CN104258463A - Medical stainless steel material and preparation method thereof - Google Patents
Medical stainless steel material and preparation method thereof Download PDFInfo
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- CN104258463A CN104258463A CN201410522169.3A CN201410522169A CN104258463A CN 104258463 A CN104258463 A CN 104258463A CN 201410522169 A CN201410522169 A CN 201410522169A CN 104258463 A CN104258463 A CN 104258463A
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Abstract
The invention discloses a medical stainless steel material and a preparation method thereof. The medical stainless steel material has the functions of bacterial adhesion resistance, sterilization and promotion of osteoblast growth and comprises a medical stainless steel matrix, a first adhesive layer and a second adhesive layer, wherein the first adhesive layer is a 3-(trifluoromethyl)benzyl mercaptan adhesive layer; the second adhesive layer is a polyethylene glycol compound adhesive layer; the surface of the second adhesive layer adsorbs silver nanoparticles. The medical stainless steel material disclosed by the invention can improve the adhesive capability of the polyethylene glycol compound on the surface of the stainless steel to prevent falling off; furthermore, the polyethylene glycol compound is utilized to improve the bacterial adhesion resistance of the stainless steel, so as to avoid that the bacteria are adhered and proliferate on the surface of the stainless steel; in addition, the silver nanoparticles are utilized to improve the bactericidal ability of the stainless steel. According to the invention, a molecular imprinting method is utilized during the process that the polyethylene glycol compound adhesive layer is introduced, so as to enable the stainless steel material to achieve a function of effectively identifying the osteoblast molecules and help the osteoblast to be fixed and grow on the surface of the material.
Description
Technical field
The present invention relates to the field of surface modification of bio-medical material, particularly relate to a kind of medical stainless steel material and preparation method thereof.
Technical background
Medical stainless steel hardness is high, average weight, low price, easy to make, but rustless steel itself does not have ability that is antibacterial and sterilization, so be easy to infect when medical stainless steel being used as dental implant and the plantation of bone kind, the life of entail dangers to people time serious, therefore, need to improve stainless anti-microbial property and sterilizing ability.
Antibacterial can be divided into three steps in the growth of material surface: be first the adhesion of antibacterial at material surface, is then the breeding of antibacterial, finally forms stable biomembrane.Therefore possess the material of opposing bacterial adhesion function and sterilizing ability, can effectively avoid bacteriogenic infection problems.Ethylene glycol compounds is modified material surface and is considered to the most effective method of opposing bacterial adhesion, and introduces silver at material surface and also have good sterilizing ability.But these two kinds of methods all can not be provided as osteocyte in the fixing of material surface and growth.
Silver ion has good bactericidal effect, and silver nano-grain is incorporated into material surface, will improve the sterilizing ability of material greatly.
First Southern proposed in 1975: can form multiple action point when template molecule (microsphere) contacts with polymer monomer, will be memorized by this effect of polymerization process, when after template molecule removing, just define the hole with multiple action point matched with template molecule steric configuration in polymer, such hole will to template molecule and analog thereof the molecular imprinting that has that selective recognition characteristic-Here it is.
Therefore, develop a kind of new medical stainless steel material and preparation method thereof, introduce ethylene glycol compounds and improve material anti-bacterial attachment ability, introduce the sterilizing ability that silver nano-grain improves material, utilize the technology of molecular engram to improve material at the application point that surface of stainless steel is introduced and osteoblast matches simultaneously and facilitate bone cell function extremely important.
Summary of the invention
The object of the invention is to the deficiency for general bio-medical material, a kind of medical stainless steel material and preparation method thereof is provided, first 3-(trifluoromethyl) benzyl mercaptan adhesive layer is introduced, introduce Polyethylene Glycol compounds adhesive layer again, both the adhesive ability of Polyethylene Glycol compounds at stainless steel surfaces had been improved, prevent from coming off, utilize again Polyethylene Glycol compounds to improve rustless steel anti-bacterial attachment ability, avoid antibacterial to adhere to breeding at stainless steel surfaces.
Another object of the present invention is, utilizes molecular engram method, make stainless steel material have the function effectively identified osteoblast molecule, be conducive to osteoblast in this material surface fixed growth in the process introducing Polyethylene Glycol compounds adhesive layer.
The present invention also introduces silver nano-grain on the surface of poly-diethanol compounds adhesive layer, improves the sterilizing ability of material, avoids bacteriological infection occurs.
Technical scheme provided by the invention is:
A kind of medical stainless steel material, it has anti-bacterial attachment, sterilizes and facilitates bone cell growth function, wherein, described in comprise:
Medical stainless steel matrix;
First adhesive layer, it is 3-(trifluoromethyl) benzyl mercaptan adhesive layer, and it is attached to the surface of described matrix, is connected with described matrix by coordinate bond;
Second adhesive layer, it is Polyethylene Glycol compounds adhesive layer, it is attached to the surface of described matrix, and described Polyethylene Glycol compounds under alcoholic environment, cross-linking reaction occurs by methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate and obtains;
Wherein, also there is in the second adhesive layer the hole matched with osteoblast steric configuration;
In addition, the second adhesive layer surface is also adsorbed with silver nano-grain.
Preferably, in described medical stainless steel material, described hole uses molecular engram method, in the process of described methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate generation cross-linking reaction, introduce osteoblast, and then using collagenase to remove, osteoblast obtains.
A preparation method for medical stainless steel material, wherein, described in comprise the following steps:
Step one, pretreatment: stainless steel substrates surface carborundum paper No. 400 to No. 1000 sequential polish, then acetone, ethanol and distilled water is used ultrasonic 30 minutes successively, 80 DEG C of vacuum dryings, then titanium sheet being soaked in volume fraction is in the alcoholic solution of 3-(methacryloxypropyl) propyl trimethoxy silicane of 5%, soak 1 hour, distilled water cleaning, drying, obtain product 1;
Step 2, molecular engram methods combining surface Atom Transfer Radical Polymerization method carries out surface modification: add in water and ethanol by methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate, stir, add osteoblast, stir; Adding area is 1 × 1cm
2product 1, adds CuBr and alpha-brominated methyl phenylacetate, and the lower 100 DEG C of reactions of logical nitrogen protection obtained product 2 after 30 minutes;
Step 3, introducing silver nano-grain: add in the aqueous solution of silver nano-grain by product 2, soak 30-60 minute, distilled water is cleaned and obtained product 3;
Step 4, post processing: product 3 is put into the bottle that 1%I Collagenase Type 2ml is housed, digest and take out after 30 seconds, and distilled water cleans latter ultrasonic 10 minutes, takes out in 80 DEG C of vacuum dryings, obtains product of the present invention.
Preferably, in the preparation method of described medical stainless steel material, the particle diameter of described silver nano-grain is 10-50 nanometer, and consumption is 0.5-1mol.
Preferably, in the preparation method of described medical stainless steel material, the consumption of described methacrylic acid 2-(2-methoxy ethoxy) ethyl ester is 1.7-1.9mol, described ethyleneglycol dimethyacrylate consumption is 0.2-0.4mol, the consumption of described water is 40-60ml, and described ethanol consumption is 10-20ml.
Preferably, in the preparation method of described medical stainless steel material, described osteoblastic density is 2 × 10
4/ ml, consumption is 0.5-2ml.
Preferably, in the preparation method of described medical stainless steel material, the consumption of described CuBr and alpha-brominated methyl phenylacetate is respectively 0.05-0.1mol and 0.01-0.05mol.
The present invention has following beneficial effect: first, first the present invention introduces 3-(trifluoromethyl) benzyl mercaptan adhesive layer on stainless steel base, introduce Polyethylene Glycol compounds adhesive layer again, both improve the adhesive ability of Polyethylene Glycol compounds at stainless steel surfaces, prevent from coming off, utilize again Polyethylene Glycol compounds to improve rustless steel anti-bacterial attachment ability, avoid antibacterial to adhere to breeding at stainless steel surfaces.
Secondly, the present invention utilizes molecular engram method in the process introducing Polyethylene Glycol compounds adhesive layer, first osteoblast is introduced in Polyethylene Glycol compounds adhesive layer, osteoblast is removed by recycling collagenase, make Polyethylene Glycol compounds adhesive layer form the hole matched with osteoblast steric configuration, make stainless steel material have the function effectively identified osteoblast molecule.The material utilizing the method to prepare, after implanting as implant, is conducive to osteoblast in this material surface fixed growth.
Again, the present invention introduces silver nano-grain on the surface of Polyethylene Glycol compounds adhesive layer, improves the sterilizing ability of material, avoids bacteriological infection.
Finally, the present invention utilizes sand paper to polish to stainless steel-based surface, with deionized water and acetone, ultrasonic cleaning is carried out to stainless steel-based surface, reduce stainless steel watch surface oxidation film to the impact of adhesive layer, increase the contact area of adhesive layer and stainless steel surfaces, improve the adhesion of adhesive layer and stainless steel base.
Accompanying drawing explanation
Fig. 1 is that stainless steel material of the present invention contrasts figure with traditional rustless steel to the comparative result that activity of osteoblast proliferation affects.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is elaborated, can implement according to this after consulting this description to make those of ordinary skill in the art.
As shown in Fig. 1, table 1 and table 2,
A kind of medical stainless steel material, it has anti-bacterial attachment, sterilizes and facilitates bone cell growth function, wherein, described in comprise:
Medical stainless steel matrix, its surface, after sand paper polishing, recycles acetone, ethanol and distilled water ultrasonic cleaning;
First adhesive layer, it is 3-(trifluoromethyl) benzyl mercaptan adhesive layer, and it is attached to the surface of described matrix, is connected with described matrix by coordinate bond;
Second adhesive layer, it is Polyethylene Glycol compounds adhesive layer, it is attached to the surface of described matrix, and described Polyethylene Glycol compounds under alcoholic environment, cross-linking reaction occurs by methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate and obtains;
Wherein, also there is in the second adhesive layer the hole matched with osteoblast steric configuration;
In addition, second adhesive layer surface is also adsorbed with silver nano-grain, carboxyl absorption in described silver nano-grain methacrylic acid 2-(2-methoxy ethoxy) ethyl ester is pasted on described adhesive layer surface, be 10-50 nanometer by the particle diameter of described granule, in its surface being distributed in the second adhesive layer and hole;
In described medical stainless steel material, described hole uses molecular engram method, osteoblast is introduced in the process of described methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate generation cross-linking reaction, and then use collagenase removal osteoblast to obtain, a kind of more stable polyethylene glycols compound molecule of network structure is formed after described second fat and alcohol ester cross-linking reaction, osteoblast is introduced in the process of reaction, after question response, osteoblast is removed, the hole that can match with osteoblast steric configuration can be left in the structure of Polyethylene Glycol compounds.
A preparation method for medical stainless steel material, wherein, described in comprise the following steps:
Step one, pretreatment: stainless steel substrates surface carborundum paper No. 400 to No. 1000 sequential polish, then acetone, ethanol and distilled water is used ultrasonic 30 minutes successively, 80 DEG C of vacuum dryings, then titanium sheet being soaked in volume fraction is in the alcoholic solution of 3-(methacryloxypropyl) propyl trimethoxy silicane of 5%, soak 1 hour, distilled water cleaning, drying, obtain product 1;
Step 2, molecular engram methods combining surface Atom Transfer Radical Polymerization method carries out surface modification: add in water and ethanol by methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate, stir, add osteoblast, stir; Adding area is 1 × 1cm
2product 1, adds CuBr and alpha-brominated methyl phenylacetate, and the lower 100 DEG C of reactions of logical nitrogen protection obtained product 2 after 30 minutes;
Step 3, introducing silver nano-grain: add in the aqueous solution of silver nano-grain by product 2, soak 30-60 minute, distilled water is cleaned and obtained product 3;
Step 4, post processing: product 3 is put into the bottle that 1%I Collagenase Type 2ml is housed, digest and take out after 30 seconds, and distilled water cleans latter ultrasonic 10 minutes, takes out in 80 DEG C of vacuum dryings, obtains product of the present invention.
In the preparation method of described medical stainless steel material, the particle diameter of described silver nano-grain is 10-50 nanometer, and consumption is 0.5-1mol.
In the preparation method of described medical stainless steel material, the consumption of described methacrylic acid 2-(2-methoxy ethoxy) ethyl ester is 1.7-1.9mol, described ethyleneglycol dimethyacrylate consumption is 0.2-0.4mol, the consumption of described water is 40-60ml, and described ethanol consumption is 10-20ml.
In the preparation method of described medical stainless steel material, described osteoblastic density is 2 × 10
4/ ml, consumption is 0.5-2ml.
In the preparation method of described medical stainless steel material, the consumption of described CuBr and alpha-brominated methyl phenylacetate is respectively 0.05-0.1mol and 0.01-0.05mol.
Embodiment 1
One, product preparation
Stainless steel substrates surface carborundum paper No. 400 to No. 1000 sequential polish, then acetone, ethanol and distilled water is used ultrasonic 30 minutes successively, 80 DEG C of vacuum dryings, then titanium sheet being soaked in volume fraction is in the alcoholic solution of 3-(methacryloxypropyl) propyl trimethoxy silicane of 5%, soak 1 hour, distilled water cleaning, drying, obtain product 1; 1.8mol methacrylic acid 2-(2-methoxy ethoxy) ethyl ester, 0.3mol ethyleneglycol dimethyacrylate are added in 50ml water and 15ml ethanol, stir, adding 1mL density is 2 × 10
4the osteoblast of/ml, stirs; Adding area is 1 × 1cm
2product 1, adds 0.05mol CuBr, the alpha-brominated methyl phenylacetate of 0.01mol, and the logical lower 100 DEG C of reactions of nitrogen protection obtain product 2 in 30 minutes; Add in the aqueous solution of silver nano-grain by product 2, soak 30-60 minute, distilled water is cleaned and is obtained product 3, product 3 is put into the bottle that 1%I Collagenase Type 2ml is housed, digest and take out after 30 seconds, distilled water cleans latter ultrasonic 10 minutes, take out in 80 DEG C of vacuum dryings, obtain product of the present invention.
Two, anti-bacterial attachment performance test
Experimentation: each specimen surface inoculation 1ml concentration is 10
5bacterium liquid cultivate after 1d at 37 DEG C, culture fluid is used for the sterilizing rate of test samples; Sample taking-up PBS gently rinsing 3 times to remove the antibacterial do not adhered to, then antibacterial sample adhered to sonic oscillation (40W) 5min is eluted in 1ml distilled water, eluent is used for the viable count in test samples surface adhesion antibacterial, calculates the anti-bacterial attachment performance of sample; Viable count is detected with doubling dilution and spread plate method.
Experimental result:
Table 1 product of the present invention is to the sterilizing rate of each antibacterial and anti-adhesive rate
| Antibacterial | ATCC?6538 | ATCC?25922 | ATCC?10231 | ATCC?9372 |
| Sterilizing rate | 100％ | 99.98％ | 100％ | 100％ |
| Anti-bacterial attachment rate | 100％ | 99.99％ | 99.98％ | 99.96％ |
Table 1 shows, the product obtained by the present invention all has very strong sterilizing rate and anti-bacterial attachment performance to staphylococcus aureus (ATCC 6538), colon bacillus (ATCC 25922), candida albicans (ATCC 10231), Bacillus subtilis endophyticus (ATCC 9372), and its sterilizing rate and anti-bacterial attachment rate all reach more than 99.9%.
Three, cytoactive detects
Experimentation: sample is placed in 24 orifice plates (often group establishes three parallel holes); 1ml density is 2 × 10
4the cell suspension inoculation of/ml, in specimen surface, then cultivates 1,4 and 7d; After predetermined point of time, transfer in 24 new orifice plates after the soft rinsing of phosphate buffer of sample pH=7.4 three times; Every hole adds 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide bromine salt and 800 μ l serum-frees that 200 μ l concentration are 5mg/ml without phenol red DMEM culture medium; 37 DEG C hatch 4h after inhale abandon supernatant, add 1ml dmso solution generate crystal, every hole is got 200 μ l lysates and is forwarded 96 well culture plates to, with spectrophotometer in 490nm place survey its OD value; Get traditional rustless steel and repeat above-mentioned experiment and the value recording its OD.
Experimental result: as shown in Figure 1, the OD value of product of the present invention and the OD value of traditional stainless steel substrates all increase with the growth of natural law, but clearly the amplitude of the OD value increase of product of the present invention is larger, illustrates that this product has stronger cell-proliferation activity.
Four, cytotoxicity analysis
Experimentation: the cytotoxicity size assessing product of the present invention using the activity of lactic acid dehydrogenase (LDH) as cytotoxicity index.Sample is placed in 24 orifice plates, by 1 × 10
4the osteoblast of individual cell is inoculated in 24 orifice plates, and cultivates 1 day.Collect culture fluid, get supernatant after centrifugal and detect for LDH activity.
Experimental result:
Table 2 product of the present invention and the stainless LDH activity of tradition (U/L)
| ? | Stainless steel substrates | Product of the present invention |
| LDH activity | 236 | 232 |
As shown in table 2, the LDH activity of product of the present invention is lower than traditional stainless LDH activity, illustrates that products upon cell of the present invention does not have toxicity.
Although embodiment of the present invention are open as above, but it is not restricted to listed in description and embodiment utilization, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other amendment, therefore do not deviating under the general concept that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the legend described.
Claims (7)
1. a medical stainless steel material, it has anti-bacterial attachment, sterilizes and facilitates bone cell growth function, it is characterized in that, described in comprise:
Medical stainless steel matrix;
First adhesive layer, it is 3-(trifluoromethyl) benzyl mercaptan adhesive layer, and it is attached to the surface of described matrix, is connected with described matrix by coordinate bond;
Second adhesive layer, it is Polyethylene Glycol compounds adhesive layer, it is attached to the surface of described matrix, and described Polyethylene Glycol compounds under alcoholic environment, cross-linking reaction occurs by methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate and obtains;
Wherein, also there is in the second adhesive layer the hole matched with osteoblast steric configuration;
In addition, the second adhesive layer surface is also adsorbed with silver nano-grain.
2. medical stainless steel material as claimed in claim 1, it is characterized in that, described hole uses molecular engram method, in the process of described methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate generation cross-linking reaction, introduce osteoblast, and then using collagenase to remove, osteoblast obtains.
3. a preparation method for medical stainless steel material as claimed in claim 1, is characterized in that, described in comprise the following steps:
Step one, pretreatment: stainless steel substrates surface carborundum paper No. 400 to No. 1000 sequential polish, then acetone, ethanol and distilled water is used ultrasonic 30 minutes successively, 80 DEG C of vacuum dryings, then titanium sheet being soaked in volume fraction is in the alcoholic solution of 3-(methacryloxypropyl) propyl trimethoxy silicane of 5%, soak 1 hour, distilled water cleaning, drying, obtain product 1;
Step 2, molecular engram methods combining surface Atom Transfer Radical Polymerization method carries out surface modification: add in water and ethanol by methacrylic acid 2-(2-methoxy ethoxy) ethyl ester and ethyleneglycol dimethyacrylate, stir, add osteoblast, stir; Adding area is 1 × 1cm
2product 1, adds CuBr and alpha-brominated methyl phenylacetate, and the lower 100 DEG C of reactions of logical nitrogen protection obtained product 2 after 30 minutes;
Step 3, introducing silver nano-grain: add in the aqueous solution of silver nano-grain by product 2, soak 10-60 minute, distilled water is cleaned and obtained product 3;
Step 4, post processing: product 3 is put into the bottle that 1%I Collagenase Type 2ml is housed, digest and take out after 30 seconds, and distilled water cleans latter ultrasonic 10 minutes, takes out in 80 DEG C of vacuum dryings, obtains product of the present invention.
4. the preparation method of medical stainless steel material as claimed in claim 3, it is characterized in that, the particle diameter of described silver nano-grain is 10-50 nanometer, and consumption is 0.5-1mol.
5. the preparation method of medical stainless steel material as claimed in claim 3, it is characterized in that, the consumption of described methacrylic acid 2-(2-methoxy ethoxy) ethyl ester is 1.7-1.9mol, described ethyleneglycol dimethyacrylate consumption is 0.2-0.4mol, the consumption of described water is 40-60ml, and described ethanol consumption is 10-20ml.
6. the preparation method of medical stainless steel material as claimed in claim 3, it is characterized in that, described osteoblastic density is 2 × 10
4/ ml, consumption is 0.5-2ml.
7. the preparation method of medical stainless steel material as claimed in claim 3, it is characterized in that, the consumption of described CuBr and alpha-brominated methyl phenylacetate is respectively 0.05-0.1mol and 0.01-0.05mol.
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Citations (4)
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| US20040115239A1 (en) * | 2002-09-20 | 2004-06-17 | Shastri Venkatram P. | Engineering of material surfaces |
| CN1775313A (en) * | 2005-10-14 | 2006-05-24 | 浙江大学 | Polymer brush for improving biomedical material compatibility and its preparing method |
| CN101812265A (en) * | 2009-02-24 | 2010-08-25 | 赛诺医疗科学技术有限公司 | Hydrophilic coat solution applied to surface of medicinal instrument and preparation method thereof |
| CN101892475A (en) * | 2010-06-24 | 2010-11-24 | 苏州大学 | Method for grafting polymer chain on stainless steel surface |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040115239A1 (en) * | 2002-09-20 | 2004-06-17 | Shastri Venkatram P. | Engineering of material surfaces |
| CN1775313A (en) * | 2005-10-14 | 2006-05-24 | 浙江大学 | Polymer brush for improving biomedical material compatibility and its preparing method |
| CN101812265A (en) * | 2009-02-24 | 2010-08-25 | 赛诺医疗科学技术有限公司 | Hydrophilic coat solution applied to surface of medicinal instrument and preparation method thereof |
| CN101892475A (en) * | 2010-06-24 | 2010-11-24 | 苏州大学 | Method for grafting polymer chain on stainless steel surface |
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