CN108066815B

CN108066815B - Bone implant material and preparation method and application thereof

Info

Publication number: CN108066815B
Application number: CN201611022649.9A
Authority: CN
Inventors: 童丽萍; 喻学锋; 王怀雨
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2016-11-18
Filing date: 2016-11-18
Publication date: 2020-07-28
Anticipated expiration: 2036-11-18
Also published as: CN108066815A

Abstract

The invention provides a bone implant material and a preparation method and application thereof. The preparation method of the bone implant material comprises the following steps: ultrasonically dispersing black phosphorus in a first solvent to obtain a dispersion liquid A; dissolving a high polymer material in a second solvent to obtain a solution B; mixing the dispersion liquid A and the solution B, and then carrying out ultrasonic treatment to obtain a uniformly dispersed BP @ high polymer material solution; and then injecting the mixture into a mold for molding or performing 3D printing molding, and volatilizing the solvent to prepare the bone implant material. The bone implant material has the function of promoting bone formation by light and heat, can be degraded in a physiological environment, can be used as an implant of a bone defect filling part to be applied to clinical treatment of bone defects, can improve the efficiency of penetrating biological tissues by near infrared to thermally promote bones, can be combined with clinical physiotherapy to achieve better curative effect of promoting bone defect healing, and can participate in the processes of bone healing and regeneration because the degradation product of the bone implant material is a substance necessary for a human body.

Description

Bone implant material and preparation method and application thereof

Technical Field

The invention belongs to the technical field of biomedical polymer materials, and relates to a bone implant material, and a preparation method and application thereof.

Background

With the coming of aging of the population of the society in China, the number of people suffering from bone trauma or bone tumor is greatly increased. Clinically, cases of bone defects, fractures of parts difficult to heal, and bone nonunion caused by bone wounds, bone tumors, and the like are gradually increased year by year, and the treatment is difficult.

Bone defects are caused by a variety of factors, and fracture, external trauma (such as car accidents, war, accidental injury and the like) and bone tumor surgery are common. Wherein the bone defect caused by fracture and external trauma caused by bone loss of the elderly accounts for over 75 percent of the total factors. The aging of population is a common problem in developed countries and developing countries, China enters the aging society in advance as early as 1999, and the fracture or bone defect of the old people caused by osteoporosis is increased greatly. Furthermore, the incidence of bone tumors is also increasing year by year, especially in the adolescent population, this trend being more pronounced.

The orthopedic material implantation is a common method clinically used for treating patients with bone defects at present, and has the greatest advantage of avoiding secondary damage to patients caused by autologous bone transplantation and virus infection risks and severe immunological rejection reactions caused by allogeneic bone transplantation. As a new medical direction, although bone implant materials have a high success rate in surgery, implant failure has been frequently reported in recent years. The failure causes mainly include: poor tissue compatibility, infection, impaired healing, etc. The failure or partial function loss of the implant is two key problems of the application of the implant material at the present stage, the improvement of the compatibility of the implant and the surrounding tissues and the improvement or enhancement of the bone healing capability of the damaged part are main ways for solving the problems.

The use of bone implant materials is currently the common method for clinical treatment of bone defects, and therefore the development of new bone repair materials is of great importance. Bone implant materials can be classified into three categories, metal, ceramic and polymer, according to the material. Among them, the degradable high molecular bone implant material has been a research hotspot in recent years due to its advantages of low elastic modulus, controllable degradation, no toxic and side effects of the degradation products, etc. Both the medical and materials communities have attempted various methods to improve the ability of bone to heal around implants to reduce implant failure. The method mainly comprises the following two ideas:

(1) through systemic or local administration, the rejection reaction of a patient body to the implant is reduced, and the healing of the damaged part is promoted; or by external aids such as physical therapy to stimulate bone healing at the site of injury.

The growth factors adopted at present mainly comprise Bone Morphogenetic Protein (BMP) and Insulin Growth Factor (IGF), other growth factors still under research also comprise Fibroblast Growth Factor (FGF), Platelet Derived Growth Factor (PDGF), Vascular Endothelial Growth Factor (VEGF), transforming growth factor β (TGF- β) and the like.

Physical therapy, for example: electrical stimulation, mechanical stimulation, ultrasonic stimulation, laser stimulation, and the like are generally used as auxiliary means for promoting bone healing. The electrical stimulation promotes local blood circulation mainly through micro-current stimulation, and induces the generation of various bone growth factors, thereby accelerating the connection of bone defect areas and the formation and reconstruction of new bones. Traction is the most commonly used mechanical stimulation means, and by means of a bone tractor, traction force in a certain speed and direction is applied to bone segments to gradually separate the bone segments, new bone is generated in the generated gap, and surrounding soft tissues are expanded to achieve the purpose of expanding, widening or repairing bone defects. The low-intensity pulse ultrasonic stimulation can not only increase the synthesis of angiogenesis-related cytokines and promote the formation of blood vessels, but also cause the change of cell membrane permeability, promote the expression of related genes and promote the formation of new bones and the reconstruction of callus. Laser stimulation of infrared or near infrared spectra can promote angiogenesis, fibrosis proliferation and collagen deposition, not only accelerate the process of bone healing, but also increase bone density and elastic modulus of bone. In addition, the photochemical and photobiological properties of laser light have been reported to slow down inflammation at the treatment site, clinically manifested as reduced discomfort and edema in the post-operative patient.

(2) The implant is modified and adapted to enhance the compatibility of the implant with human tissue.

Currently, the research on surface and bulk treatment of implants mainly includes two aspects: the surface of the implant is loosened and roughened by applying mechanical or physical and chemical methods, so that the implant has better biological adhesion, surface tension and bone tissue affinity; the biological energy of the material is improved by changing the surface energy, the surface charge and the surface composition of the material. Active groups are introduced on the surface or inside of the implant material by a biochemical method, and bioactive molecules or drugs are loaded, so that the implant has the functions of promoting osteogenesis and locally carrying drugs on the basis of the mechanical support function of the implant.

Many bone defect schemes are currently treated clinically or under study, such as drug intervention, physical therapy, tissue engineering scaffolds, etc. Each method has certain limitations.

The systemic or local administration mode, especially the growth factor type drugs, mostly protein or polypeptide, are easy to degrade in vivo, have short biological half-life and short effective time, and the diffusion and metabolism of the drug occur in the whole body, actually reach the target site and have effective dose far lower than the administration dose. When the dose is increased, the side effects on the liver, kidney and other organs may be increased. In addition, such drugs are costly and frequent use can place a heavy economic burden on the patient.

The traction period of the distraction osteogenesis is long, and the condition that new bones are formed badly or healing is delayed can occur; the placement of the retractor not only risks rejection, infection, loosening, but also causes some damage to the nerve and muscle function at the location of the retractor. Ultrasound stimulation is still controversial at present, and although studies have shown that ultrasound can accelerate the formation and remodelling of new bone, its effects are not significant in the middle and late stages. Laser photo-thermal stimulation osteogenesis usually needs to increase the irradiation intensity of laser to obtain better treatment effect because the penetrating capability of infrared/near-infrared laser is weak, and the increase of the irradiation intensity can burn skin or muscle tissues on the surface layer.

At present, most of the common degradable high polymer bone implant materials are inert materials, and insufficient bone activity is a key factor for restricting the application of the degradable high polymer bone implant materials. Therefore, how to improve the osteogenesis promoting ability of degradable polymer bone implant materials is important for research and development.

Disclosure of Invention

In order to solve the above technical problems, an object of the present invention is to provide a bone implant material and a method for preparing the same, which can improve the ability of bone healing around an implant.

The invention also aims to provide application of the bone implant material as an implant of a bone defect filling part in clinical treatment of bone defects.

The purpose of the invention is realized by the following technical scheme:

the invention provides a preparation method of a bone implant material, which comprises the following steps:

step one, ultrasonically dispersing black phosphorus in a first solvent to obtain a dispersion liquid A;

dissolving a high polymer material in a second solvent to obtain a solution B;

mixing the dispersion liquid A and the solution B, and then carrying out ultrasonic treatment to obtain a uniformly dispersed BP @ high polymer material solution;

step four, injecting the solution of the BP @ high polymer material obtained in the step three into a mold for molding or performing 3D printing molding, and volatilizing the solvent to prepare the bone implant material;

wherein the mass ratio of the black phosphorus to the high polymer material is (0.1-1.0%): 1.

in the above preparation method, preferably, the first solvent may include one or a combination of more of anhydrous ethanol, deionized water, dimethyl sulfoxide, halogenated hydrocarbon solvents, dimethylformamide, tetrahydrofuran, and the like.

In the above production method, preferably, the second solvent includes one or a combination of more of a halogenated hydrocarbon solvent, dimethylformamide, tetrahydrofuran, and the like;

more preferably, the halogenated hydrocarbon solvent may include dichloromethane and/or chloroform, but is not limited thereto.

In the preparation method, the first solvent is used for cleaning and assisting the dispersion of the black phosphorus, so that the addition amount of the first solvent can be carried out according to actual operation, and the black phosphorus precipitate is covered.

In the above preparation method, preferably, the polymer material may include one or a combination of more of polylactic acid-glycolic acid copolymer (P L GA), polycaprolactone (PC L), polylactic acid (P L a), polybutylene succinate (PBS), polytrimethylene carbonate (PTMC), and the like.

In the above preparation method, preferably, in the polylactic acid-glycolic acid copolymer, the mass ratio of polylactic acid to glycolic acid may be 75: 25. 85: 15 or 90: 10. different mass ratios can adjust the parameters of the hardness, Young modulus and the like of the final bone implant material film or scaffold.

In the preparation method, the addition amount of the second solvent is carried out according to actual operation, so that the polymer can be completely dissolved, the viscosity of the solution is not high, the water sample fluidity is ensured, the uniform black phosphorus ultrasonic dispersion can be ensured, and preferably, the dosage ratio of the polymer material to the second solvent can be (25mg-50 mg): 1m L.

In the above production method, preferably, the volatilization of the solvent is carried out at room temperature or at low temperature; more preferably, the low temperature means 4 ℃ or lower.

In the above preparation method, preferably, the black phosphorus may include black phosphorus nanosheets and/or black phosphorus quantum dots;

more preferably, the black phosphorus nanosheet is a two-dimensional layered black phosphorus having a thickness of less than 100 nm;

more preferably, the black phosphorus quantum dots are ultra-small black phosphorus nanosheets having a thickness of less than 10 nm.

In the above preparation method, preferably, the preparation method of the two-dimensional layered black phosphorus comprises:

weighing blocky black phosphorus, gradually adding N-methyl pyrrolidone, fully grinding, performing ice-water bath ultrasonic treatment, performing first centrifugation, and taking supernatant to obtain a two-dimensional layered black phosphorus suspension; centrifuging the suspension of the two-dimensional layered black phosphorus for the second time and taking the precipitate to obtain the two-dimensional layered black phosphorus;

wherein the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is less than 2 mg: 1m L, the time of ultrasonic treatment is 4-12h, the first centrifugation rotating speed is 4000-14000 rpm, the centrifugation time is 10-20min, the second centrifugation rotating speed is 12000-14000rpm, and the centrifugation time is 10-20 min;

more preferably, the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1 mg: 1m L, the ultrasonic treatment time is 6h, the first centrifugation rotating speed is 4000rpm, the centrifugation time is 15min, the second centrifugation rotating speed is 12000rpm, and the centrifugation time is 15 min.

In the above preparation method, preferably, the preparation method of the ultra-small black phosphorus nanosheet is:

weighing massive black phosphorus, gradually adding N-methyl pyrrolidone, fully grinding, placing on ice, performing ultrasonic treatment by using a probe, performing first centrifugation, and taking supernatant to obtain a suspension of ultra-small black phosphorus nanosheets; centrifuging the suspension of the ultra-small black phosphorus nanosheets for the second time and taking the precipitate to obtain the ultra-small black phosphorus nanosheets;

wherein the dosage ratio of the block black phosphorus to the N-methyl pyrrolidone is less than 2 mg: 1m L, the time of ultrasonic treatment is 4-12h, the first centrifugation rotating speed is 7000-14000 rpm, the centrifugation time is 10-20min, the second centrifugation rotating speed is 12000-14000rpm, and the centrifugation time is 10-20 min;

more preferably, the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1 mg: 1m L, the ultrasonic treatment time is 10h, the first centrifugation rotating speed is 10000rpm, the centrifugation time is 15min, the second centrifugation rotating speed is 12000rpm, and the centrifugation time is 15 min.

In the preparation method, the preparation methods of the two-dimensional layered black phosphorus nanosheet and the ultra-small black phosphorus nanosheet are different in that: difference in ultrasonic power and post-ultrasonic centrifugal velocity. The two-dimensional layered black phosphorus is substantially stripped from the black phosphorus block by applying external mechanical force. From the ultrasonic power, the power of the water bath ultrasonic is generally 300W, and the power of the probe ultrasonic is larger and can reach 700-900W. From the centrifugal condition, the centrifugal speed of the two-dimensional layered black phosphorus is mainly 4000-7000rpm, and the centrifugal speed of the ultra-small black phosphorus nanosheet is mainly 7000-12000 rpm.

In the preparation method, because the black phosphorus is active in nature, and is easily oxidized and degraded when meeting oxygen and moisture, and the special properties of the black phosphorus are lost, the two-dimensional layered black phosphorus and the ultra-small black phosphorus nanosheet which are generally prepared are stored in NMP (N-methyl pyrrolidone) to play a role in stabilization and protection.

However, the black phosphorus dispersion prepared by the above method has a relatively low concentration, and can be concentrated by centrifugation. NMP has a high boiling point and is difficult to volatilize, and the subsequent preparation of a film or a bracket is not facilitated if NMP is not removed; and NMP is highly cytotoxic, so it needs to be removed by centrifugation. And (3) dispersing the centrifuged BP in an organic solvent, on one hand, cleaning the BP precipitate to completely remove NMP, and on the other hand, helping the BP to be uniformly dispersed in a high molecular solution.

In the above-mentioned preparation method, the shape of the mold used may be a film shape, a three-dimensional scaffold shape, or a cylindrical hollow shape, but is not limited thereto.

In the preparation method, the mass ratio of the black phosphorus to the high polymer material is (0.1-1.0%): 1, by adopting the proportion, firstly, the temperature can be raised to the required temperature (40.5 +/-0.5 ℃) after the near-infrared laser irradiation, secondly, the high polymer material and the black phosphorus are both acidic after degradation, a strong acidic environment has certain negative influence on the osteogenesis induction of stem cells, and the mass ratio of the black phosphorus to the high polymer material is controlled to be (0.1-1.0%): 1, the negative effect can be overcome, and a good practical effect is achieved.

In the preparation method, the solution of the BP @ high polymer material is poured into a mold, and the organic solvent is volatilized. The volatilization rate of the organic solvent can be adjusted by controlling the ambient temperature, which further affects the surface topography of the prepared film or stent. Adopting normal temperature and/or low temperature conditions, if the film is placed in a lower temperature environment (such as a refrigerator at 4 ℃), the volatilization speed of the organic solvent can be slowed down, and the film or the bracket is placed for 3-10 days to slowly volatilize the solvent, so that the surface of the obtained film or the bracket is flat and smooth; if the environmental temperature is about 20 ℃, the organic solvent can be volatilized quickly, the surface of the prepared film or the bracket is rough, and a hundred-nanometer or even micron-scale pore structure is generated. According to previous experiments, the lower ambient temperature and the flat and smooth surface are more conducive to osteogenic induction under the existing conditions. In order to ensure sufficient volatilization of the organic solvent and reduce the cytotoxicity possibly caused by the solvent residue, the sample which is close to the forming can be placed in a vacuum drying oven and dried overnight (about 10-12 hours) by vacuum pumping at room temperature in the last step of solvent volatilization. And taking out the sample to be the finally molded sample.

The invention also provides the bone implant material prepared by the preparation method, and the bone implant material preferably has a structure of a thin film structure, a three-dimensional scaffold structure or a columnar solid structure, but is not limited thereto.

The invention also provides application of the bone implant material as an implant of a bone defect filling part in clinical treatment of bone defects.

The bone implant material provided by the invention has the function of promoting osteogenesis by photo-thermal, and the black phosphorus can be used as a built-in photo-thermal converter in the implant, so that the efficiency of penetrating the near-infrared laser into biological tissues is greatly improved, and the curative effect is enhanced; meanwhile, the proportion of the black phosphorus mixed in the high polymer material is controlled, and the black phosphorus is slowly and gradually released along with the high polymer material, meets water in tissues and is oxidized to produce phosphate ions to reach bone defect parts. The phosphate is an essential substance in the bone healing and regeneration process, can participate in the bone healing of the defect part, and achieves the treatment effect of 'one dose and two effects'.

Considering that the implant needs to exist in the body as a foreign body for a long time and has a risk of rejection, the degradable implant material is a research hotspot of the current orthopedic implant material. The most fundamental starting point for the selection of degradable implant materials is the hope that with the formation of new bone, the implant will gradually degrade and eventually be completely metabolized or absorbed by the body. Therefore, the ideal degradable implant material can provide necessary elements for local bone regeneration while being gradually degraded, and promotes the bone regeneration.

The novel two-dimensional photothermal material black phosphorus is innovatively applied to the preparation of the bone implant material, and the stable photothermal effect of the black phosphorus in a near infrared region is utilized to stimulate the regulation of stem cells of a human to the osteogenesis direction, so that the effect of promoting osteogenesis of the bone implant material is achieved. Compared with other two-dimensional materials capable of generating photothermal effect, such as graphene, gold rods and the like, the black phosphorus is a degradable two-dimensional material, and the degradation product is phosphate ions, so that the generation of bones is promoted to a certain extent. Meanwhile, phosphorus is also one of the essential elements widely existing in the human body. The black phosphorus is used as a heat-generating medium, other substances harmful to human bodies are not introduced into the internal environment, and potential hazards caused by using other media are effectively avoided.

The bone implant material provided by the invention has a photo-thermal osteogenesis promoting function, can be degraded in a physiological environment, can be used as an implant of a bone defect filling part to be applied to clinical treatment of bone defects, can improve the efficiency of penetrating biological tissues by near infrared rays to thermally promote bones, can be combined with clinical physiotherapy to achieve a better curative effect of promoting bone defect healing, and meanwhile, the bone implant material degradation product is a substance necessary for a human body, can participate in bone healing and regeneration processes, and does not leave harmful substances in the human body.

Drawings

FIG. 1 is a graph comparing the weight loss tests of BP @ P L GA film and P L GA degradation experiments in example 2 of the present invention;

FIG. 2 is a graph comparing the "degradation time-mass retention" of BP @ P L GA films and P L GA degradation experiments in example 2 of the present invention;

FIG. 3 is a graph of "degradation time versus phosphate ion concentration" of black phosphorus in BP @ P L GA film in example 2 of the present invention;

FIG. 4 is a schematic diagram showing the expression of osteocalcin in which the BP @ P L GA film promotes osteogenesis under the stimulation of near-infrared laser in example 3 of the invention;

FIG. 5 is a schematic diagram showing the expression of osteocalcin in which the BP @ P L GA film promotes osteogenesis under the stimulation of near-infrared laser in example 3 of the invention;

FIG. 6 is a schematic representation of a pig muscle tissue model in the experiment of BP @ P L GA membrane near-infrared laser penetration into biological tissue in example 4 of the present invention;

FIG. 7 is a graph showing the effect of BP @ P L GA film near-infrared laser penetrating biological tissues in example 4 of the present invention.

Detailed Description

The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention is not limited to the practical scope of the present invention.

The block black phosphorus, P L GA, PC L, P L a, PBS, and PTMC provided in this example are all commercially available.

Example 1

The embodiment provides a preparation method of a bone implant material BP @ P L GA film, which comprises the following steps:

firstly, preparing two-dimensional layered black phosphorus, namely weighing blocky black phosphorus in a mortar, gradually adding N-methyl pyrrolidone, wherein the using amount ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1 mg: 1m L, performing ice-water bath ultrasound after sufficient grinding, performing ultrasound for 6 hours, centrifuging at the speed of 4000rpm for 15min, and taking supernatant to obtain a suspension of the two-dimensional layered black phosphorus;

ultrasonically dispersing the two-dimensional layered black phosphorus in a small amount of absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol can cover the two-dimensional layered black phosphorus precipitate to obtain a dispersion liquid A.

Step two, weighing a polylactic acid-glycolic acid copolymer (P L GA), wherein the mass ratio of polylactic acid to glycolic acid in P L GA is 75: 25, dissolving P L GA in dichloromethane, and the dosage ratio of P L GA to dichloromethane is 50 mg: 1m L to obtain a solution B.

And step three, mutually dissolving the dispersion liquid A and the solution B, uniformly mixing, and then carrying out water bath ultrasonic treatment to obtain a uniformly dispersed BP @ P L GA high polymer solution, wherein the mass ratio of the two-dimensional layered black phosphorus to the P L GA is 0.2: 1.

And step four, injecting the BP @ P L GA high polymer solution obtained in the step three into a mold, wherein the mold is in a film shape, placing the mold in a low-temperature environment at 4 ℃, standing for 3-4 days, and preparing the bone implant material BP @ P L GA film after the solvent is slowly volatilized.

Example 2 degradation experiment of BP @ P L GA film as bone implant material in example 1

The degradation experiment is carried out by adopting the bone implant material BP @ P L GA film prepared in example 1, the BP @ P L GA film is taken as a degradation research object, weight loss measurement is carried out by sampling every week, and photographing observation is carried out once every two weeks, and the results are shown in a figure 1, a figure 2 and a figure 3, wherein the figure 1 is a weight loss test comparison graph of the BP @ P L GA film and the P L GA degradation experiment of the embodiment, the figure 2 is a degradation time-mass retention rate comparison graph of the BP @ P L GA film and the P L GA degradation experiment of the embodiment, and the figure 3 is a degradation time-phosphate ion concentration graph of black phosphorus in the BP @ P L GA film of the embodiment.

As shown in FIG. 1, from week four, it was observed that the film had partially ruptured and foamed, and the film was significantly ruptured by week six and had degraded a large portion by week eight. the specific weight loss was plotted as "degradation time-mass retention" (as shown in FIG. 2), and it was observed that both conventional P L GA and BP @ P L GA in the first four weeks were slightly degraded, but the degradation rate of BP @ P L GA was gradually increased from week five (as shown in FIG. 3).

With the degradation of the P L GA film, after moisture enters the inside of the film, the Black Phosphorus (BP) wrapped inside the film is gradually degraded, and finally phosphate ions are generated, as shown in the following reaction formula:

the "degradation time-phosphate ion concentration" measurements (as shown in fig. 3) were performed on the solution after soaking the BP @ P L GA film, and the results showed that the phosphate ion concentration gradually increased with the increase of the degradation time, and showed the characteristics of "slow degradation, slow release".

The experimental results of the example show that the P L GA and the BP in the BP @ P L GA film prepared in the example can be degraded under the physiological environment, and when the BP @ P L GA film is used for an implant at a bone defect filling part, the implant is gradually metabolized along with the growth of new bone and the healing of the defect part.

Example 3 near infrared laser assisted experiment of BP @ P L GA film promoting new bone formation in example 1

A near-infrared laser-assisted new bone formation promotion experiment was performed by using the bone implant material BP @ P L GA film prepared in example 1.

The optimal osteogenesis promoting condition of the embodiment is that 808nm near infrared laser irradiation is heated to 40.5 +/-0.5 ℃, the temperature is kept for 1 minute, then the temperature is naturally reduced, the treatment period is 4 days/time and lasts for 3 weeks, the expression of stem cell osteogenesis related proteins such as Osteocalcin (OCN) and Osteopontin (OPN) in 7 days, 14 days and 21 days is respectively measured, and the results are shown in figures 4 and 5, figure 4 is a schematic diagram of the BP @ P L GA film promoting osteogenesis osteocalcin expression under the near infrared laser stimulation, figure 5 is a schematic diagram of the BP @ P L GA film promoting osteogenesis under the near infrared laser stimulation, and figures 4 and 5 show that the protein expression of human bone marrow stem cells GA growing on the BP @ P L film is remarkably improved on the 14 days and 21 days through the auxiliary near infrared laser treatment under the conditions, which means that the generation speed of new bone is remarkably increased compared with the ordinary P L GA.

EXAMPLE 4 BP @ P L GA film near-infrared laser penetration experiment in example 1

The bone implant material BP @ P L GA thin film prepared in example 1 is adopted, and pig muscle tissue is taken as a tissue model (shown in figure 6), blocks with the thickness of 1mm to 7mm are respectively cut out, and in-vitro near infrared penetration effect verification is carried out, under normal conditions, a muscle layer with the thickness of 2 mm to 3mm can block laser emitted by a near infrared laser with the thickness of 808nm, and in the invention, the BP @ P L GA thin film blocked by the muscle layer with the thickness of 7mm can be heated by the laser with the thickness of 808nm to raise the temperature by 5 ℃, and the temperature of 3 ℃ to 4 ℃ can reach 40 ℃ to 41 ℃ required by optimal treatment (shown in figure 7) calculated according to the normal human body temperature of 37 ℃.

Example 5

The embodiment provides a preparation method of a bone implant material BP @ PC L columnar solid implant material, which comprises the following steps:

firstly, preparing two-dimensional layered black phosphorus, namely weighing blocky black phosphorus in a mortar, gradually adding N-methyl pyrrolidone, wherein the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1.5 mg: 1m L, carrying out ice-water bath ultrasound after sufficient grinding, carrying out ultrasound for 5h, centrifuging at the speed of 5000rpm for 10min, and taking supernatant to obtain suspension of the two-dimensional layered black phosphorus;

ultrasonically dispersing the two-dimensional layered black phosphorus in deionized water, wherein the addition amount of the deionized water can cover the two-dimensional layered black phosphorus precipitate to obtain a dispersion liquid A.

Step two, weighing polycaprolactone (PC L), dissolving PC L in dimethylformamide, wherein the dosage ratio of PC L to dimethylformamide is 25 mg: 1m L, and obtaining a solution B.

And step three, mutually dissolving the dispersion liquid A and the solution B, uniformly mixing, and then carrying out water bath ultrasonic treatment to obtain a uniformly dispersed BP @ PC L high polymer solution, wherein the mass ratio of the two-dimensional layered black phosphorus to the PC L is 0.1: 1.

And step four, injecting the BP @ PC L high-molecular solution in the step three into a mold, wherein the mold is a cylindrical hollow mold, placing the mold in a low-temperature environment at 4 ℃, standing for 3-4 days, and after the solvent is slowly volatilized, preparing the bone implant material BP @ PC L columnar solid implant material.

Example 6

The embodiment provides a preparation method of a bone implant material BP @ P L A three-dimensional scaffold, which comprises the following steps:

firstly, preparing the ultra-small black phosphorus nanosheet, namely weighing blocky black phosphorus in a mortar, gradually adding N-methyl pyrrolidone, wherein the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1.5 mg: 1m L, fully grinding, placing the blocky black phosphorus on ice, performing ultrasound treatment for 5h by using a probe, centrifuging at the speed of 10000rpm for 15min, taking supernatant fluid, and obtaining suspension of the ultra-small black phosphorus nanosheet;

ultrasonically dispersing the ultra-small black phosphorus nanosheets in absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol can cover the precipitates of the ultra-small black phosphorus nanosheets, so as to obtain a dispersion liquid A.

Step two, weighing polylactic acid (P L A), dissolving P L A in dimethylformamide, wherein the dosage ratio of P L A to dimethylformamide is 50 mg: 1m L, and obtaining solution B.

And step three, mutually dissolving the dispersion liquid A and the solution B, uniformly mixing, and then carrying out water bath ultrasonic treatment to obtain a uniformly dispersed BP @ P L A high polymer solution, wherein the mass ratio of the two-dimensional layered black phosphorus to the P L A is 0.5 to 1.

And step four, injecting the BP @ P L A high polymer solution in the step three into a mold, wherein the mold is in the shape of a three-dimensional porous scaffold, placing the mold in a low-temperature environment at 4 ℃, standing for 3-4 days, and preparing the bone implant material BP @ P L A three-dimensional scaffold after the solvent is slowly volatilized.

Example 7

The embodiment provides a preparation method of a bone implant material BP @ P L GA three-dimensional scaffold, which comprises the following steps:

firstly, preparing the ultra-small black phosphorus nanosheet, namely weighing blocky black phosphorus in a mortar, gradually adding N-methyl pyrrolidone, wherein the using amount ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1.5 mg: 1m L, fully grinding the blocky black phosphorus, placing the blocky black phosphorus on ice, performing ultrasonic treatment for 10 hours by using a probe, centrifuging the blocky black phosphorus and the N-methyl pyrrolidone at 8000rpm for 15 minutes, taking supernatant to obtain suspension of the ultra-small black phosphorus nanosheet, centrifuging the suspension of the ultra-small black phosphorus nanosheet at 12000rpm for 15 minutes, and taking precipitate to obtain the ultra-small black phosphorus nanosheet;

Step two, weighing a polylactic acid-glycolic acid copolymer (P L GA), wherein the mass ratio of polylactic acid to glycolic acid in P L GA is 85: 15, dissolving P L GA in tetrahydrofuran, and the dosage ratio of P L GA to tetrahydrofuran is 40 mg: 1m L, so as to obtain a solution B.

And step three, mutually dissolving the dispersion liquid A and the solution B, uniformly mixing, and then carrying out water bath ultrasonic treatment to obtain a uniformly dispersed BP @ P L GA high polymer solution, wherein the mass ratio of the ultra-small black phosphorus nanosheets to the P L GA is 0.3% to 1.

And step four, injecting the BP @ P L GA high polymer solution in the step three into a mold, wherein the mold is in a three-dimensional support shape, placing the mold in a low-temperature environment at 4 ℃, standing for 3-4 days, and preparing the bone implant material BP @ P L GA three-dimensional support after the solvent is slowly volatilized.

Example 8

The embodiment provides a preparation method of a bone implant material BP @ PBS three-dimensional scaffold, which comprises the following steps:

firstly, preparing two-dimensional layered black phosphorus, namely weighing blocky black phosphorus in a mortar, gradually adding N-methyl pyrrolidone, wherein the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 1.0 mg: 1m L, fully grinding, placing the blocky black phosphorus in an ice water bath for 6h by ultrasound, centrifuging at the speed of 6000rpm for 20min, taking supernatant fluid, obtaining suspension of the two-dimensional layered black phosphorus, centrifuging the suspension of the two-dimensional layered black phosphorus at the speed of 12000rpm for 15min, taking sediment, and obtaining the two-dimensional layered black phosphorus;

ultrasonically dispersing two-dimensional layered black phosphorus in dimethyl sulfoxide (DMSO), wherein the addition amount of the DMSO can cover the precipitate of the super-two-dimensional layered black phosphorus, and thus obtaining a dispersion liquid A.

Step two, weighing polybutylene succinate (PBS), dissolving the PBS in tetrahydrofuran, wherein the dosage ratio of the PBS to the tetrahydrofuran is 40 mg: 1m L, and obtaining a solution B.

Dissolving the dispersion liquid A and the solution B mutually, uniformly mixing, and performing water bath ultrasonic treatment to obtain a uniformly dispersed BP @ PBS high polymer solution; wherein the mass ratio of the two-dimensional layered black phosphorus to the PBS is 0.4%: 1.

and step four, injecting the BP @ PBS high molecular solution in the step three into a mold, wherein the mold is in the shape of a three-dimensional porous scaffold, placing the mold in a room temperature environment for 7-10 days, placing the mold in a vacuum drying oven for vacuumizing, and preparing the bone implant material BP @ PBS three-dimensional scaffold after the solvent is slowly volatilized.

Example 9

The embodiment provides a preparation method of a bone implant material BP @ PTMC three-dimensional scaffold, which comprises the following steps:

firstly, preparing two-dimensional layered black phosphorus, namely weighing blocky black phosphorus in a mortar, gradually adding N-methyl pyrrolidone, wherein the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is 0.5 mg: 1m L, fully grinding, placing the blocky black phosphorus in an ice water bath for ultrasonic treatment for 8h, centrifuging at the speed of 7000rpm for 20min, taking supernatant fluid to obtain a suspension of the two-dimensional layered black phosphorus, centrifuging the suspension of the two-dimensional layered black phosphorus at the speed of 12000rpm for 15min, taking precipitate and obtaining the two-dimensional layered black phosphorus;

ultrasonically dispersing the two-dimensional layered black phosphorus in absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol can cover the precipitation of the super-two-dimensional layered black phosphorus, and obtaining a dispersion liquid A.

Step two, weighing polytrimethylene carbonate (PTMC), and dissolving the PTMC in dichloromethane, wherein the dosage ratio of the PTMC to the dichloromethane is 30 mg: 1m L, so as to obtain a solution B.

Dissolving the dispersion liquid A and the solution B mutually, and performing water bath ultrasonic treatment after uniform mixing to obtain uniformly dispersed BP @ PTMC high molecular solution; wherein, the mass ratio of the super two-dimensional layered black phosphorus to the PTMC is 0.3%: 1.

and step four, constructing the BP @ PTMC high polymer solution in the step three by using a stent, wherein the stent construction method is a 3D printing method. After a porous structure support is designed by a high polymer 3D printer, printing is completed under a low temperature condition, the porous structure support is placed in a low temperature environment of 4 ℃ for 3-4 days, and after a solvent slowly volatilizes, the bone implant material BP @ PTMC three-dimensional printing porous support is prepared.

In conclusion, the bone implant material provided by the invention has the function of promoting bone formation by light and heat, can be degraded in a physiological environment, can be used as an implant of a bone defect filling part to be applied to clinical treatment of bone defects, can improve the efficiency of penetrating biological tissues by near infrared to thermally promote bones, can be combined with clinical physical therapy to achieve better curative effect of promoting bone defect healing, and meanwhile, the degradation product of the bone implant material is a substance essential to a human body, can participate in the bone healing and regeneration processes, and does not leave harmful substances in the human body.

Claims

1. A preparation method of a bone implant material comprises the following steps:

dissolving a high polymer material in a second solvent to obtain a solution B;

2. the method of claim 1, wherein: the first solvent comprises one or more of absolute ethyl alcohol, deionized water, dimethyl sulfoxide, halogenated hydrocarbon solvents, dimethylformamide and tetrahydrofuran;

the second solvent comprises a combination of one or more of a halogenated hydrocarbon solvent, dimethylformamide, and tetrahydrofuran.

3. The method of claim 2, wherein: the halogenated hydrocarbon solvent includes dichloromethane and/or trichloromethane.

4. The method of claim 1, wherein: the high molecular material comprises one or more of polylactic acid-glycolic acid copolymer, polycaprolactone, polylactic acid, poly butylene succinate and poly trimethylene carbonate.

5. The method of claim 4, wherein: in the polylactic acid-glycolic acid copolymer, the mass ratio of polylactic acid to glycolic acid is 75: 25. 85: 15 or 90: 10.

6. the method according to any one of claims 1 to 5, wherein the ratio of the amount of the polymeric material to the amount of the second solvent is (25mg to 50 mg): 1m L.

7. The method of claim 1, wherein: the solvent volatilization is carried out at room temperature or low temperature.

8. The method of claim 7, wherein: the low temperature is less than or equal to 4 ℃.

9. The method of claim 1, wherein: the black phosphorus comprises black phosphorus nanosheets and/or black phosphorus quantum dots.

10. The method of claim 9, wherein: the black phosphorus nanosheet is two-dimensional layered black phosphorus with the thickness of less than 100 nm;

the black phosphorus quantum dots are ultra-small black phosphorus nanosheets with the thickness of less than 10 nm.

11. The preparation method according to claim 10, wherein the two-dimensional layered black phosphorus is prepared by:

wherein the dosage ratio of the blocky black phosphorus to the N-methyl pyrrolidone is less than 2 mg: 1m L, the time of ultrasonic treatment is 4-12h, the first centrifugation rotating speed is 4000-14000 rpm, the centrifugation time is 10-20min, the second centrifugation rotating speed is 12000-14000rpm, and the centrifugation time is 10-20 min.

12. The preparation method of claim 11, wherein the dosage ratio of the block black phosphorus to the N-methylpyrrolidone is 1 mg: 1m L, the ultrasonic treatment time is 6h, the first centrifugation speed is 4000rpm, the centrifugation time is 15min, the second centrifugation speed is 12000rpm, and the centrifugation time is 15 min.

13. The preparation method according to claim 10, wherein the preparation method of the ultra-small black phosphorus nanosheet is:

wherein the dosage ratio of the block-shaped black phosphorus to the N-methyl pyrrolidone is less than 2 mg: 1m L, the time of ultrasonic treatment is 4-12h, the first centrifugation rotating speed is 7000-14000 rpm, the centrifugation time is 10-20min, the second centrifugation rotating speed is 12000-14000rpm, and the centrifugation time is 10-20 min.

14. The preparation method of claim 13, wherein the dosage ratio of the black phosphorus block to the N-methylpyrrolidone is 1 mg: 1m L, the ultrasonic treatment time is 10h, the first centrifugation rotation speed is 10000rpm, the centrifugation time is 15min, the second centrifugation rotation speed is 12000rpm, and the centrifugation time is 15 min.

15. The bone implant material prepared by the preparation method according to any one of claims 1 to 14, characterized in that: the structure of the bone implant material is a film structure, a three-dimensional bracket structure or a columnar solid structure.

16. Use of the bone implant material according to claim 15 for the preparation of an implant for the filling of a bone defect.