阅读说明：本技术 种植体液态存储方法 (Method for storing liquid state of implant ) 是由 邱憬 邵水易 张文思 于 2020-04-01 设计创作，主要内容包括：种植体液态存储方法,依次用SiC防水砂纸同方向逐级打磨钛片至表面呈金属光泽,表面划痕一致,再依次以双蒸水、无水乙醇和双蒸水超声清洗；将上步所得钛片经喷砂,再依次置于丙酮、乙醇、蒸馏水中超声清洗并干燥；硝酸,氢氟酸混合液室温处理,双蒸水超声,干燥；盐酸硫酸混合酸80℃水浴处理至起泡后,双蒸水超声清洗；将维生素C溶解于生理盐水中；将所得钛片经双蒸水超声清洗后密闭、避光储存于配制好的维生素C溶液中。本发明提供的植体液态存储方法,钛表面能够被储存液提前活化,具有促进成骨细胞黏附、增殖和分化的作用,更好的促进新骨形成。同时具有良好的润湿性和生物相容性,有广泛的应用价值。(Sequentially polishing titanium sheets step by step in the same direction by using SiC waterproof abrasive paper until the surface presents metal luster and the surface scratches are consistent, and then sequentially ultrasonically cleaning by using double distilled water, absolute ethyl alcohol and double distilled water; sand blasting the titanium sheet obtained in the previous step, then placing the titanium sheet in acetone, ethanol and distilled water in sequence, ultrasonically cleaning and drying; treating the mixed solution of nitric acid and hydrofluoric acid at room temperature, performing double-distilled water ultrasound treatment, and drying; carrying out water bath treatment on hydrochloric acid and sulfuric acid mixed acid at 80 ℃ until foaming, and ultrasonically cleaning by double distilled water; dissolving vitamin C in physiological saline; and (3) carrying out ultrasonic cleaning on the obtained titanium sheet by double distilled water, and then storing the titanium sheet in a prepared vitamin C solution in a closed and lightproof manner. According to the plant fluid storage method provided by the invention, the titanium surface can be activated in advance by the storage fluid, so that the method has the effects of promoting the adhesion, proliferation and differentiation of osteoblasts and better promoting the formation of new bones. Meanwhile, the composite material has good wettability and biocompatibility and has wide application value.)

1. The method for storing the implant liquid state is characterized by comprising the following steps of:

(1) sequentially polishing titanium sheets step by step in the same direction by using SiC waterproof abrasive paper until the surfaces of the titanium sheets are in metal luster and consistent in surface scratch, and then sequentially ultrasonically cleaning by using double distilled water, absolute ethyl alcohol and double distilled water;

(2) using Al (OH) to the titanium sheet obtained in the previous step₃Sand blasting, then placing the mixture in acetone, 75vt.% ethanol and distilled water in sequence, ultrasonically cleaning and drying; treating with mixed solution of nitric acid and hydrofluoric acid at room temperature for 10min, ultrasonic treating with double distilled water for 15min, and drying; mixing hydrochloric acid and sulfuric acid, performing water bath treatment at 80 ℃ until foaming, and performing ultrasonic cleaning by double distilled water;

(3) preparing a normal saline solution containing vitamin C;

(4) and (3) carrying out ultrasonic cleaning on the titanium sheet obtained in the step (2) by double distilled water, and then storing the titanium sheet in a prepared vitamin C solution in a closed and dark manner.

2. The implant fluid storage method according to claim 1, wherein the SiC waterproof abrasive paper is used for polishing in stages with 320, 600, 800, 1200 and 1500 meshes.

3. The method for storing vegetation fluid according to claim 1, wherein said Al (OH)₃Pressure of sand blasting: 0.3MPa, particle size: 80 mesh, distance: 1.5cm, angle: 80 degrees, time: 15 s.

4. The method for storing the liquid state of the planting body according to claim 1, wherein the volume ratio of the mixed solution of the nitric acid and the hydrofluoric acid is double distilled water, 0.22wt.% of HF and 0.57 wt.% of HNO₃=1000:2:4。

5. The method for storing the liquid state of the planting body according to claim 1, wherein the mixed acid of hydrochloric acid and sulfuric acid is double distilled water 37 wt.% HCl 98 wt.% H according to the volume ratio₂SO₄=6:1:1。

6. The method for liquid storage of plant growth according to claim 1, wherein the vitamin C solution is prepared by dissolving vitamin C powder in 0.9 vt.% physiological saline to prepare a vitamin C-containing solution of 100mM or 10 mM.

Technical Field

The invention belongs to the field of implant storage, and particularly relates to an implant liquid storage method.

Background

The oral implanting systems Straumann, 3i, Nobel Biocare all pure titanium implants. The systems respectively adopt technologies such as large-particle sand blasting-acid etching (SLA), double acid etching, micro-arc oxidation and the like to construct micron-scale rough shapes on the surfaces so as to promote the adhesion, proliferation and differentiation of osteoblasts and increase the contact area and mechanical embedding force between an implant and bones. However, pure titanium itself lacks osteoinductive properties, fails to form chemical bonds with the surrounding bone tissue, and requires a long time to form strong osseointegration after implantation. In view of this, scholars at home and abroad have conducted a lot of studies on surface modification and activation of titanium implants, and it is expected that the osteogenesis activity of the implants can be improved, the bone formation speed after implantation can be increased, the bone union strength can be increased, and the bone formation cycle can be shortened by a proper modification method. Therefore, the modification and activation technology of titanium surface is one of the research hotspots in the field of oral implantation.

However, the recent research shows that the biological activity aging phenomenon which depends on the time exists on the pure titanium surface and the modified titanium surface. Compared with the newly prepared titanium surface, the adhesion quantity, the proliferation activity and the osteogenic function phenotype of the osteoblasts on the old titanium surface are obviously reduced. Mechanical polishing, sand blasting-acid etching and physical deposition of titanium surfaces all suffer from aging, and the bone conductivity of the prepared titanium surfaces at the periphery is only less than half of that of newly prepared titanium surfaces. The biocompatibility of the titanium surface after aging is reduced, and the reason for this may be related to the reduction of hydrophilicity. Freshly prepared titanium surfaces have excellent hydrophilicity, while aged titanium surfaces tend to be hydrophobic. The research proves that: the newly prepared titanium surface can be aged after four weeks, and the surface contact angle can reach as high as 60 degrees. The hydrophilicity of the titanium surface is one of the key properties of the titanium implant in forming osseointegration. Moreover, the carbon content on the titanium surface also increases along with the prolonging of the time after the preparation, and the carbon is mainly accumulated on the titanium surface in the form of hydrocarbon continuously, so that the carbon percentage can be increased from 20 percent to more than 60 percent. The higher the carbon content on the titanium surface is, the less osteoblasts are adhered to the titanium surface, and the two are in obvious negative correlation. Therefore, the carbon accumulation effect is also an important factor influencing the biocompatibility of the titanium after the surface of the titanium is aged.

Vitamin C is essential for collagen synthesis and normal skeletal development. Research has shown that vitamin C can reduce the process of bone resorption caused by oxidative stress caused by active oxygen. In addition, vitamin C plays a major role in the formation of hydroxyproline residues, which stabilize the triple helix structure of collagen. It is worth mentioning that about 90-95% of the bone is composed of collagen. Therefore, vitamin C deficiency can block collagen synthesis, which in turn leads to the inhibition of new bone formation. Studies have shown that vitamin C also inhibits the bone resorption process by down-regulating RANKL expression. Subject group early experiments store the smooth titanium in a normal saline solution containing vitamin C in a sealed and lightproof manner, and are beneficial to adhesion, proliferation and differentiation of osteoblasts. Based on the reasons, the titanium implant is stored in the solution containing the vitamin C in a sealed and lightproof manner, so that the surface of the titanium implant is activated in advance, and the vitamin C reserved on the surface of the titanium implant can better promote the process of osseointegration.

With the explosive growth of the implantation number of the implants in China in recent years, the clinical importance of preventing the titanium implants from aging is increasingly shown. The key for solving the problem lies in the storage link of the titanium implant. However, the targeted study of implant storage is rather poor compared to the well-established study of surface modification of titanium implants. Therefore, research on the technology of activating the surface of the titanium implant in advance to prevent the surface of the titanium implant from aging enables the surface of the rough titanium implant to have a hydrophilic surface, so that vitamin C elements can be released at the beginning of implantation of the implant, and the process of osseointegration can be better promoted.

Disclosure of Invention

The technical problem to be solved is as follows: the invention provides an implant liquid state storage method aiming at the defects of the existing titanium implant storage mode.

The technical scheme is as follows: the implant liquid storage method comprises the following steps: (1) sequentially polishing titanium sheets step by step in the same direction by using SiC waterproof abrasive paper until the surfaces of the titanium sheets are in metal luster and consistent in surface scratch, and then sequentially ultrasonically cleaning by using double distilled water, absolute ethyl alcohol and double distilled water; (2) using Al (OH) to the titanium sheet obtained in the previous step₃Sand blasting, then placing the mixture in acetone, 75vt.% ethanol and distilled water in sequence, ultrasonically cleaning and drying; treating with mixed solution of nitric acid and hydrofluoric acid at room temperature for 10min, ultrasonic treating with double distilled water for 15min, and drying; mixing hydrochloric acid and sulfuric acid, performing water bath treatment at 80 ℃ until foaming, and performing ultrasonic cleaning by double distilled water; (3) preparing a normal saline solution containing vitamin C; (4) and (3) carrying out ultrasonic cleaning on the titanium sheet obtained in the step (2) by double distilled water, and then storing the titanium sheet in a prepared vitamin C solution in a closed and dark manner.

The SiC waterproof abrasive paper has the mesh number of 320 meshes, 600 meshes, 800 meshes, 1200 meshes and 1500 meshes for gradual grinding.

Preferably, the above-mentioned Al (OH)₃Pressure of sand blasting: 0.3MPa, particle size: 80 mesh, distance: 1.5cm, angle: 80 degrees, time: 15 s.

Preferably, the volume ratio of the mixed solution of nitric acid and hydrofluoric acid is double distilled water, 0.22wt.% HF and 0.57 wt.% HNO₃=1000:2:4。

Preferably, the mixed acid of hydrochloric acid and sulfuric acid is double distilled water, 37 wt.% HCl and 98 wt.% H according to volume ratio₂SO₄=6:1:1。

Preferably, the vitamin C solution is prepared by dissolving vitamin C powder in 0.9 vt.% physiological saline to prepare a vitamin C-containing solution of 100mM or 10 mM.

Has the advantages that: according to the liquid storage method of the implant, the titanium surface can be activated in advance by the storage liquid, so that the liquid storage method has the effects of promoting adhesion, proliferation and differentiation of osteoblasts and better promoting formation of new bones. Meanwhile, the composite material has good wettability and biocompatibility and has wide application value.

Drawings

FIG. 1 is a fluorescent image of adhesion morphology of smooth titanium sheet surface MC3T3-E1 after osteoblast inoculation for 2h and 4 h;

FIG. 2 is a graph showing the comparison of cell proliferation activity of smooth titanium sheets stored in different storage solutions for 10 days, surface-seeded with MC3T3-E1 osteoblasts 1, 3, and 6 days cck-8;

FIG. 3 is an electrophoresis diagram of the expression of OPN, OSX, OCN and Runx2 bone formation related proteins detected by inoculating MC3T3-E1 osteoblast western blot on the surface of a smooth titanium sheet stored in different storage solutions for 10 days;

FIG. 4 is a comparison of water contact angles on the surface of a smooth titanium sheet stored in different storage solutions for 10 days;

FIG. 5 is a fluorescence image of the adhesion morphology of SLA titanium plate surface MC3T3-E1 osteoblasts inoculated for 2h and 4 h;

FIG. 6 shows that the SLA titanium sheets are stored in different storage solutions for 10 days, and are surface-inoculated with MC3T3-E1 osteoblasts 1, 3 and 6 days cck-8 to detect the cell proliferation activity;

FIG. 7 is an electrophoresis diagram of expression of OPN, OSX, OCN and Runx2 osteogenesis related proteins detected by surface inoculation of MC3T3-E1 osteoblast western blot on SLA titanium sheets stored in different storage solutions for 10 days;

FIG. 8 is a comparison of water contact angles on the surface of SLA titanium sheets stored in different storage solutions for 10 days.

Detailed Description

The process is further illustrated by the following examples.