt, the lithium salt, the sodium salt or the potassium salt is 100: (0 to 12).

And 3) drying the sol in the step 3), specifically discharging the moisture in the sol, preferably drying or vacuum drying.

The multifunctional mesoporous biomaterial is applied to hemostasis and healing repair of wounds on the body surface and in vivo.

The multifunctional mesoporous biomaterial is applied to hemostasis of body surface and internal artery and vein damage, healing of soft tissue wound surfaces, repairing and reconstructing of articular cartilage calcified layers, tendons, ligaments and periosteum, regeneration and repair of various bone wounds and the like.

The multifunctional mesoporous biomaterial not only contains a common mesoporous structure, but also contains an internal porous structure in the form of cabbage, and the gap pitch scale is larger than the external hexagonal mesoporous pore scale, so that a large amount of water, active molecules, protein and the like can be quickly adsorbed, the adhesion and the growth matrix surface activity of tissue repair cells in injury can be improved, and the composition of various inorganic ions released by the material degradation can efficiently regulate inflammatory immune reaction, prevent and treat bacterial biofilm formed after pathogenic bacteria reside in the wound, therefore, the hemostatic and healing agent can be applied to the fields of rapid and efficient hemostasis and healing repair of body surface and internal wounds, particularly and preferably applied to the fields of hemostasis of various body surfaces and internal artery and vein damages, healing of soft tissue wound surfaces, repairing and reconstruction of articular cartilage calcified layers, tendons, ligaments and periosteum, and regeneration and repair of various bone wounds.

The chemical components of the multifunctional mesoporous bioglass are not strictly limited by the above components, for example, the components such as silicon dioxide, boron trioxide and calcium oxide are taken as essential components, copper oxide, zinc oxide, strontium oxide and magnesium oxide can not be 0 at the same time, the contents of sodium oxide and potassium oxide are not strictly limited, and other inorganic oxides of lithium, selenium and iron which are necessary for physiological metabolism can be added into the multifunctional material.

The chemical composition of the secondary granulated particles with the granularity of 300 mu m-3 mm is not strictly limited, the secondary granulated particles can be particles which are granulated and dried by hyaluronic acid, chitosan, sodium alginate and the like and are not calcined, and the secondary granulated particles can be directly used for hemostasis, and can also be used for promoting bone wound filling and repairing after being calcined at low temperature again to form mesoporous bioglass particles.

The multifunctional mesoporous biomaterial has no strict limitation on hybrid inorganic ions in the mesoporous framework, and all inorganic ions which are beneficial to maintaining and improving the biocompatibility, the hemostatic efficiency and the bacteriostatic efficiency, controlling inflammatory reaction and promoting tissue repair can be added into the mesoporous framework.

The granularity of the multifunctional mesoporous biomaterial is not strictly limited, the multifunctional mesoporous biomaterial can be ultrafine particles from nano-scale to hundred-micron scale, and also can be large particles from hundreds of microns to millimeter scale, the form and the structure of large particles are not strictly limited, and the multifunctional mesoporous biomaterial can be loose particles formed by stacking ultrafine particles, so that the requirements of dust removal, filling, implantation and the like can be favorably met.

The specific surface area level of the multifunctional mesoporous biomaterial is not a key factor for determining biological functions.

The multifunctional mesoporous biomaterial provided by the invention is beneficial to maintaining and improving the biocompatibility of the mesoporous biomaterial and modifying organic and inorganic molecules or ions which promote the repair efficiency of damaged tissues into the walls of the pore channels.

The multifunctional mesoporous biomaterial is prepared by coating, drying and calcining ultrafine particles on organic viscous liquid drops, or stirring the ultrafine particles in an organic viscous solution, and then drying, crushing and calcining the organic viscous solution.

The multifunctional mesoporous biomaterial is prepared into superfine particles by using organic dispersing agents or combustion improvers and other auxiliary substances.

The multifunctional mesoporous biomaterial is added into other organic and inorganic matrixes to form a film, a sponge, a bandage, a colloid or a paste for hemostasis or wound repair and the like.

The multifunctional mesoporous biomaterial has no strict limitation on the application range, and can be applied to the fields of body surface and in-vivo wound hemostasis, soft/hard tissue wound repair, inflammation regulation, infection prevention and control and the like.

The scheme of the invention is that the microstructure guiding agent is used as the microstructure guiding agent, the sol-gel method is adopted to prepare the particle material with the unique mesoporous pattern, the preparation process is simple, the particle material is used for artery and vein hemostasis and the promotion healing or regeneration and repair of soft tissue and bone tissue wounds, the tissue compatibility of the material is good, the inflammatory reaction in the wounds can be adjusted, and the biomedical material for preventing and treating pathogenic bacteria infection is prepared.

The invention has the advantages that:

1) in composition, the amorphous glassy mesoporous material ultrafine particles or aggregated particles subjected to secondary granulation prepared based on low-temperature calcination treatment contain multiple oxides which can be rapidly degraded when contacting body fluid, the released inorganic ion composition and the dosage level thereof are the dosage level of mineral substances or trace elements necessary for human physiological metabolism, the problem of seriously influencing cell and tissue compatibility does not exist, and meanwhile, the inorganic ion compositions also remarkably show the characteristics of inhibiting the activity of common pathogenic bacteria of a wound surface, promoting the apoptosis of the pathogenic bacteria and promoting the vascularization effect, so that the wound surface does not need to be thoroughly debrided and post-treated.

2) On the (micro) structure, the mesoporous structure wrapped outside and the superfine particles formed by the cabbage structure inside have small pore channel size (the pore diameter is 5-25 nm), the cabbage structure inside has large gap pore distance (30-200 nm) so as to form a unique dual porous structure, and although the specific surface area (15-120 m2/g) of the superfine particles is one order of magnitude lower than that of zeolite and mesoporous silicon spheres, the specific microstructure and chemical composition show superior hemostatic efficiency than Quikclot and the like, and show a hemostatic mechanism different from the conventional high specific surface area; meanwhile, the unique microstructure which does not depend on high specific surface area completely does not cause a severe thermal effect when contacting with a wound, and does not cause secondary damage to wound tissues; in addition, the loose large particles formed by secondary granulation are more beneficial to control when larger wound surface artery hemostasis and soft tissue or bone defect filling are carried out, thereby realizing ideal fit of biological function and operation implantation process.

3) In terms of biological effect, when the mesoporous material is contacted with a wound, the mesoporous material is constructed by the synergy of human body minerals and trace elements with specific biological effects of promoting vascularization, preventing infection, controlling inflammation, quickly coagulating blood, promoting regeneration and repair of soft/hard tissues and the like, and the synergistic effect of multiple mechanisms of quickly enriching platelets, quickly coagulating blood, quickly absorbing water and the like is utilized to realize the excellent performances of quickly stopping bleeding of the aorta and promoting adhesion and growth of tissue regeneration cells, thereby realizing the multifunctionality of stopping bleeding, resisting infection, controlling inflammatory reaction and promoting regeneration and repair of the wound, and fundamentally solving the problems of wound emergency treatment and synchronous repair of soft-hard tissue wound.

4) In operability, different series of multifunctional mesoporous biomaterials with wide scale distribution, which are manufactured by the technologies of a conventional sol-gel method, low-temperature calcination, secondary granulation and the like, can realize the implants required by any wound hemostasis, wound healing or regeneration and repair in vivo and in vitro.

Therefore, the degradable glassy mesoporous granular material which has the advantages of high-efficiency hemostasis, promotion of tissue repair and cooperative prevention and control of various complications and side reactions is remarkably characterized in that the preparation process of the material is simple, materials with different granularities can be selected according to the size of a wound to be stuffed or coated on the surface of the damaged wound, the coagulation and hemostasis can be realized efficiently, the inorganic ion composition released by the degradation of the granules can effectively kill germs on the wound surface and promote the healing of the wound surface, the problems of wound infection and the like are solved, and the treatment strategy of wound emergency treatment and tissue repair is greatly improved.

The multifunctional mesoporous biomaterial can be applied to the fields of hemostasis emergency treatment of wounds of soft and hard tissues on body surfaces and in vivo and tissue regeneration and repair.

Drawings

FIG. 1 is a transmission electron microscope photograph of ultrafine particles of multifunctional mesoporous biomaterial.

FIG. 2 is a photo of the multi-functional mesoporous biomaterial with ultrafine particles having a particle size of 300-400 μm, B with a particle size of 700-900 μm, C with a particle size of 2000-3000 μm after secondary granulation.

FIG. 3 is a photograph of the multifunctional mesoporous biomaterial degraded to release ion leaching solution to kill Staphylococcus aureus.

FIG. 4 is a photograph of multifunctional mesoporous biomaterial for promoting the healing of the skin wound of a diabetic rat.

FIG. 5 is a photograph of multifunctional mesoporous biomaterial repairing bone defect.

Detailed Description

The present invention will be further illustrated with reference to the following examples, which are not intended to limit the scope of the present invention, and all the techniques and materials prepared based on the above-mentioned contents of the present invention shall fall within the protection scope of the present invention. The purity of the reagents used in the examples is no lower than the purity of the reagents used in the analytical tests.

The examples of the invention are as follows:

example 1: SiO 2₂-B₂O₃-CaO-P₂O₅-CuO-MgO-ZnO-SrO-Li₂O-Na₂O-K₂O mesoporous material

1) Adding 0.02mol of P123 microstructure directing agent into 3000ml of deionized water according to the proportion of 1:20, mechanically stirring for 4 hours at 40 ℃ to fully dissolve, adjusting the pH value to 1.0 by using a hydrochloric acid solution, and continuously stirring until the solution is clear; adding 0.75mol of ethyl orthosilicate into the acidic solution, fully hydrolyzing, adding 0.06mol of triethyl phosphate, hydrolyzing, and then sequentially stirring 0.57mol of boric acid, 0.74mol of calcium nitrate, 0.03mol of copper nitrate, 0.01mol of magnesium nitrate, 0.01mol of zinc nitrate, 0.05mol of strontium nitrate, 0.01mol of lithium nitrate, 0.01mol of sodium nitrate and 0.01mol of potassium nitrate at 40 ℃ for 12 hours, and stirring at 95 ℃ for 24 hours to obtain a sol solution;

2) drying the sol solution obtained in the step 1) at 120 ℃, fully drying, and calcining at 700 ℃ for 8 hours to obtain mesoporous glass state inorganic powder;

3) ball-milling the powder obtained in the step 2) for 8 hours, adding the superfine powder into a 12.5% calcium chloride solution according to a solid/liquid ratio of 1:20, soaking at 100 ℃ for 12 hours, then carrying out suction filtration on the solution, washing with deionized water and absolute ethyl alcohol, and drying to obtain mesoporous bioglass powder with the granularity of 200 nm-120 mu m;

4) and (3) uniformly dispersing 10g of the powder obtained in the step 3) into 30 g of sodium alginate solution with the mass percentage of 3% under continuous stirring, dropwise adding the paste into 5% of chitosan solution with the mass percentage by using a micro sample injection peristaltic pump after uniform stirring, drying the separated micro particles, and calcining at 550 ℃ for 60 minutes to obtain the particles with the particle size of 2.0 mm.

As shown in the attached figure 1A, the transmission electron microscope shows that the ultrafine particles are approximate to spherical particles, the outside is a wrapping mesoporous structure, the inside is a cabbage structure, the outer mesoporous aperture is 10-15 nm, and the gap pitch of the inner cabbage structure is 60-120 nm; as shown in FIG. 2A, the secondary granulated particles photographed by the digital camera were approximately spherical, with a particle size of 2.0mm level.

Example 2: SiO 2₂-B₂O₃-CaO-P₂O₅-CuO-MgO-ZnO mesoporous material

1) Adding 0.02mol of P123 microstructure guiding agent into 3000ml of deionized water according to the proportion of 1:30, mechanically stirring for 4 hours at 40 ℃ for full dissolution, adjusting the pH value to 2.0 by using hydrochloric acid solution, and continuously stirring until the solution is clear; adding 0.89mol of ethyl orthosilicate into the acidic solution, fully hydrolyzing, adding 0.04mol of triethyl phosphate, sequentially adding 0.23mol of boric acid, 0.74mol of calcium nitrate, 0.03mol of copper nitrate, 0.01mol of magnesium nitrate and 0.01mol of zinc nitrate, stirring at 40 ℃ for 12 hours, and stirring at 90 ℃ for 24 hours to obtain a sol solution;

2) drying the sol solution obtained in the step 1) at 120 ℃, fully drying, and calcining at 650 ℃ for 6 hours to obtain glassy mesoporous powder;

3) ball-milling the powder obtained in the step 2) for 5 hours, adding the superfine powder into a 10% calcium chloride solution according to a solid-to-liquid ratio of 1:20, soaking at 85 ℃ for 12 hours, then carrying out suction filtration on the solution, washing with deionized water and absolute ethyl alcohol, and drying to obtain mesoporous bioglass powder with the granularity of 500 nm-120 mu m;

4) and (3) uniformly dispersing 10g of the powder obtained in the step 3) into 60 g of carboxymethyl chitosan solution with the mass percentage of 4% under continuous stirring, dropwise adding the paste into a sodium alginate solution with the mass percentage of 4% by using a micro-injection peristaltic pump after uniform stirring, drying the micro-particles after separation, and calcining the particles at 600 ℃ for 30 minutes to obtain the particles with the particle size of 1.5 mm.

As shown in attached figures 1B and C, the ultrafine particles are similar to spherical particles observed by a high-resolution transmission electron microscope, the outer part of the ultrafine particles is of a wrapping hexagonal mesoporous structure, the inner part of the ultrafine particles is of a cabbage structure, the outer mesoporous aperture is 25nm, and the gap pitch of the inner cabbage structure is 80-150 nm; as shown in FIG. 2B, the secondary granulated particles photographed by the digital camera were approximately spherical, with a particle size of the level of-1.5 mm.

Example 3: SiO 2₂-B₂O₃-CaO-P₂O₅-CuO-ZnO mesoporous material

1) Adding 0.02mol of F127 microstructure directing agent into 2500ml of deionized water according to the proportion of 1:25, mechanically stirring for 4 hours at 40 ℃ to fully dissolve, adjusting the pH value to 1.5 by using a nitric acid solution, and continuously stirring until the solution is clear; then adding 0.89mol of ethyl orthosilicate and 0.02mol of triethyl phosphate into the acidic solution, fully hydrolyzing, sequentially adding 0.19mol of boric acid, 0.52mol of calcium nitrate, 0.005mol of copper nitrate and 0.02mol of zinc nitrate, stirring for 12 hours at 40 ℃, and stirring for 24 hours at 90 ℃ to obtain a sol solution;

2) drying the sol solution obtained in the step 1) at 120 ℃, fully drying, and calcining at 700 ℃ for 8 hours to obtain glassy mesoporous powder;

3) ball-milling the powder obtained in the step 2) for 3 hours, adding the superfine powder into 15% calcium chloride solution according to the solid-to-liquid ratio of 1:20, soaking at 90 ℃ for 8 hours, then carrying out suction filtration on the solution, washing with deionized water and absolute ethyl alcohol, and drying to obtain mesoporous bioglass powder with the granularity of 100-120 mu m;

4) uniformly dispersing 20 g of the powder obtained in the step 3) into 90 g of hyaluronic acid solution with the mass percentage of 1% under continuous stirring, dropwise adding the paste into 5% of chitosan solution with the mass percentage by using a micro-sampling peristaltic pump after uniform stirring, drying after micro-particles are separated, and calcining at 650 ℃ for 30 minutes to obtain the particles with the particle size of 1.2-2.5 mm.

Example 4: SiO 2₂-B₂O₃Mesoporous material of-CaO-ZnO-SrO bioglass

1) Adding 0.02mol of P123 microstructure directing agent into 2500ml of deionized water according to the proportion of 1 (20-30), mechanically stirring for 4 hours at 40 ℃ for full dissolution, adjusting the pH value to 1.0 by using a hydrochloric acid solution, and continuously stirring until the solution is clear; adding 0.75mol of ethyl orthosilicate into the acidic solution, fully hydrolyzing, sequentially adding 0.19mol of boric acid, 0.87mol of calcium nitrate, 0.01mol of zinc nitrate and 0.005mol of strontium nitrate, stirring at 40 ℃ for 12 hours, and stirring at 90 ℃ for 24 hours to obtain a sol solution;

2) drying the sol solution obtained in the step 1) at 120 ℃, fully drying, and calcining at 700 ℃ for 8 hours to obtain mesoporous bioglass powder;

3) ball-milling the powder obtained in the step 2) for 6 hours, adding the superfine powder into a 12.5% calcium chloride solution according to a solid-to-liquid ratio of 1:20, soaking at 100 ℃ for 2-12 hours, then carrying out suction filtration on the solution, washing with deionized water and absolute ethyl alcohol, and drying to obtain glass-state mesoporous inorganic powder with the granularity of 600 nm-40 mu m;

4) adding 20 g of the powder obtained in the step 3) into 200 g of 2% sodium alginate solution, then uniformly stirring to form extremely thin slurry, then spraying the thin slurry into hot air at 80 ℃ through a high-pressure sprayer to rapidly dry slurry particles, and then calcining at 580 ℃ for 45 minutes to obtain particles with the particle size of 0.2-1.0 mm.

Example 5: SiO 2₂-B₂O₃Mesoporous material of-CaO-CuO-MgO

1) Adding 0.02mol of P123 microstructure directing agent into 2500ml of deionized water according to the proportion of 1:30, mechanically stirring for 4 hours at 40 ℃ to fully dissolve, adjusting the pH value to 1.2 by using a hydrochloric acid solution, and continuously stirring until the solution is clear; adding 0.85mol of ethyl orthosilicate into the acidic solution, fully hydrolyzing, sequentially adding 0.25mol of boric acid, 0.56mol of calcium nitrate, 0.02mol of copper nitrate and 0.01mol of magnesium nitrate, stirring at 40 ℃ for 12 hours, and stirring at 90 ℃ for 24 hours to obtain a sol solution;

2) drying the sol solution obtained in the step 1) at 120 ℃, fully drying, and calcining at 600 ℃ for 8 hours to obtain glassy mesoporous powder;

3) ball-milling the powder obtained in the step 2) for 2 hours, adding the superfine powder into a 12% calcium chloride solution according to a solid-to-liquid ratio of 1:20, soaking for 4 hours at an ambient temperature of 100 ℃, then carrying out suction filtration on the solution, washing with deionized water and absolute ethyl alcohol, and drying to obtain glassy mesoporous powder with the granularity of 50-180 mu m;

4) adding 20 g of the powder obtained in the step 3) into 150g of 2% sodium alginate solution, stirring to form uniform slurry, spraying the thin slurry into hot air at 100 ℃ through a high-pressure sprayer to rapidly dry slurry particles, and calcining at 620 ℃ for 45 minutes to obtain particles with the particle size of 1.0-2.5 mm.

Example 6: SiO 2₂-B₂O₃Mesoporous material of-CaO-ZnO

1) Adding 0.02mol of P123 microstructure directing agent into 2500ml of deionized water according to the proportion of 1 (20-30), mechanically stirring for 4 hours at 40 ℃, fully dissolving, adjusting the pH value to 2.0 by using nitric acid solution, and continuously stirring until the solution is clear; adding 0.96mol of ethyl orthosilicate into the acidic solution, fully hydrolyzing, sequentially adding 0.19mol of boric acid, 0.85mol of calcium nitrate and 0.01mol of zinc nitrate, stirring at 40 ℃ for 12 hours, and stirring at 90 ℃ for 24 hours to obtain a sol solution;

2) drying the sol solution obtained in the step 1) at 120 ℃, fully drying, and calcining at 700 ℃ for 4 hours to obtain glassy mesoporous powder;

3) ball-milling the powder obtained in the step 2) for 2 hours, adding the superfine powder into a calcium chloride solution with the concentration of 8% according to the solid-to-liquid ratio of 1:20, soaking for 10 hours at the ambient temperature of 100 ℃, then carrying out suction filtration on the solution, washing with deionized water and absolute ethyl alcohol, and drying to obtain mesoporous powder with the granularity of 5-120 mu m;

4) adding 10g of the powder obtained in the step 3) into 180 g of 2% chitosan solution, then uniformly stirring to form extremely thin slurry, then spraying the thin slurry into hot air at 90 ℃ through a high-pressure sprayer to quickly dry slurry particles, and then calcining at 620 ℃ for 30 minutes to obtain particles with the particle size of 0.9-1.5 mm.

The experiments of the examples verify that:

1. bacteriostasis test

The bioactive glass multifunctional mesoporous biomaterial powder of the embodiment 1, the embodiment 2, the embodiment 3 and the embodiment 6 is respectively soaked in 10ml of PBS solution according to 0.10g of powder and shaken in a shaking table at 37 ℃ for 24 hours, then the supernatant is filtered and filtered by a 0.22-mum pore membrane, then the supernatant is respectively added into agar inoculated with staphylococcus aureus (S, Aureus), the culture is continued for 24 hours under the anaerobic environment, and the condition of the bacterial content in the surface dish agar is observed.

The results show that as shown in fig. 3, high-density bacteria exist in the petri dish of the biological control group without adding the powder leaching liquor, but no bacteria are found in the agar of the petri dish added with the leaching liquor of examples 1 to 3, and only very rare bacteria are found in the agar of the petri dish added with the powder leaching liquor of example 6, which indicates that the multifunctional mesoporous biomaterial of the mesoporous bioglass has an obvious inhibiting effect on the activity of clinical dry pathogenic bacteria, namely staphylococcus aureus, and can even kill the growing bacteria.

2. Biosafety testing

1) Hemolysis test

According to the test method specified in the national medical apparatus biological safety evaluation standard GB/T16886.4, 120ml of fresh New Zealand white rabbit blood is divided into 6 equal parts to prepare 2% erythrocyte suspension for test. Sequentially adding 2% of erythrocyte suspension, distilled water and the physiological saline leaching liquor on the multifunctional mesoporous biomaterial of the embodiments 1-6 into a clean test tube, uniformly mixing, immediately placing into a 37 ℃ constant temperature water bath box, incubating, detecting OD values at different time points, and calculating the hemolysis rate. The result shows that the hemolysis rate of the multifunctional mesoporous biomaterial is less than 5 percent, and the hemolysis rate of less than 5 percent is qualified according to the national standard.

2) Acute systemic toxicity test

20 SD rats with cleaning grade about 8 weeks are selected, the weight of each SD rat is controlled between 130 g and 150g, the tail veins of the SD rats are injected with multifunctional mesoporous biomaterial physiological saline leaching liquor (experimental group), the growth condition of experimental animals is monitored according to GB/T16886.11 standard, no poisoning symptom given by standard is seen, no death occurs, the weights of the experimental group and the control group are weighed at each time point, the average weight of the two groups of animals is gradually increased, and no significant difference (p is greater than 0.05) exists.

3) Pyrogen test

Taking healthy adult New Zealand white rabbits, wherein the male and female parts are unlimited, injecting the multifunctional mesoporous biomaterial physiological saline leaching liquor with mesoporous channels from the ear edge vein, detecting the anal temperature of the experimental rabbits for 1 time/hour, continuously measuring for 4 hours, and the experimental result shows that the temperature of the experimental animals rises to 0.24 ℃ and is lower than 1.4 ℃, thereby meeting the regulation of the national biosafety evaluation standard GB/T16886.4 of medical instruments.

4) Intradermal reaction test

Taking healthy adult New Zealand rabbits, shaving backs, designing 5 injection points in test skin areas on two sides of a spine, enabling the distance between the injection points and the spine to be 2cm, enabling the interval between each injection point and the spine to be 2cm, injecting about 0.2ml of multifunctional mesoporous biomaterial physiological saline leaching liquor with mesoporous channels, observing and recording the conditions of the injection points at 24 h, 48 h and 72h time points after injection, and grading according to the grading standard in GB/T16886.10.

The experimental result shows that the skin of each animal test area has no erythema and edema and has no any stimulation reaction, and the average primary stimulation index PII at each time point is 0 according to the standard calculation, which indicates that the material has no stimulation effect on the skin.

5) Muscle implantation test

Taking SD rats, after depilation, designing 4 skin incisions of 1cm at the thick part of muscles of 1.5cm away from the spine at the back, performing aseptic operation, implanting a multifunctional mesoporous biomaterial wafer with mesoporous channels of 6mm in diameter and prepared by tabletting, observing the recovery condition of wounds in appearance, and simultaneously taking muscle tissues around the implanted part for HE dyeing histological observation in 1 week, 2 weeks and 12 weeks respectively.

According to the GB/T16886.6 standard, the general condition of experimental animals after observation is good, the wound is free from red swelling and exudation, the wound is well healed after 1 week, the experimental group can touch a mass of 3mm, the mass of 1mm can touch after 2 weeks, the mass disappears after 12 weeks, the tissue HE staining result at each time point shows that acute inflammation is presented at the early stage of back burying (1-2 weeks), new capillary vessels can be formed among loose fibrous tissues around the material after 12 weeks, part of the material is divided and wrapped by fibrous connective tissues, the wrapping material is greatly proliferated in fiber, the arrangement is regular, and a small amount of lymphocyte infiltration can be seen at the junction of the material and the tissue.

3. Hemostasis test of femoral artery

Taking a healthy adult New Zealand white rabbit, weighing about 2.5kg, injecting sodium pentobarbital intravenously, and fixing on an operating table on the back after anesthesia. The femoral artery was dissected and the artery was cut transversely through an incision occupying diameter 1/3. Immediately applying the multifunctional mesoporous biomaterial or particle product of examples 1, 2 and 3 of the present invention to the bleeding site respectively

Observing and recording bleeding time, blood loss and monitoring the hemostatic processThe local temperature of the wound surface in (1) and the survival condition of the rabbit within 72 hours after the operation.

Test results show that the blood loss of tested rabbits made of the materials of examples 1, 2 and 3 is 10-20 ml, the hemostasis time is 0.5-2 min, no temperature change exists in the testing process, and 3 days after the operation, the tested rabbits have good wound healing, free movement, good vital signs and a survival rate of 100%; use of hemostatic particulate products

The blood loss of the tested rabbit is 16-23 ml, the hemostasis time is 2-4 min, the temperature rises to 65 ℃ in the testing process, the inflammatory reaction of the tested rabbit wound occurs 3 days after the operation, 45% of the life signs of the tested rabbit are good, and the survival rate is 70%. Experiments show that the mesoporous multifunctional mesoporous biomaterial has excellent hemostatic effect and is obviously superior to hemostatic particle products

4. Hemostasis test of femoral vein

Taking a healthy adult New Zealand white rabbit, weighing about 2.5kg, injecting sodium pentobarbital intravenously, and fixing on an operating table on the back after anesthesia. The femoral artery was dissected and the artery was cut transversely through an incision occupying diameter 1/3. Immediately applying the multifunctional mesoporous biomaterial of examples 1, 2 and 3 of the present invention to the bleeding site, observing and recording the bleeding time, the amount of blood loss and monitoring the local temperature of the wound surface during hemostasis, and the survival condition of rabbits within 72h after the operation.

Test results show that the blood loss of a tested rabbit is 2-3 ml, the hemostasis time is 0.5-1.5 min, no temperature change exists in the test process, the wound of the tested rabbit is well healed 3 days after an operation, the tested rabbit freely moves, the vital sign is good, the survival rate is 100%, and experiments show that the mesoporous multifunctional mesoporous biomaterial has an excellent venous hemostasis effect.

5. Hepatic artery hemostasis test

Taking a healthy adult New Zealand white rabbit, weighing about 2.5kg, injecting sodium pentobarbital intravenously, fixing on the operating table on the back after anesthesiaThe above. The femoral artery was dissected and the artery was cut transversely through an incision occupying diameter 1/3. Immediately applying the multifunctional mesoporous biomaterial or hemostatic particle product of examples 1, 2 and 3 of the present invention to the bleeding site

The bleeding time, the amount of blood lost and the local temperature of the wound surface during the hemostasis process and the survival condition of the rabbit within 72h after the operation are observed and recorded.

Test results show that the tested rabbits using the materials of examples 1, 2 and 3 have the blood loss of 7-13 ml, the hemostasis time of 0.5-2 min, no temperature change in the testing process, good wound healing, free movement, good vital signs and the survival rate of 100% 3 days after the operation; use of hemostatic particulate products

The blood loss of the tested rabbit is 12-17 ml, the hemostasis time is 1-3 min, the temperature rises by about 25 ℃ in the testing process, and the tested rabbit has obvious inflammation on the wound and the survival rate is 60% after 3 days after the operation. Experiments show that the multifunctional mesoporous biomaterial has excellent hemostatic effect.

6. Hemostasis test of rectal vein

Taking a healthy adult New Zealand white rabbit, weighing about 2.5kg, injecting sodium pentobarbital intravenously, and fixing on an operating table on the back after anesthesia. The femoral artery was dissected and the artery was cut transversely through an incision occupying diameter 1/3. Immediately applying the multifunctional mesoporous biomaterial of examples 1, 2 and 3 of the present invention to the bleeding sites, respectively, observing and recording the bleeding time, the amount of blood loss and monitoring the local temperature of the wound surface during hemostasis and the survival condition of rabbits within 72h after the operation.

Test results show that the blood loss of a tested rabbit is 10-20 ml, the hemostasis time is 0.5-2 min, no temperature change exists in the test process, the wound of the tested rabbit is well healed 3 days after an operation, the tested rabbit freely moves, the vital sign is good, the survival rate is 100%, and experiments show that the multifunctional mesoporous biomaterial has an excellent hemostasis effect.

7. Hemostasis experiment of carotid artery of large animal

Taking a healthy adult beagle dog with the weight of about 18kg, carrying out intravenous injection of sodium pentobarbital, and fixing the beagle dog on an operating table on the back after anesthesia. The femoral artery was dissected and the artery was cut transversely through an incision occupying diameter 1/3. Immediately and respectively applying the multifunctional mesoporous biomaterial with mesoporous channels or the hemostatic particle products of the embodiments 1, 2 and 3 of the invention to the bleeding part

And observing and recording bleeding time, blood loss and monitoring the local temperature of the wound surface in the hemostasis process and the survival condition of the beagle dog within 72 hours after the operation.

The test results show that the tested beagle dogs using the materials of examples 1, 2 and 3 have the blood loss of 30-50 ml, the hemostasis time of 1-3 min, no temperature change in the test process, and 3 days after the operation, the tested beagle dogs have good wound healing, free activity, good vital signs and the survival rate of 100%. Use of hemostatic particulate products

The blood loss of the tested beagle dog is 35-65 ml, the hemostasis time is 2-4 min, no temperature change exists in the testing process, the wound of the tested beagle dog is inflamed 3 days after the operation, 60% of vital signs are good, and the survival rate is 100%. Experiments show that the multifunctional mesoporous biomaterial has excellent hemostatic effect.

8. White rabbit diabetes model skin wound repair experiment

Taking a healthy adult New Zealand white rabbit with the weight of about 2.5kg, injecting a medicament alloxan to establish a diabetes model, anesthetizing the rabbit after 6 weeks of medicament injection, removing back hairs, performing full-thickness peeling by using surgical scissors to establish a wound model, respectively coating the wound surface with the multifunctional mesoporous bioglass powder material, the conventional 45S5 glass powder material or the conventional 58S mesoporous bioglass powder material in the embodiment 1 of the invention, observing the inflammatory reaction of the wound surface, and periodically replacing the wound surface powder dressing.

The test result is shown in figure 4, the tested rabbit wound healing efficiency using the material of the example 1 is the highest, the wound inflammatory reaction degree is lower than that of the wound of the conventional 45S5 and mesoporous 58S glass dressing, the wound healing rate reaches over 90% when a wound model is established for 21 days, but the healing area of other dressing groups is only about 50% -65%. Experiments show that the multifunctional mesoporous biomaterial has excellent effect of promoting healing and repairing of soft tissue wounds.

9. Experiment for repairing femur defect of osteoporosis model of white rabbit

Taking a female healthy adult New Zealand white rabbit with the weight of about 2.5kg, carrying out an ovary extirpation operation, feeding for 12 weeks, injecting sodium pentobarbital intravenously, and fixing the rabbit on an operating table on the back after anesthesia. The soft tissues of the femoral condyle part are incised, a bone defect with the diameter of 6mm and the depth of 8mm is established by using a bone drill, then secondary granulation particles of the multifunctional mesoporous biomaterial prepared in the example 1 are filled in the bone defect, then the bone defect is sutured, the bone defect is bred for 4 weeks, 8 weeks and 12 weeks under the standard condition, then the animal is respectively treated by adopting an excessive anesthesia method, the femoral specimen is taken out, and the Micro-CT two-dimensional reconstruction analysis is carried out on the defect part.

As shown in FIG. 5, the femoral defects using the secondary granulated material of example 1 all observed new bone growth into the interparticle spaces at 4 weeks after implantation of the material, and did not have any inflammatory response; at 8 and 12 weeks post-surgery, significant degradation of the particulate material occurred and new bone in-growth increased significantly. Experiments show that the multifunctional mesoporous biomaterial has excellent bone regeneration effect.

Claims

1. A multifunctional mesoporous biomaterial is characterized in that: the multifunctional mesoporous biomaterial is a mesoporous structure particulate matter, and has a cabbage structure positioned inside and a wrapping mesoporous structure positioned outside.

2. The multifunctional mesoporous biomaterial according to claim 1, wherein:

The pore diameter of the outer wrapping mesopore is 5-25 nm, and the pore structure chemical substance formed by the outer wrapping mesopore and the inner cabbage is in an amorphous glass state.

3. The multifunctional mesoporous biomaterial according to claim 1, wherein:

4. The multifunctional mesoporous biomaterial according to claim 3, wherein:

5. The preparation method of the multifunctional mesoporous biomaterial is characterized by comprising the following steps of:

3) drying the sol, calcining to obtain powder, ball-milling the powder to obtain glassy mesoporous powder, and performing secondary granulation on the glassy mesoporous powder to obtain particles serving as the multifunctional mesoporous biomaterial.

6. The preparation method of the multifunctional mesoporous biomaterial according to claim 5, wherein the preparation method comprises the following steps: the method comprises the following specific steps:

3) drying the sol obtained in the step 2) at 60-160 ℃, calcining at 550-750 ℃, ball-milling to obtain glassy mesoporous powder, and performing secondary granulation on the glassy mesoporous powder to obtain particles serving as the multifunctional mesoporous biomaterial.

7. The preparation method of the multifunctional mesoporous biomaterial according to claim 5, wherein the preparation method comprises the following steps:

8. The preparation method of the multifunctional mesoporous biomaterial according to claim 5, wherein the preparation method comprises the following steps:

The viscous solution spray wrapping specifically comprises the following steps:

9. The preparation method of the multifunctional mesoporous biomaterial according to claim 5, is characterized by comprising the following steps of:

10. The use of the multifunctional mesoporous biomaterial according to any one of claims 1 to 4 or the multifunctional mesoporous biomaterial obtained by the method according to any one of claims 6 to 9, wherein the multifunctional mesoporous biomaterial is characterized in that: