Bone filling stent material composition for osteomyelitis and preparation method thereof
Technical Field
The application relates to the field of biomedical materials, in particular to a bone filling stent material composition for osteomyelitis and a preparation method thereof.
Background
Osteomyelitis is a serious bone infection disease, brings great pain to patients, and faces a plurality of challenges in treatment. At present, methods for treating osteomyelitis comprise surgical debridement, antibiotic treatment and the like, but the traditional treatment mode has a plurality of limitations, such as antibiotic resistance problem, difficulty in thoroughly clearing infection focus and the like. In particular, since the lesions of chronic osteomyelitis are often surrounded by ischemic tissue, a biofilm layer is formed on the surface of bacteria after colonization, and antibiotics in the blood are difficult to enter the lesions. Therefore, it is generally difficult to thoroughly cure chronic osteomyelitis with antibiotics alone, and combination surgical treatment is often required.
At present, a clinically common surgical debridement treatment strategy is mainly a maxkoy technology comprising two stages, which is also called an induced membrane technology, and is characterized in that after a specific bone defect reconstruction material is implanted, the material stimulates surrounding tissues to generate a membrane structure with biological activity. The induced membrane has good vascularization, contains a large number of mature mesenchymal stem cells and can secrete various growth factors, so that a microenvironment favorable for tissue regeneration is formed locally. Wherein the first stage comprises debridement and placement of a Polymethylmethacrylate (PMMA) spacer containing an antibiotic to effect sterilization and induce vascular membrane formation, and the second stage involves removal of the spacer and bone grafting within the induced membrane. PMMA implantation can stimulate the formation of an induction membrane and provide a favorable environment for secondary bone grafting, but the nondegradable property of PMMA implantation can lead antibiotics to be not released effectively, so that the concentration of local antibiotics is insufficient, infection relapse and the like exist in a long-term indwelling body, and the PMMA implantation has certain toxic and side effects, does not have bone conductivity and needs secondary treatment, so that a plurality of students are promoted to focus research hot spots on degradable biological materials in recent years.
The polymer material becomes a research hot spot for repairing bone defects due to excellent biocompatibility, degradability and mechanical properties. Along with the development of material science and biomedical engineering, the application of the high polymer material in bone defect repair is continuously broken through, and a new solution is provided for clinical treatment. However, bone repair materials are also to be improved in terms of anti-infection and mechanical properties. Therefore, the development of a novel bone repair material with antibacterial and self-reinforcing properties is of great importance for the treatment of osteomyelitis.
Disclosure of Invention
In view of the above, the application provides the bone filling stent material, which is cured in situ by a cross-linking agent, effectively fills the defect part and gradually degrades along with bone healing, and the second component can generate a Fenton-like reaction with H 2O2 in an inflammatory microenvironment to generate hydroxyl free radicals (OH), so that the antibacterial performance is realized, the cross-linking of residual monomers in the first component is initiated, the modulus and the bonding strength of the bone filling material are enhanced, and the bone healing can be promoted.
The application provides a bone filling support material composition for osteomyelitis, which comprises a first component and a second component, wherein the first component comprises 1 part of cyclic monomer and 1 to 10 parts of double bond-containing monomer in parts by mass, and the second component comprises 0.0011 to 12 parts of metal ion compound in parts by mass. The bone filling stent material composition for osteomyelitis provided by the application responds to Reactive Oxygen Species (ROS) in an environment in inflammation, releases hydroxyl free radicals capable of strongly sterilizing, enhances the crosslinking of olefin monomers, reduces the residual quantity of the monomers, reduces toxicity, enhances the modulus matching property of a material and a hard bone part, and has application potential as a bone defect filling repair material for osteomyelitis. The bone filling stent material composition for osteomyelitis has the advantages of simple preparation method, convenient use along with the preparation, convenient use process, ideal implementation operation time window of 1-3 min and strong clinical operability.
The bone filling stent material composition for osteomyelitis comprises a first component and a second component, wherein the first component comprises a cyclic monomer. In some embodiments, the cyclic monomers include, but are not limited to, norbornene, cyclic ketoacetals or lipoic acid compounds, and the present application does not require any particular choice of cyclic monomer, preferably norbornene. In some specific embodiments, the lipoic acid compounds include one or more of lipoic acid, amidated lipoic acid compounds, aminated lipoic acid, or 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, cyanated lipoic acid, or N-hydroxysuccinimide amidated lipoic acid compounds. The mass part of the cyclic monomer is 1 to 5 parts, and can be 1 part, 2 parts, 3 parts, 4 parts and 5 parts. The cyclic monomer includes amidated lipoic acid compounds such as aminoethanol and ethylenediamine, lipoic acid modified with amination such as ethylenediamine, or lipoic acid compounds having other groups introduced through amidation reaction of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The cyclic enones include, but are not limited to, 2-methylene-1, 3-dioxepan and/or 5, 6-benzo-2-methylene-1, 3-dioxepan, and the application is not particularly limited to the selection of cyclic enones.
The first component comprises a monomer containing double bonds. In some specific embodiments, the double bond containing monomers include, but are not limited to, one or more of acrylic acid, methacrylic acid, acrylamide, methacrylamide, hydroxyethyl acrylate, 2-hydroxy methacrylate, methacrylated chitosan, hydroxyethyl methacrylate, or acrylated gelatin, and the selection of the double bond containing monomers is not particularly critical to the present application. The double bond-containing monomer is 1 to 10 parts by mass, and can be 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts and 10 parts. The molar ratio of the cyclic monomer to the double bond-containing monomer is 1 (0.1-10), preferably 1:0.5.
The first component of the present application includes an initiator. In some specific embodiments, the initiator includes, but is not limited to, dibenzoyl peroxide and/or N, N-dimethyl-p-toluidine, and the application is not limited to the choice of initiator. The mass part of the initiator is 0.001 to 0.1 part, and can be 0.001 part, 0.005 part, 0.01 part, 0.02 part, 0.03 part, 0.04 part, 0.05 part, 0.08 part, 0.09 part, 0.095 part and 0.1 part.
The first component of the present application includes a cross-linking agent. In some specific embodiments, the cross-linking agent includes, but is not limited to, one or more of ethylene glycol dimethacrylate, maleic anhydride or trimethylolpropane triacrylate, and the present application does not impose particular requirements on the selection of the cross-linking agent. The cross-linking agent is 0.001 to 0.1 part by mass, and can be 0.001 part, 0.005 part, 0.01 part, 0.02 part, 0.03 part, 0.04 part, 0.05 part, 0.08 part, 0.09 part, 0.095 part and 0.1 part.
The second component of the present application comprises a metal ion compound. In some specific embodiments, the metal ion compound includes, but is not limited to, one or more of cobalt acetylacetonate, copper acetylacetonate, cobalt sulfide, copper sulfide, calcium sulfide, iron sulfide, cobalt chloride, copper chloride, calcium chloride, or iron chloride, and the application is not particularly limited to the selection of the metal ion compound. The mass fraction of the metal ion compound is 0.0011 to 12 parts, and can be 0.0011 part, 0.01 part, 0.02 part, 0.05 part, 0.1 part, 0.5 part, 1 part, 2 parts, 5 parts, 8 parts, 10 parts, 11 parts and 12 parts. In some embodiments, the mass ratio of the first component to the second component is 1 (0.001-1), preferably 1:0.01. In some specific embodiments, the osteomyelitis bone filler scaffold material composition has a volume fraction of metal ion compounds of 3% to 8%, preferably 5%.
The second component of the present application comprises an organic compound. In some specific embodiments, the organic compound includes, but is not limited to, one or more of phenol, glucose, polyvinyl alcohol, chitosan, cellulose, tannic acid, citric acid, polyacrylic acid, ethylenediamine tetraacetic acid, vitamin C, oxalic acid, thioglycolic acid, ethylenediamine, or sodium alginate, and the present application does not require any particular choice of organic compound. The mass fraction of the organic compound is 0.01 to 10 parts, and can be 0.01 part, 0.05 part, 0.1 part, 0.5 part, 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts and 10 parts. The second component comprises a functional component which can be coordinated and complexed with metal ions to form nano groups and contains mono/polyhydroxy groups and carboxyl groups, preferably phenol and other components which can be coordinated and synthesized with metal ions to form metal organic frameworks, glucose, polyvinyl alcohol, chitosan, cellulose, tannic acid and other polyhydroxy compounds and other components which can be coordinated and complexed with metal ions to form nano structures, and citric acid, polyacrylic acid, ethylenediamine tetraacetic acid, vitamin C, sodium alginate and other components which can be coordinated and complexed with metal ions to form nano structures. The second component also comprises 0.1 to 20 parts of solvent by mass, wherein the solvent comprises but is not limited to dimethyl sulfoxide and/or ethanol, and the selection of the solvent is not particularly required. The solvent is 0.1 to 20 parts by mass, and can be 0.1 part, 0.5 part, 1 part, 2 parts, 3 parts, 5 parts, 8 parts, 10 parts, 12 parts, 15 parts, 18 parts and 20 parts.
The bone filling stent material composition for osteomyelitis provided by the application utilizes hydrogen peroxide in the microenvironment of osteomyelitis to generate hydroxyl free radicals through Fenton reaction, so that effective killing of infectious bacteria is realized, and the risk of infection recurrence is reduced. The self-reinforcement is realized in the polymerization process, the mechanical strength of the material is improved, the mechanical requirement in the bone repair process is better met, and the stable support is provided for the fracture part. The addition of low-concentration metal ions or bioactive substances promotes the repair and regeneration of bone tissues, is beneficial to improving the treatment success rate and reducing the occurrence of complications such as bone nonunion and the like. The material is gradually degraded in the body, so that adverse reactions caused by long-term retention in the body are avoided, and degradation products are nontoxic and harmless and can be metabolized and discharged by the human body.
The application also provides a preparation method of the bone filling stent material composition for osteomyelitis, which comprises the following steps:
mixing a cyclic monomer with a monomer containing double bonds to obtain a first component, wherein a metal ion compound is used as a second component;
the first component is mixed with the second component to obtain the bone filling stent material composition for osteomyelitis.
In some specific embodiments, the first component is mixed with the second component for a period of time ranging from 1s to 100s, preferably from 3s to 60s, and more preferably from 5s to 20s. In some specific implementations, the mixing of the first component with the second component includes ultrasound for a period of time ranging from 5 to 60 seconds.
The bone filling stent material composition for osteomyelitis performs polymerization reaction with hydrogen peroxide in an osteomyelitis site environment to form a bone filling material;
In some specific embodiments, the polymerization is carried out at a temperature of 20 ℃ to 50 ℃, preferably 30 ℃ to 40 ℃, in the presence of a crosslinking agent and an initiator, for a time of 1s to 100s, preferably 3s to 60s, more preferably 5s to 20s. In some specific implementations, the hydrogen peroxide may be provided directly from the osteomyelitis environment or generated by the addition of calcium peroxide, glucose oxidase or peroxidase to react with the osteomyelitis environment.
When the coating is coated on broken bone fracture surfaces or filled in bone defect positions, the first component and the second component are mixed and then are rapidly coated or filled, free radical ring-opening polymerization reaction rapidly occurs, and rapid bonding closure of broken bones or perfect filling and solidification of bone defect positions are realized. The curing time of the osteomyelitis bone filling scaffold material composition is 1s to 300 s, preferably 30s to 120 s.
In the high ROS environment of excessive inflammation, the metal ions can react with H 2O2 to generate hydroxyl free radicals with higher oxidation potential energy, and due to the energy generated by the Fenton reaction and the action of the free radicals, the polymer chains are crosslinked and entangled, so that the mechanical property of the bone filling stent material composition for osteomyelitis is improved, self-reinforcement is realized, the monomer residual quantity in the material is effectively reduced, and the biocompatibility of the material is improved.
Drawings
Fig. 1 is a schematic view showing the effect of the bone filler scaffold material composition for osteomyelitis according to example 1 of the present application.
Detailed Description
It should be understood that one or more of the expressions ". The expressions" individually include each of the objects recited after the expressions and various combinations of two or more of the recited objects unless otherwise understood from the context and usage. The expression "and/or" in combination with three or more recited objects should be understood as having the same meaning unless otherwise understood from the context.
The use of the terms "comprising," "having," or "containing," including grammatical equivalents thereof, should generally be construed as open-ended and non-limiting, e.g., not to exclude other unrecited elements or steps, unless specifically stated otherwise or otherwise understood from the context.
It should be understood that the order of steps or order of performing certain actions is not important so long as the application remains operable. Furthermore, two or more steps or actions may be performed simultaneously.
The use of any and all examples, or exemplary language such as "e.g." comprising "or" including "in this document is intended merely to better illuminate the application and does not pose a limitation on the scope of the application. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the application.
Furthermore, the numerical ranges and parameters setting forth the present application are approximations that may vary as precisely as possible in the exemplary embodiments. However, any numerical value inherently contains certain standard deviations found in their respective testing measurements. Accordingly, unless explicitly stated otherwise, it is to be understood that all ranges, amounts, values and percentages used in this disclosure are modified by "about". As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a particular value or range.
The application provides a bone filling support material composition for osteomyelitis, which comprises a first component and a second component, wherein the first component comprises 1 to 5 parts of cyclic monomers and 0.1 to 10 parts of double bond-containing monomers in parts by mass, and the second component comprises 0.0011 to 12 parts of metal ion compounds in parts by mass.
The effective schematic diagram of the bone filling stent material composition for osteomyelitis is shown in figure 1, and in the high ROS environment of excessive inflammation, metal ions can react with H 2O2 to generate hydroxyl free radicals with higher oxidation potential energy, and due to the energy generated by Fenton reaction and the action of the free radicals, polymer chains are crosslinked and entangled, so that the mechanical property of the material is improved, self-reinforcement is realized, the monomer residual quantity in the bone filling stent material composition for osteomyelitis is effectively reduced, and the biocompatibility of the material is improved.
The application is further illustrated by the following examples. The scope of the application is not limited by the following examples.
Example 1
This example provides a bone filler scaffold material composition for osteomyelitis, comprising a first component comprising 1 part cyclic ketoacetal monomer (2-methylene-1, 3-dioxepane, MDO), 1 part double bond-containing monomer (hydroxyethyl methacrylate HEMA), 0.02 part initiator (0.01 part dibenzoyl peroxide and 0.01 part N, N-dimethyl-p-toluidine) and 0.02 part cross-linker (ethylene glycol dimethacrylate), and a second component comprising 0.2 part metal ion compound (cobalt acetylacetonate).
The preparation method of the bone filling stent material composition for osteomyelitis comprises the following steps:
Mixing a cyclic monomer, a monomer containing double bonds, a cross-linking agent and an initiator to obtain a first component;
Cobalt acetylacetonate is added into the first component, and is blown or mixed evenly by ultrasound, so as to prepare the bone filling bracket material composition containing metal ions, wherein the volume fraction of the bone filling bracket material composition is 5 percent.
Example 2
The present example provides a bone filler scaffold material composition for osteomyelitis, comprising a first component comprising 1 part cyclic ketoacetal Monomer (MDO), 1 part double bond containing monomer (HEMA), 0.02 part initiator (0.01 part dibenzoyl peroxide and 0.01 part N, N-dimethyl-p-toluidine) and 0.02 part cross-linker (ethylene glycol dimethacrylate), and a second component comprising 0.2 part metal ion compound (CoCl 2).
The preparation method of the bone filling scaffold material composition for osteomyelitis is the same as that of example 1.
Example 3
This example provides a bone filler scaffold material composition for osteomyelitis, comprising a first component comprising 1 part cyclic ketoacetal Monomer (MDO), 1 part double bond-containing monomer (HEMA), 0.02 part initiator (0.01 part dibenzoyl peroxide and 0.01 part N, N-dimethyl-p-toluidine) and 0.02 part cross-linking agent (ethylene glycol dimethacrylate), and a second component comprising 0.2 part metal ion compound (CoCl 2) and 0.1 part carboxyl-containing compound (alginic acid, carboxymethyl chitosan).
The preparation method of the bone filling scaffold material composition for osteomyelitis is the same as that of example 1.
Example 4
This example provides a bone filler scaffold material composition for osteomyelitis comprising a first component comprising 1 part of N-hydroxysuccinimide amidated lipoic acid, 1 part of double bond containing monomers (HEMA, methacrylated chitosan), 0.01 part of initiator (0.005 part of dibenzoyl peroxide and 0.005 part of N, N-dimethyl-p-toluidine) and 0.01 part of cross-linker (ethylene glycol dimethacrylate), and a second component comprising 0.2 part of metal ion compound (cobalt acetylacetonate, copper acetylacetonate).
The preparation method of the bone filling scaffold material composition for osteomyelitis is the same as that of example 1.
Example 5
This example provides a bone filler scaffold material composition for osteomyelitis comprising a first component comprising 1 part of N-hydroxysuccinimide amidated lipoic acid, 1 part of double bond containing monomers (HEMA, methacrylated chitosan), 0.01 part of initiator (0.005 part of dibenzoyl peroxide and 0.005 part of N, N-dimethyl-p-toluidine) and 0.01 part of cross-linker (ethylene glycol dimethacrylate), and a second component comprising 0.2 part of metal ion compound (CoCl 2、FeCl3).
The preparation method of the bone filling scaffold material composition for osteomyelitis is the same as that of example 1.
Example 6
This example provides a bone filler scaffold material composition for osteomyelitis, which comprises a first component and a second component, wherein the first component comprises 1 part of N-hydroxysuccinimide amidated lipoic acid, 1 part of double bond-containing monomer (HEMA, methacryloylated chitosan), 0.01 part of initiator (0.005 part of dibenzoyl peroxide and 0.005 part of N, N-dimethyl-p-toluidine) and 0.01 part of cross-linking agent (ethylene glycol dimethacrylate), and the second component comprises 0.2 part of metal ion compound (CoCl 2).
The preparation method of the bone filling scaffold material composition for osteomyelitis is the same as that of example 1.
Comparative example 1
This example provides a bone filler scaffold composition for osteomyelitis, which differs from example 1 in that no metal ionizer is added.
Comparative example 2
This example provides a bone filler scaffold composition for osteomyelitis, which differs from example 1 in that the metal ion compound is replaced with the commercial antibiotic vancomycin.
The bone filling scaffold material compositions for osteomyelitis provided in examples 1 to 6 and comparative examples 1 to 2 were subjected to performance test as follows:
The antibacterial performance test comprises the steps of respectively culturing staphylococcus aureus and escherichia coli to a logarithmic phase, washing bacteria by using a sterile PBS solution, then adding a bone filling bracket material composition sample for osteomyelitis into a bacterial suspension with the concentration of 2 mL adjusted, adding different amounts of H 2O2 solution, diluting the bone filling bracket material composition for osteomyelitis and common pathogenic bacteria for osteomyelitis such as staphylococcus aureus, escherichia coli and the like by adopting a plate colony counting method, coating the mixture on an LB solid culture medium, culturing the mixture in a 37 ℃ incubator for 16H, observing and counting the number of colonies, evaluating the antibacterial performance of the material, and the test result is shown in a table 1;
The self-reinforcing performance test comprises the steps of using a universal tester to test the mechanical property and the bonding strength of the bone filling support material composition for osteomyelitis to bone tissue, uniformly coating 100uL of bone filling support material composition precursor solution for osteomyelitis on the surface of a bone block (25 mm multiplied by 25 mm), curing at room temperature for 3min, soaking in 10mM H 2O2 solution for 30min, and finally carrying out a combination tensile test by using the universal tester, or measuring the body performance of a preformed bone filling material by using a compression test, and the like, wherein the test results are shown in Table 2.
The results in Table 1 show that, compared with the control group without metal ions and antibiotics, the bone filling stent material composition for osteomyelitis provided by the examples 1-6 has obviously reduced colony numbers around the bone filling stent material composition, has a sterilization rate of more than 83% for Escherichia coli and more than 85% for Staphylococcus aureus, and has good antibacterial effect. The results in Table 2 show that the mechanical properties of the bone filling stent material composition for osteomyelitis in examples 1-6 containing metal ions are better than those of comparative examples 1-2, and the existence of H 2O2 gradually enhances the mechanical properties of the bone filling stent material composition for osteomyelitis in the application, so that the mechanical self-enhancement under the environment of high inflammation of osteomyelitis is realized, and the mechanical requirements of bone on materials at filling positions are better matched.
The test result shows that the bone filling support material composition for osteomyelitis has lap shear tensile strength of 0.3-10 MPa to bone tissue, lamination adhesive strength of 0.5-20 MPa to bone tissue, good bone tissue adhesive property, compression modulus of 10-100 MPa, good matching with the adhered bone tissue modulus, H 2O2 in the inflammatory microenvironment in ROS, release of OH with sterilization property, sterilization rate of E.coli being more than or equal to 90%, sterilization rate of staphylococci being more than or equal to 92%, excellent antibacterial property, release of OH, further crosslinking reaction of residual double bond-containing monomers, crosslinking agent and cyclic monomers in a material system, so that residual monomer amount in the system is reduced, residual monomer amount is less than or equal to 5%, cytotoxicity is less than or equal to 15%, bacterial residual amount is 10% after the implantation of a mouse osteomyelitis 7 days, bacterial residual amount is less than or equal to 5% after 14 days, and degradable property is good matching with bone tissue growth performance that the material is good after implantation of 30 days. The degradation rate can also be controlled by adjusting the type and proportion of the ring-opening monomer, the concentration of metal ions and the like, and is matched with the repair process of the bone tissue, so that the bone tissue gradually degrades and disappears after the regeneration of the bone tissue is finished, and the trouble of taking out in a secondary operation is avoided.
The foregoing is only a preferred embodiment of the present application, but the scope of the present application is not limited thereto, and any person skilled in the art, who is within the scope of the present application, shall cover the scope of the present application by equivalent substitution or modification according to the technical scheme of the present application and the application concept thereof.