CN117442775A

CN117442775A - Polyether-ether-ketone-based composite material and preparation method and application thereof

Info

Publication number: CN117442775A
Application number: CN202311116500.7A
Authority: CN
Inventors: 李国林; 谢宁; 韩妲丽; 连周燕; 迟惠熔; 郭净洁; 李凤前; 魏杰
Original assignee: SHANGHAI EIGHTH PEOPLE'S HOSPITAL
Current assignee: SHANGHAI EIGHTH PEOPLE'S HOSPITAL
Priority date: 2023-08-31
Filing date: 2023-08-31
Publication date: 2024-01-26

Abstract

The invention provides a polyether-ether-ketone-based composite material, and a preparation method and application thereof. The preparation method comprises the following steps: performing hot press molding on the mixture of tantalum oxide and polyether-ether-ketone PEEK; wherein the tantalum oxide is prepared by any one of the following methods: the method comprises the following steps: reacting tantalum salt with organic alcohol, dissolving the obtained mixture in a solvent, mixing and stirring with water, and calcining the stirred precipitate TO obtain TO particles; the second method is as follows: stirring and aging tantalum salt in an acidic solution containing a template agent to prepare a precipitate; calcining the precipitate to obtain MTO particles; the calcining temperature is 500-700 ℃. The invention regulates and controls the mechanical property of the material to be matched with bone tissue through blending modification and surface active treatment, and the prepared PT, PTF and PTFE have multiple effects of promoting regeneration and integration of bone/gum tissue, resisting bacteria and inflammation and the like, thereby realizing the repair and functional reconstruction of tooth defects.

Description

Polyether-ether-ketone-based composite material and preparation method and application thereof

Technical Field

The invention relates to a polyether-ether-ketone-based composite material, and a preparation method and application thereof.

Background

High-performance implant materials/implant instruments are required for repairing and functionally reconstructing large bone/tooth defects caused by diseases such as wounds, tumors and the like. For the bone repair material, the biological activity and osteoinductive property of the material have key effects on promoting new bone regeneration, osseointegration and realizing successful bone repair after implantation. By optimally designing the surface composition, structure and the like of the bone repair material, osteoblasts generate benign cell reaction, which is favorable for guiding the subsequent regeneration and integration of bone tissues and has great research significance for realizing successful bone repair.

Dental implant materials need to be integrated not only with bone to achieve fixation, but also with soft tissue of the gums to achieve biological sealing, thus preventing bacterial attack. For a large bone/tooth defect, the repair material should have the following essential characteristics: (1) good biocompatibility: no immunological rejection reaction is formed with the organism, and no harmful substances are formed; (2) biological Activity: stimulating osteoblast response and new bone tissue formation; (3) good mechanical properties: the mechanical property is matched with host bone, and the mechanical strength is high and the fatigue resistance and the wear resistance are excellent.

Polyether ether ketone (PEEK) has excellent biocompatibility and mechanical properties, and has been widely used for orthopedic implant devices; however, PEEK is biologically inert and has poor osteogenic properties, and is difficult to form osseointegration with bone tissue, resulting in loosening/failure after implantation. Therefore, improving the PEEK body and the surface bioactivity is a research hot spot in the field of bone repair materials. The modes for modifying PEEK include blending modification and surface modification. In the prior art, surface modification often cannot regulate and control the mechanical property of PEEK matrix material to be perfectly matched with bone tissue; in the existing modification modes, the osteogenesis activity/antibacterial performance of the PEEK material is mostly improved, and no improvement measures for improving the material and promoting soft tissue healing are aimed at. In recent years, the bioactive modification of PEEK is an important and difficult point of research in the field of bone repair materials.

Tantalum Oxide (TO) and epigallocatechin gallate (EGCG) are good modifiers. The TO has extremely good plasticity and ductility, is easy TO process and design the shape, has excellent biocompatibility and osteogenic activity, can stimulate osteoblast response (such as adhesion, proliferation and osteogenic differentiation), but has certain brittleness and lower strength, and limits the mechanical properties. EGCG is a natural functional small molecule with antibacterial, anti-inflammatory and osteoclast inhibiting functions.

The femtosecond laser (Femtosecond Laser) refers to a laser having a pulse width on the order of femtosecond (fs). The femtosecond laser technology is an emerging micro-nano structure manufacturing technology, has the advantages of high accuracy, low energy consumption, small thermal damage, wide processing range and the like, and has wide application prospect in micromachining the surface of a dental implant material by adopting the femtosecond laser technology.

In summary, aiming at the challenges of large number of patients with large bone/tooth defect, high treatment difficulty, long treatment period and the like, development of a high-performance bone repair material is urgently needed to realize effective repair of the bone defect part, recover normal physiological functions of the patients and relieve pressure of the patients.

Disclosure of Invention

The invention solves the technical problems that the bone repair material in the prior art has poor bioactivity or mechanical property which is not matched with human bone and is difficult to promote regeneration of bone/gum tissue, and provides a polyether-ether-ketone-based composite material and a preparation method and application thereof. The invention regulates and controls the mechanical property of the material to be matched with bone tissue through blending modification and surface active treatment, and the prepared PT, PTF and PTFE have multiple effects of promoting regeneration and integration of bone/gum tissue, resisting bacteria and inflammation and the like, thereby realizing the repair and functional reconstruction of tooth defects.

The invention solves the technical problems through the following technical proposal.

The invention provides a preparation method of a polyether-ether-ketone-based composite material, which comprises the following steps: performing hot press molding on a mixture of tantalum oxide and polyether ether ketone (PEEK);

wherein the tantalum oxide is prepared by any one of the following methods:

the method comprises the following steps: reacting tantalum salt with organic alcohol, dissolving the obtained mixture in a solvent, mixing and stirring with water, and calcining the stirred precipitate TO obtain TO particles;

the second method is as follows: stirring and aging tantalum salt in an acidic solution containing a template agent to prepare a precipitate; calcining the precipitate to obtain MTO particles; the calcining temperature is 500-700 ℃.

In either method one or method two, the species of tantalum salt may be independently selected from one or more of tantalum ethoxide, tantalum chloride and tantalum oxalate, such as tantalum ethoxide.

In process one, the organic alcohol may be a hydroxyl-containing material conventional in the art, preferably ethylene glycol and/or ethanol.

In one method, the mass to volume ratio of the tantalum salt to the organic alcohol may be 1g (8-12 mL), for example 1g:10mL.

In one method, the temperature of the reaction may be 70 to 90 ℃, for example 80 ℃.

In method one, the reaction time may be 1 to 3 hours, for example 2 hours.

In process one, preferably, the reaction is generally carried out by passing an inert atmosphere to remove water and oxygen. The time for introducing the inert atmosphere may be 20 to 40 minutes, for example 30 minutes.

Wherein, after heating, the method can further comprise the operation of naturally cooling to room temperature. The room temperature may be 20 to 25 ℃ as is conventionally understood in the art.

In the first method, the solvent is preferably a ketone solvent and/or an alcohol solvent. The ketone solvent is preferably acetone. The alcohol solvent is preferably ethanol. After the solvent is added, tantalum alkoxide generated by the reaction of the tantalum salt and the organic alcohol can be dissolved.

In one method, the mass to volume ratio of the tantalum salt to the solvent may be 1g (30-50 mL), such as 1g:40mL.

In method one, the water may be deionized water as is conventional in the art. The water acts as an anti-solvent in the system to precipitate out the tantalum alkoxide.

In one method, the mass to volume ratio of the tantalum salt to the water may be 1g (0.3-0.5) mL, such as 1g:0.4mL.

In one method, the stirring time may be 6 to 10 hours, for example 8 hours.

In the first method, the precipitate can also comprise centrifugal washing and drying operations before calcination. Centrifugal washing can remove byproducts.

Wherein the solvent for centrifugal washing can be one or more of ethanol, acetone and water. The water may be deionized water as is conventional in the art.

Wherein the temperature of the drying may be 80 to 120 ℃, for example 100 ℃.

In one method, the calcination temperature may be 600 to 800 ℃, such as 700 ℃.

In one method, the calcination time may be 2 to 4 hours, for example 3 hours.

In the first method, the grain diameter of TO is 170-260 nm; preferably, the TO has an average particle size of 217nm.

In the second method, the acid solution containing the template agent can be prepared by dissolving the template agent in the acid solution and stirring.

Wherein the templating agent may be an easily removable templating agent conventional in the art, such as PEO-PPO-PEO.

Wherein the acidic solution can be an inorganic acid solution or an organic acid solution. The mineral acid solution is preferably a hydrochloric acid solution.

When the acidic solution is a hydrochloric acid solution, the concentration of the hydrochloric acid solution may be 1 to 3mol/L, for example, 2mol/L.

Wherein the mass to volume ratio of the template agent to the acidic solution may be 1g (20-40) mL, for example 1g:30mL.

Wherein, when the template agent is dissolved in the acid solution, stirring until the solution is clear; preferably, the stirring is for a period of time ranging from 5 to 7 hours, for example 6 hours.

In method two, the mass to volume ratio of the tantalum salt to the acidic solution containing the template agent may be 1g (20-30) mL, for example 1g:24mL.

In method two, the temperature of the agitation may be 40 to 60 ℃, for example 50 ℃.

In the second method, the stirring time may be 4 to 6 hours, for example, 5 hours.

In method two, the temperature of the aging may be 50 to 70 ℃, for example 60 ℃.

In method two, the aging time may be 2 to 4 days, for example 3 days.

In the second method, the precipitate can also comprise centrifugal washing and drying operations before calcination. Centrifugal washing can remove byproducts.

In the second method, the temperature of the calcination is preferably 550 to 650 ℃, for example 600 ℃. If the calcining temperature is too low, the template agent is difficult to remove cleanly, and the precipitate cannot be completely converted into tantalum oxide, so that the purity of the product is low; if the calcining temperature is too high, the formed tantalum oxide particles are easy to melt, so that the mesoporous structure collapses or the particles are agglomerated, and the final effect is affected.

In method two, the calcination time may be 5 to 7 hours, for example 6 hours.

In the second method, the particle size of the MTO can be 170-300 nm; preferably, the TO has an average particle size of 274nm.

In the second method, preferably, the MTO is a mesoporous material. The aperture of the MTO can be 9-10 nm; preferably, the pore size of the MTO is 9.69nm.

In the present invention, the mass ratio of the tantalum oxide to the Polyetheretherketone (PEEK) may be (5 to 10): 1, preferably (7 to 9): 1, for example, 8.72:1.

In the present invention, preferably, the hot press molding is to heat the mixture to a molding temperature, keep the temperature, press the mixture to a molding pressure, and keep the pressure, and press the mixture to obtain the composite material.

Wherein the molding temperature may be 340 to 370 ℃, for example 355 ℃.

Wherein the time of the incubation may be 0.2 to 1.5 hours, for example 0.5 hours or 1 hour.

Wherein the molding pressure may be 2 to 6MPa, for example 4MPa.

Wherein the dwell time may be 0.2 to 1.5 hours, for example 0.5 hours or 1 hour.

In the present invention, preferably, a femtosecond laser leveling treatment is performed on the surface of the composite material prepared by the hot press molding. According to the invention, through a femtosecond laser leveling mode, a nanometer petal-shaped structure can be formed on the surface of the tantalum oxide while the bioactive tantalum oxide is exposed, so that the specific surface area of the tantalum oxide is increased, the bioactive sites on the surface of the composite material are increased, and the bioactivity is further improved.

The pulse frequency of the femtosecond laser leveling sweep can be 900-1100 Hz, for example 1000Hz.

The pulse width of the femtosecond laser leveling scan can be 100-140 fs, for example 120fs.

Wherein, the pulse power of the femtosecond laser leveling sweep can be 30-50 mW, for example 40mW.

Wherein the single pulse energy of the femtosecond laser leveling sweep can be 150-250 muj, for example 200 muj.

Wherein the scanning speed of the femtosecond laser leveling scanning can be 300-500 μm/s, for example 400 μm/s.

Wherein, the processing environment of the femtosecond laser leveling scanning can be 1 standard atmosphere.

Wherein the femtosecond laser leveling process can be performed a plurality of times. Multiple passes can result in a more uniform surface, finer and more uniform particles, and removal of oxide films from the particle surface.

Preferably, the surface of the composite material subjected to femtosecond laser leveling treatment is loaded with epigallocatechin gallate (EGCG).

Preferably, the loading is performed by immersing the composite material subjected to the femtosecond laser leveling treatment in an EGCG aqueous solution.

The concentration of the EGCG aqueous solution may be 3-10 mg/mL, for example 5mg/mL.

The mass to volume ratio of the femtosecond laser swept composite material to the EGCG aqueous solution can be (0.1-0.5 g): 1mL, for example, 0.3g:1mL.

The manner of impregnation may be by shaking.

The temperature of the impregnation may be 20 to 40 ℃, for example 37 ℃.

The time of the impregnation may be 1 to 48 hours, for example 48 hours.

The invention provides a polyether-ether-ketone-based composite material, which is prepared by the preparation method.

The invention provides application of the polyether-ether-ketone-based composite material in preparing bone implant materials/dental implants.

In the present invention, the bone implant material is preferably a large-segment bone implant material.

On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.

The reagents and materials used in the present invention are commercially available.

The invention has the positive progress effects that:

(1) The TO particles and the MTO particles prepared by the invention have uniform submicron sizes, and the surface roughness, hydrophilicity, surface energy and protein adsorption capacity of the PEEK/TO composite material (PT) and the PEEK/MTO composite material (PMT) prepared by the invention are obviously improved compared with those of PEEK. In particular, because of the large number of mesoporous structures in the MTO particles, the specific surface area is larger, the active sites are more, and the prepared PMT has more excellent performance than PT, not only can promote bone tissue and integration, but also can promote regeneration of soft tissues (gum tissues), has multiple effects of resisting bacteria, resisting inflammation and the like, and can greatly widen the application fields of the composite material (not only can be applied to the field of bone repair of large-section bone defects and the like, but also can realize repair and functional reconstruction of tooth defects).

(2) Further, the PTF is prepared by the composite material after femto-second laser modification, a submicron-nanometer structure can be constructed on the surface, a nanometer petal structure is formed, more active sites are exposed, and meanwhile, the loading of EGCG is facilitated.

(3) Furthermore, in the invention, EGCG molecules are combined with PTF through hydrogen bonds to synthesize PTPE, so that the combination force is stronger, the sustained release of EGCG molecules can be realized, and the effect is better; meanwhile, as more tantalum oxide active sites are exposed by surface modification, the EGCG loading regulation range is larger. PTPE has good antibacterial property, stimulates BMSC/GEC/BMM response, promotes regeneration and integration of bone/gum tissue in vivo, has multiple biological functions of antibacterial, anti-inflammatory and the like, and has wide application prospect in the field of dental implant materials.

Drawings

Fig. 1 is an SEM image of TO submicron particles (part (a) of fig. 1), a TEM image (part (b) of fig. 1), an EDS spectrum (part (c) of fig. 1), and DLS analysis (part (d) of fig. 1).

FIG. 2 is an SEM image of MTO particles (FIG. 2 part (a) and FIG. 2 part (b)), and EDS spectra (FIG. 2 part (c)) and DLS analysis (FIG. 2 part (d))

FIG. 3 is a TEM image of MTO particles (part (a) of FIG. 3 and part (b) of FIG. 3), and N ₂ Adsorption/desorption isotherm (part (c) of fig. 3) and pore size distribution curve (part (d) of fig. 3)

FIG. 4 shows water contact angles (part (a) of PEEK, PT and PMT, diiodomethane contact angle (part (b) of FIG. 4), surface energy (part (c) of FIG. 4) and protein adsorption (part (d) of FIG. 4) (. P < 0.05, vs. PEEK;.. P < 0.05, vs. PT).

Fig. 5 shows FTIR (part (a) of fig. 5) and XRD (part (b) of fig. 5) spectra (+.s representing characteristic peaks of TO;) representing characteristic peaks of PEEK) of TO, PEEK, PMT and PTF.

Fig. 6 is an XPS high resolution spectrum of peaks of C1s (part (a) of fig. 6 and part (b) of fig. 6) and Ta4f (part (C) of fig. 6 and part (d) of fig. 6) of PMT and PTF.

Fig. 7 is an SEM image and EDS spectrum of PEEK (part (a) of fig. 7, part (b) of fig. 7, part (c) of fig. 7, and part (d) of fig. 7), PMT (part (e) of fig. 7, part (f) of fig. 7, part (g) of fig. 7, and part (h) of fig. 7) and PTF (part (i) of fig. 7, part (j) of fig. 7, part (k) of fig. 7, and part (l) of fig. 7).

Fig. 8 is an EDS element plane distribution image of PEEK (part (a) of fig. 8, part (b) of fig. 8, and part (C) of fig. 8), PMT (part (d) of fig. 8, part (e) of fig. 8, and part (f) of fig. 8) and PTF (part (g) of fig. 8, part (h) of fig. 8, and part (i) of fig. 8) for C (part (a) of fig. 8, part (d) of fig. 8, and part (g) of fig. 8), O (part (b) of fig. 8, part (e) of fig. 8, and part (h) of fig. 8), and Ta (part (C) of fig. 8, part (f) of fig. 8, and part (i) of fig. 8).

Fig. 9 is LCM and AFM images of PEEK (part (a) of fig. 9, part (d) of fig. 9, and part (g) of fig. 9), PMT (part (b) of fig. 9, part (e) of fig. 9, and part (h) of fig. 9), and PTF (part (c) of fig. 9, part (f) of fig. 9, and part (i) of fig. 9).

FIG. 10 shows water contact angles (part (a) of FIG. 10), diiodomethane contact angles (part (b) of FIG. 10), surface energies (part (c) of FIG. 10) and protein adsorption (part (d) of FIG. 10) for PEEK, PMT and PTF (p < 0.05, vs. PEEK; # p < 0.05, vs. PMT).

Fig. 11 is FTIR (part (a) and XRD (part (b) of fig. 11) spectra of EGCG, PTE and PTFE, and loading amount (part (c) of fig. 8) and cumulative release rate (part (d) of fig. 11) of PTE and PTFE to EGCG.

FIG. 12 is a photograph of PEEK, PMT, PTF and PTFE surface isolated bacteria after 24 hours of incubation on agar plates (FIG. 12 part (a)) and the bacteriostasis to E.coli (FIG. 12 part (b)) and S.aureus (FIG. 12 part (c)) (< 0.05 for p < 0.05 for vs.PMT; # p < 0.05 for vs.PTF.

Fig. 13 is an SEM image of BMSC after surface culture 12 (part (a) of fig. 13 to part (d) of fig. 13) and 48 (part (e) of fig. 13 to part (h) of fig. 13) for PEEK (part (a) of fig. 13 and part (e) of fig. 13), PMT (part (b) of fig. 13 and part (f) of fig. 13), PTF (part (c) of fig. 13 and part (g) of fig. 13) and PTFE (part (d) of fig. 13 and part (h) of fig. 13) for hours.

Fig. 14 is an SEM image of GEC after surface culture 6 (part (a) of fig. 14 to part (d) of fig. 14) and 24 (part (e) of fig. 14 to part (h) of fig. 14) for PEEK (part (a) of fig. 14 and part (e) of fig. 14), PMT (part (b) of fig. 14 and part (f) of fig. 14), PTF (part (c) of fig. 14 and part (g) of fig. 14) and PTFE (part (d) of fig. 14 and part (h) of fig. 14) for hours.

Fig. 15 shows CLSM images of BMSC after 1 (part a of fig. 15, part e of fig. 15 and part i of fig. 15), PMT (part b of fig. 15, part f of fig. 15 and part j of fig. 15), PTF (part c of fig. 15, part g of fig. 15 and part k of fig. 15) and PTFE (part d of fig. 15, part h of fig. 15 and part l of fig. 15) are surface-cultured on PEEK (part a of fig. 15, part e of fig. 15 and part i of fig. 15), part 3 (part e of fig. 15) to part h of fig. 15 and part 7 (part i of fig. 15 to part l of fig. 15) days.

Fig. 16 is a CLSM image of GEC after surface culturing 1 (part a of fig. 16 to part d of fig. 16, part e of fig. 16, and part i of fig. 16), PMT (part b of fig. 16, part f of fig. 16, and part j of fig. 16), PTF (part c of fig. 16, part g of fig. 16, and part k of fig. 16), and PTFE (part d of fig. 16, part h of fig. 16, and part l of fig. 16) on PEEK (part a of fig. 16, part e of fig. 16, and part i of fig. 16 to part h of fig. 16), and part 7 (part i of fig. 16 to part l of fig. 16) days.

FIG. 17 shows the OD values (FIG. 17 part (a)) and ALP activities (FIG. 17 part (b)) of BMSCs after various times of surface culture with PEEK, PMT, PTF and PTFE, and the cell adhesion rates (FIG. 17 part (c)) and OD values (FIG. 17 part (d)) of GECs after various times of surface culture with PEEK, PMT, PTF and PTFE (.p < 0.05, vs. PMT; #p < 0.05, vs. PTF).

FIG. 18 shows the expression of the osteogenic genes ALP (FIG. 18 part (a)), colI (FIG. 18 part (b)), OPN (FIG. 18 part (c)) and OCN (FIG. 18 part (d)) after 7 and 14 days of BMSC surface culture with PEEK, PMT, PTF and PTFE (.p < 0.05, vs. PMT; # p < 0.05, vs. PTF).

FIG. 19 is a TRAP staining image of BMM after 4 days of co-culture of extracts of PEEK (FIG. 19 part (a)), PMT (FIG. 19 part (b)), PTF (FIG. 19 part (d)) and PTFE (FIG. 19 part (e)), osteoclast number (FIG. 19 part (c)) and area (FIG. 19 part (f) (. P < 0.05, vs. PMT;.. #p < 0.05, vs. PTF).

FIG. 20 shows the expression of the osteoclast-associated genes TRAP (FIG. 20 part (a)), CTSK (FIG. 20 part (b)), OPG (FIG. 20 part (c)) and RANKL (FIG. 20 part (d)) after 4 and 7 days of BMM surface culture with PEEK, PMT, PTF and PTFE (< 0.05 for p, 0.05 for vs. PMT; # p, 0.05 for vs. PTF).

FIG. 21 shows the expression of inflammatory factor-related genes IL-6 (FIG. 21 part (a)) and TNF-. Alpha. (FIG. 21 part (b)) and Western blot images of inflammatory factor expression (< 0.05, vs. PMT; #p < 0.05, vs. PTF) of BMM after 1 and 3 days of surface culture with PTFE and PEEK, PMT, PTF.

Fig. 22 is a stained image of a hard tissue section of the implanted region (VG stained in part (a) of fig. 22 and HE stained in part (b) of fig. 22) after PEEK, PMT, PTF and PTFE implantation in vivo for 4 and 12 weeks.

Fig. 23 shows the bone contact rate (part (a) of fig. 23), gum contact rate (part (b) of fig. 23), push-out strength (part (c) of fig. 23) and pull-out strength (part (d) of fig. 23) after implantation of PEEK, PMT, PTF and PTFE in the body for 4 and 12 weeks (p < 0.05, vs. pmt; # p < 0.05, vs. ptf).

Detailed Description

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

Example 1 preparation of PEEK/TO (PT) composite

(1) TO synthesis

TO submicron particles were synthesized by anti-solvent precipitation. First, 5g of tantalum ethoxide (99%, shanghai Ala Biotechnology Co., ltd., china) was charged into a flask containing 50mL of ethylene glycol (AR, shanghai Milin Biotechnology Co., ltd., china) and magnetically stirred, and then nitrogen gas was introduced into the flask for 30 minutes to remove water and oxygen gas. Then, the flask was heated in an oil bath at 80 ℃ for 2 hours under magnetic stirring, after naturally cooling to room temperature, the mixture was quickly poured into a flask with 200mL of acetone, 2mL of deionized water was added and the flask was capped and stirring was continued for 8 hours. The resulting precipitate was collected by centrifugation and washed with ethanol, acetone and deionized water, respectively, and filtered to remove byproducts. Finally, the sample was dried in a 100 ℃ oven and calcined in a muffle furnace at 700 ℃ for 3 hours TO give TO submicron particles.

(2) Composite material

The PEEK/TO (PT) composite material is prepared by adopting a physical mixing-hot press forming method, wherein the TO content is optimized TO be 50vt percent, and the PEEK/TO (PT) composite material has two sizes (in vitro physicochemical, drug loading, antibacterial, cell experiment: phi 18 multiplied by 2mm, in vivo implantation experiment: phi 5 multiplied by 6 mm), and comprises the following specific steps:

100.25g of TO particles and 11.5g of PEEK powder were weighed and mixed, and the composite powder was ground in a planetary ball mill for 24 hours TO obtain a uniformly dispersed composite powder. The temperature of the press was raised from room temperature to 355 ℃, and the PEEK powder and the uniformly dispersed composite powder were respectively put into a mold of the press to be preheated for 30 minutes, and then the powder was hot-press-molded under a pressure of 4MPa for 30 minutes. And after the flat vulcanizing machine is cooled under constant pressure, cutting the obtained PEEK and composite board by using a numerical control cutting machine to obtain the material with the size of phi 18 multiplied by 2 mm. Finally, the material surface was polished using a polisher, and then washed with ethanol, acetone and deionized water for 3 times, respectively, to remove impurities, and dried.

Example 2 preparation of PEEK/MTO (PMT) composite

(1)MTO

Mesoporous Tantalum Oxide (MTO) is synthesized by a sol-gel method. First, template PEO-PPO-PEO (P123, 4g, sigma-Aldrich, USA) was dissolved in hydrochloric acid solution (2 mol/L,120 mL) and stirred for 6 hours until the solution was clear. Then, 5g of tantalum ethoxide (AR, shanghai Meilin Biochemical Co., ltd., china) was added, and after stirring at 50℃for 5 hours, the reaction system temperature was kept at 60℃and left for 3 days. And (3) filtering and centrifuging the milky white precipitate in the system, and respectively washing with ethanol, acetone and deionized water to remove impurities. Finally, the precipitate was dried in an oven at 100 ℃ and then calcined in a muffle furnace at 600 ℃ for 6 hours to remove the residual templating agent, yielding MTO particles.

(2) Composite material

The PEEK/MTO (PMT) composite material is prepared by adopting a physical mixing-hot press forming method, wherein the MTO content is optimized to be 50vt percent, and the PEEK/MTO (PMT) composite material has two sizes (in vitro physicochemical, drug loading, antibacterial, cell experiment: phi 18 multiplied by 2mm, in vivo implantation experiment: phi 5 multiplied by 6 mm), and comprises the following specific steps:

100.25g of MTO particles and 11.5g of PEEK powder were weighed and mixed, and milled in a planetary ball mill for 24 hours to obtain a uniformly dispersed composite powder. The temperature of the press vulcanizer was raised from room temperature to 355 ℃, the PEEK powder and the uniformly dispersed composite powder were respectively put into a mold of the press vulcanizer to be preheated for 30 minutes, and then the powder was hot-pressed for 30 minutes under a pressure of 4 MPa. And after the flat vulcanizing machine is cooled under constant pressure, cutting the obtained PEEK and composite board by using a numerical control cutting machine to obtain the material with the size of phi 18 multiplied by 2 mm. Finally, the material surface was polished using a polisher, and then washed with ethanol, acetone and deionized water for 3 times, respectively, to remove impurities, and dried.

Example 3 preparation of surface modified PEEK/MTO composite (PTF)

The PMT composite surface produced in example 2 was subjected to a swipe process using a femtosecond laser (GLX-200 HP-1053, time-Bandwidth Products AG, belgium). Wherein the pulse frequency is 1000Hz, the pulse width is 120fs, the pulse power is 40mW, the single pulse energy is 200 mu J, the scanning speed is 400 mu m/s, and the processing environment is 1 standard atmosphere air. After the processing is completed, the material is respectively washed by ethanol, acetone and deionized water, and a PMT surface femtosecond laser scanning (PTF) material is obtained.

Example 4 preparation of EGCG-loaded surface modified PEEK/MTO composite (PTFE)

50mg EGCG (95%, sigma, USA) was dissolved in 10mL of PBS buffer to prepare EGCG solution (5 mg/mL), and the PMT prepared in example 2 and the PTF material prepared in example 4 (Φ18×2mm, mass about 0.3 g) were placed in 12-well plates (mass about 0.3g per well of sample) respectively, and an equal volume of EGCG solution (1 mL) was added per well. The entire EGCG load experiment was performed in a thermostatted shaker and maintained at 37 ℃ for 48 hours.

Comparative example 1PEEK

The difference compared TO example 1 is that no TO treatment was performed.

Effect example 1TO and MTO characterization

The part (a) of fig. 1 is an SEM image of TO submicron particles, and it can be seen that the TO submicron particles have a uniform particle size and are spherical. The TEM image of TO submicron particles in section (b) of FIG. 1 shows that the TO particles are in the submicron range (170 nm TO 260 nm). The EDS spectrum of the TO submicron particles in part (c) of FIG. 1, in which characteristic peaks of Ta and O elements can be detected, and Ta ₂ O ₅ The chemical composition of (a) is the same. DLS analysis of the TO submicron particles as part (d) of FIG. 1 shows that the average size of the TO submicron particles is about 217nm.

The portions (portion (a) of fig. 2 and (portion b) of fig. 2) are SEM images of MTO particles, and it can be seen that the MTO particles are spherical and have a uniform particle size. The EDS spectrum of the MTO particle in part (c) of FIG. 2 shows characteristic peaks of Ta and O elements. DLS analysis of MTO particles in part (d) of FIG. 2 shows that the average size of MTO particles is about 274nm.

The portions (portion (a) of fig. 3 and (portion b) of fig. 3) are TEM images of MTO particles, and it can be seen that the MTO particles are uniform in size and have a uniform, regular mesoporous channel structure with uniform pore diameters. Part (c) of FIG. 3N being MTO particle ₂ Adsorption and desorption isotherms can be seen that MTO particles have N type IV with H1 hysteresis loop ₂ Adsorption and desorption isotherms indicate that the pore size of the MTO particles is highly uniform. FIG. 3 (d) is a graph showing the pore size distribution of MTO particles having a pore size of 9.69nm and a specific surface area of 313.18m according to BET and BJH analyses ² Per gram, pore volume of 0.48cm ³ And/g. In conclusion, the prepared MTO particle has high specific surface area and pore volume, and the mesoporous pore canal structure is uniform and regular, and has the potential of loading small molecules and medicines.

Effect example 2 characterization of physical and chemical Properties of materials

1. Contact angle, surface energy and protein adsorption analysis

The high hydrophilicity and surface energy of the implant material facilitate blood contact, protein adsorption, cell adhesion and diffusion, proliferation and differentiation, and promote new bone formation. Contact angles of water and diiodomethane on the surface of the material were measured separately using a contact angle meter. In addition, the surface energy of the material was calculated according to the Owen-Wendt formula. Protein adsorption is the initial event that occurs after implantation of the implant material, and cell adhesion is later than protein adsorption. BSA and FN are selected as model proteins, and the protein adsorption capacity of the material surface is analyzed.

Parts (a) and (b) of fig. 4 are water and diiodomethane contact angles of PEEK, PT and PMT surfaces. As can be seen from the figure, the PEEK surface has the highest water/diiodomethane contact angle. After blending the TO submicron particles, the water/diiodomethane contact angle of PT was slightly reduced. After blending the MTO particles, the water/diiodomethane contact angle of the PMT surface was further reduced. The water contact angles of the PEEK, PT and PMT surfaces were 84.5±1.2°, 75.2±2.2° and 65.3±1.6° respectively, and the diiodomethane contact angles of the surfaces were 68.3±1.7°, 61.3±1.5° and 48.6±1.0°, respectively.

Part (c) of fig. 4 is the surface energy of PEEK, PT, and PMT. It can be seen that the surface energy of PEEK is the lowest and PT is elevated after blending TO submicron particles. After blending the MTO particles, the surface energy of PMT was further increased than PEEK and PT. The surface energies of PEEK, PT and PMT were 26.7.+ -. 3.3mJ/m, respectively ² 、33.7±2.7mJ/m ² And 43.4.+ -. 2.5mJ/m ² 。

In summary, PT has significantly increased hydrophilicity and surface energy compared TO PEEK having hydrophobicity and low surface energy, probably due TO PT surface exposing more TO submicron particles having high hydrophilicity and surface energy. In addition, in view of the existence of a large number of mesoporous structures in the MTO particles, the specific surface area is larger, the exposed active tantalum oxide is more, and the hydrophilicity and the surface energy of PMT are further improved compared with PT.

The part (d) of FIG. 4 shows the adsorption results of PEEK, PT and PMT on both BSA and FN proteins. As can be seen from the figure, PEEK exhibits the weakest protein adsorption capacity. The protein adsorption capacity of PT surface was improved when TO submicron particles were blended. After blending the MTO particles, the protein adsorption capacity of PMT is obviously improved. Protein adsorption rates of PEEK, PT and PMT to BSA were 3.90.+ -. 0.71%, 7.32.+ -. 0.39% and 12.44.+ -. 0.63%, respectively, and protein adsorption rates to FN were 3.32.+ -. 0.49%, 6.48.+ -. 0.64% and 10.84.+ -. 0.70%, respectively.

Studies have shown that the surface of biological materials with suitable roughness, high hydrophilicity, surface energy and hydroxyl (-OH) groups is favorable for protein adsorption, and the improvement of protein adsorption can promote subsequent cell adhesion and diffusion. In the present invention, the adsorption amount of proteins (BSA and FN) on PT surface is significantly increased compared TO PEEK due TO the exposure of part of TO submicron particles on the surface. In addition, PMT adsorption to proteins (BSA and FN) is further enhanced by the higher biological activity of MTO particles, exposed to the material surface, showing the potential to induce cellular responses.

Based on the effects, the PMT is adopted for further modification and characterization in the follow-up process.

2. FTIR and XRD analysis

The FTIR spectra of part (a) of fig. 5 are TO, PEEK, PMT and PTF. Analysis of functional groups of materials using FTIR, detection wavenumber ranges from 2000 to 650cm ^-1 . For TO, the stretching vibration peak of Ta-O-Ta is 930cm ^-1 The in-plane bending vibration peak of Ta-OH is 1638cm ^-1 . Characteristic peaks of PEEK and MTO were found in PMT, demonstrating successful compounding of the two materials. After the femtosecond laser treatment, the characteristic peak intensity of PEEK in the PTF was significantly reduced.

The XRD patterns of part (b) of FIG. 5 are MTO, PEEK, PMT and PTF. XRD was used to analyze the phase composition and crystalline state of the material, with the degree of detection ranging from 10 to 70. Characteristic peak position of MTO particle synthesized by sol-gel method corresponds to beta phase Ta of orthorhombic system ₂ O ₅ . Characteristic peaks of PEEK and tantalum oxide were found in PMT, demonstrating successful compounding of the two materials.

3. XPS analysis

XPS is used for analyzing the element composition and the functional group of the material, and the detection binding energy range is 0-800 eV.

The XPS high-resolution spectra of the C1s peaks of the PMT and PTF are shown in FIG. 6 (a) and FIG. 6 (b). For PMT, the peak for C-C/C-H (92.44 at%) appears at 284.60eV, while the peaks for C-O (5.43 at%) and c=o (2.13 at%) appear at 286.80eV and 288.00eV, respectively. For PTF, the peak intensities of C-O (20.36 at%) and c=o (3.41 at%) were significantly higher than PMT, the peak intensity of C-C/C-H (74.12 at%) was significantly lower than PMT, and the peak of C (2.11 at%) appeared at 284.40eV, and the peak intensity was weaker.

The XPS high-resolution spectra of the Ta4f peaks of FIG. 6, part (c) and FIG. 6, part (d) are PMT and PTF. For PMT, ta4f _7/2 The peak at (56.42 at%) appears at 25.95eV, and Ta4f _5/2 The peak at (43.58 at%) appears at 27.75eV, with Ta ₂ O ₅ Corresponding to the peak of (c). For PTF, ta4f _7/2 (56.11 at%) and Ta4f _5/2 The peaks at (43.89 at%) appear at 25.95eV and 27.75eV, respectively, indicating no change in the valence state of Ta element compared to PMT.

4. SEM observation and EDS analysis

Adhering the material on conductive adhesive, spraying metal for 40 seconds, observing the microscopic morphology of the material surface by using SEM, and analyzing the element composition, content and distribution of the material surface by using EDS.

Fig. 7 is an SEM image of PEEK, PMT, and PTF surfaces, and EDS spectra. As can be seen from the SEM images, PEEK has a flat, uniform surface (part (a) of fig. 7, part (b) of fig. 7, and part (c) of fig. 7), and the PMT surface presents uniformly distributed MTO particles compared to PEEK, the particles being spherical (about 200nm in size) (part (e) of fig. 7, part (f) of fig. 7, and part (g) of fig. 7). After the femtosecond laser treatment, the PTF surface exposed a large amount of MTO particles, and the MTO particle surface had a "nano petal" structure due to high temperature ablation, the size was about 20nm (part (i) of FIG. 7, part (j) of FIG. 7, and part (k) of FIG. 7).

It can be seen from the EDS spectrum that (part (d) of fig. 7, part (h) of fig. 7, part (l) of fig. 7), C, O element was detected on the surface of each of the three materials, and that characteristic peaks of Ta element were present on the surfaces of PMT and PTF. After femtosecond laser treatment, the peak intensity of Ta element on the surface of the PTF is obviously improved compared with PMT.

FIG. 8 is an EDS elemental plane distribution image of PEEK, PMT, and PTF surfaces. As can be seen from part (a) of fig. 8, part (b) of fig. 8, part (c) of fig. 8, part (d) of fig. 8, part (e) of fig. 8, part (f) of fig. 8, part (g) of fig. 8, part (h) of fig. 8, and part (i) of fig. 8, the C, O and Ta elements have a uniform distribution on all material surfaces. The PEEK surface has a more distributed C element and less distribution on the PMT and PTF surfaces blended with MTO particles. The distribution of Ta elements is detected on the surfaces of the PMT and the PTF, and the distribution of Ta elements on the surface of the PTF after the femtosecond laser treatment is obviously increased compared with that of the PMT.

5. Analysis of surface roughness

The surface of the dental implant material is the interface that is in direct contact with the tissue after implantation, and the surface properties of the dental implant material play a critical role in regulating the cell/tissue response. The two-dimensional and three-dimensional topography of the material surface was observed using LCM and AFM, and the micro/nano roughness (Ra) of the material surface was measured.

Part (a) to part (f) of fig. 9 are LCM images of PEEK, PMT, and PTF surfaces. As can be seen from the figure, PEEK has a smooth and even surface and PMT surface roughness is improved after blending the MTO particles. After the femtosecond laser treatment, the PTF surface roughness was significantly improved and pit-like structures were present. The micrometer roughness (Ra) of PEEK, PMT and PTF surfaces were 1.63.+ -. 0.07. Mu.m, 2.29.+ -. 0.10. Mu.m, and 5.40.+ -. 0.20. Mu.m, respectively.

Parts (g) of fig. 9, part (h) of fig. 9, and part (i) of fig. 9 are AFM images of PEEK, PMT, and PTF surfaces. As can be seen from the figure, the PEEK surface is smoother and flatter. After blending the MTO particles, the surface appeared to have a nano-peak-like structure and was coarser. After the femtosecond laser treatment, the peak-like structure of the PTF surface is increased and the roughness is remarkably improved. The nano roughness (Ra) of PEEK, PMT and PTF surfaces were 10.42.+ -. 0.39nm, 18.66.+ -. 0.73nm and 29.24.+ -. 0.41nm, respectively.

In summary, PMT surfaces appeared to be rougher and exposed a large number of MTO particles compared to PEEK with smooth surfaces; compared with PMT, the surface roughness of PTF is significantly improved, more MTO particles are exposed on the surface, and the surface of MTO particles forms a nano petal structure. The result is probably that due to the high-temperature ablation effect of the femtosecond laser, the local high temperature generated by the femtosecond laser not only ablates and carbonizes PEEK, MTO particles in the material are exposed, and the ablation effect etches a nanometer petal structure on the surface of the MTO particles, so that the micrometer and nanometer roughness of the surface of the material are greatly improved.

6. Contact angle, surface energy and protein adsorption analysis

Part (a) of fig. 10 and part (b) of fig. 10 are water and diiodomethane contact angles of PEEK, PMT, and PTF surfaces. As can be seen from the figure, the water/diiodomethane contact angle of the PTF surface was significantly reduced from PMT after femtosecond laser treatment. The water contact angles of the PEEK, PMT, and PTF surfaces were 83.25 ±0.75°, 66.50±1.50° and 22.00±0.50°, respectively, and the diiodomethane contact angles of the surfaces were 70.00±2.00°, 50.50±1.00° and 8.75±0.25°, respectively.

Part (c) of fig. 10 is the surface energy of PEEK, PMT, and PTF. After the femtosecond laser treatment, the surface energy of the PTF is significantly improved compared with PEEK and PMT. PEEK, PMT and PTF have surface energies of 27.246 + -4.563 mJ/m, respectively ² 、41.598±3.250mJ/m ² And 71.962.+ -. 0.313mJ/m ² 。

In summary, the hydrophilicity and surface energy of the PTF are further improved compared with PMT, probably because a large number of MTO particles are exposed from the interior of the material under the high temperature ablation effect of the femtosecond laser, and the surfaces of the MTO particles form a nano petal structure after the femtosecond laser treatment, so that Ta-OH is easier to form on the surfaces, and hydrogen bonds are further formed with water molecules, so that the PTF surface shows stronger hydrophilicity and surface energy.

FIG. 10 (d) shows the adsorption results of PEEK, PMT and PTF on both BSA and FN proteins. The protein adsorption capacity of the PTF was significantly further improved after femtosecond laser treatment. Protein adsorption rates of PEEK, PMT and PTF to BSA were 3.60.+ -. 0.96%, 11.20.+ -. 1.00% and 25.20.+ -. 1.24%, respectively, and protein adsorption rates to FN were 3.00.+ -. 0.78%, 9.40.+ -. 0.98% and 22.60.+ -. 1.20%, respectively.

In the present invention, since more MTO particles are exposed to the surface of the material and the particle surface forms a nano petal structure, the adsorption of PTF to proteins (BSA and FN) is further enhanced. In conclusion, the PTF with the micro-nano structure on the surface has good adsorption capacity to proteins, and shows potential of inducing cell response.

Effect example 3 drug loading experiments

(1) Drug loading

EGCG (95%, sigma, usa) was dissolved in PBS buffer to make EGCG solution, and PMT and PTF materials were placed in 12-well plates, with equal volumes of EGCG solution added to each well. The entire EGCG load experiment was performed in a thermostatted shaker and maintained at 37 ℃. The material was removed at different time points, the OD of the remaining solution was measured by an enzyme-labeled instrument, and the OD of EGCG solution was measured. The loading of EGCG on the material surface was calculated by the following formula:

where C represents the concentration of EGCG solution, V represents the volume of EGCG solution, ODE represents the OD of EGCG solution, ODR represents the OD of the remaining solution, and M represents the initial mass of material.

(2) Drug release

Equal volumes of EGCG solution were added to the PMT and PTF material surfaces, and then naturally dried at room temperature to obtain PMT (PTE) and PTF (PTFE) materials loaded with equal mass EGCG. The PTE and PTFE materials were then placed in a 12-well plate and an equal volume of PBS buffer was added to each well. The entire EGCG release experiment was performed in a thermostatted shaker and maintained at 37 ℃. Supernatants were removed at various time points and OD values (optical density) of the supernatants were measured by a microplate reader. The cumulative release rate of EGCG from the material surface was calculated by the following formula:

Where C represents the concentration of EGCG solution, V represents the volume of PBS buffer, ODS represents the OD of the supernatant, ODE represents the OD of EGCG solution, and M represents the mass of EGCG added.

(3) Experimental results

After drug loading experiments, FTIR was used (detection wavenumber range 4000-650cm ^-1 ) And XRD (detection degree range of 10-60 DEG) for analysis of the material.

Part (a) of fig. 11 shows FTIR spectra of EGCG, PTE and PTFE. For EGCG, the stretching vibration peak of Ar-OH is 3372cm ^-1 The C=O and the stretching vibration peak of the benzene ring are 1696cm respectively ^-1 And 1458cm ^-1 . Characteristic peaks of EGCG are observed in both PTE and PTFE, and the strength of the characteristic peaks of EGCG in PTFE is higher than that of PTE.

Part (b) of fig. 11 is the XRD pattern of EGCG, PTE and PTFE. From the figure, it can be seen that the characteristic peak positions of EGCG are 12.1 °, 15.6 °, 17.0 °, 19.5 °, 20.7 °, 21.4 °, 24.5 °, 25.9 °, 28.1 °, 28.8 °, 29.5 ° and 37.0 °. Furthermore, the characteristic peak intensity of EGCG in PTFE is higher than PTE.

Part (c) of fig. 11 is the loading of PTE and PTFE to EGCG. The EGCG loading efficiency of PTE in the first 12 days is weak, and the final EGCG loading amount is 2.01+/-0.79 mg/g. PTFE exhibits higher EGCG loading efficiency over the whole drug loading period compared to PTE, with a final EGCG loading of 5.73+ -0.08 mg/g.

Part (d) of fig. 11 is the cumulative release rate of PTE and PTFE for EGCG. PTE showed EGCG burst release behavior within the initial 7 days, with a release rate up to 70.61 ±0.79% and an accumulated release rate up to 71.97±1.22% after 30 days. PTFE shows slow sustained EGCG release behavior and cumulative release rates after 30 days reach 68.51 ±1.42%.

Effect example 4 in vitro antibacterial experiment

Bacteria easily attack the surface of dental implant materials and form permanent implants, form stable bacterial films through continuous proliferation, release inflammatory factors to cause infection, and finally cause implantation failure. Therefore, as a dental implant material, a function of having an antibacterial and inflammatory reaction reducing function on the surface is important for the dental implant material. According to the result of the drug load experiment, four groups of materials of PEEK, PMT, PTF and PTFE are optimized for in-vitro antibacterial experiment, and the specific steps are as follows:

first, the surface of the material was wiped clean with cotton wool containing 75% alcohol and dried in the air. Then collecting a small amount of bacteria from the activated slant strain on an ultra clean bench using a sterile inoculating loopDiluted with physiological saline and arranged as a uniform bacterial suspension (2 to 5X 10 ⁶ CFU/mL). 50. Mu.L of the bacterial suspension was pipetted onto the surface of the material, and the material was then placed in a bacterial incubator for incubation for 24 hours (temperature 37 ℃ C., relative humidity) >90%). Finally, bacterial liquid on the surface of the material is washed off and evenly coated on an agar plate, and the agar plate is placed in a bacterial incubator again for culturing for 24 hours, and the number of bacterial colonies on the agar plate is photographed and counted. The bacteriostasis rate of the material was calculated by using a blank agar plate coated with the original bacterial suspension as a control group by the following formula:

Antibacterial ratio(％)＝(M-N)/M×100％

wherein M represents the colony count of a blank agar plate, and N represents the colony count of a plating and material co-culture bacteria liquid agar plate.

FIG. 12 (a) is a photograph showing the bacteria isolated from the PTFE surface after PEEK, PMT, PTF and 24 hours of culturing on an agar plate. As can be seen from the figure, the PEEK group has a large number of colonies on both E.coli and S.aureus agar plates, demonstrating that PEEK does not possess antimicrobial activity. After blending the MTO particles, the colony count on the agar plates of the PMT group was slightly reduced, demonstrating that the MTO particles had antibacterial activity. After femtosecond laser treatment, the colony count on the agar plates of the PTF group was not significantly different from that of the PMT group. After EGCG is loaded, the colony number on the agar plate of the PTFE group is obviously reduced, and the EGCG medicine with the surface load has stronger antibacterial performance.

The bacteriostasis rates of the materials on e.coli and s.aureus are shown in fig. 12 (b) and fig. 12 (c), respectively. According to quantitative analysis, the bacteriostasis rates of PEEK, PMT, PTF and PTFE on E.coli are 3.17+/-1.01%, 9.83+/-2.26%, 14.17+/-1.93% and 100%, respectively, and the bacteriostasis rates on S.aureus are 4.42+/-1.36%, 11.36+/-1.99%, 15.95+/-2.44% and 100%, respectively.

In conclusion, PEEK, PMT and PTF have no significant inhibition of both e.coli and s.aureus, showing weak antibacterial properties. However, after EGCG loading, PTFE showed strong inhibition on both E.coli and S.aureus, showing strong antibacterial properties. From a mechanism point of view, ar-OH in EGCG can react with nitrogen atoms in basic amino acid side chains and further intercalate into bacteria, thereby inhibiting the functions of bacterial cell membrane proteins. In addition, EGCG is negatively charged and is capable of electrostatic repulsion with the bacterial cell walls, which are also negatively charged, thereby impeding bacterial adhesion. Therefore, PTFE having a good antibacterial activity can effectively inhibit initial adhesion of bacteria and formation of bacterial films as a dental implant material.

Effect example 5 in vitro cell experiment

The good proliferation behavior of cells on the surface of dental implant material plays a key role in subsequent cell migration and differentiation. Cell culture:

the influence of PEEK, PMT, PTF and PTFE four groups of materials on osteoblasts is evaluated by using rat bone marrow mesenchymal stem cells (BMSC), and primary cells are separated from rat femur bone marrow cavities on an ultra-clean bench. Cells were cultured in alpha-MEM medium (Gibco, thermo Fisher Scientific, U.S.) containing 10vt% fetal bovine serum and 1vt% penicillin/streptomycin. The culture medium is placed in a cell culture box, wherein CO ₂ The concentration was 5%, the humidity was 100% and the temperature was 37 ℃. The medium was changed every 3 days and cells cultured to 3-5 passages were selected for subsequent experiments.

Furthermore, gingival Epithelial Cells (GECs) were selected to evaluate the effect of the material on gingival cells, and primary GECs (medical college, shanghai university of transportation) were cultured in DMEM medium (Gibco, thermo Fisher Scientific, usa) containing 10vt% fetal bovine serum, 1vt% penicillin/streptomycin. The culture medium is placed in a cell culture box, wherein CO ₂ The concentration was 5%, the humidity was 100% and the temperature was 37 ℃. The medium was changed every 3 days and cells cultured to 2-4 passages were selected for subsequent experiments. In addition, bone marrow derived macrophages (BMM) were selected to evaluate the effect of the material on osteoclasts. Dislocation was sacrificed after 4 weeks of C57BL/6 mice were anesthetized on an ultra clean bench, femur and tibia were removed and bone marrow cavity was flushed with PBS buffer until the flushed PBS buffer was clear from red. The rinse solution was collected for centrifugation, then the supernatant was discarded and 5 volumes of red blood cell lysate were added for lysis in ice for 10 minutes. The supernatant was then centrifuged again, discarded and the cells were cultured in a medium containing 10vt% fetal bovine serum, 1vt% penicillin/streptomycin and 50ng/mL RANKL (Gibco, Thermo Fisher Scientific, usa). The culture medium is placed in a cell culture box, wherein CO ₂ The concentration was 5%, the humidity was 100%, the temperature was 37℃and 4 days later adherent cells were selected for subsequent experiments.

1. Cell adhesion

The material was co-cultured with BMSC for 12, 24, 48 hours, and the adhesion and spreading morphology of the cells on the surface of the material was observed using SEM. In addition, the material was co-cultured with GEC for 6, 12, 24 hours, the adhesion and spreading morphology of the material surface cells was observed using SEM, and the adhesion of the material surface cells was quantitatively analyzed by CCK-8 detection.

Fig. 13 is an SEM image of BMSCs after 12 and 48 hours of surface culture in PEEK, PMT, PTF and PTFE. In part (a) of fig. 13 and part (e) of fig. 13, it can be seen that the PEEK surface cell adhesion state is poor and spherical. In fig. 13 (b) and fig. 10 (f), PMT surface cell adhesion status was significantly better than PEEK, the number was significantly increased and filopodia were protruded after 48 hours. In part (c) of fig. 13 and part (g) of fig. 13, the PTF surface cell adhesion state was better after the femtosecond laser treatment, the cells exhibited a flat adhesion morphology, and the spreading area was further increased than PMT. In part (d) of fig. 13 and part (h) of fig. 13, after EGCG loading, the cell was further promoted in the PTFE surface adhesion state, and after 48 hours, the cell covered most of the material surface.

FIG. 14 is an SEM image of GEC after 6 and 24 hours of surface culture at PEEK, PT, PTF and PTFE. In part (a) of fig. 14 and part (e) of fig. 14, PEEK surface cells exhibit poor adhesion, and are spherical. In FIG. 14 (b) and FIG. 14 (f), PMT had significantly better cell adhesion than PEEK, increased numbers and filopodia extension throughout the culture period. In fig. 14 (c) and 14 (g), the cell adhesion state on the PTF surface was better after the femtosecond laser treatment, and the cell spreading area was further increased than PMT. In part (d) of fig. 14 and part (h) of fig. 14, after EGCG is loaded, the adhesion state of cells on the PTFE surface is further improved, and after 24 hours, the entire material surface is covered.

2. Cell proliferation and osteogenic differentiation

(1) Cell proliferation: the material was co-cultured with BMSC for 1, 3, 7 days, proliferation of material surface cells and cell nucleus/skeleton were observed by CLSM, and proliferation of material surface cells was quantitatively analyzed by CCK-8 detection. In addition, the material was co-cultured with GEC for 1, 3, 7 days, proliferation of material surface cells and nuclei/skeletons were observed by CLSM, and proliferation of material surface cells was quantitatively analyzed by CCK-8 detection.

(2) ALP activity: the material was co-cultured with BMSCs for 7, 10, 14 days and cells were quantitatively assayed for early osteogenic differentiation capacity by ALP activity assay.

(3) Cell osteoclast differentiation: first, the material extract was co-cultured with BMM for 4 days, and then the medium was aspirated using a pipette. Cells were then fixed with 2.5% glutaraldehyde solution for 2 hours and washed 3 times with PBS buffer. Then, cells were stained with a tartrate-resistant acid phosphatase (TRAP) staining kit (TaKaRa, japan), and osteoclasts (nuclei ≡3) were observed with a binocular biomicroscope and photographed, and the osteoclast differentiation was qualitatively analyzed. Finally, the Image-Pro Plus6.0 software matched to the microscope was used to quantitatively analyze the number and area of osteoclasts in the Image.

(4) Gene expression: the material was co-cultured with BMSCs for 7, 14 days, and the intracellular osteogenic related genes (ALP, colI, OPN and OCN) were quantitatively analyzed by RT-qPCR. In addition, the material was co-cultured with BMM for 4 and 7 days, and the expression of intracellular osteoclast-related genes (TRAP, cathepsin K (CTSK), osteoclast inhibitor (OPG) and nuclear factor κb receptor activator ligand (RANKL)) was quantitatively analyzed by RT-qPCR. Finally, the material was co-cultured with BMM for 1, 3 days, and the expression of intracellular inflammatory factor related genes (interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha)) was quantitatively analyzed by RT-qPCR. The following table shows the primer sequences of the osteoclast-related gene and inflammatory factor-related gene used.

Primer sequences for RT-qPCR

(5) Protein expression: the material was co-cultured with BMSCs for 7 and 14 days, and the expression of intracellular inflammatory factor related proteins (IL-6 and TNF-alpha) was qualitatively analyzed by Western blot.

(6) Experimental results

FIG. 15 is a CLSM image of BMSCs after 1, 3 and 7 days of surface culture with PEEK, PMT, PTF and PTFE. The surface cell numbers of PEEK (part (a) of fig. 15, part (e) of fig. 15, and part (i) of fig. 15) were not significantly changed throughout the culture period, showing a weaker proliferation state, whereas the surface cell proliferation state of PMT (part (b) of fig. 15, part (f) of fig. 15, and part (j) of fig. 15) was significantly improved over PEEK. After femtosecond laser treatment, the cells on the surface of the PTF (part (c) of FIG. 15, part (g) of FIG. 15 and part (k) of FIG. 15) proliferated significantly, and the cell number was significantly increased, and cell pseudopodia and skeletons were significantly spread on the surface of the material after 7 days. After loading EGCG, the cell proliferation state of the PTFE surface (part (d) of fig. 15, part (h) of fig. 15, and part (l) of fig. 15) was further improved, and the cytoskeleton almost completely covered the material surface after 7 days.

FIG. 16 is a CLSM image of GEC after 1, 3 and 7 days of surface culture on PEEK, PMT, PTF and PTFE. As can be seen from the figure, the GEC takes on a polygonal morphology and is smaller than the BMSC. The surface cell numbers of PEEK (part (a) of fig. 16, part (e) of fig. 16, and part (i) of fig. 16) were not significantly changed throughout the culture period, showing a weak proliferation state. Compared with PEEK, PMT (part (b) of fig. 16, part (f) of fig. 16, and part (j) of fig. 16) surface cell proliferation status and cell number of the blended MTO particles were significantly improved over PEEK. After femtosecond laser treatment, the PTF (part (c) of FIG. 16, part (g) of FIG. 16 and part (k) of FIG. 16) surface cells proliferated significantly, and the cell number was significantly increased, cell pseudopodia and skeleton spread significantly throughout the culture period. After EGCG loading, the cell proliferation state on the surface of PTFE (part (d) of FIG. 16, part (h) of FIG. 16 and part (l) of FIG. 16) was significantly promoted, and after 7 days, the cells covered almost the entire material surface.

From the above, it is clear that the proliferation of BMSC and GEC on the PEEK surface is the weakest, and that the proliferation of BMSC and GEC does not significantly change with time due to the bio-inertness of PEEK. Proliferation of PMT surface BMSC and GEC was significantly enhanced compared to PEEK, and PTF surface BMSC and GEC proliferation was further enhanced due to more MTO particle exposure and the nano petal structure of the particle surface. In addition, the proliferation of BMSCs and GECs is strongest on PTFE surfaces compared to PTF, since EGCG is able to stimulate osteoblast proliferation and differentiation by activating the NF-. Kappa.B pathway, thus promoting new bone formation. Thus, in comparison to PTF, the PTFE surface is promoted by the sustained release of EGCG, which promotes cell proliferation of BMSC and GEC on the surface.

The OD values of BMSCs after 1, 3 and 7 days of surface culture with PEEK, PMT, PTF and PTFE are shown in part (a) of FIG. 17. Along with the extension of the culture time, the OD value of PEEK surface cells does not change obviously, but the OD value of PMT surface cells is improved obviously, which indicates that the blending of MTO particles promotes the proliferation of cells. In addition, after femtosecond laser treatment, the OD value of PTF surface cells is obviously improved compared with PMT, and the OD value is obviously higher than PMT and PEEK in the whole culture period. After EGCG loading, the OD of PTFE surface cells was significantly higher than PTF throughout the culture period. Details are shown in Table 1.

TABLE 1

Group (BMSC)	PEEK	PMT	PTF	PTFE
					OD value (1 day)	0.25±0.03	0.40±0.04	0.42±0.06	0.51±0.05
OD value (3 day)	0.28±0.03	0.51±0.03	0.62±0.04	0.78±0.03
					OD value (7 day)	0.31±0.02	0.58±0.02	0.78±0.02	1.01±0.03

FIG. 17 part (b) shows ALP activity of BMSCs after culturing on PEEK, PMT, PTF and PTFE surfaces for 7, 10 and 14 days. ALP of PEEK surface cells does not change significantly during the whole culture period, while ALP activity of PMT surface cells blended with MTO particles is significantly improved compared with PEEK. The ALP activity of PTF surface cells slightly increased after femtosecond laser treatment, and the ALP activity of PTFE surface cells significantly increased compared to PTF after EGCG loading. Details are shown in Table 2.

TABLE 2

Group (BMSC)	PEEK	PT	PTF	PTFE
					ALP Activity (7 day)	0.28±0.05	0.73±0.03	0.77±0.03	1.24±0.06
ALP Activity (10 day)	0.36±0.05	1.15±0.05	1.32±0.05	2.20±0.04
					ALP Activity (14 day)	0.61±0.06	1.68±0.05	2.01±0.05	3.08±0.04

Part (c) of FIG. 17 is the cell adhesion rate of GEC after 6, 12 and 24 hours of surface culture at PEEK, PMT, PTF and PTFE. Along with the extension of the culture time, the adhesion rate of PEEK surface cells is not obviously changed, and the adhesion rate of PMT surface cells is obviously improved compared with PEEK. After femtosecond laser treatment, the cell adhesion rate of the PTF surface was significantly higher than that of PMT, and significantly higher than that of PMT and PEEK throughout the whole culture period. After EGCG loading, the PTFE surface cell adhesion rate was significantly higher than that of PTF throughout the whole culture period. Details are shown in Table 3.

TABLE 3 Table 3

Group (GEC)	PEEK	PMT	PTF	PTFE
					Cell adhesion Rate (6 Hour)	5±1％	18±2％	30±2％	32±2％
Cell adhesion Rate (12 Hour)	9±1％	33±3％	54±2％	62±3％
					Cell adhesion Rate (24 Hour)	14±2％	53±2％	69±3％	87±2％

FIG. 17 (d) shows the OD of the GEC after 1, 3 and 7 days of surface culture with PEEK, PMT, PTF and PTFE. The OD of PEEK surface cells did not change significantly throughout the culture period, whereas PMT surface cells had higher OD than PEEK. In addition, after femtosecond laser treatment, the OD value of PTF surface cells is further improved than PMT. After EGCG is loaded, the OD value of PTFE surface cells is obviously improved compared with PTF in the whole culture period. Details are shown in Table 4.

TABLE 4 Table 4

Group (GEC)	PEEK	PMT	PTF	PTFE
					OD value (1 day)	0.25±0.04	0.32±0.03	0.53±0.05	0.68±0.05
OD value (3 day)	0.29±0.03	0.52±0.02	0.63±0.04	0.88±0.04
					OD value (7 day)	0.33±0.03	0.64±0.06	0.78±0.03	1.13±0.02

FIG. 18 shows the expression of bone formation related genes ALP (FIG. 18 (a)), colI (FIG. 18 (b)), OPN (FIG. 18 (c)) and OCN (FIG. 18 (d)) after BMSC was surface-cultured for 7 and 14 days on PEEK, PMT, PTF and PTFE. As can be seen from the figure, the PEEK surface has no promoting effect on cell bone-related gene expression. After blending the MTO particles, PMT surface promoted cell osteogenesis-related gene expression, and ALP and OCN genes were expressed significantly higher than PEEK. The PTF surface further promoted the osteogenic related gene expression after femtosecond laser treatment, and PTFE promoted the osteogenic related gene expression most strongly after EGCG loading, i.e., significantly promoted cell osteogenic differentiation. Details are shown in Table 5.

TABLE 5

Osteoblast differentiation is generally manifested as an increase in ALP activity and is associated with an early stage of bone regeneration. From the above, BMSC showed that ALP activity on PEEK was the lowest, indicating that PEEK was not effective in promoting cell osteogenic differentiation. Compared to PEEK, BMSC has higher ALP activity on PMT due to MTO particles on the surface, and significantly enhanced ALP activity on PTF due to more surface exposed more MTO particles and the nano petal structure of the particle surface. Furthermore, the ALP activity of BMSC on PTFE was further increased, indicating that sustained release of EGCG on the PTFE surface further promoted osteogenic differentiation of BMSC.

Bone remodeling is a process in which osteoblasts, which play a key role in bone regeneration, and osteoclasts, which cause bone resorption, are in equilibrium. In the present invention, it can be seen from TRAP staining images after 4 days of co-culture of extracts of PEEK (part (a) of fig. 19), PMT (part (b) of fig. 19), PTF (part (d) of fig. 19) and PTFE (part (e) of fig. 19), PTFE significantly inhibited osteoclast differentiation of BMM, and significantly down-regulated expression of osteoclast-related genes (TRAP, CTSK and RANKL) (osteoclast number (part (c) of fig. 19) and area (part (f) of fig. 19)). FIG. 20 shows the expression of the osteoclast-associated genes TRAP (FIG. 20 part (a)), CTSK (FIG. 20 part (b)), OPG (FIG. 20 part (c)) and RANKL (FIG. 20 part (d)) after 4 and 7 days of BMM surface culture with PEEK, PMT, PTF and PTFE. From the figure, it is clear that the sustained release of EGCG on the PTFE surface enables PTFE to significantly inhibit osteoclast activity, thereby inhibiting bone resorption and promoting new bone regeneration from the point of view of osteogenic-osteoclast balance. See tables 6 and 7 for details.

TABLE 6

	PEEK	PMT	PTF	PTFE
					Number of broken bones	363±12	349±10	340±8	151±10
Area of	86.9±4.1％	77.1±2.8％	72.3±3.2％	20.2±2.9％

TABLE 7

Gene expression	PEEK	PMT	PTF	PTFE
					TRAP(4day)	1.00±0.03	0.92±0.05	0.96±0.07	0.57±0.04
TRAP(7day)	1.94±0.04	1.47±0.05	1.23±0.06	0.46±0.05
					CTSK(4day)	1.00±0.05	0.91±0.09	0.85±0.06	0.65±0.07
CTSK(7day)	2.28±0.06	1.61±0.05	1.59±0.07	0.53±0.06
					OPG(4day)	1.0±0.2	1.8±0.3	2.1±0.2	5.2±0.3
OPG(7day)	1.9±0.3	4.2±0.4	4.8±0.3	7.9±0.3
					RANKL(4day)	1.00±0.04	0.97±0.07	0.91±0.06	0.72±0.06
RANKL(7day)	2.17±0.06	1.62±0.05	1.56±0.07	0.54±0.06

Inflammation is a common condition after implantation of dental implant materials, usually caused by foreign body reactions or bacterial invasion, and inflammatory factors can up-regulate the activity of osteoclasts, causing bone resorption. In the present invention, FIG. 21 shows the expression of inflammatory factor-related genes IL-6 (FIG. 21 part (a)) and TNF-. Alpha. (FIG. 21 part (b)) and Western blot images of inflammatory factor expression (FIG. 21 part (c)) of BMM after 1 and 3 days of surface culture with PEEK, PMT, PTF and PTFE. PTFE significantly inhibits the expression of BMM inflammatory factor-related genes (IL-6, TNF- α) compared to PMT and PTF, and as can also be seen from the Western blot results, PTFE significantly inhibits the expression of inflammatory factors (IL-6, TNF- α). Details are shown in Table 8.

TABLE 8

Gene expression	PEEK	PMT	PTF	PTFE
					IL-6(1day)	1.00±0.06	0.82±0.07	0.87±0.03	0.37±0.05
IL-6(3day)	1.04±0.03	0.71±0.05	0.75±0.04	0.45±0.03
					TNF-α(1day)	1.00±0.00	1.08±0.07	1.12±0.07	0.73±0.05
TNF-α(3day)	2.24±0.07	1.68±0.05	1.37±0.07	0.59±0.05

Effect example 6 in vivo implantation experiments

(1) In vivo implantation experiments:

beagle dogs were selected as experimental animals to create a tooth defect model, and the entire in vivo implantation experimental procedure was approved by the institutional animal ethics committee of Shanghai university of transportation medical college. Four groups of PEEK, PMT, PTF and PTFE materials were gamma-ray radiation sterilized prior to the experiment. First, male healthy beagle dogs with a body weight of 20.0.+ -. 1.5kg and an age of 18 months were selected, and then the beagle dogs were subjected to general anesthesia by means of intravenous injection of 1% pentobarbital sodium solution into the hind limbs at a dose of 80mg/kg. Then, the double-sided mandibular teeth of each dog were removed and ground flat using a rasp, and then sutured. After 2 weeks, the mucoperiosteum was cut and a bone hole having a diameter of 5mm and a depth of 6mm was drilled in the alveolar bone using a medical drill, and then the material was filled into the bone hole, the mucoperiosteum was reset and tightly sutured. After 4 and 12 weeks of implantation, beagle dogs were sacrificed by injection of excess sodium pentobarbital solution, alveolar bone with implant material was removed, washed clean, and then immersed in 2.5% glutaraldehyde solution for preservation.

(2) Histological evaluation

Alveolar bone samples were dehydrated, embedded, sectioned and polished after SR μct analysis, histological sections were stained with VG and hematoxylin-eosin (Hematoxylin Eosin, HE) staining solutions, respectively, and the stained sections were observed by binocular biomicroscopy. Finally, the BIC and gingival contact rate (GIC) of the material in the slice images were quantitatively analyzed using Image-Pro Plus 6.0 software matched to the microscope.

(3) Analysis of biomechanical properties

The push-out strength between the material and the alveolar bone was measured using a universal material testing system. First, the femur specimen was fixed on a test bed of a test system, and then the test force was automatically loaded (loading speed: 5 mm/min). Load-time curves were recorded using the software kit, the peaks in the curves being defined as the push-out strength of the material. In addition, the pull-off strength between the material and gum tissue was measured using a universal material testing system. First, the top and side surfaces of the alveolar bone sample were fixed on a test bed of a test system, respectively, and then the gingival tissue was fixed with a jig and a pulling force (pulling speed: 1 mm/min) was gradually applied. The maximum pulling force at which the interface between the gum tissue and the material completely tears was recorded using the software kit, and the pull-off strength of the material was calculated by measuring the surface area of the material in contact with the gum tissue.

(4) Experimental results

Part (a) of fig. 22 is PEEK, PMT, PTF and a hard tissue section stained image (VG stain) of the implanted region after 4 and 12 weeks of PTFE implantation in the body. During the entire implantation cycle, the new bone composition around PEEK is less and there is significant clearance with the material. More new bone tissue formed around PMT and the gap between material was reduced compared to PEEK. The new bone tissue around the PTF is significantly increased, the gap between the PTF and the material is smaller, and the contact is tighter. EGCG loaded PTFE forms a large amount of new bone around the entire implantation cycle and osseointegration between the material and bone tissue occurs.

Part (b) of fig. 22 is PEEK, PMT, PTF and a hard tissue section staining image (HE staining) of the implanted region after 4 and 12 weeks of PTFE implantation in the body. As can be seen, the gum tissue forms less around PEEK throughout the implant cycle, with significant gaps. PMT significantly promotes formation and attachment of gum tissue compared to PEEK, and PTF after surface femtosecond laser modification further promotes formation and attachment of gum tissue. PTFE significantly promotes the formation of gingival tissue throughout the implant cycle with little to no gap seen between the material and the gingival tissue, forming a gingival seal.

Part (a) of fig. 23 is PEEK, PMT, PTF and bone contact rate (BIC) after PTFE implantation in the body for 4 and 12 weeks. PTFE has significantly higher bone contact rates than PTF, PMT, and PEEK after 4 weeks of implantation, and PTF has significantly higher bone contact rates than PMT and PEEK. After 12 weeks of implantation, the bone contact rates of PTF and PTFE were significantly higher than those of PMT and PEEK, and the bone contact rates exhibited by PTF and PTFE were similar, indicating good osseointegration properties for both materials.

Part (b) of fig. 23 is PEEK, PMT, PTF and gingival contact rate (GIC) after PTFE implantation in the body for 4 and 12 weeks. After 4 weeks of implantation, PTFE has higher gum contact rates than PTF, PMT, and PEEK, and PTF has higher gum contact rates than PMT and PEEK. After 12 weeks of implantation, the gingival contact rates of PTF and PTFE were significantly higher than PMT and PEEK, and the gingival contact rates of PTF and PTFE were similar, showing good binding ability of the material to gingival tissue.

Part (c) of fig. 23 is PEEK, PMT, PTF and Push-out strength (Push-out strength) after 4 and 12 weeks of PTFE implantation in the body. After 4 weeks of implantation, the push-out strength of PTF and PTFE was higher than PMT and PEEK, and the push-out strength of PTFE was higher than PTF. After 12 weeks of implantation, the push-out strength of PTF and PTFE was significantly higher than PMT and PEEK, and the push-out strength of PTFE was significantly higher than PTF. Over time, the push-out strength of the material increased, indicating that the material was more tightly bound to bone tissue.

Part (d) of FIG. 23 is PEEK, PMT, PTF and Pull-off strength (Pull-off strength) after 4 and 12 weeks of PTFE implantation in a body. The results show that PTF and PTFE have significantly higher pull-off strength than PMT and PEEK after 4 weeks of implantation, and that PTFE has higher pull-off strength than PTF. After 12 weeks of implantation, both PTF and PTFE had significantly higher pull-off strength than PMT and PEEK, and PTFE had significantly higher pull-off strength than PTF, indicating the most intimate bond of PTFE to gingival tissue. See Table 9 for details.

TABLE 9

For the dental implant material, factors such as surface chemical composition, micro-nano structure, roughness, hydrophilicity and the like have obvious regulation and control effects on inducing osteoblast/gingival cell response and promoting bone/gingival tissue integration. In the present invention, the osteogenic activity of the implant material was first evaluated by SR μct. At weeks 4 and 12, PEEK had minimal formation of new bone around it due to its bio-inertness. The amount of new bone formation around PMT was significantly increased compared to PEEK, as PMT surface exposed MTP particles with bioactivity. In addition, the amount of new bone formation around the PTF is significantly increased compared with PMT, because the PTF surface exposes a large number of MTO particles with nano petal structure, and the PTF surface shows more excellent in vivo osteogenesis activity. Due to the sustained release of EGCG, the most amount of new bone formation around PTFE was found to have the best osteogenic activity. In addition, the bone/gum tissue integration ability of the material was evaluated by tissue sections. The results show that at weeks 4 and 12, the amount of bone/gum tissue production around PT was significantly increased compared to PEEK and the gap between PMT and surrounding bone/gum tissue was smaller than PEEK. The bone/gum tissue production was higher around PTF and PTFE than PMT and PEEK, and at week 12, PTF and PTFE formed a direct bond with bone/gum tissue with little clearance, showing good bone/gum tissue integration. BIC quantitative analysis showed that PTFE (99.17±2.65%) showed the highest bone contact rate at week 12 compared to PMT (48.26±2.41%) and PTF (83.19 ±2.33%); the GIC quantitative analysis results showed that PTFE (94.36 ±3.22%) showed the highest bone contact rate compared to PMT (46.65 ±1.87%) and PTF (75.24±2.47%) at week 12. The above results suggest that PTFE having a micro-nano structure on the surface and sustained release EGCG has excellent ability to promote regeneration of bone/gum tissue, showing superior bone/gum tissue integration effects compared to PEEK, PMT and PTF.

The surface of a dental implant material is the interface that is in direct contact with surrounding tissue after implantation, and thus the surface characteristics of the material (e.g., chemical composition, micro-nano morphology, roughness, hydrophilicity, etc.) are key factors in determining its surface biological response. The bioactive micro-nano structure can stimulate cell response due to the proper surface roughness and high hydrophilicity, and the protein adsorbed on the surface can provide more binding sites for cell adhesion, so that the subsequent cell functions are activated. In addition, EGCG has biological functions such as antioxidation, anti-tumor, anti-inflammatory and antibacterial as a natural bioactive molecule, and also has promotion effect on proliferation and differentiation of osteoblasts. In the present invention, PTFE promotes not only adhesion, proliferation and osteogenic differentiation of BMSCs, but also adhesion and proliferation of GECs, and significantly inhibits osteoclast differentiation and inflammatory factor expression of BMMs. In addition, PTFE significantly promotes regeneration of and integration with bone/gum tissue, exhibiting good in vivo bioactivity. In conclusion, PTFE with the bioactive micro-nano structure and the surface EGCG sustained release can inhibit bacterial activity in vitro and stimulate BMSC/GEC/BMM cell response, and can promote regeneration and integration of bone/gum tissues in vivo, so that the PTFE has great application potential in the field of tooth defect repair as a tooth implant material.

All the experiments above were performed independently in 5 times, and the raw data for all experiments were processed by the OriginPro 9.0 (origin lab, usa) software and expressed in the form of mean±sd. Comparison of statistical differences between data sets using analysis of variance (ANOVA), p <0.05 indicated significant differences between the data sets obtained from the experiments.

Claims

1. The preparation method of the polyether-ether-ketone-based composite material is characterized by comprising the following steps of: performing hot press molding on the mixture of tantalum oxide and polyether-ether-ketone PEEK;

wherein the tantalum oxide is prepared by any one of the following methods:

2. The method of preparing a polyetheretherketone-based composite as claimed in claim 1, wherein the mass ratio of tantalum oxide to polyetheretherketone PEEK is (5-10): 1, preferably (7-9): 1, for example 8.72:1;

And/or, the hot press molding is to heat the mixture to a molding temperature, keep the temperature, pressurize to a molding pressure, and pressurize to prepare a composite material by press molding;

wherein the molding temperature is preferably 340-370 ℃, such as 355 ℃;

the time of the incubation is preferably 0.2 to 1.5 hours, for example 0.5 hours or 1 hour;

the molding pressure is preferably 2 to 6MPa, for example 4MPa;

the dwell time is preferably 0.2 to 1.5 hours, for example 0.5 hours or 1 hour.

3. The method for preparing a polyether-ether-ketone-based composite material according to claim 1, wherein in the second method, the acidic solution containing the template agent is prepared by dissolving the template agent in the acidic solution and stirring;

and/or, in the second method, the mass-volume ratio of the tantalum salt to the acidic solution containing the template agent is 1g (20-30 mL;

and/or, in the second method, the temperature of stirring is 40-60 ℃;

and/or in the second method, the stirring time is 4-6 hours;

and/or, in the second method, the aging temperature is 50-70 ℃;

and/or, in the second method, the aging time is 2-4 days;

and/or, in the second method, the calcining temperature is 550-650 ℃;

and/or in the second method, the calcination time is 5-7 h;

And/or, in the second method, the particle size of the MTO is 170-300 nm;

and/or, in the second method, the MTO is a mesoporous material; the pore size of the MTO is preferably 9-10 nm.

4. A method of preparing a polyether ether ketone composite material as claimed in claim 3, wherein the templating agent is PEO-PPO-PEO;

and/or the acidic solution is an inorganic acid solution or an organic acid solution; the mineral acid solution is preferably hydrochloric acid solution; when the acid solution is hydrochloric acid solution, the concentration of the hydrochloric acid solution is preferably 1-3 mol/L;

and/or the mass volume ratio of the template agent to the acidic solution is 1g (20-40) mL;

and/or stirring for 5-7 h when the template agent is dissolved in the acidic solution.

5. The method of preparing a polyetheretherketone-based composite material according to claim 1, wherein in method one or method two, the kind of tantalum salt is independently selected from one or more of tantalum ethoxide, tantalum chloride and tantalum oxalate, such as tantalum ethoxide;

and/or, in method one, the organic alcohol is ethylene glycol and/or ethanol;

and/or, in the first method, the mass-volume ratio of the tantalum salt to the organic alcohol is 1g (8-12) mL;

And/or, in the first method, the temperature of the reaction is 70-90 ℃;

and/or, in the first method, the reaction time is 1-3 h;

and/or, in method one, the reaction is carried out under an inert atmosphere;

and/or, in the method one, the solvent is a ketone solvent and/or an alcohol solvent; the ketone solvent is preferably acetone; the alcohol solvent is preferably ethanol;

and/or, in the first method, the mass-volume ratio of the tantalum salt to the solvent is 1g (30-50) mL;

and/or, in the first method, the mass volume ratio of the tantalum salt to the water is 1g (0.3-0.5) mL;

and/or, in the first method, the stirring time is 6-10 h;

and/or, in the first method, the calcining temperature is 600-800 ℃;

and/or, in the first method, the calcination time is 2-4 h;

and/or in the first method, the particle size of the TO is 170-260 nm.

6. The method for preparing a polyether-ether-ketone-based composite material according to claim 1, wherein the surface of the composite material prepared by hot press molding is subjected to femtosecond laser leveling treatment;

preferably, the surface of the composite material subjected to femtosecond laser leveling treatment is loaded with epigallocatechin gallate EGCG.

7. The method for preparing a polyether-ether-ketone composite material according to claim 6, wherein the pulse frequency of the femtosecond laser leveling sweep is 900-1100 Hz;

and/or the pulse width of the femtosecond laser leveling sweep is 100-140 fs;

and/or the pulse power of the femtosecond laser leveling sweep is 30-50 mW;

and/or the single pulse energy of the femtosecond laser leveling sweep is 150-250 mu J;

and/or the scanning speed of the femtosecond laser leveling scanning is 300-500 mu m/s;

and/or the processing environment for the femtosecond laser leveling is typically 1 atm.

8. The method for preparing a polyether-ether-ketone-based composite material according to claim 6, wherein the loading mode is achieved by immersing the femtosecond laser swept composite material in an EGCG aqueous solution;

wherein the concentration of the EGCG water solution is preferably 3-10 mg/mL;

the impregnation mode is preferably oscillation;

the temperature of the impregnation is preferably 20-40 ℃;

the time of the impregnation is preferably 1 to 48 hours.

9. A polyether ether ketone based composite material prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the polyether ether ketone based composite material according to claim 9 for the preparation of bone implant materials/dental implants; preferably, the bone implant material is a large section of bone implant material.