CN108751126B - The method for preparing three-dimensional self-supporting film based on lysozyme nano thin-film - Google Patents

The method for preparing three-dimensional self-supporting film based on lysozyme nano thin-film Download PDF

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CN108751126B
CN108751126B CN201810587315.9A CN201810587315A CN108751126B CN 108751126 B CN108751126 B CN 108751126B CN 201810587315 A CN201810587315 A CN 201810587315A CN 108751126 B CN108751126 B CN 108751126B
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lysozyme
micro
supporting film
dimensional self
nano granules
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CN108751126A (en
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杨鹏
刘瑞瑞
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Shaanxi Normal University
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B81—MICROSTRUCTURAL TECHNOLOGY
    • B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
    • B81C99/00—Subject matter not provided for in other groups of this subclass
    • C—CHEMISTRY; METALLURGY
    • C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
    • C23C—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; SURFACE TREATMENT OF METALLIC MATERIAL BY DIFFUSION INTO THE SURFACE, BY CHEMICAL CONVERSION OR SUBSTITUTION; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL
    • C23C18/00—Chemical coating by decomposition of either liquid compounds or solutions of the coating forming compounds, without leaving reaction products of surface material in the coating; Contact plating
    • C23C18/16—Chemical coating by decomposition of either liquid compounds or solutions of the coating forming compounds, without leaving reaction products of surface material in the coating; Contact plating by reduction or substitution, e.g. electroless plating
    • C23C18/18—Pretreatment of the material to be coated
    • C23C18/20—Pretreatment of the material to be coated of organic surfaces, e.g. resins
    • C23C18/28—Sensitising or activating
    • C23C18/30—Activating or accelerating or sensitising with palladium or other noble metal
    • C—CHEMISTRY; METALLURGY
    • C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
    • C23C—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; SURFACE TREATMENT OF METALLIC MATERIAL BY DIFFUSION INTO THE SURFACE, BY CHEMICAL CONVERSION OR SUBSTITUTION; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL
    • C23C18/00—Chemical coating by decomposition of either liquid compounds or solutions of the coating forming compounds, without leaving reaction products of surface material in the coating; Contact plating
    • C23C18/16—Chemical coating by decomposition of either liquid compounds or solutions of the coating forming compounds, without leaving reaction products of surface material in the coating; Contact plating by reduction or substitution, e.g. electroless plating
    • C23C18/31—Coating with metals
    • C23C18/38—Coating with copper
    • C23C18/40—Coating with copper using reducing agents
    • C23C18/405—Formaldehyde
    • C—CHEMISTRY; METALLURGY
    • C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
    • C23C—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; SURFACE TREATMENT OF METALLIC MATERIAL BY DIFFUSION INTO THE SURFACE, BY CHEMICAL CONVERSION OR SUBSTITUTION; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL
    • C23C18/00—Chemical coating by decomposition of either liquid compounds or solutions of the coating forming compounds, without leaving reaction products of surface material in the coating; Contact plating
    • C23C18/16—Chemical coating by decomposition of either liquid compounds or solutions of the coating forming compounds, without leaving reaction products of surface material in the coating; Contact plating by reduction or substitution, e.g. electroless plating
    • C23C18/31—Coating with metals
    • C23C18/42—Coating with noble metals
    • C23C18/44—Coating with noble metals using reducing agents

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Materials Engineering (AREA)
  • Mechanical Engineering (AREA)
  • Metallurgy (AREA)
  • Organic Chemistry (AREA)
  • Microelectronics & Electronic Packaging (AREA)
  • Medicinal Preparation (AREA)
  • Manufacturing Of Micro-Capsules (AREA)

Abstract

The invention discloses a kind of methods for preparing three-dimensional self-supporting film based on lysozyme nano thin-film, this method is using micro-nano granules as template, one layer of lysozyme nano thin-film is coated on its surface based on lysozyme phase transition, directly removing template can be obtained lysozyme three-dimensional self-supporting film；Also one layer of metallic can be deposited on the micro-nano granules surface that lysozyme coats by electroless deposition processes or one layer of hydroxyapatite is grown on the micro-nano granules surface that lysozyme coats by biomineralization, template is removed again, obtains metal three-dimensional self-supporting film or hydroxyapatite three-dimensional self-supporting film.Preparation process of the present invention is of short duration and simple easily operated, it is low in cost, environmentally protective simultaneously, it can be used for preparing organic, inorganic and metal class three-dimensional self-supporting film, with extensive universality, easily realize large-scale industrial application, and the size of gained three-dimensional self-supporting film, shape can be regulated and controled by template.

Description

The method for preparing three-dimensional self-supporting film based on lysozyme nano thin-film
Technical field
The present invention relates to it is a kind of it is environmentally protective, be simple and efficient and the system of the three-dimensional self-supporting film with extensive universality Preparation Method.
Background technique
Metal, inorganic or protein tridimensional capsule with special colloform texture and property have the space of isolation due to it With the multi-functional and potential application such as controllable permeability, in medicine/gene delivering, catalysis, sensor, medicine, cosmetics etc. Have in different field and is widely applied and is particularly concerned.Manufacture can encapsulate the micron of a variety of materials and the capsule of nano-scale (or shell) has the double meaning of Science and Technology.Up to the present, developed extensive hollow structure, as condensate, Multilayer capsule and tiny balloon.The structure and form of capsule play a key role in terms of the property and function that determine capsule. Although having there is the various methods for preparing three-dimensional capsule, most preparation method is complicated, with high costs, biocompatibility It is poor, such as layer-by-layer (LBL), be difficult to meet it is industrial there is an urgent need to.In another example in solid shell Grain (core-shell particles) be widely used as microcapsules, for drug control release and targeting and for protect such as enzyme and The sensitive agents such as protein.Such particle can be made by interfacial polymerization or by the mutually separation of polymer-solvent mixture It is standby.However, their manufacture faces many common problems, including the covering of polydispersity, non-uniform shell and core solidification. The urgent need therefore it provides a kind of method that can be designed and make various types of other three-dimensional capsule seems.
Summary of the invention
It is a kind of easy to operate, low in cost, green technical problem to be solved by the present invention lies in view of the above deficiencies, providing The method of the three-dimensional self-supporting film of the preparation that colour circle is protected.
Technical solution used by above-mentioned technical problem is solved to be made of following step:
1, the modification of micro-nano granules
The 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 1~100mmol/L tri- (2- carboxyethyl) phosphine is adjusted to NaOH PH value is 5.0~10.0, then mixes it with the 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 1~50mg/mL lysozyme It is even, then micro-nano granules are immersed in gained mixed liquor, it cultivates 20~60 minutes at room temperature, forms micro-nano granules surface One layer of lysozyme nano thin-film, centrifugation, washing, obtains the micro-nano granules of lysozyme cladding.
2, three-dimensional self-supporting film is prepared
Micro-nano granules inside the micro-nano granules of lysozyme cladding are removed, it is thin to obtain lysozyme three-dimensional self-supporting Film；Or through electroless deposition processes after micro-nano granules surface one layer of metallic of deposition that lysozyme coats, then remove it Internal micro-nano granules, obtain metal three-dimensional self-supporting film；Or it is coated by biomineralization in lysozyme micro-nano After particle surface grows one layer of hydroxyapatite, then its internal micro-nano granules is removed, obtains hydroxyapatite three-dimensional from branch Support film.
In above-mentioned preparation method, the micro-nano granules are CaCO3When particle, in step 2, by lysozyme cladding Micro-nano granules are immersed in 0.01~1mol/L disodium ethylene diamine tetra-acetic acid aqueous solution, are cultivated 30~90 minutes, are obtained at room temperature To lysozyme three-dimensional self-supporting film.
In above-mentioned preparation method, when the micro-nano granules are polystyrene microsphere, in step 2, by lysozyme packet After the micro-nano granules area load palladium ion covered, then by electroless deposition processes one layer of metallic of deposition, then roasting is removed Micro-nano granules obtain metal three-dimensional self-supporting film, wherein the metal is copper or silver.
In above-mentioned preparation method, when the micro-nano granules are polystyrene microsphere, in step 2, by lysozyme packet The micro-nano granules surface chelating Ca covered2+Afterwards, it immerses in simulated body fluid, is kept at 37 DEG C 7~10 days, then roasting removes micro- Nano particle obtains hydroxyapatite three-dimensional self-supporting film.
Beneficial effects of the present invention are as follows:
1, the present invention coats one layer of lysozyme nanometer on its surface based on lysozyme phase transition using micro-nano granules as template Film, directly removing template can be obtained lysozyme three-dimensional self-supporting film.The present invention can also be by electroless deposition processes in bacteriolyze The micro-nano granules surface of enzyme cladding deposits one layer of metallic or coat by biomineralization in lysozyme micro-nano Grain surface grows one layer of hydroxyapatite, then removes template, and metal three-dimensional self-supporting film or hydroxyapatite three can be obtained Tie up self-supporting film.
2, preparation process of the present invention is of short duration and simple easily operated while low in cost, environmentally protective, can be used for being prepared with Machine class, inorganic and metal class three-dimensional self-supporting film have extensive universality, Yi Shixian large-scale industrial application, and institute Size, the shape for obtaining three-dimensional self-supporting film can be regulated and controled by template.
Detailed description of the invention
Fig. 1 is the optical microscopy map for the lysozyme three-dimensional self-supporting film that embodiment 1 obtains.
Fig. 2 is the scanning electron microscopy of the polystyrene microsphere for the surface deposition silver particles that embodiment 2 obtains.
Fig. 3 is the scanning electron microscopy for the three-dimensional self-supporting film of silver that embodiment 2 obtains.
Fig. 4 is the energy spectrum diagram for the three-dimensional self-supporting film of silver that embodiment 2 obtains.
Fig. 5 is the scanning electron microscopy of the polystyrene microsphere for the copper-depositing on surface particle that embodiment 3 obtains.
Fig. 6 is the scanning electron microscopy for the copper three-dimensional self-supporting film that embodiment 3 obtains.
Fig. 7 is the energy spectrum diagram for the copper three-dimensional self-supporting film that embodiment 3 obtains.
Fig. 8 is the scanning electron microscopy for the hydroxyapatite three-dimensional self-supporting film that embodiment 4 obtains.
Specific embodiment
The present invention is described in more detail with reference to the accompanying drawings and examples, but protection scope of the present invention is not limited only to These embodiments.
Embodiment 1
1、CaCO3The modification of particle
The 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 5mmol/L tri- (2- carboxyethyl) phosphine is adjusted to NaOH PH value is 7.0, is then uniformly mixed it with the 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 1mg/mL lysozyme, then The CaCO for being 10 μm by partial size3Particle is immersed in gained mixed liquor, is cultivated 50 minutes at room temperature, is made CaCO3Particle surface is formed One layer of lysozyme nano thin-film obtains the CaCO of lysozyme cladding finally by centrifugation, deionized water washing3Particle.
2, lysozyme three-dimensional self-supporting film is prepared
The CaCO that lysozyme is coated3Particle is immersed in 0.1mol/L disodium ethylene diamine tetra-acetic acid aqueous solution, at room temperature It cultivates 60 minutes, obtains lysozyme three-dimensional self-supporting film, as shown in Figure 1.
Embodiment 2
1, the modification of polystyrene microsphere
The 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 50mmol/L tri- (2- carboxyethyl) phosphine is adjusted to NaOH PH value is 5.0, is then uniformly mixed it with the 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 2mg/mL lysozyme, then The polystyrene microsphere that partial size is 10 μm is immersed in gained mixed liquor, cultivates 40 minutes at room temperature, makes polystyrene microsphere Surface forms one layer of lysozyme nano thin-film, finally by centrifugation, deionized water washing, obtains the polystyrene of lysozyme cladding Microballoon.
2, the three-dimensional self-supporting film of preparation silver
The polystyrene microsphere that lysozyme coats is immersed in 0.007g/mL tetrachloro-palladium acid aqueous ammonium, is trained at room temperature It educates 15 minutes, centrifugation, deionized water washing obtain the polystyrene microsphere of area load palladium ion；It is 1% by mass fraction Silver ammino solution is uniformly mixed in equal volume with the aqueous sodium potassium tartrate that mass fraction is 4%, then by area load palladium ion Polystyrene microsphere submergence wherein, at room temperature cultivate 15 minutes, centrifugation, deionized water washing, obtain surface deposition silver granuel The polystyrene microsphere of son, as shown in Figure 2.The polystyrene microsphere of surface deposition silver particles is placed in Muffle furnace, 350 DEG C of roastings It burns 1 hour, removes polystyrene microsphere, obtain silver-colored three-dimensional self-supporting film, as shown in Figure 3.The power spectrum of Fig. 4 further illustrates The three-dimensional self-supporting film of fine silver is arrived.
Embodiment 3
1, the modification of polystyrene microsphere
The 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 10mmol/L tri- (2- carboxyethyl) phosphine is adjusted to NaOH PH value is 6.0, is then uniformly mixed it with the 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 2mg/mL lysozyme, then The polystyrene microsphere that partial size is 10 μm is immersed in gained mixed liquor, cultivates 40 minutes at room temperature, makes polystyrene microsphere Surface forms one layer of lysozyme nano thin-film, finally by centrifugation, deionized water washing, obtains the polystyrene of lysozyme cladding Microballoon.
2, copper three-dimensional self-supporting film is prepared
The polystyrene microsphere that lysozyme coats is immersed in 0.007g/mL tetrachloro-palladium acid aqueous ammonium, is trained at room temperature It educates 15 minutes, centrifugation, deionized water washing obtain the polystyrene microsphere of area load palladium ion；12g/L hydroxide will be contained Sodium, 13g/L Salzburg vitriol, 29g/L sodium potassium tartrate tetrahydrate aqueous solution be uniformly mixed in equal volume with 10mL/L formalin, Then wherein by the polystyrene microsphere submergence of area load palladium ion, cultivate 15 minutes at room temperature, centrifugation, deionization washing It washs, obtains the polystyrene microsphere of copper-depositing on surface particle, as shown in Figure 5.By the polystyrene microsphere of copper-depositing on surface particle It is placed in Muffle furnace, 350 DEG C roast 1 hour, remove polystyrene microsphere, obtain copper three-dimensional self-supporting film, as shown in Figure 6. The power spectrum of Fig. 7 further illustrates to have obtained the three-dimensional self-supporting film of fine copper.
Embodiment 4
1, the modification of polystyrene microsphere
The 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 1mmol/L tri- (2- carboxyethyl) phosphine is adjusted to NaOH PH value is 7.0, is then uniformly mixed it with the 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 10mL 1mg/mL lysozyme, then The polystyrene microsphere that partial size is 10 μm is immersed in gained mixed liquor, cultivates 40 minutes at room temperature, makes polystyrene microsphere Surface forms one layer of lysozyme nano thin-film, finally by centrifugation, washing, obtains the polystyrene microsphere of lysozyme cladding.
2, three-dimensional self-supporting film is prepared
The polystyrene microsphere that lysozyme coats is immersed in 20mmol/L CaCl2In aqueous solution, impregnate 2 days at room temperature, Centrifugation, deionized water washing, remove the Ca of non-chelating2+, obtain chelating Ca2+Polystyrene microsphere；By chelating Ca2+Polyphenyl Ethylene microballoon immerses in simulated body fluid, is kept for 10 days at 37 DEG C, obtains the polystyrene microsphere of Coated With Hydroxyapatite；By hydroxyl The polystyrene microsphere of base apatite cladding is placed in Muffle furnace, and 350 DEG C roast 1 hour, is removed polystyrene microsphere, is obtained hydroxyl Base apatite three-dimensional self-supporting film, as shown in Figure 8.

Claims (4)

1. a kind of method for preparing three-dimensional self-supporting film based on lysozyme nano thin-film, it is characterised in that this method is by following steps Rapid composition:
(1) modification of micro-nano granules
The 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 1~100 mmol/L tri- (2- carboxyethyl) phosphine is adjusted to pH value with NaOH It is 5.0~10.0, is then uniformly mixed it with the 4- hydroxyethyl piperazineethanesulfonic acid buffer solution of 1~50 mg/mL lysozyme, Micro-nano granules are immersed in gained mixed liquor again, are cultivated 20~60 minutes at room temperature, micro-nano granules surface is made to form one Layer lysozyme nano thin-film, centrifugation, washing, obtains the micro-nano granules of lysozyme cladding；
(2) three-dimensional self-supporting film is prepared
After depositing one layer of metallic on the micro-nano granules surface that lysozyme coats by electroless deposition processes, then remove inside it Micro-nano granules, obtain metal three-dimensional self-supporting film；
Or through biomineralization after one layer of hydroxyapatite of micro-nano granules surface growth that lysozyme coats, then remove it Internal micro-nano granules, obtain hydroxyapatite three-dimensional self-supporting film.
2. the method according to claim 1 for preparing three-dimensional self-supporting film based on lysozyme nano thin-film, feature exist In: when the micro-nano granules are polystyrene microsphere, in step (2), by the micro-nano granules surface of lysozyme cladding After loading palladium ion, then by electroless deposition processes one layer of metallic of deposition, then roasting removes micro-nano granules, obtains metal Three-dimensional self-supporting film.
3. the method according to claim 2 for preparing three-dimensional self-supporting film based on lysozyme nano thin-film, feature exist In: the metal is copper or silver.
4. the method according to claim 1 for preparing three-dimensional self-supporting film based on lysozyme nano thin-film, feature exist In: when the micro-nano granules are polystyrene microsphere, in step (2), by the micro-nano granules surface of lysozyme cladding Chelating Ca2+Afterwards, it immerses in simulated body fluid, is kept at 37 DEG C 7~10 days, then roasting removes micro-nano granules, obtains hydroxyl Apatite three-dimensional self-supporting film.
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