Preparation method of titanium implant baicalin salt storage solution

Abstract

The invention discloses a preparation method of a titanium implant baicalin salt storage solution, and belongs to the field of biomedical materials. The method comprises the following steps: carrying out normal-temperature acid etching on dried pure titanium by using a hydrofluoric acid and nitric acid mixed solution, sequentially washing and drying after acid etching, carrying out secondary acid etching by using a concentrated hydrochloric acid and concentrated sulfuric acid mixed solution at a water bath temperature of 75-85 DEG C, washing and drying again after secondary acid etching, so that a micron structure is formed on the titanium surface, and a sand blasting acid etching (SLA) titanium surface is constructed; and the titanium sheet subjected to sand blasting and acid etching is placed in a baicalin salt solution to be stored in a sealed mode. Baicalin salt storage solutions with different components are prepared, the advantages of baicalin and salt solutions are expected to be combined, surface activation of the titanium implant is promoted, faster and better osseointegration is achieved, and the success rate of implant repair is increased.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical materials. Specifically, it relates to a method for preparing a storage solution of baicalin salt for titanium implants. Background Art

[0002] Although artificial dental implants have been applied clinically for nearly half a century and have been widely proven to have reliable clinical effects, there are still the following problems: (1) It still takes a long healing time from implanting the implant into the body to clinical loading, at least 6 - 12 weeks, which to a certain extent limits the clinical application of implant restoration; (2) The bonding rate between the implant and bone is still relatively low, only 50 - 65%, which is not conducive to the long-term stability of the implant. Accelerating the speed of implant-bone bonding not only means shortening the repair time, but more importantly, making artificial dental implants have higher safety. Research shows that most implant failures occur during the early healing process of the implant. The longer the healing time, the greater the influence of various risk factors on the implant, and the higher the risk of implant failure. Therefore, faster and better bone bonding is still the goal pursued by implant restoration.

[0003] A large number of studies have shown that the speed and strength of the formation of the bond between the implant and bone tissue are closely related to the bioactivity of the implant surface, and the bioactivity is closely linked to the physical and chemical properties of the implant surface. Due to the large differences in the structure and properties between titanium and its alloys and bone tissue, and the presence of a passive oxide film on the surface, titanium materials generally cannot form direct chemical bonds with bone tissue like other bioactive materials. Therefore, titanium metal is generally considered a bio-inert material, and various methods are sought to carry out surface activation modification. Some commonly used surface treatment techniques currently include sandblasting and acid etching, micro-arc oxidation, hydroxyapatite coating, etc. Compared with the smooth implant surface, these surface treatment techniques have significantly improved the surface activity of the implant, which is beneficial to promoting implant-bone bonding. However, further research results show that the high bioactivity effect obtained through surface modification has a certain timeliness. After being stored for a period of time, this effect gradually decreases, resulting in a significant decrease in the bioactivity of the titanium material surface, that is, the aging phenomenon of titanium surface bioactivity.

[0004] At present, the reasons for the decline of the physical and chemical properties and biological activity of titanium surface over time are still unclear. Some studies believe that this may be related to the contamination of the titanium surface by hydrocarbons in the air during storage, which leads to changes in the surface element composition and a decrease in surface energy. Since the conventional storage method of implants is to store them in sealed bottles at room temperature and pressure, they may be contaminated by hydrocarbons. Therefore, during the storage process, the physical and chemical properties and biological activity of the titanium surface may have changed. Since in actual applications, clinicians cannot obtain newly prepared titanium implants, the implants used in clinical practice have been stored for a long or short time, resulting in different degrees of aging of the implants, which has an adverse effect on bone integration. In order to delay the aging of the biological activity of the titanium implant surface and improve the bone integration rate, the optimization of titanium implant storage media has become a research hotspot. Domestic and foreign studies have proposed a variety of anti-aging methods, such as ultraviolet light functionalization, inert gas storage, plasma treatment, low vacuum storage, etc., but due to the cumbersome processing process and high processing cost, they have not been put into use on a large scale. Summary of the invention

[0005] Technical problem to be solved: In view of the shortcomings of the above-mentioned existing implant storage technology, the present invention provides a method for preparing a titanium implant baicalin salt solution, attempting a convenient, fast and inexpensive implant storage method. From the perspective of optimizing the implant storage solution, this study develops a new type of baicalin salt storage solution, in order to maintain the physical and chemical properties and biological activity of the titanium implant surface as much as possible, so that the implant used in clinical practice has better bone bonding performance.

[0006] The object of the present invention can be achieved by the following technical solutions:

[0007] A method for preparing a titanium implant baicalin salt storage solution comprises the following steps:

[0008] 1) The pure titanium sample was polished, sandblasted, ultrasonically cleaned and dried;

[0009] 2) The pure titanium dried in step 1) is etched at room temperature with a mixture of hydrofluoric acid and nitric acid, washed with water and dried in sequence after etching, and then etched for the second time with a mixture of concentrated hydrochloric acid and concentrated sulfuric acid at a water bath temperature of 75-85° C., and washed with water and dried again after the second etching to form a micron structure on the titanium surface, thereby constructing a sandblasted acid-etched (SLA) titanium surface;

[0010] 3) preparing a baicalin salt storage solution;

[0011] 4) The titanium sheet after sandblasting and acid etching is placed in a baicalin salt solution for airtight storage.

[0012] In the technical solution of the present invention, in step 1), the pure titanium sample is polished successively with sandpapers numbered 600#, 800#, 1200#, and 1500# from coarse to fine to polish the pure titanium step by step.

[0013] In the technical solution of the present invention, in step 1), the pure titanium sample is sandblasted with 70-100 mesh Al2O3 sand, at a pressure of 0.3-0.5 MPa, a distance of 1-2 cm, and sandblasted for 10-20 s.

[0014] In the technical solution of the present invention, in step 2), the volume ratio of the mixed solution of hydrofluoric acid and nitric acid is double-distilled water: 0.1-0.5 wt.% HF: 0.1-1 wt.% HNO3 = 1000: 1-10: 1-10.

[0015] In the technical solution of the present invention, in step 2), the volume ratio of the mixed acid of concentrated hydrochloric acid and concentrated sulfuric acid is double-distilled water: 35-40 wt.% HCl: 98 wt.% H2SO4 = 4-8: 0.5-1.5: 0.5-1.5.

[0016] In the technical solution of the present invention, in step 3), the concentration of the prepared baicalin salt storage solution is 0.01-1 mM.

[0017] In the technical solution of the present invention, in step 3), the prepared 0.01-1 mM baicalin salt storage solution is the experimental group, and air storage and 0.9% NaCl solution storage are the control groups.

[0018] In the technical solution of the present invention, in step 4), the storage time described is 3-5 weeks.

[0019] In the technical solution of the present invention, in step 4), the storage temperature described is room temperature.

[0020] Beneficial effects: Baicalin is a flavonoid compound present in the roots, rhizomes, and leaves of Scutellaria baicalensis plants. It not only has a bone protection effect but also has biological activities such as antibacterial, anti-inflammatory, and immunomodulatory effects, and can be an excellent choice for implant storage solutions. Another study shows that storing titanium implants in salt solutions can reduce carbon element pollution and maintain their hydrophilicity, which is beneficial for protein adsorption on the titanium material surface and cell adhesion, proliferation, and differentiation. In summary, the present invention prepares baicalin salt storage solutions with different components in order to combine the advantages of baicalin and salt solutions, promote the activation of the titanium implant surface, achieve faster and better bone bonding, and improve the success rate of implant restoration. Description of the Drawings

[0021] Figure 1 It is a scanning electron microscope image (100,000 times) of the sample prepared in Comparative Example 1 of the present invention;

[0022] Figure 2SEM image (100,000 times magnification) of the sample prepared in Comparative Example 2 of the present invention;

[0023] Figure 3 SEM image (100,000 times magnification) of the sample prepared in Example 1 of the present invention;

[0024] Figure 4 SEM image (100,000 times magnification) of the sample prepared in Example 2 of the present invention;

[0025] Figure 5 SEM image (100,000 times magnification) of the sample prepared in Example 3 of the present invention;

[0026] Figure 6 XPS spectra of 5 groups of titanium materials measured with Examples 1, 2, and 3 of the present invention as the experimental groups and Comparative Examples 1 and 2 as the control groups;

[0027] Figure 7 Water contact angles of 5 groups of titanium materials measured with Examples 1, 2, and 3 of the present invention as the experimental groups and Comparative Examples 1 and 2 as the control groups;

[0028] Figure 8 Roughness of 5 groups of titanium materials measured with Examples 1, 2, and 3 of the present invention as the experimental groups and Comparative Examples 1 and 2 as the control groups;

[0029] Figure 9 3D topography maps of the roughness of 5 groups of titanium materials measured with Examples 1, 2, and 3 of the present invention as the experimental groups and Comparative Examples 1 and 2 as the control groups;

[0030] Figure 10 Bar graph of CCK-8 values of cell proliferation measured after inoculating the MC3T3-E1 osteoblast cell line on the surfaces of Examples 1, 2, and 3 of the present invention as the experimental groups and Comparative Examples 1 and 2 as the control groups for 1, 3, and 6 days, with * indicating significant differences. Detailed implementation manners

[0031] The following examples can enable those skilled in the art to understand the present invention more comprehensively, but do not limit the present invention in any way.

[0032] Example 1

[0033] A preparation method of a baicalin salt solution for titanium implants, comprising the following steps:

[0034] Step A) Polish pure titanium (successively polish pure titanium from coarse to fine using sandpapers with numbers 600#, 800#, 1200#, and 1500#), perform sandblasting (use 80-mesh Al2O3 sand, pressure 0.4 MPa, distance 1.5 cm, sandblast for 15 s), ultrasonically clean and dry for later use;

[0035] Step B) The pure titanium material treated in Step A) is etched with a hydrofluoric acid / nitric acid mixture at room temperature (the volume ratio of the hydrofluoric acid and nitric acid mixture is double-distilled water: 0.2 wt.% HF: 0.5 wt.% HNO3 = 1000:5:5), ultrasonically cleaned and dried with double-distilled water, and then etched with a concentrated hydrochloric acid / concentrated sulfuric acid mixture (the volume ratio of the concentrated hydrochloric acid and concentrated sulfuric acid mixture is double-distilled water: 38 wt.% HCl: 98 wt.% H2SO4 = 6:1:1) at a water bath temperature of 80 °C, and ultrasonically cleaned and dried with double-distilled water;

[0036] Step C) Prepare a 10 μM baicalin salt solution and transfer it into a sterile well plate;

[0037] Step D) Place the sandblasted and acid-etched titanium material prepared in Step B) into the sterile well plate in Step C), and store it airtight at room temperature for 4 weeks.

[0038] Example 2

[0039] Other conditions are the same as in Example 1, except that in Step C), a 100 μM baicalin salt solution is prepared and transferred into a sterile well plate.

[0040] Example 3

[0041] Other conditions are the same as in Example 1, except that in Step C), a 1 mM baicalin salt solution is prepared and transferred into a sterile well plate.

[0042] Comparative Example 1

[0043] Other conditions are the same as in Example 1, except that in Step C), a dry sterile well plate (air storage) is prepared.

[0044] Comparative Example 2

[0045] Other conditions are the same as in Example 1, except that in Step C), a 0.9% NaCl solution is prepared and transferred into a sterile well plate.

[0046] Performance detection:

[0047] The scanning electron microscope images of Comparative Examples 1 and 2 and Examples 1, 2, and 3 were observed as shown in Figures 1 to 5 (100,000 times). The results show that the surfaces of the five groups of materials are all uneven. The surface of Comparative Example 2 is dense salt crystals, and micron-sized particles are formed on the surfaces of Examples 1, 2, and 3. Among them, there are fewer particles on the surface of Example 1, and the diameter differences are large. The diameters of the particles on the surface of Example 2 are basically the same and are evenly distributed on the SLA titanium material surface. The diameters of the particles on the surface of Example 3 not only have large differences but also tend to aggregate into flakes.

[0048] Examples 1, 2, and 3 were the experimental groups, and Comparative Examples 1 and 2 were the control groups. The main elements contained in the 5 groups were measured using an X-ray energy spectrometer, and the XPS spectra were obtained as shown in Figure 6 . The results showed that Comparative Examples 1 and 2 and Examples 1, 2, and 3 all contained elements such as C, O, and Ti. Among them, Example 3 contained the highest C element and the lowest Ti element.

[0049] Examples 1, 2, and 3 were the experimental groups, and Comparative Examples 1 and 2 were the control groups. The water contact angles of the titanium sheets in the 5 groups were measured. Figure 7 The results showed that the water contact angles of Examples 1, 2, and 3 were significantly smaller than those of Comparative Examples 1 and 2, indicating that Examples 1, 2, and 3 had better surface hydrophilicity than Comparative Examples 1 and 2. However, there were no significant differences in the water contact angles of Examples 1, 2, and 3.

[0050] Examples 1, 2, and 3 were the experimental groups, and Comparative Examples 1 and 2 were the control groups. The roughness of the titanium sheets in the 5 groups was measured, and the results were as shown in Figure 8 and Figure 9 . It can be seen from the figure that the surface roughness of Examples 1 and 2 was significantly smaller than that of Comparative Example 1, but there was no obvious difference compared with Comparative Example 2; the surface roughness of Example 3 had no obvious difference from that of Comparative Example 1, but there was a significant increase compared with the surface roughness of Comparative Example 2 and Examples 1 and 2.

[0051] Examples 1, 2, and 3 were the experimental groups, and Comparative Examples 1 and 2 were the control groups. After culturing the MC3T3-E1 osteoblast cell line on their surfaces for 1, 3, and 6 days, a CCK-8 kit was used to detect cell proliferation. Figure 10 The results showed that the cell proliferation of Examples 1, 2, and 3 was significantly better than that of Comparative Examples 1 and 2, indicating that Examples 1, 2, and 3 had better biocompatibility, and Example 2 had the best biocompatibility.

Claims

1. A method for preparing a titanium implant baicalin salt storage solution, characterized in that The steps include: 1) The pure titanium sample was polished, sandblasted, ultrasonically cleaned and dried; 2) The pure titanium dried in step 1) is etched at room temperature with a mixture of hydrofluoric acid and nitric acid, washed with water and dried in sequence after etching, and then etched for the second time with a mixture of concentrated hydrochloric acid and concentrated sulfuric acid at a water bath temperature of 75-85° C., and washed with water and dried again after the second etching to form a micron structure on the titanium surface, thereby constructing a sandblasted acid-etched (SLA) titanium surface; 3) preparing a baicalin salt storage solution; 4) The titanium sheet after sandblasting and acid etching is placed in a baicalin salt solution for airtight storage.

2. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that In step 1), the pure titanium sample is ground and polished using sandpapers numbered 600#, 800#, 1200#, and 1500#, from coarse sand to fine sand.

3. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that In step 1), the pure titanium sample is sandblasted using 70-100 mesh Al2O3 sand, with a pressure of 0.3-0.5 MPa, a distance of 1-2 cm, and a sandblasting time of 10-20 seconds.

4. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that In step 2), the volume ratio of the mixed solution of hydrofluoric acid and nitric acid is double distilled water: 0.1-0.5wt.% HF: 0.1-1wt.% HNO3=1000: 1-10: 1-10.

5. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that Step 2) The volume ratio of the mixed acid of concentrated hydrochloric acid and concentrated sulfuric acid is double distilled water: 35-40wt.% HCl: 98wt.% H2SO4=4-8: 0.5～1.5：0.5～1.5。 6. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that The concentration of the baicalin salt storage solution prepared in step 3) is 0.01-1 mM.

7. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that The storage temperature in step 4) is room temperature.

8. The method for preparing the titanium implant baicalin salt storage solution according to claim 1, characterized in that The storage period in step 4) is 3 to 5 weeks.

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