Selenium-containing polymer compound, preparation method therefor, metal material modified therewith, and use thereof

Abstract

The present invention belongs to the technical field of biomedical engineering, and relates to a selenium-containing polymer compound, a preparation method therefor, a metal material modified therewith, and the use thereof. The selenium-containing polymer compound is prepared from raw materials containing R1-R2-Se-R3-R4, a compound of formula (I), a diisocyanate and mPEG-R5 by means of a reaction. The selenium-containing polymer compound of the present invention can be used for modifying a metal material, and the modified metal material can be used as an oral implant or an orthopedic implant, which can reduce the inflammation caused by implantation after being implanted into the body, thereby realizing a synergistic effect of resisting inflammation and promoting osteogenesis.

Description

Selenium-containing polymer compound, preparation method thereof, modified metal material and use thereof Technical Field

[0001] The present invention belongs to the technical field of biomedical engineering, and relates to a selenium-containing polymer compound, a preparation method thereof, a modified metal material and applications thereof. Background Art

[0002] Metal materials, such as titanium, titanium-based materials, and zinc alloys, are currently widely or potentially used in oral implants and orthopedic implants. However, implant osseointegration malabsorption and peri-implantitis remain unresolved clinical issues, with most cases resulting from implant loosening or loss due to inflammation or infection in the implant area. Furthermore, the increasing incidence of chronic systemic diseases such as diabetes and osteoporosis, coupled with persistent inflammation and decreased bone formation, poses a greater challenge to controlling peri-implant inflammation. Furthermore, the prognosis of oral implants is closely linked to smoking, a key risk factor. Long-term inhalation of harmful substances in tobacco (such as nicotine) increases free radical production and reduces antioxidant enzyme activity, leading to chronic low-grade inflammation. Furthermore, smoking not only inhibits osteoblast activity but also promotes osteoclast activity, thereby impairing bone repair and regeneration. Consequently, smokers face a higher risk of implant failure, postoperative infection, and marginal bone loss compared to non-smokers. Therefore, designing and developing surface-modified metal materials with synergistic anti-inflammatory and osteopromoting properties can alleviate inflammation in the implant area in the early stage of implantation, promote bone formation around the implant, reduce the risk of implant loss due to inflammation, and enable them to be used in people with diabetes, osteoporosis, and smokers, thereby broadening clinical indications and having great clinical significance and application prospects.

[0003] Existing surface modification methods for titanium and titanium-based materials focus on the co-regulation of osteogenesis and immunity. For example, the patent application "A titanium surface modification method with osteogenesis and immunomodulatory properties" (application number: CN202211496632.2) improves the osteoimmunoregulatory ability of titanium-based implants by constructing a bismuth trioxide coating on the surface. However, it only preliminarily verified the role of the coating in osteogenesis and osteoimmunity.

[0004] Selenium is an essential trace element for the human body. At low doses, it can act as an antioxidant, supporting cell survival and growth, reducing inflammation, and having certain antibacterial effects. Currently, research on selenium-containing polymers focuses on targeted tumor therapy. For example, the patent application "A targeted reduction-sensitive polymer micelle, preparation method, and application" (Application Number: CN202211516175.9) uses high-dose levels of selenium to kill tumor cells. However, the optimal concentration of low-dose selenium in inflammatory environments remains undetermined, and its effect on osteogenesis is also unclear. Summary of the Invention

[0005] The primary purpose of the present invention is to provide a selenium-containing polymer compound so that the metal material modified by the selenium can be used as an oral implant or orthopedic implant, which can reduce the inflammation caused by the implant after implantation into the body, thereby achieving a synergistic effect of anti-inflammatory and osteogenesis.

[0006] To achieve this object, in a basic embodiment, the present invention provides a selenium-containing polymer compound, which is prepared by reacting raw materials comprising R1-R2-Se-R3-R4, a compound of formula (I), a diisocyanate, and mPEG-R5.

[0007] in:

[0008] R1 and R4 are independently selected from hydroxyl or amino;

[0009] R2 and R3 are independently selected from C 2-20 alkyl;

[0010] R5 is selected from hydroxyl or amino;

[0011] R6 and R7 are independently selected from C 1-5 alkyl.

[0012] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein the average molecular weight of the selenium-containing polymer compound is 10,000-40,000.

[0013] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein R1 and R4 are the same, and / or R2 and R3 are the same, and / or R6 and R7 are the same.

[0014] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein R2 and R3 are independently selected from C 3-12 alkyl.

[0015] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein the diisocyanate is selected from 2,4-toluene diisocyanate, hexamethylene diisocyanate, diphenylmethane diisocyanate or isophorone diisocyanate.

[0016] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein the selenium-containing polymer compound is prepared by reacting raw materials comprising R1-R2-Se-R3-R4, 1,4-di(2-hydroxyethyl)piperazine, diisocyanate, and polyethylene glycol monomethyl ether.

[0017] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein the R1-R2-Se-R3-R4 is H2N-(CH2)3-Se-(CH2)3-NH2 or HO-(CH2) 11 -Se-(CH2) 11 -OH.

[0018] In a preferred embodiment, the present invention provides a selenium-containing polymer compound, wherein the structure of the selenium-containing polymer compound is shown in the following formula (II) or formula (III):

[0019] wherein m and n are independently selected from natural numbers of 1-250, and x and y are independently selected from natural numbers of 1-100.

[0020] The second object of the present invention is to provide a method for preparing the selenium-containing polymer compound as described above, so as to better prepare the selenium-containing polymer compound as described above.

[0021] To achieve this object, in a basic embodiment, the present invention provides a method for preparing a selenium-containing polymer compound as described above, the preparation method comprising: mixing R1-R2-Se-R3-R4, a compound of formula (I), a diisocyanate, mPEG-R5, and a catalyst in an organic solvent for reaction, and recrystallizing and washing the reaction product.

[0022] In a preferred embodiment, the present invention provides a method for preparing the selenium-containing polymer compound as described above, wherein:

[0023] The organic solvent is selected from one or more of tetrahydrofuran, N,N-dimethylformamide, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate and n-butyl acetate;

[0024] and / or the molar ratio of R1-R2-Se-R3-R4, the compound of formula (I), the diisocyanate, and mPEG-R5 is x:1:(1.1x+1.1):(0.2x+0.2), wherein x is 0.1 to 10;

[0025] And / or the catalyst is dibutyltin dilaurate.

[0026] In a preferred embodiment, the present invention provides a method for preparing the selenium-containing polymer compound as described above, wherein the reaction temperature is 30-100° C. and the reaction time is 1-48 hours.

[0027] In a preferred embodiment, the present invention provides a method for preparing the selenium-containing polymer compound as described above, wherein the recrystallization is performed using ether, and the washing is performed using ethanol having a volume percentage concentration of 5-50%.

[0028] The third object of the present invention is to provide a metal material modified with a selenium-containing polymer compound as described above, which can be used as an oral implant or an orthopedic implant to reduce inflammation caused by the implant after implantation into the body, thereby achieving a synergistic effect of anti-inflammatory and osteogenesis.

[0029] To achieve this object, in a basic embodiment, the present invention provides a metal material modified with a selenium-containing polymer compound as described above, wherein the metal material modified with a selenium-containing polymer compound is obtained by complexing the selenium-containing polymer compound described above with a metal material.

[0030] In a preferred embodiment, the present invention provides a metal material modified with a selenium-containing polymer compound as described above, wherein the metal material is selected from one or more of pure titanium, titanium alloy, zinc alloy, magnesium alloy, and cobalt-chromium alloy.

[0031] In a preferred embodiment, the present invention provides a metal material modified with a selenium-containing polymer compound as described above, wherein the zinc alloy is selected from zinc-lithium alloy and / or zinc-lithium-magnesium alloy, preferably Zn0.8Li and / or Zn0.8Li0.4Mg.

[0032] The fourth object of the present invention is to provide a method for preparing the metal material modified with the selenium-containing polymer compound as described above, so as to better prepare the metal material modified with the selenium-containing polymer compound as described above.

[0033] To achieve this object, in a basic embodiment, the present invention provides a method for preparing a metal material modified with a selenium-containing polymer compound as described above, the preparation method comprising: dissolving the selenium-containing polymer compound as described above in an organic solvent, and then immersing the metal material in the organic solvent for a modification reaction.

[0034] In a preferred embodiment, the present invention provides a method for preparing a metal material modified with a selenium-containing polymer compound as described above, wherein:

[0035] The organic solvent is dimethyl sulfoxide and / or N,N-dimethylformamide;

[0036] And / or the concentration of the selenium-containing polymer compound after being dissolved in an organic solvent is 1-10 mg / mL.

[0037] In a preferred embodiment, the present invention provides a method for preparing a metal material modified with a selenium-containing polymer compound as described above, wherein:

[0038] The metal material is a metal sheet or a metal rod;

[0039] And / or the metal material is polished and ultrasonically cleaned before use.

[0040] In a preferred embodiment, the present invention provides a method for preparing a metal material modified with a selenium-containing polymer compound as described above, wherein the modification reaction time is 1-3 hours.

[0041] The fifth object of the present invention is to provide the use of the metal material modified with the selenium-containing polymer compound as described above for the preparation of oral implants or orthopedic implants, so as to reduce the inflammation caused by the implantation after the metal material modified with the selenium-containing polymer compound as described above is implanted into the body, thereby achieving a synergistic effect of anti-inflammatory and osteogenesis.

[0042] To achieve this object, in a basic embodiment, the present invention provides use of the metal material modified with the selenium-containing polymer compound as described above for preparing an oral implant or an orthopedic implant.

[0043] In a preferred embodiment, the present invention provides the use of the metal material modified with the selenium-containing polymer compound as described above for preparing oral implants or orthopedic implants, wherein the oral implants or orthopedic implants are targeted at one or more of diabetic patients, osteoporosis patients, and smokers.

[0044] The beneficial effect of the present invention is that the metal material modified by the selenium-containing polymer compound of the present invention can be used as an oral implant or orthopedic implant, which can reduce the inflammation caused by the implant after implantation into the body, thereby achieving a synergistic effect of anti-inflammatory and osteogenesis.

[0045] The beneficial effects of the present invention are specifically embodied in:

[0046] (1) The present invention introduces selenium into the surface of titanium and titanium-based materials, utilizes selenium-containing polymer compounds to coordinate and compound with metallic titanium, and after implantation into the body, reduces the inflammation caused by implantation through the effect of selenium on surrounding tissues, thereby realizing the multifunctionality of titanium-based implant materials that are synergistic in anti-inflammatory and osteogenic properties and their applicability in diseases. The titanium material modified with the selenium-containing polymer compound of the present invention can promote the osteogenesis of hBMMSCs on its surface, and can regulate the polarization of THP-1-induced macrophages to the anti-inflammatory M2 type, thereby inhibiting their polarization to the pro-inflammatory M1 type; the SD rat femoral defect model shows that the titanium material modified with the selenium-containing polymer compound of the present invention can promote bone regeneration around the implant; in an in vitro high-sugar environment, the titanium material modified with the selenium-containing polymer compound of the present invention is more likely to promote osteogenesis than pure titanium, suggesting that the titanium material modified with the selenium-containing polymer compound of the present invention can be used in a diabetic environment. The titanium material modified with the selenium-containing polymer compound in the present invention has the advantages of easy application and convenient surface treatment; it also has broad application prospects. Because of its synergistic anti-inflammatory and osteogenesis effects, it can be used for early inflammation control of implants, promoting bone formation around implants, and controlling inflammation around implants. It has good application prospects even in the pathological environment of systemic chronic diseases including diabetes.

[0047] (2) The present invention modifies the surface of the degradable zinc alloy with a selenium-containing polymer by chemical bonding, and utilizes the coordination bonding between selenium and zinc and the wear resistance of the polyurea compound to achieve efficient and reliable modification of the degradable zinc alloy. This modification can alleviate the wear of Zn 2+ The invention can solve the problem of biological toxicity of existing zinc alloys in vivo and in vitro, and expand its clinical application range. After the zinc alloy material modified with the selenium-containing polymer compound of the present invention is implanted into the body, the effect of selenium on the surrounding tissues can reduce the inflammation caused by the implantation, and can effectively promote the osteogenesis of mesenchymal stem cells hBMMSCs, induce THP-1-derived macrophages to polarize toward the anti-inflammatory M2 type, and inhibit their polarization toward the pro-inflammatory M1 type, thereby realizing the multifunctionality of the implant material with synergistic anti-inflammatory and osteogenesis and its applicability in diseases, and promoting bone regeneration around the implant.

[0048] (3) The metal material modified with the selenium-containing polymer compound of the present invention, when used as an oral implant or orthopedic implant, can effectively alleviate inflammation in the implant area in the early post-implantation period in smokers, regulate bone metabolism around the implant, and thereby reduce the risk of implant loosening and dislodgement. Therefore, the metal material modified with the selenium-containing polymer compound of the present invention, designed for implants for smokers, not only has significant clinical significance but also has broad prospects in practical applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 shows the characterization results of the selenium-containing small molecule prepared in Example 1, wherein Figure 1a shows the structure of the selenium-containing small molecule; Figure 1b shows the H NMR spectrum; Figure 1c shows the C NMR spectrum; Figure 1d shows the characterization results of the Se NMR spectrum; Figure 1e shows the experimental results of electrospray ionization mass spectrometry; and Figure 1f shows the peak shape of the theoretical simulation of the electrospray ionization mass spectrometry. Predicted molecular weight [M+Na] + :445.26, experimentally determined molecular weight [M+Na] + :445.25.

[0050] FIG2 is the nuclear magnetic resonance hydrogen spectrum detection result of the selenium-containing polymer compound prepared in Example 1, wherein SePU1 is a 25% Se selenium-containing polymer compound, SePU2 is a 50% Se selenium-containing polymer compound, and SePU3 is a 75% Se selenium-containing polymer compound.

[0051] FIG3 is the gel permeation chromatography test results of the selenium-containing polymer compound prepared in Example 1, wherein SePU1 is a 25% Se selenium-containing polymer compound, SePU2 is a 50% Se selenium-containing polymer compound, and SePU3 is a 75% Se selenium-containing polymer compound.

[0052] Figure 4 is a characterization result diagram of the titanium material modified with a selenium-containing polymer compound, wherein Figure 4a is a characteristic peak diagram of selenium ions on the surface of the titanium material modified with a selenium-containing polymer compound; Figure 4b is a surface scan of selenium ions on the surface of pure titanium (Ti) and a titanium material modified with a selenium-containing polymer compound (SePU-Ti).

[0053] Figure 5 is a graph showing the contact angle test results of the titanium material surface modified with selenium-containing polymer compounds, wherein Figure 5a is a graph showing the contact angle test results of the titanium material surface modified with pure titanium (Ti) and selenium-containing polymer compounds with different hydrophilic group ratios and different concentrations; Figure 5b is a graph showing the comparison of the contact angle sizes of the titanium material surface modified with pure titanium (Ti) and selenium-containing polymer compounds with different hydrophilic group ratios and different concentrations.

[0054] Figure 6 shows the live / dead cell staining results of titanium materials modified with selenium-containing polymer compounds, wherein Figure 6a shows the live / dead cell staining results of pure titanium (Ti) and titanium materials modified with selenium-containing polymer compounds with different ratios of hydrophilic groups and different concentrations on human bone marrow mesenchymal stem cells (hBMMSCs); Figure 6b shows the live / dead cell staining results of pure titanium (Ti) and titanium materials modified with selenium-containing polymer compounds with different ratios of hydrophilic groups and different concentrations on human monocytic leukemia cells (THP-1).

[0055] Figure 7 shows the results of the in vitro mesenchymal stem cell osteogenic effect test of titanium materials modified with selenium-containing polymer compounds, wherein Figure 7a shows the alkaline phosphatase (ALP) staining results of pure titanium (Ti) and titanium materials modified with selenium-containing polymer compounds of different ratios and concentrations on human bone marrow mesenchymal stem cells (hBMMSCs); Figure 7b shows the quantitative results of alkaline phosphatase (ALP); Figure 7c shows the expression detection results of osteogenesis-related genes ALP, OCN, and RUNX2 detected by qPCR; Figure 7d shows the expression detection results of osteogenesis-related protein RUNX2 detected by Western Blot; and Figure 7e shows the immunofluorescence image detection results of human bone marrow mesenchymal stem cells (hBMMSCs).

[0056] Figure 8 shows the results of in vitro detection of the effect of titanium materials modified with selenium-containing polymer compounds on macrophage polarization, wherein Figure 8a shows the results of qPCR detection of the effects of pure titanium (Ti) and titanium materials modified with selenium-containing polymer compounds of different ratios and concentrations on the expression of pro-inflammatory phenotype polarization genes TNFα and iNOS in macrophages induced by human monocytic leukemia cells (THP-1); Figure 8b shows the results of qPCR detection of the expression of anti-inflammatory phenotype polarization genes ARG1 and TGFβ in macrophages; Figure 8c shows the results of Western Blot detection of the expression of proteins iNOS related to macrophage polarization toward pro-inflammatory phenotype and ARG-1 related to macrophage polarization toward anti-inflammatory phenotype; Figure 8d shows the results of fluorescence staining of reactive oxygen species (ROS) expression in macrophages; and Figure 8e shows the quantitative results of fluorescence staining of reactive oxygen species (ROS) expression in macrophages.

[0057] FIG9 is a graph showing the test results of the effect of titanium materials modified with selenium-containing polymer compounds on the implantation and repair of femoral defects in SD rats in vivo.

[0058] Figure 10 shows the results of the in vitro osteogenic effect test of titanium materials modified with selenium-containing polymer compounds on mesenchymal stem cells under a high-glucose environment, wherein Figure 10a shows the ARS staining results of pure titanium (Ti) and 25% Se 3 mg / mL selenium-containing polymer modified titanium materials on human bone marrow mesenchymal stem cells (hBMMSCs); Figure 10b shows the expression detection results of osteogenesis-related genes ALP, OCN, and RUNX2 detected by qPCR; and Figure 10c shows the expression detection results of osteogenesis-related protein RUNX2 detected by Western Blot.

[0059] Figure 11 is a comparison of the in vitro mesenchymal stem cell-promoting effects of titanium materials modified with selenium-containing polymer compounds and titanium materials modified with sodium selenite, wherein Figure 11a is a graph showing the ALP staining results of human bone marrow mesenchymal stem cells (hBMMSCs) by titanium materials modified with 1 μM sodium selenite and titanium materials modified with 25% Se 3 mg / mL selenium-containing polymer compounds; Figure 11b is a graph showing the expression detection results of osteogenesis-related genes ALP, OCN, and RUNX2 detected by qPCR.

[0060] Figure 12 is a characterization result diagram of the selenium-containing small molecule prepared in Example 9, wherein a is the structural formula of the selenium-containing small molecule; b is the molecular weight result of the selenium-containing small molecule determined by electrospray mass spectrometry; and c is the nuclear magnetic resonance hydrogen spectrum, selenium spectrum, and carbon spectrum of the selenium-containing small molecule.

[0061] Figure 13 shows the structural formulas of the selenium-containing polymer compounds prepared in Example 1 and Example 9 and the characterization results of their successful modification on degradable zinc alloys, wherein a is the structural formula of the selenium-containing polyurea polymer compound SePUA prepared in Example 9 and the selenium-containing polyurethane polymer compound SePU prepared in Example 1; b is the nuclear magnetic resonance hydrogen spectrum result of the selenium-containing polyurea polymer compound prepared in Example 9; c is the gel permeation chromatography characterization result diagram of the selenium-containing polyurea polymer compound prepared in Example 9; d is the contact angle diagram of the degradable zinc alloy surface modified with the selenium-containing polymer compound prepared in Example 9; and e is the selenium ion surface scan diagram of the zinc alloy surface by secondary time-of-flight ion mass spectrometry.

[0062] Figure 14 shows the degradation effect of the degradable zinc alloy modified with the selenium-containing polymer prepared in Example 9 on delaying the degradation of the degradable zinc alloy, where a is the weight loss of the alloy during the degradation process; b is the pH change of the degradation solution; and c is the surface micromorphology of the degradable zinc alloy before and after modification with the selenium-containing polymer under an electron microscope.

[0063] FIG15 is the biocompatibility test results of the selenium-containing polymer modified degradable zinc alloys prepared in Example 1 and Example 9, wherein a is the Zn content in the 24-hour leachate of the selenium-containing polymer modified zinc alloys prepared in Example 1 and Example 9 and the unmodified zinc alloy group. 2+ concentration; b is the cck-8 result of culturing hBMMSCs with degradable zinc alloy extracts of different concentrations; c is the result of cellular selenium uptake when culturing human bone marrow mesenchymal stem cells (hBMMSCs) in vitro with the selenium-containing polymer modified zinc alloy prepared in Example 9; d is the live / dead cell staining result of the degradable zinc alloy extract before and after modification with the selenium-containing polymer compound prepared in Example 9 with different hydrophilic group ratios and different concentrations.

[0064] Figure 16 shows the effect of the selenium-containing polymer modified degradable zinc alloy prepared in Example 1 and Example 9 on promoting osteogenesis of human bone marrow mesenchymal stem cells (hBMMSCs) in vitro, where a and b are the alkaline phosphatase (ALP) staining and quantitative results of hBMMSCs of the unmodified and zinc alloys modified with selenium-containing polymers of Example 1 and Example 9 at different concentrations; c is the expression detection result of osteogenesis-related genes OSX and OCN detected by qPCR; d is the expression detection result of osteogenesis-related protein RUNX2 detected by Western Blot; e is the immunofluorescence image result of hBMMSCs cultured in the extract of the degradable zinc alloy before and after modification with the selenium-containing polymer.

[0065] Figure 17 shows the polarization of THP-1-derived macrophages cultured in vitro with the selenium-containing polymer compound-modified degradable zinc alloy prepared in Example 1 and Example 9, wherein a is the expression detection result of related genes iNOS and ARG-1 detected by qPCR; b is the expression result of inflammation-related proteins iNOS and ARG-1 detected by Western Blot; c is the detection result of intracellular reactive oxygen species ROS level; d is the expression detection result of polarization-related proteins iNOS and ARG-1 detected by immunofluorescence.

[0066] FIG18 shows the in vivo osteogenic effect of the selenium-containing polymer modified degradable zinc alloy prepared in Example 1 and Example 9 after implantation into femoral defects of SD rats, wherein a is the Micro-CT scanning result; b is the hard tissue grinding section staining result.

[0067] Figure 19 is a graph showing the inhibitory effect of CSE exposure on osteogenic differentiation of mesenchymal stem cells in vitro in Example 16, wherein a is the alkaline phosphatase (ALP) staining result; b is the Alizarin Red (ARS) staining result.

[0068] FIG20 is a graph showing the alkaline phosphatase (ALP) staining results of the titanium material modified with the selenium-containing polymer compound prepared in Example 2 under CSE exposure in Example 16 to improve the osteogenic differentiation of mesenchymal stem cells in vitro.

[0069] Figure 21 is a graph showing the detection results of the in vivo osteogenic effect of the titanium material modified with a selenium-containing polymer compound prepared in Example 2 under CSE exposure in Example 16, wherein a is a graph showing the Micro-CT scanning result under CSE exposure; b is a graph showing the ELISA detection result of inflammatory factors IL-1β, IL-6 and TNF-α under CSE exposure; and c is a graph showing the Micro-CT scanning result of the in vivo osteogenic effect of the titanium material modified with a selenium-containing polymer compound. DETAILED DESCRIPTION

[0070] The specific embodiments of the present invention are further described below through examples.

[0071] Example 1: Preparation and Characterization of Selenium-Containing Polymer Compounds (I)

[0072] 1.0 g of selenium powder (0.0114 mol) and 2.0 g of sodium borohydride (0.0528 mol) were added to a 250 mL round-bottom flask. 30 mL of water was slowly added dropwise in an ice-water bath. After the reaction gradually subsided and gas production ceased, the flask was sealed with a rubber stopper fitted with a balloon to obtain a colorless, transparent solution. This solution was then brought to room temperature and used for further processing. 5.7 g of 11-bromo-1-undecanol (0.0227 mol) was dissolved in 40 mL of tetrahydrofuran to obtain a solution. The prepared 11-bromo-1-undecanol solution was added to the colorless, transparent solution using a syringe and syringe with stirring. The reaction was heated at 50°C for 6 h. After the reaction was completed, the reaction apparatus was cooled to room temperature. Dichloromethane and deionized water were added to the remaining mixture, and the organic phase was separated and collected. The aqueous phase was washed with dichloromethane until colorless, and the organic phase solutions were combined. Anhydrous sodium sulfate was added for drying, and the anhydrous sodium sulfate was removed by filtration. Most of the dichloromethane was removed by rotary evaporation to obtain a concentrated solution, which was then added dropwise into vigorously stirred petroleum ether for recrystallization. Filtering afforded a pale yellowish-white solid product. The molecular structure of the resulting selenium-containing small molecule is shown in Figure 1a. The H NMR, C NMR, and Se NMR spectra are shown in Figures 1b, 1c, and 1d, respectively. The electrospray ionization mass spectrometry experimental results and the peak shapes simulated by electrospray ionization mass spectrometry are shown in Figures 1e and 1f, respectively.

[0073] Results of H NMR spectrum of selenium-containing small molecules: 1 H NMR(400MHz,Chloroform-d)δ3.63(dt,J1=J2=6.1Hz,4H,-CH2-OH),2.54(t,J=7.5Hz,4H,-Se-CH2-),1.70-1.60(p, J=6.8Hz,4H,-CH2CH2-OH),1.56(p,J=6.8Hz,4H,-Se-CH2CH2-),1.42-1.22(m,28H,-Se-CH2CH2(CH2)7CH2CH2-OH).

[0074] Results of C-NMR spectrometry of selenium-containing small molecules: 13C NMR(101MHz,Chloroform-d)δ63.22(-CH2-OH),32.93(-CH2CH2-OH),30.80(-CH2CH2CH2-OH),30.11(-CH2CH2CH2CH2-OH),29 .71-29.55(-Se-CH2CH2CH2-(CH2)4-CH2CH2CH2CH2-OH),29.29(-Se-CH2CH2CH2-),25.86(-Se-CH2CH2-),24.13(-Se-CH2-).

[0075] Results of NMR selenium spectrum test of selenium-containing small molecules: 77 Se NMR(76MHz,Chloroform-d)δ154.90.

[0076] 1.0 mmol of 1,4-bis(2-hydroxyethyl)piperazine and 1.0 mmol of the selenium-containing small molecule HO-(CH2) prepared above were added to 8 mL of tetrahydrofuran. 11 -Se-(CH2) 11 -OH, and 383.2 mg of 2,4-diisocyanatotoluene (2,4-TDI, 2.2 mmol) (see Table 1 below for specific proportions and amounts), along with 20 μL of dibutyltin dilaurate, were added. The reaction vessel was sealed with a rubber stopper and nitrogen was purged for 5 minutes to remove air from the system. The reaction was then heated at 50°C overnight. Subsequently, 0.8 g of dried polyethylene glycol monomethyl ether (weight-average molecular weight = 2000, 0.4 mmol) was weighed and dissolved in 5 mL of tetrahydrofuran. This was added to the reaction system via syringe and needle, and the reaction was continued at 50°C for 24 hours. After the reaction, most of the tetrahydrofuran was removed by rotary evaporation to obtain a concentrated solution. The solution was then added dropwise into vigorously stirred icy ether for recrystallization. The product, an egg yolk-white viscous solid, was collected by filtration, washed five times with a 10% ethanol solution, and lyophilized to remove moisture. The structure of the purified polymer is shown in the lower figure of FIG13a, namely, formula (III), and was characterized by H NMR spectroscopy and gel permeation chromatography, with the results shown in FIG2 and FIG3, respectively.

[0077] The results of H NMR spectrum of selenium-containing polymer compounds, taking SePU2 (50% Se) as an example: 1H NMR(400MHz,Chloroform-d)δ4.33(b,-NH2COO-CH2CH2-N),4.11(b,-NH2COO-CH2(CH2)9CH2-Se-),3.64(b,-OCH2CH2-of mPEG),3.40(b,CH3-OCH2CH2-of mPEG),2.86(b,-NH2COO-CH2CH2-N)2.54(b,-CH2-Se-CH2-),2.18(b,-N(CH2CH2)2N-),1.64( b,-NH2COO-CH2CH2(CH2)7CH2CH2-Se-),1.30-1.25(b,-NH2COO-CH2CH2(CH2)7CH2CH2-Se-).

[0078] Table 1

[0079] Example 2: Preparation and characterization of titanium materials modified with selenium-containing polymer compounds

[0080] (1) Preparation of titanium materials modified with selenium-containing polymer compounds

[0081] 50 mg of the selenium-containing polymer compound prepared in Example 1 was weighed and dissolved in 5 mL of dimethyl sulfoxide (DMSO) to obtain a clear and transparent solution to prepare a 10 mg / mL selenium-containing polymer solution. According to the ratio of the hydrophilic group and the selenium group in the structure of the selenium-containing polymer compound, the selenium-containing polymer solution was divided into 75% Se (hydrophilic group: selenium group = 1:3), 50% Se (hydrophilic group: selenium group = 1:1),

[0082] Three selenium-containing polymer solutions (25% Se (hydrophilic group: selenium group = 3:1)) were prepared. Based on experimental requirements, the 75% Se, 50% Se, and 25% Se solutions were diluted with DMSO to 1 mg / mL, 3 mg / mL, and 5 mg / mL, respectively, for subsequent experiments.

[0083] Pure titanium was processed into 0.2 mm × 10 mm × 10 mm, 0.2 mm × 20 mm × 20 mm titanium sheets, and Φ1.5 mm × 5 mm titanium rods. Each sheet was polished to a smooth surface without noticeable lines and then used for preparation. Before modification with the selenium-containing polymer compound, the titanium sheet and rod were gently polished with 5000- and 10000-grit silicon carbide sandpaper to remove the dense oxide layer on the titanium surface. The sheet and rod were then ultrasonically cleaned with anhydrous ethanol, acetone, and deionized water for 10 minutes each. After polishing and cleaning, the remaining moisture on the titanium sheet and rod was wiped dry. The titanium sheet and rod were then immersed in selenium-containing polymer solutions of varying concentrations for 1.5 hours for surface modification. Excess selenium-containing polymer compounds were then eluted with anhydrous ethanol. The titanium sheet and rod were then placed, with the modified surface facing up, in a well plate of appropriate size for preparation.

[0084] (2) Characterization of titanium materials modified with selenium-containing polymer compounds

[0085] The surfaces of pure titanium and titanium modified with selenium-containing polymers were tested using a time-of-flight secondary ion mass spectrometer (TOF-SIMS). The results are shown in Figure 4. Selenium ions (Se - ) and, compared to the unmodified pure titanium surface, surface scan results indicate that the selenium-containing polymer is more abundant on the modified titanium surface. TOF-SIMS results confirm the successful modification of the titanium with the selenium-containing polymer, and that the titanium metal can be chemically bonded by immersing it in a solution of the selenium-containing polymer.

[0086] (3) Detection of static contact angle of titanium material surface modified with selenium-containing polymer compounds

[0087] A static contact angle meter (Dataphysics, Germany) was used to measure the static contact angles of pure titanium and titanium surfaces modified with different hydrophilic group ratios and different concentrations of selenium-containing polymer compounds. Two replicate samples were set up for each group, and the static contact angles of the center and two random corners of the titanium sheet were measured. The prepared titanium sheet was placed on the stage, and the optical lens was adjusted to make the sample image clear and the needle visible. After determining the position of the needle, deionized water was slowly injected outward, and the stage height was adjusted so that the water droplet contacted the sample surface, and then the image was captured immediately. The volume and speed of the droplet added each time were determined to be constant, and the contact angle of water with the titanium surface was measured. GraphPad was used to analyze the differences in contact angles between the groups and evaluate the hydrophilicity of the titanium sheet surface.

[0088] The results, shown in Figure 5, show that compared to the unmodified pure titanium surface, the contact angles of the titanium surfaces modified with the selenium-containing polymer increased to varying degrees, indicating that the successful modification with the selenium-containing polymer reduced their hydrophilicity. Due to the varying ratios of selenium groups to hydrophilic groups in the three selenium-containing polymers, as the hydrophilic group ratio decreased, the hydrophilicity gradually decreased, and the contact angle increased. It can be seen that 25% Se exhibited the highest hydrophilicity, followed by 50% Se, and the lowest by 75% Se.

[0089] Example 3: Cytocompatibility testing of titanium materials modified with selenium-containing polymer compounds

[0090] The cytocompatibility of titanium modified with selenium-containing polymers was evaluated using a live / dead cell staining kit (Keygen Biosciences, China). Human bone marrow mesenchymal stem cells (hBMMSCs) were seeded on pure and modified titanium sheets and cultured in cell proliferation medium (α-MEM, 10% FBS, 1% penicillin-streptomycin). On day 3, cell viability was assessed using live / dead cell staining after adhesion to titanium surfaces modified with different hydrophilic group ratios and concentrations of selenium-containing polymers. Human monocytic leukemia cells (THP-1) were cultured in suspension by centrifugation (600 rpm, 3 minutes), the supernatant discarded, and the pellet resuspended in cell proliferation medium (RPMI 1640, 10% FBS, 1% penicillin-streptomycin) before addition of phorbol methyl ester (PMA) to a PMA concentration of 100 ng / mL. The cell suspension was inoculated on pure titanium and modified titanium sheets. After 24 hours, THP-1 was induced to adhere to the wall and grow under a microscope by PMA. The culture medium was replaced with cell proliferation medium without PMA and continued to be cultured until the third day. Live / dead cell staining was used to detect cell viability after adhesion to the surface of titanium materials modified with different hydrophilic group ratios and different concentrations of selenium-containing polymer compounds.

[0091] A TE2000-U inverted fluorescence microscope (Nikon, Japan) was used with F3.0 digital photography system software (NIS-Elements, Japan) to photograph cells on the surface of the titanium sheet. The fluorescence color of live cells was green and the fluorescence color of dead cells was red. As shown in Figure 6a, the red fluorescence intensity of hBMMSCs on the surface of the titanium material modified with selenium-containing polymer compounds was significantly weaker than that on the pure titanium surface. At the same time, there was no significant difference in the effect of titanium materials modified with different hydrophilic group ratios and different concentrations of selenium-containing polymer compounds on cell viability. As shown in Figure 6b, the red fluorescence intensity of THP-1 on the surface of the titanium material modified with selenium-containing polymer compounds was significantly weaker than that on the pure titanium surface. At the same time, there was no significant difference in the effect of titanium materials modified with different hydrophilic group ratios and different concentrations of selenium-containing polymer compounds on cell viability.

[0092] Example 4: Detection of the osteogenic effect of mesenchymal stem cells on titanium materials modified with selenium-containing polymer compounds in vitro

[0093] The in vitro osteogenic effect of mesenchymal stem cells on titanium materials modified with selenium-containing polymer compounds was evaluated using alkaline phosphatase (ALP) staining and quantification, quantitative real-time PCR (qPCR), Western blot, and immunofluorescence techniques.

[0094] An alkaline phosphatase (ALP) staining kit (Biyuntian, China) and an alkaline phosphatase quantification kit (Biyuntian, China) were used to detect and quantify the ALP activity of hBMMSCs cultured for 7 days on pure titanium and titanium surfaces modified with different hydrophilic group ratios and different concentrations of selenium-containing polymers. As shown in Figure 7a, ALP staining of hBMMSCs cultured on titanium surfaces modified with three different concentrations of 25% Se and 1 mg / mL of 50% Se was significantly deeper than that of pure titanium and titanium surfaces modified with other hydrophilic group ratios and other concentrations of selenium-containing polymers. The highest ALP activity was observed in the 25% Se 3 mg / mL group. As shown in Figure 7b, the ALP quantification results were consistent with the ALP staining results. Therefore, subsequent experiments were performed using five groups: pure titanium (Ti), 25% Se 1 mg / mL, 25% Se 3 mg / mL, 25% Se 5 mg / mL, and 50% Se 1 mg / mL.

[0095] qPCR was used to examine the effects of pure titanium and titanium modified with varying hydrophilic group ratios and concentrations of selenium-containing polymers on the expression of genes related to osteogenesis in hBMMSCs. As shown in Figure 7c, the expression levels of ALP, OCN, and RUNX2 in the titanium modified with varying hydrophilic group ratios and concentrations were all upregulated compared to the pure titanium group, with the most significant increase observed in the 25% Se 3 mg / mL group.

[0096] Western blot and immunofluorescence techniques were used to examine the effects of pure titanium and titanium modified with varying hydrophilic group ratios and concentrations of selenium-containing polymers on the expression of proteins involved in the osteogenic development of hBMMSCs. As shown in Figure 7d, the expression of RUNX2 protein increased in the titanium modified with varying hydrophilic group ratios and concentrations of selenium-containing polymers, with the most significant increase in protein expression in the 25% Se 3mg / mL group. As shown in Figure 7e, the immunofluorescence results were consistent with the Western blot results, indicating that selenium-containing polymers can promote the osteogenic differentiation of hBMMSCs.

[0097] Example 5: In vitro detection of the polarization of macrophages toward anti-inflammatory phenotypes promoted by titanium materials modified with selenium-containing polymer compounds

[0098] Real-time fluorescence quantitative PCR (qPCR), Western Blot and reactive oxygen species (ROS) fluorescence staining kit were used to evaluate the effect of titanium material modified with selenium-containing polymer compounds on the polarization of PMA-induced attached THP-1.

[0099] qPCR was used to examine the expression levels of genes associated with macrophage polarization toward pro-inflammatory and anti-inflammatory phenotypes. As shown in Figures 8a and 8b, the expression levels of pro-inflammatory genes TNFα and iNOS were downregulated in the titanium material group modified with selenium-containing polymers compared to the pure titanium group, while the expression levels of anti-inflammatory genes ARG1 and TGFβ were significantly upregulated. Furthermore, as shown in Figure 8c, Western Blot results showed that the expression of the pro-inflammatory protein iNOS decreased with increasing titanium modification concentration, while the expression of the anti-inflammatory protein ARG-1 was significantly increased in all three 25% Se concentration groups, indicating that selenium-containing polymers can promote macrophage polarization toward an anti-inflammatory phenotype in vitro.

[0100] The ROS level in THP-1 cells was detected, and the results are shown in Figures 8d and 8e. The fluorescence intensity of the titanium material surface modified with selenium-containing polymer compounds was significantly lower than that of the pure titanium group, indicating that selenium-containing polymer compounds can clear ROS generated by oxidative stress in macrophages, thereby inhibiting the secretion of their pro-inflammatory related factors.

[0101] Example 6: In vivo osteogenic effect testing of titanium materials modified with selenium-containing polymer compounds

[0102] Bilateral femoral defect models were created using 6-8 week old male Sprague-Dawley rats. The femoral defects were located approximately 5 mm proximal to the lateral epicondyle on both sides of the femur, with a diameter of 1.5 mm and a depth of 5 mm. Holes were drilled perpendicular to the long axis of the femur, and titanium rods modified with a selenium-containing polymer were implanted in the defects. Based on the in vitro results, in vivo experiments were divided into four groups: Ti, 25% Se 3 mg / mL, 25% Se 5 mg / mL, and 50% Se 1 mg / mL. Micro-CT analysis was performed after 4 and 8 weeks. As shown in Figure 9, micro-CT scans revealed varying degrees of new bone formation around the implanted titanium rods. The titanium rod modified with a selenium-containing polymer exhibited greater new bone formation than the pure titanium group, and the 25% Se 3 mg / mL group exhibited significantly more new bone tissue than the other experimental groups, consistent with the in vitro results. These results indicate that titanium rods modified with a selenium-containing polymer exhibit superior osseointegration and promote new bone formation around the implanted material. The effect of 25% Se 3 mg / mL group was the best, indicating that selenium-containing polymers can more effectively promote bone regeneration at appropriate concentrations.

[0103] Example 7: Detection of the osteogenic effect of mesenchymal stem cells on titanium materials modified with selenium-containing polymers in a high-glucose environment

[0104] Alizarin red (ARS) staining, quantitative real-time PCR (qPCR) and Western blot were used to evaluate the in vitro mesenchymal stem cell-induced osteogenic effect of titanium materials modified with selenium-containing polymer compounds under a high glucose environment.

[0105] hBMMSCs cultured for 14 days on pure titanium and titanium modified with 25% 3 mg / mL selenium-containing polymer were stained for mineralized nodules using 1% Alizarin Red (ARS) solution (Sigma-Aldrich). As shown in Figure 10a, hBMMSCs cultured in a high-glucose environment on titanium modified with 25% Se (3 mg / mL) selenium-containing polymer (HG-Se) exhibited significantly deeper ARS staining than those on pure titanium (HG-Ti).

[0106] qPCR was used to examine the expression of genes related to osteogenesis in hBMMSCs in a high-glucose environment using pure titanium and titanium modified with 25% (3 mg / mL) selenium-containing polymers. As shown in Figure 10b, the expression of ALP, OCN, and RUNX2 in the titanium modified with 25% (3 mg / mL) Selenium-containing polymers was upregulated compared to the pure titanium group.

[0107] Western Blot analysis was used to examine the effects of pure titanium and titanium modified with 25% (3 mg / mL) selenium-containing polymer on the expression of hBMMSCs' osteogenesis-related proteins under high glucose conditions. As shown in Figure 10c, the expression of RUNX2 protein increased in the titanium modified with 25% (3 mg / mL) selenium-containing polymer.

[0108] Example 8: Comparative test of the osteogenic effect of mesenchymal stem cells in vitro on titanium materials modified with selenium-containing polymer compounds and titanium materials modified with sodium selenite

[0109] Alkaline phosphatase (ALP) staining and real-time fluorescence quantitative PCR (qPCR) were used to evaluate the in vitro mesenchymal stem cell-induced osteogenic effect of titanium materials modified with selenium-containing polymer compounds and titanium materials modified with sodium selenite.

[0110] An alkaline phosphatase (ALP) staining kit (Beyotime, China) was used to stain hBMMSCs cultured for 7 days on titanium surfaces modified with either 25% Se (3 mg / mL) or 1 μM sodium selenite. As shown in Figure 11a, ALP staining was significantly deeper in hBMMSCs cultured on the titanium surface modified with 25% Se (3 mg / mL) than on the titanium surface modified with 1 μM sodium selenite.

[0111] qPCR was used to examine the effects of titanium materials modified with 3 mg / mL of a 25% Se-containing polymer and 1 μM sodium selenite on the expression of genes related to osteogenesis in hBMMSCs. As shown in Figure 11b, the expression levels of ALP, OCN, and RUNX2 were all upregulated in the titanium material modified with 3 mg / mL of a 25% Se-containing polymer compared to the titanium material modified with 1 μM sodium selenite.

[0112] Example 9: Preparation and Characterization of Selenium-Containing Polymer Compounds (II)

[0113] 1.0g selenium powder (0.0127mol) and 2.0g sodium borohydride (0.0529mol) are added to a 250mL round-bottom flask, and 30mL of water is slowly added dropwise thereto under ice-water bath conditions. After the degree of question reaction gradually tends to ease and stops generating gas, the flask is sealed with a rubber stopper with a balloon to obtain a colorless, transparent solution, which is then returned to room temperature for standby use. 5.6g 3-bromo-1-propylamine hydrobromide (0.0254mol) is dissolved in 10mL of water to obtain a solution. Under stirring, the prepared 3-bromo-1-propylamine hydrobromide solution is added to the above-mentioned colorless, transparent solution using a syringe and a needle, and the reaction is carried out at room temperature for 24h. After the reaction is completed, dichloromethane and deionized water are added to the remaining mixture, and the organic phase is separated and collected. The aqueous phase is washed with dichloromethane until colorless, and the organic phase solution is combined. Anhydrous sodium sulfate was added for drying, and the anhydrous sodium sulfate was removed by filtration. Most of the dichloromethane was removed by rotary evaporation to obtain a concentrated solution, which was then drained to obtain a pale yellowish-white liquid. The molecular structure of the resulting selenium-containing small molecule H2N-(CH2)3-Se-(CH2)3-NH2 is shown in Figure 12a. The molecular weight of the selenium-containing small molecule determined by electrospray ionization mass spectrometry is shown in Figure 12b. The hydrogen, selenium, and carbon nuclear magnetic resonance spectra of the selenium-containing small molecule are shown in Figure 12c.

[0114] To 10 mL of N,N-dimethylformamide (DMF), 1,4-bis(2-hydroxyethyl)piperazine, the selenium-containing small molecule H2N-(CH2)3-Se-(CH2)3-NH2, and 1.3 mmol of 2,4-toluene diisocyanate (2,4-TDI) were added in the various molar ratios shown in Table 2. 20 μL of dibutyltin dilaurate was also added. The reaction was sealed with a rubber stopper, and nitrogen was purged through the system for 5 minutes to remove air from the apparatus. The reaction was then heated at 90°C for 5 hours. Subsequently, 0.4 g of dried polyethylene glycol monomethyl ether (weight-average molecular weight = 2000, 0.2 mmol) was weighed and dissolved in 5 mL of DMF. This was added to the reaction system via syringe and needle, and the reaction was continued by heating at 90°C for 12 hours. After the reaction, the resulting product was added dropwise to vigorously stirred icy ether for precipitation. The resulting product, an egg-yolk-white viscous solid, was collected by filtration, washed five times with a 10% (v / v) ethanol solution in water, and lyophilized to remove moisture. The purified polymer was characterized by proton nuclear magnetic resonance spectroscopy (results shown in Figure 13b) and gel permeation chromatography (results shown in Figure 13c). The results showed that the desired selenium-containing polymer compound (structure shown in the upper panel of Figure 13a, i.e., Formula (II)) had been successfully synthesized.

[0115] Table 2

[0116] Example 10: Preparation and characterization of zinc alloy materials modified with selenium-containing polymer compounds

[0117] (1) Preparation of zinc alloy materials modified with selenium-containing polymer compounds

[0118] 50 mg of the selenium-containing polymer compound prepared in Example 9 was weighed separately and dissolved in 5 mL of dimethyl sulfoxide (DMSO) to obtain a clear and transparent solution to prepare a 10 mg / mL selenium-containing polymer solution. According to the ratio of the hydrophilic group and the selenium group in the structure of the selenium-containing polymer compound, three selenium-containing polymer solutions were prepared: 75% Se (hydrophilic group: selenium group = 1:3), 50% Se (hydrophilic group: selenium group = 1:1), and 25% Se (hydrophilic group: selenium group = 3:1). According to the experimental requirements, the selenium-containing polymer solutions with different selenium group ratios were diluted with DMSO to three concentrations of 1 mg / mL, 3 mg / mL, and 5 mg / mL for subsequent experiments.

[0119] To prepare a Zn0.8Li alloy (containing 0.8wt% Li and 99.2wt% Zn), 99.99% pure metallic Zn and 99.9% pure metallic Li were first melted at 520°C and then mixed and cast into cylindrical shapes. The zinc alloy was cut using molybdenum wire into thin sheets measuring 20mm x 0.5mm and 7mm x 0.5mm, as well as rods measuring 1.5mm x 5mm. Before use, all zinc alloys were polished to a 10,000-grit finish using silicon carbide sandpaper to obtain a smooth, oxide-free surface. The samples were then ultrasonically cleaned with acetone, anhydrous ethanol, and deionized water for 10 minutes each to remove any surface residue. After removal, the remaining surface moisture was dried. The zinc alloys were then tweezered and immersed in selenium-containing polymer solutions of varying concentrations and selenium group ratios for 2 hours for surface modification. After removal, the samples were rinsed with anhydrous ethanol, air-dried, and placed in appropriately sized well plates for later use. For ease of description, the zinc alloy modified with the selenium-containing polyurea polymer of Example 9 prepared by this method will be abbreviated as SePUA-Zn0.8Li in the following text, and the selenium group ratio and solution concentration of the selenium-containing polymer used for modification will be noted. Those not specifically noted are modified with 25% Se 3 mg / mL selenium-containing polymer.

[0120] Similarly, the selenium-containing polyurethane polymer compound prepared in Example 1 was used to prepare a zinc alloy material modified with a selenium-containing polymer compound. Hereinafter, the prepared selenium-containing polyurethane-modified zinc alloy will be abbreviated as SePU-Zn0.8Li, and the selenium group ratio and solution concentration of the selenium-containing polymer used for modification will be noted. Those not specifically noted are modified with 25% Se 3 mg / mL selenium-containing polymer.

[0121] (2) Detection of surface modification effect of zinc alloy materials modified with selenium-containing polymer compounds

[0122] A static contact angle meter (Dataphysics, Germany) was used to measure the static contact angles of the surface of the degradable zinc alloy material before and after modification with different concentrations of selenium-containing polymers to compare their hydrophilicity. The groups of degradable zinc alloy sheets prepared in advance were placed horizontally on the sample table and adjusted to a suitable position. A deionized water droplet was slowly released from the needle so that the droplet was spherical in contact with the sample surface. The camera parameters were adjusted and the imaging system of the measuring instrument was used to capture the contact angle image of the droplet and the solid surface. The volume and speed of the droplet added each time were determined to be constant, the morphology of the droplet on the sample surface was tested, and the contact angle was measured. To ensure the reliability of the results, 5 test points were randomly selected for each group, and the average value of their contact angles was calculated as the final result. As shown in Figure 13d, it can be seen that the selenium-containing polyurea of ​​Example 9 is more hydrophilic than the selenium-containing polyurethane of Example 1 after modification of the zinc alloy surface, and the hydrophilicity decreases as the proportion of selenium groups in the selenium-containing polymer increases.

[0123] A time-of-flight secondary ion mass spectrometer (ION-TOF GmbH, Germany) was used to detect the surfaces of the degradable zinc alloys (represented as Zn-0.8Li, SePUA-Zn0.8Li and SePU-Zn0.8Li respectively in the figure) before and after modification with selenium-containing polyurea and selenium-containing polyurethane. The results are shown in Figure 13e, indicating that the selenium-containing polymers prepared in Example 1 and Example 9 can be uniformly modified on the metal surface, and the Se element concentration on the metal surface after modification with the selenium-containing polyurea in Example 9 is significantly higher.

[0124] Example 11: Degradability test of zinc alloy material modified with selenium-containing polymer compound

[0125] Zinc alloy sheets with a diameter of 7 mm and a thickness of 0.5 mm were polished in stages using 500-10,000 grit sandpaper. The polished zinc alloys in the experimental group were then modified in a 25% Se solution (3 mg / mL) containing a selenium-containing polymer (Example 9) for 2 hours. The control group consisted of zinc alloy sheets not modified with the selenium-containing polymer. Three samples were collected from each group at each time point. The experimental and control group samples were each immersed in simulated body fluid (SBF, purchased from Beijing Coolbo Technology Co., Ltd., catalog number SL6710) at a solution volume to zinc alloy surface area ratio of 20 mL / cm. 2 The system was placed in a shaker at 37°C. The zinc alloys were removed at 1, 3, 5, 7, 14, 21, 28, 42, and 56 days, and the simulated body fluid was collected and replaced. The zinc alloy sheets were rinsed with 200g / L CrO₃, deionized water, and anhydrous ethanol, respectively, to remove surface degradation products. After drying in an oven at room temperature for 2 hours, the samples were weighed on an electronic balance. The weight changes of the samples are shown in Figure 14a. During the first few days of immersion, the zinc alloy modified with the selenium-containing polymer from Example 9 exhibited significantly slower corrosion compared to the unmodified group. Throughout the degradation process, the weight loss of the modified group was less than that of the unmodified group at each time point, indicating that the selenium-containing polymer modification slowed the degradation rate of the zinc alloy, particularly the early burst of degradation. The pH values ​​of the solutions collected at these time points were measured, and the results are shown in Figure 14b. Selenium-containing polymer modification resulted in a more alkaline extraction solution, which is more conducive to the osteogenic differentiation of hBMMSCs.

[0126] After sampling on the 56th day of degradation, the microstructure and corrosion morphology of the surfaces of the selenium-polymer-modified and unmodified groups were observed using a scanning electron microscope (JSM-7900F, Japan). The results are shown in Figure 14c. It can be seen that the granular degradation products are more evenly distributed on the surface of the zinc alloy modified with the selenium-containing polymer than on the surface of the unmodified zinc alloy.

[0127] Example 12: In vitro biocompatibility testing of zinc alloy materials modified with selenium-containing polymer compounds

[0128] The degradable zinc alloy extract was prepared according to ISO 10993. The specific method was as follows: the zinc alloy metal discs of Φ20 mm × 0.5 mm before and after modification with the selenium-containing polymer of Example 9 were immersed in a solution with a specific surface area of ​​1.25 mL / cm 2 Cells were extracted in a cell proliferation medium (PM: α-MEM + 10% FBS + 1% penicillin-streptomycin) at 37°C for 24 hours on a shaker. The extract was then filtered using a sterile filter in a clean bench. The extract was mixed with PM at different volume ratios to obtain cell culture media containing 10%, 25%, 50%, and 100% extract.

[0129] The Zn content in the zinc alloy extracts before and after modification with selenium-containing polymers prepared in Example 1 and Example 9 was detected by inductively coupled plasma mass spectrometry (Agilent 7700 / 7800, USA). 2+ The results are shown in Figure 15a. It can be seen that after 24 hours, the Zn content in the extract of the selenium-containing polymer modified group 2+ The concentration was significantly lower than that of the unmodified group extract, and the modification of selenium-containing polymers could significantly alleviate the 2+ Burst release, and the SePUA group had a better effect in alleviating burst release. hBMMSCs cells were cultured using cell proliferation medium (PM), cell proliferation medium containing different volume ratios of degradable zinc alloy extract (represented in the figure as 10%, 25%, 50%, and 100% Zn0.8Li), and PM with 10% DMSO. The cells were cultured in a 5% CO2, 37°C constant temperature cell incubator, and the culture medium was changed every 2 to 3 days. Cell viability was assessed using a CCK-8 kit (Biyuntian, China) on days 0, 1, 3, 5, and 7. The results are shown in Figure 15b. It can be seen that the cell culture medium with 10% and 25% extract did not affect the proliferation of the cultured cells, while the cell proliferation level in the 50% and 100% extract concentration groups decreased significantly. Therefore, the cell culture medium with a 25% extract concentration was used in subsequent experiments.

[0130] As shown in Figure 15c, when hBMMSCs cells were cultured in the cell proliferation medium containing 25% extract for 24 and 48 hours, the cellular uptake of Se in the experimental group was significantly increased compared with the control group, indicating that selenium can be effectively taken up by cells after modification with selenium-containing polymers to exert subsequent effects.

[0131] The live / dead cell staining kit (Keygene Biotechnology, China) was used to evaluate the cell compatibility of the selenium-containing polymer modified degradable zinc alloy prepared in Example 9. Human bone marrow mesenchymal stem cells (hBMMSCs) were seeded in a well plate. On the second day, the culture medium was replaced with a 25% extract of a selenium-containing polymer modified degradable zinc alloy with different hydrophilic group ratios and different selenium concentrations. Cell viability was detected by live / dead cell staining on the third day of culture. A TE2000-U inverted fluorescence microscope (Nikon, Japan) was used with F3.0 digital photography system software (NIS-Elements, Japan) to photograph the cells on the surface of the zinc sheet. The fluorescence color of live cells was green and the fluorescence color of dead cells was red. As shown in Figure 15d, there was no significant difference in the effect of the 25% concentration culture medium prepared with different hydrophilic group ratios and different concentrations of selenium-containing polymer modified zinc alloy extracts on cell viability.

[0132] Example 13: Detection of the osteogenic effect of mesenchymal stem cells in vitro using zinc alloy materials modified with selenium-containing polymers

[0133] The in vitro osteogenic effect of mesenchymal stem cells on zinc alloy materials modified with selenium-containing polymer compounds was evaluated using alkaline phosphatase (ALP) staining and quantification, alkaline phosphatase (ARS) staining and quantification, quantitative real-time PCR (qPCR), Western blot, and immunofluorescence technique.

[0134] Here, a 25% Se-containing selenium polymer with a 3:1 ratio of hydrophilic groups to selenium groups, as described above, was selected. hBMMSCs were seeded in 24-well plates, 0.5 mL per well, at a cell density of approximately 3,000 cells / mL. Cells were incubated in PM cell proliferation medium until 80% confluency was reached. The medium was then replaced with the aforementioned medium containing the 25% extract by volume. The medium was changed every two days. After seven days of incubation at 37°C and 5% CO₂, the cells were removed from the incubator and washed three times with PBS. The cells were stained and quantitatively analyzed using an alkaline phosphatase (ALP) staining kit (Biyuntian, China) and an alkaline phosphatase quantification kit (Biyuntian, China). The results are shown in Figure 16a. As can be seen, ALP staining of hBMMSCs cultured in the SePUA-Zn0.8Li extract (Example 9) was generally deeper than that in the SePU-Zn0.8Li extract (Example 1), with the highest ALP activity observed in the 25% Se 3 mg / mL group.

[0135] FIG16 b shows the quantitative ALP data of hBMMSCs cultured in SePUA-Zn0.8Li (Example 9) extract medium with different selenium-containing group ratios and concentrations. Similarly, the 25% Se 3 mg / mL group showed the highest ALP activity.

[0136] The qPCR method was used to analyze the expression mechanism of osteogenic genes in hBMMSCs. The cells were seeded and cultured using the same method as the above experiment. When the confluence reached 80%, the culture medium was replaced with different 25% extract-containing medium. The sheet used for extraction was unmodified zinc alloy Zn0.8Li in the control group, and the experimental groups were 25% Se group ratio, 1, 3, and 5 mg / mL of selenium-containing polyurethane (Example 1) and selenium-containing polyurea (Example 9) modified degradable zinc alloys, which are abbreviated as 25-1SePU, 25-3SePU, 25-5SePU, 25-1SePUA, 25-3SePUA, and 25-5SePUA in the figure. The medium was changed every 48 hours, and samples were collected 7 days after induction to measure the mRNA expression levels of two marker genes, osterix (OSX) and osteocalcin (OCN). The results are shown in Figure 16c. As can be seen from Figure 16c, the expression levels of various marker genes of degradable zinc alloy after surface modification with selenium-containing polymer compounds are upregulated compared with the unmodified group, and the expression levels of marker genes of SePUA-Zn0.8Li in Example 9 are more increased than those of SePU-Zn0.8Li in Example 1.

[0137] Western Blot and immunofluorescence techniques were used to measure the RUNX2 protein expression levels of hBMMSCs cultured on degradable zinc alloys before and after modification with selenium-containing polymer compounds. hBMMSCs cells were seeded in 24-well plates, incubated with proliferation medium until 80% confluence, and then replaced with 25% extract cell culture medium. After 7 days of induction, the cells were centrifuged and the supernatant was collected for detection. The results are shown in Figures 16d and 16e. As can be seen from Figure 16d, the expression of the osteogenic-related protein RUNX2 in the degradable zinc alloy group modified with selenium-containing polymer compounds (Example 9) increased; as shown in Figure 16e, the immunofluorescence results showed that the fluorescence intensity of RUNX2 and OCN in the group modified with selenium-containing polymer compounds (Example 9) was significantly stronger than that in the unmodified group, which is consistent with the immunofluorescence results, indicating that selenium-containing polymers can promote the osteogenic differentiation of hBMMSCs.

[0138] Example 14: In vitro detection of the polarization of macrophages toward anti-inflammatory phenotype promoted by zinc alloy materials modified with selenium-containing polymer compounds

[0139] The effect of degradable zinc alloy modified with selenium-containing polymer compounds on the polarization of THP-1-derived macrophages was evaluated. The suspension cultured human monocytic leukemia cells (THP-1) were centrifuged (600 rpm, 3 minutes) and the supernatant was discarded. The pellet was resuspended in cell proliferation medium (RPMI 1640, 10% FBS, 1% penicillin-streptomycin) to obtain a cell suspension, and 100 ng / mL phorbol ester (PMA) was added. The cell suspension was inoculated into a well plate, and after 24 hours, the adherent growth was induced under a microscope and replaced with a cell proliferation medium containing an extract without PMA. Samples were collected after 4 days, and qPCR was used to detect the expression levels related to the polarization of THP-1-derived macrophages to pro-inflammatory and anti-inflammatory phenotypes. The results of the modification of the selenium-containing polymers in Example 1 and Example 9 on the surface of the zinc alloy are shown in Figure 17a. After modification with all selenium-containing polymer compounds, the expression levels of the pro-inflammatory gene iNOS were downregulated, and the expression levels of the anti-inflammatory gene ARG-1 were upregulated, and Example 9 showed a more significant regulatory effect than Example 1.

[0140] The level of reactive oxygen species (ROS) in macrophages was detected (Solaibao, China). The results are shown in Figure 17c. The fluorescence intensity of macrophages cultured with degradable zinc alloy modified with selenium-containing polymer compounds was significantly lower than that of the unmodified group, indicating that selenium-containing polymer compounds can eliminate ROS generated in macrophages due to oxidative stress in the zinc alloy environment, thereby inhibiting the secretion of pro-inflammatory related factors.

[0141] The Western Blot and immunofluorescence in Figures 17b and 17d used the 25% Se 3mg / mL SePUA-Zn0.8Li with the best osteogenic effect as the experimental group, which also showed the regulatory effect of selenium-containing polymer modification on the expression levels of iNOS and ARG-1, indicating that selenium-containing polymer compound modification of degradable zinc alloy can better promote the polarization of human monocytes toward an anti-inflammatory phenotype in vitro.

[0142] Example 15: In vivo osteogenic effect testing of zinc alloy materials modified with selenium-containing polymer compounds

[0143] Bilateral femoral defects were created in 6-8 week old male Sprague-Dawley rats. After anesthetizing the rats, a 1.5 mm diameter defect was created perpendicular to the long axis of the femur using a grinder at a fixed point 5 mm proximal to the lateral epicondyle. Three rats were implanted into the bone defects with either the previously polished unmodified zinc alloy rod or a degradable zinc alloy rod modified with 25% Se (3 mg / mL) and various selenium-containing polymers. Four weeks after surgery, the rats were sacrificed and the femurs were removed for micro-CT scanning. The results are shown in Figure 18a. In the unmodified group, no significant hyperdensity was observed around the implant, indicating almost no new bone formation. In the SePU polymer-modified zinc alloy rod from Example 1, only a small amount of hyperdensity was observed around the implant. In the SePU polymer-modified zinc alloy rod from Example 9, a significant amount of hyperdensity was observed around the implant, indicating greater new bone formation compared to the SePU polymer-modified group from Example 1. This demonstrates that the SePU polymer-modified zinc alloy from Example 9 exhibits superior in vivo osteogenic efficacy, consistent with the in vitro results. The femurs of the unmodified zinc alloy group and the selenium-containing polymer modified group of Example 9 were subjected to bone tissue grinding and methylene blue-acid fuchsin staining. The results are shown in Figure 18b, indicating that at 4 weeks, the unmodified zinc alloy group had a small amount of discontinuous new bone formation, while the new bone formed by the modified group of Example 9 was more, more continuous, and more closely aligned with the implant material; at 8 weeks, although the unmodified zinc alloy group had increased bone mass compared to that at 4 weeks, the new bone formed was still discontinuous and fragmentary, while the new bone formed by the modified group of Example 9 was more, more continuous, and more tightly integrated with the implant. The above results indicate that modification with selenium-containing polymer compounds enhances the bone integration ability of metal materials, and that the selenium-containing polyurea in Example 9, which is modified on the surface of the degradable zinc alloy, can have a better osteogenic effect than the selenium-containing polyurethane of Example 1, and can enhance bone regeneration around the metal material.

[0144] Example 16: Testing the osteogenic effect of titanium materials modified with selenium-containing polymer compounds on smokers

[0145] (1) Preparation of cigarette smoke extract (CSE)

[0146] A cigarette (Taishan White General, Shandong) was connected to one end of a microvacuum pump (Qilin Bell, GL-802) and the cigarette was lit to initiate combustion. Each cigarette contained 12 mg of tar and 1.1 mg of nicotine. The microvacuum pump was turned on and the main stream of smoke was drawn into the pump at a power of 0.01 MPa into a centrifuge tube containing 1 mL of phosphate-buffered saline (PBS) at the other end. The CSE was filtered through a 0.22 μm pore size filter and labeled as 100% CSE (100% CSE).

[0147] (2) Inhibitory effect of CSE exposure on osteogenic differentiation of mesenchymal stem cells in vitro

[0148] Alkaline phosphatase (ALP) staining and Alizarin red (ARS) staining were used to evaluate the inhibitory effect of CSE exposure on the osteogenic differentiation of human BMMSCs (hBMMSCs) in vitro.

[0149] An alkaline phosphatase (ALP) staining kit (Beyotime, China) was used to detect the ALP activity of hBMMSCs cultured for 7 days in medium without CSE (control) or with 0.2% CSE. For example, 50 mL of culture medium was used: the medium without CSE consisted of 5 mL FBS, 500 μL penicillin-streptomycin, and 44.5 mL α-MEM; the medium with 0.2% CSE consisted of 5 mL FBS, 500 μL penicillin-streptomycin, 100 μL 100% CSE, and 44.4 mL α-MEM. As shown in Figure 19a, ALP staining of hBMMSCs cultured in medium with 0.2% CSE was significantly lighter than that in the control, indicating that CSE exposure inhibited ALP activity in hBMMSCs.

[0150] The mineralized nodules of hBMMSCs cultured for 14 days in culture medium containing either 0.2% CSE or without CSE (control) were stained using 1% Alizarin Red (ARS) solution (Sigma-Aldrich). As shown in Figure 19b, hBMMSCs cultured in the 0.2% CSE medium produced significantly fewer mineralized nodules than the control, indicating that CSE exposure reduced the formation of mineralized nodules in hBMMSCs. These results demonstrate that exposure to 0.2% CSE inhibits the osteogenic differentiation of hBMMSCs in vitro.

[0151] (3) Titanium materials modified with selenium-containing polymers improve osteogenic differentiation of mesenchymal stem cells in vitro under CSE exposure

[0152] An alkaline phosphatase (ALP) staining kit (Biyuntian, China) was used to measure the ALP activity of hBMMSCs cultured for 7 days on pure titanium and titanium materials modified with a 25% Se 3 mg / mL selenium-containing polymer (prepared in Example 2, labeled Se) in a medium containing 0.2% CSE. As shown in Figure 20, under CSE exposure, ALP staining of hBMMSCs cultured on the selenium-containing polymer-modified titanium surface was significantly deeper than that of pure titanium, indicating that the selenium-containing polymer-modified titanium material increases ALP activity and can improve the decreased ALP activity of hBMMSCs caused by CSE exposure.

[0153] (4) In vivo osteogenic effect testing of titanium materials modified with selenium-containing polymers under CSE exposure

[0154] A rat smoking model was established using 8-week-old male Sprague-Dawley rats (Weitong Lihua Laboratory Animal Technology Co., Ltd., Beijing). Three rats were included in each experimental group. The control group (denoted as the PBS group) received an intraperitoneal injection of 4 mL / kg PBS, while the experimental group (denoted as the CSE group) received an intraperitoneal injection of 4 mL / kg 100% CSE every two days for a total of four weeks. After four weeks of modeling, the rat femurs were collected and fixed in 4% paraformaldehyde for 24 hours, followed by microcomputed tomography (Micro-CT) analysis. As shown in Figure 21a, compared with the PBS-only group, the CSE-exposed group had reduced bone mass, with more sparse and discontinuous trabeculae, indicating that CSE exposure disrupts bone homeostasis in rats and leads to bone loss in the femurs. Blood was collected from the rat tail vein, allowed to stand at room temperature for 30 minutes, and centrifuged at 1000g for 15 minutes. The supernatant was then collected and the levels of IL-1β (KE20005), IL-6 (KE20024), and TNF-α (KE20018) were measured using the corresponding ELISA kits (Proteintech Group Inc.) according to the manufacturer's instructions. As shown in Figure 21b, the concentrations of IL-1β, IL-6, and TNF-α in the serum of the PBS control group were all below the minimum detectable values ​​of the kits (31.25 pg / mL for IL-1β; 62.5 pg / mL for IL-6; and 4.7 pg / mL for TNF-α), and therefore were recorded as minimum values. Furthermore, the concentrations of IL-1β, IL-6, and TNF-α in the CSE-exposed group were significantly increased compared to the control group, indicating that continuous CSE exposure induces chronic inflammation in rats. The combined results of Micro-CT and ELISA demonstrated that CSE caused damaged osteogenesis and increased inflammation levels in rats, and the rat smoking model was successfully established.

[0155] Then, 8-week-old SD male rats (Weitong Lihua Experimental Animal Technology Co., Ltd., Beijing) were used to build a rat smoking model. The rats were intraperitoneally injected with 4mL / kg 100% CSE, injected once every 2 days, and the model was built for 4 weeks. After 4 weeks of modeling, a bilateral femoral defect model was prepared. The femoral defect site was located at about 5mm proximal to the lateral epicondyle of the bilateral femur, with a defect diameter of 1.5mm and a defect depth of 5mm. A hole was punched perpendicular to the long axis of the femur, and the defect site was implanted with pure titanium and a titanium rod modified with a selenium-containing polymer compound. The rat in vivo experiment was divided into a Ti control group and a 25% Se 3mg / mL experimental group (prepared in Example 2, labeled Se). After 4 weeks, the rats were sampled and analyzed by Micro-CT. As shown in Figure 21c, Micro-CT scanning showed that there was basically no new bone formation around the implanted titanium rod, and the titanium rod modified with a selenium-containing polymer compound had more new bone formation than the pure titanium group, which was consistent with the in vitro results. The results showed that titanium rods modified with selenium-containing polymer compounds can promote new bone formation around the implant material and improve bone integration ability under CSE exposure.

[0156] Therefore, the metal material modified with a selenium-containing polymer compound of the present invention can address the problem of chronic inflammation and osteogenesis inhibition in the local microenvironment caused by smoking, ultimately leading to poor implant prognosis. This material can improve the inhibition of hBMMSC osteogenic differentiation caused by CSE exposure in vitro and in vivo in a SD rat smoking model, which also reduces the inhibition of bone tissue formation around rat implants caused by CSE exposure.

[0157] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these changes and variations. The above embodiments or implementation methods are merely illustrative of the present invention, and the present invention may also be implemented in other specific ways or other specific forms without departing from the gist or essential features of the present invention. Therefore, the described implementation methods should be regarded as illustrative and not restrictive in any respect. The scope of the present invention should be described by the appended claims, and any changes that are equivalent to the intent and scope of the claims should also be included within the scope of the present invention.

Claims

1. A selenium-containing polymer compound, characterized in that: The selenium-containing high molecular compound is prepared by reacting raw materials including R1-R2-Se-R3-R4, a compound of formula (I), diisocyanate, and mPEG-R5. Wherein: R1 and R4 are each independently selected from a hydroxyl group or an amino group; R2 and R3 are each independently selected from C 2-20 alkyl; R5 is selected from a hydroxyl group or an amino group; R6 and R7 are each independently selected from C 1-5 alkyl groups.

2. The selenium-containing high molecular compound according to claim 1, wherein: R1 and R4 are the same, and / or R2 and R3 are the same, and / or R6 and R7 are the same.

3. The selenium-containing high molecular compound according to claim 1, wherein: The diisocyanate is selected from tolylene 2,4-diisocyanate, hexamethylene diisocyanate, diphenylmethane diisocyanate or isophorone diisocyanate.

4. The selenium-containing polymer compound according to claim 1, wherein: The structure of the selenium-containing polymer compound is as shown in the following formula (II) or formula (III). Wherein m and n are each independently selected from natural numbers from 1 to 250, and x and y are each independently selected from natural numbers from 1 to 100.

5. A method for preparing a selenium-containing polymer compound according to any one of claims 1-4, characterized in that, The preparation method includes: reacting R1-R2-Se-R3-R4, the compound of formula (I), diisocyanate, mPEG-R5, and a catalyst in an organic solvent, and subjecting the resulting product of the reaction to recrystallization and washing.

6. The preparation method according to claim 5, wherein: The organic solvent is selected from one or more of tetrahydrofuran, N,N-dimethylformamide, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate and n-butyl acetate; The molar ratio of R1-R2-Se-R3-R4, the compound of formula (I), diisocyanate, and mPEG-R5 is x:1:(1.1x + 1.1):(0.2x + 0.2), where x is from 0.1 to 10; The catalyst is dibutyltin dilaurate.

7. The preparation method according to claim 5, characterized in that: The reaction temperature is 30-100°C, and the reaction time is 1-48 hours.

8. A metal material modified with a selenium-containing polymer compound, characterized in that: The selenium-containing polymer compound-modified metal material is obtained by coordinating and compounding the selenium-containing polymer compound according to any one of claims 1-4 with a metal material.

9. The metal material modified with a selenium-containing polymer compound according to claim 8, wherein: The metal material is selected from one or more of pure titanium, titanium alloy, zinc alloy, magnesium alloy, and cobalt-chromium alloy.

10. A method for preparing a selenium-containing polymer compound-modified metal material according to claim 8 or 9, characterized in that, The preparation method includes: dissolving the selenium-containing polymer compound according to any one of claims 1-4 in an organic solvent, and then immersing a metal material therein for a modification reaction.

11. The preparation method according to claim 10, wherein: The organic solvent is dimethyl sulfoxide and / or N,N-dimethylformamide; The concentration of the selenium-containing polymer compound dissolved in the organic solvent is 1-10 mg / mL.

12. The preparation method according to claim 10, characterized in that: The time of the modification reaction is 1-3 hours.

13. Use of the selenium-containing polymer compound-modified metal material according to claim 8 or 9 for the preparation of an oral implant or an orthopedic implant.

14. The use according to claim 13, wherein: The oral implant or orthopedic implant is for one or more of diabetic patients, osteoporosis patients, and smoking patients.

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