DENTAL IMPLANT WITH NANO BACTERIOSTATIC STRUCTURE RING AT TRANSGINGIVAL PART AND MACHINING METHOD THEREOF

Abstract

Provided are a dental implant with a nano bacteriostatic structure ring at a transgingival part and a machining method thereof. The transgingival part of the dental implant has a three-level micro-nano composite structure, and the three-level micro-nano composite structure endows a surface of the transgingival part of the implant with functions of promoting adhesion and proliferation of a gingival fibroblast and a gingival mesenchymal stem cell and inhibiting adhesion and growth of various oral bacteria. A preparation method thereof comprises: firstly, injecting bioactive, wear-resistant or corrosion-resistant C, N, Ca and P elements into the transgingival part of the dental implant by a plasma injection method; and then, preparing the three-level micro-nano composite structure at a part in which the elements are injected.

Claims

1. A dental implant, wherein a nano bacteriostatic structure ring is prepared on a surface of a transgingival part, a surface of the nano bacteriostatic structure ring has a three-level micro-nano composite structure, the three-level micro-nano composite structure is formed by superposing structures with three levels of sizes, a second-level structure is distributed on a surface of a first-level structure, a third-level structure is distributed on a surface of the second-level structure, and the first-level structure is a micron-level groove structure with a width of 20 μm to 60 μm and a depth of 1 μm to 2 μm; the second-level structure is composed of a stripe with a width of 100 nm to 500 nm and a height of 50 nm to 200 nm, or an array protrusion with a height of 50 nm to 200 nm; and the third-level structure is composed of a nano particle, a nano rod and a nano cone with a sub-micron or nano scale.

2. A machining method of the dental implant according to claim 1, wherein the three-level micro-nano composite structure is machined by a pulse laser.

3. The machining method according to claim 2, wherein before the three-level micro-nano composite structure is machined, C, N, Ca and P elements are injected into a transgingival part of an implant tooth by a plasma injection method.

4. The machining method according to claim 2, wherein machining parameters of the pulse laser are: a laser frequency of 1 kHz to 10 kHz, pulse energy of 3,000 uJ to 8,000 uJ, a light spot diameter of 50 μm to 60 μm, a line spacing of 40 μm to 60 μm, and a scanning speed of 10 mm/s to 20 mm/s.

5. The machining method according to claim 3, wherein a depth of plasma injection is 500 nm to 2,000 nm.

Description

DESCRIPTION OF THE DRAWINGS

[0014] FIG. 1a, FIG. 1B and FIG. 1c are schematic diagrams of an implant, a plasma processing region on a surface of the implant and a femtosecond laser processing region on the surface of the implant in Embodiment 1 respectively;

[0015] FIG. 2 shows a micro-morphology of a surface of the femtosecond laser machining region in Embodiment 1, wherein a, b and c are diagrams of first-level, second-level and third-level structures respectively;

[0016] FIG. 3 is a composition analysis diagram of the surface of the femtosecond laser machining region in Embodiment 1;

[0017] FIG. 4 is a comparative diagram of adhesion conditions of a gingival fibroblast and a gingival mesenchymal stem cell on four surfaces in Comparative Example 1;

[0018] FIG. 5 is a comparative diagram of adhesion conditions of Escherichia coli and Staphylococcus aureus on four surfaces in Embodiment 1;

[0019] FIG. 6 is a comparative diagram of wear resistance performances of two surfaces in Comparative Example 2; and

[0020] FIG. 7 is a comparative diagram of adhesion conditions of oral bacteria on two surfaces in Comparative Example 3, wherein a to g are alpha streptococcus, anaerobic streptococcus, Staphylococcus epidermidis, Neisseria, lactobacillus, spirochete and candida respectively.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

[0021] The present invention is further described in detail hereinafter with reference to specific embodiments.

Embodiment 1

[0022] In this embodiment, plasma injection and femtosecond laser machining methods were used to prepare a bacteriostatic bioactive surface on a surface of a transgingival part of an implant.

[0023] Specific steps and process were as follows.

[0024] (1) Firstly, C, N, Ca and P elements were injected into the surface of the implant by the plasma injection method. Apure Ti implant was placed in an atmosphere containing the C, N, Ca and P elements, and a composition layer containing the four elements above was prepared in a range from the surface of the implant to a depth of 500 nm from the surface by a metal plasma immersion injection technology. FIG. 1a was a schematic diagram of the implant. FIG. 1B was a schematic diagram of a processing part of the plasma injection method, and a dark part was a plasma injection region.

[0025] (2) A size of the implant and a size of a machining position were measured. The implant used in this case had a length of 8 mm, wherein the transgingival part had a length of 3 mm, a bone binding part had a diameter of 3.5 mm, and the transgingival part had a widest neck part of 4.8 mm.

[0026] (3) The implant was fixed on a four-axis translation table. A laser device was turned on, and laser parameters were adjusted. A position of a light spot was adjusted, so that the light spot of the laser was irradiated on an initial position of the transgingival part. The machining process was that: the surface of the transgingival part was machined by a laser frequency of 1 kHz, pulse energy of 8,000 uJ, a light spot diameter of 50 μm, a line spacing of 50 μm and a scanning speed of 10 mm/s, a rotating axis was rotated by one circle during machining, a Y axis was translated by 0.1 mm, and an X axis was moved at the same time to ensure that a distance from a focal point of the laser to a surface of a sample remained unchanged, so that the surface of the whole transgingival part was covered repeatedly, and a three-level micro-nano composite structure was synchronously induced and generated on the surface of the transgingival part.

[0027] FIG. 2 showed a morphology of the surface of the part after machining, wherein a, b and c respectively showed a first-level structure, a second-level structure and a third-level structure in the three-level micro-nano composite structure. FIG. 3 showed chemical composition of the surface of the transgingival part of the implant obtained. FIG. 1c showed the laser machining region.

[0028] (4) The machined implant was taken down, cleaned, sterilized and packaged.

Comparative Example 1

[0029] In this comparative example, influences of a pure titanium polished surface, a polished titanium surface with a bioactive component, a titanium surface with a three-level micro-nano composite structure and a titanium surface with the bioactive component and the micro-nano composite structure on adhesion and growth of various cells and bacteria on the surfaces were compared through an in-vitro cell experiment and a bacteriostatic adhesion experiment.

[0030] A preparation method of the pure titanium polished surface (surface a) was that: a pure titanium surface was polished by a mechanical polishing method. A preparation method of the polished titanium surface with the bioactive component (surface b) was that: firstly, the polished pure titanium surface was prepared by the mechanical polishing method, and subsequently, Ca, P, C and N elements were injected into the surface by a metal plasma immersion injection technology, wherein parameters of the plasma injection process were the same as those used in Embodiment 1. A preparation method of the titanium surface with the three-level micro-nano composite structure (surface c) was that: firstly, the polished titanium surface was prepared by the mechanical polishing method, and subsequently, the micro-nano composite structure was prepared on the surface by a femtosecond laser machining method, wherein the femtosecond laser machining process was the same as that in Embodiment 1. A preparation method of the titanium surface with the bioactive component and the micro-nano composite structure (surface d) was that: the titanium surface containing Ca, P, C and N elements was prepared by the same method as that of the surface b, and subsequently, the micro-nano structure was prepared on the surface by the femtosecond laser machining method, wherein the preparation process was the same as the femtosecond laser machining process in Embodiment 1.

[0031] Firstly, an adhesion experiment of a gingival fibroblast and a gingival mesenchymal stem cell was carried out. 40 μl of 5×10.sup.4/ml gingival fibroblast suspension and 40 μl of 5×10.sup.4/ml gingival mesenchymal stem cell suspension were dropwise added on surfaces of four samples respectively, and cultured for 24 hours respectively, then the surfaces were washed with PBS, and numbers of cells adhered to the surfaces of the four samples were compared by a CCK-8 method.

[0032] FIG. 4 showed statistical results of OD values of the surfaces of the four samples. The OD value of the surface a was the lowest, and the OD values of the surfaces b and c were higher than that of the surface a, which indicated that the adhesion of the gingival fibroblast and the gingival mesenchymal stem cell could be promoted by injecting the bioactive component into the pure titanium surface by the plasma injection method and preparing the micro-nano composite structure on the surface by the femtosecond laser machining method. The OD value of the surface d was significantly higher than those of the other three surfaces, which indicated that the adhesion and proliferation of the two cells could be significantly promoted by surface active component injection combined with micro-nano composite structure preparation.

[0033] Subsequently, an adhesion experiment of Escherichia coli and Staphylococcus aureus on the four surfaces was carried out. 40 μl of 106/ml Escherichia coli liquid and 40 μl of 106/ml Staphylococcus aureus liquid were dropwise added on the surfaces of the four samples respectively, and cultured for 6 hours, then the bacterial liquids on the surfaces were washed with PBS, subjected to fluorescence staining, and observed by a laser confocal microscope, and fluorescence intensities of any 10 positions on each surface were counted.

[0034] As shown in FIG. 5, the two bacteria were adhered to the surfaces a and b in large quantities. Bacterial adhesion quantities on the surface c and the surface d were obviously smaller than those on the surface a and the surface b, which indicated that the micro-nano composite structure had an obvious inhibitory effect on the adhesion of the two bacteria. It was worth noting that the bacterial adhesion quantity on the surface b was slightly higher than that on the surface a, which indicated that the bacterial adhesion on the surface b was slightly promoted after the surface b had a high biological activity, and the bacterial adhesion quantity on the surface d was basically the same as that on the surface c, which indicated that the inhibition effect on bacterial adhesion of the micro-nano composite structure eliminated the influence of increased bacterial adhesion caused by the improved biological activity of the surface d.

[0035] Results of the experiment of the cells and the experiment of the bacteria above showed that the surface with the bioactive component and the micro-nano composite structure prepared on the surface of the implant by the plasma injection method combined with the femtosecond laser machining method had dual functions of promoting the adhesion and proliferation of the gingival fibroblast and the gingival mesenchymal stem cell and inhibiting the adhesion of the Escherichia coli and the Staphylococcus aureus.

Comparative Example 2

[0036] In this comparative example, wear resistance performances of a pure titanium surface with a micro-nano composite structure and a titanium surface with the micro-nano composite structure injected with C, N, Ca and P elements were compared. The pure titanium surface with the micro-nano composite structure was prepared according to the preparation method of the surface c in Comparative Example 1, and the titanium surface with the micro-nano composite structure injected with C, N, Ca and P elements was prepared according to the preparation method of the surface d in Comparative Example 1.

[0037] The wear resistance performances of the two surfaces were tested by a wear tester, dry friction was carried out on the surfaces by a Si3N4 grinding ball at a rotating speed of 300 r/min and a load of 200 N, and after the friction was carried out for 30 minutes, wearing capacities of the two surfaces were counted respectively. As shown in FIG. 6, after the friction was carried out for 30 minutes, the wearing capacity of the surface injected with the elements was obviously lower than that of the pure titanium surface, which indicated that the wear resistance performance of the surface injected with the elements by the plasma injection method was obviously improved.

Comparative Example 3

[0038] In this comparative example, adhesion conditions of various common oral bacteria on a smooth pure titanium surface and a titanium surface with a bioactive component and a micro-nano composite structure were compared.

[0039] A preparation method of the pure titanium surface was that: the pure titanium surface was processed by a mechanical polishing method. The surface with the bioactive component and the micro-nano composite structure was prepared by the method in Embodiment 1. Bacteria selected for a bacteriostasis experiment comprised alpha streptococcus, anaerobic streptococcus, Staphylococcus epidermidis, Neisseria, lactobacillus, spirochete and candida respectively. Numbers of bacteria adhered to the two surfaces were counted by a fluorescence intensity statistic method and a plate counting method respectively.

[0040] The fluorescence intensity statistical experiment method was that 40 μl of 106/ml Escherichia coli liquid and 40 μl of 106/ml Staphylococcus aureus liquid were dropwise added on the surfaces of the four samples respectively, and cultured for 6 hours, then the bacterial liquids on the surfaces were washed with PBS, subjected to fluorescence staining, and observed by a laser confocal microscope, and fluorescence intensities of any 10 positions on each surface were counted. As shown in FIG. 7, since the polished titanium surface had no bacteriostatic performance, the seven oral bacteria were all adhered to the surface in large quantities, and adhesion quantities of the bacteria on the bioactive surface 7 with the micro-nano composite structure were all obviously less than those on the polished titanium surface. This result showed that, compared with the smooth pure titanium implant surface, the implant surface with the micro-nano composite structure and the bioactive component proposed by the present invention had a significant inhibitory effect on the adhesion of the common oral bacteria on the surface.

Embodiment 2

[0041] In this embodiment, femtosecond laser machining was carried out twice to obtain a required structure at a transgingival part. Specific femtosecond laser machining steps and process were as follows.

[0042] (1) Alight spot of the laser was irradiated on an initial position of the transgingival part, and the transgingival part of the implant was machined. The machining process was that: a surface of the transgingival part was machined by a laser frequency of 10 kHz, pulse energy of 3,000 uJ, a light spot diameter of 60 μm, a line spacing of 50 μm and a scanning speed of 20 mm/s, a T axis was rotated by one circle during machining, a Y axis was translated by 0.2 mm, and an X axis was moved at the same time to ensure that a distance from a focal point of the laser to a surface of a sample remained unchanged, so that the surface of the whole transgingival part was covered repeatedly.

[0043] (2) The X axis was moved to make the light spot of the laser return to a machining starting position of this part for secondary machining. The secondary machining process was that: the surface of the transgingival part was machined by a laser frequency of 10 kHz, pulse energy of 200 uJ, a light spot diameter of 60 μm, a line spacing of 50 μm and a scanning speed of 20 mm/s, a T axis was rotated by one circle during machining, a Y axis was translated by 0.2 mm, and an X axis was moved at the same time to ensure that a distance from a focal point of the laser to a surface of a sample remained unchanged, so that the surface of the whole transgingival part was covered repeatedly.

[0044] Others were the same as those in Embodiment 1.

[0045] The above are only the preferred embodiments of the present invention, and it should be pointed out that those of ordinary skills in the art may further make several modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.