CN106955358A - A kind of method that utilization elastic substrates prepare phosphatide pipe - Google Patents
A kind of method that utilization elastic substrates prepare phosphatide pipe Download PDFInfo
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
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- C08J2383/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing silicon with or without sulfur, nitrogen, oxygen, or carbon only; Derivatives of such polymers
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Abstract
A kind of method that utilization elastic substrates prepare phosphatide pipe, it is related to a kind of method for preparing phosphatide pipe.The invention aims to solve the existing method for preparing phosphatide pipe to need to build complicated fluid passage and strength manipulation, complex operation and it is costly the problem of.Method：First, elastic substrates are prepared；2nd, clean；3rd, clean elastic substrates are in extended state using external force；4th, mixed liquor is prepared；5th, the immobilized artificial membrane sprawled is prepared；6th, reduce or remove the external force to elastic substrates, obtain phosphatide pipe.The present invention realizes the simple and quick preparation of phosphatide pipe using the recoverable resilient characteristic of substrate, and the blank filled up on phosphatide tube preparation method has the advantages that simple to operate, low energy consumption, required mild condition, generation phosphatide pipe controllability are good.The present invention is applied to prepare phosphatide pipe.
Description
Technical field
The present invention relates to a kind of method for preparing phosphatide pipe.
Background technology
Phosphatide pipe, as the important existence of phosphatide, is iuntercellular and cell and the extraneous important field for carrying out mass exchange
Institute, is also the good biological template for building micro Nano material.There is important application in the field such as biological, medicine and materialogy.
The preparation method of current phosphatide pipe mainly has the methods such as change solvent method, force-extension method.Change solvent method is essentially limited to
Acetylenic phosphatide, such as DC8,9PC.Force-extension method is to carry out stretching preparation to phospholipid molecule or phospholipid capsule bubble using the method for external force
It is cumbersome and costly, it is necessary to build fluid passage and the strength manipulation of complexity during phosphatide pipe.Phosphatide pipe is except being used as life
Matter transportation in object and film properties research are outer, are also used as template and build micro Nano material, such as its surface plated with nickel,
The formation metal tube such as copper deposits silicon and forms silicone tube, can also be prepared using phosphatide pipe hollow lumen insertion nano-particle one-dimensional
Nanotube.At present, the simple and quick preparation of phosphatide pipe is realized, it is most important in the application of every field for expanding phosphatide.This
The stretching for the utilization elastic substrates being related in invention and shrinkage character so that the immobilized artificial membrane in elastic substrates is re-assemblied and realized
The preparation of phosphatide pipe, this method have it is simple to operate, the advantages of quick, gentle reaction condition and good controllability.
The content of the invention
The invention aims to solve the existing method for preparing phosphatide pipe to need to build fluid passage and the strength of complexity
Manipulation, complex operation and it is costly the problem of, and provide a kind of method that utilization elastic substrates prepare phosphatide pipe.
A kind of method that utilization elastic substrates prepare phosphatide pipe, is specifically realized by the following steps：
First, elastic substrates are prepared：
The prepolymer A of dimethyl silicone polymer and curing agent B are mixed, then carry out spin coating, obtain thickness for 0.01mm~
0.3mm elastic substrates；
The prepolymer A and curing agent B of dimethyl silicone polymer described in step one mass ratio are (10~11):1；
2nd, clean：
The elastic base for the use of absolute ethyl alcohol and deionized water being respectively 0.01mm~0.3mm to the thickness obtained in step one
Bottom is cleaned by ultrasonic 10min~20min, reuses plasma cleaner cleaning 30s, obtains the elastic substrates of cleaning；
3rd, clean elastic substrates are in extended state using external force, thickness is changed into the 1/3~2/3 of original thickness, obtained
Elastic substrates in extended state；
4th, the phospholipid capsule bubble aqueous solution and concentration are mixed for the 0.5mol/L trishydroxymethylaminomethane aqueous solution, obtained
Mixed liquor；
The phospholipid capsule bubble aqueous solution and the trishydroxymethylaminomethane aqueous solution of the concentration for 0.5mol/L described in step 4
Volume ratio be 1:(2~10)；
5th, the mixed liquor obtained in step 4 is placed in the elastic substrates in extended state obtained in step 3,
Again 40min~60min, the immobilized artificial membrane sprawled in elastic substrates are stood in the case where temperature is 30 DEG C~50 DEG C；
6th, reduce or remove the external force to elastic substrates, make elastic substrates thickness be changed into original thickness 85%~
100%, phosphatide pipe is obtained in elastic substrates.
Advantages of the present invention：
First, the present invention realizes the simple and quick preparation of phosphatide pipe using elastic substrates, and the tubular state of phosphatide is good, yield compared with
It is high；It is widely used in the associated biomolecule membrane property in the fields such as cell biology, membrane biophysics, biochemistry, bionics, carefully
Born of the same parents simulate, the research such as pharmaceutical carrier and target administration and application；
2nd, the present invention is realized the simple and quick preparation of phosphatide pipe, filled up using the recoverable resilient characteristic of substrate
Blank on phosphatide tube preparation method is good etc. with simple to operate, low energy consumption, required mild condition, generation phosphatide pipe controllability
Advantage；
3rd, the length of phosphatide pipe prepared by the present invention is 2 μm~100 μm, a diameter of 0.3 μm~4 μm；
4th, the method that a kind of utilization elastic substrates of the invention prepare phosphatide pipe is with low cost, and technique is simple, is adapted to industrialization
Produce phosphatide pipe.
The present invention is applied to prepare phosphatide pipe.
Brief description of the drawings
Fig. 1 is the fluorescent microscopy images of the DOPC immobilized artificial membranes sprawled obtained in the step 5 of embodiment one；
Fig. 2 is the fluorescent microscopy images for the DOPC phosphatide pipes that the step 6 of embodiment one is obtained；
Fig. 3 is the process chart of embodiment one, and A is clean elastic substrates in Fig. 3, and B is first cylindrical drum, and C is
Second cylindrical drum, D is inflating pressure, and E is the DOPC phospholipid capsule bubble aqueous solution, and F is DOPC immobilized artificial membranes, and G is DOPC phosphatide pipes, 1
For the joint that clean elastic substrates are fixed between two identical cylindrical drums, 2 drawing is in for clean elastic substrates
State is stretched, 3 is add the phospholipid capsule bubble aqueous solution, and 4 is form the DOPC immobilized artificial membranes sprawled, and 5 be the DOPC phosphatide pipes formed.
Embodiment
Embodiment one：Present embodiment is a kind of method that utilization elastic substrates prepare phosphatide pipe, is specifically pressed
What following steps were completed：
First, elastic substrates are prepared：
The prepolymer A of dimethyl silicone polymer and curing agent B are mixed, then carry out spin coating, obtain thickness for 0.01mm~
0.3mm elastic substrates；
The prepolymer A and curing agent B of dimethyl silicone polymer described in step one mass ratio are (10~11):1；
2nd, clean：
The elastic base for the use of absolute ethyl alcohol and deionized water being respectively 0.01mm~0.3mm to the thickness obtained in step one
Bottom is cleaned by ultrasonic 10min~20min, reuses plasma cleaner cleaning 30s, obtains the elastic substrates of cleaning；
3rd, clean elastic substrates are in extended state using external force, thickness is changed into the 1/3~2/3 of original thickness, obtained
Elastic substrates in extended state；
4th, the phospholipid capsule bubble aqueous solution and concentration are mixed for the 0.5mol/L trishydroxymethylaminomethane aqueous solution, obtained
Mixed liquor；
The phospholipid capsule bubble aqueous solution and the trishydroxymethylaminomethane aqueous solution of the concentration for 0.5mol/L described in step 4
Volume ratio be 1:(2~10)；
5th, the mixed liquor obtained in step 4 is placed in the elastic substrates in extended state obtained in step 3,
Again 40min~60min, the immobilized artificial membrane sprawled in elastic substrates are stood in the case where temperature is 30 DEG C~50 DEG C；
6th, reduce or remove the external force to elastic substrates, make elastic substrates thickness be changed into original thickness 85%~
100%, phosphatide pipe is obtained in elastic substrates.
Prepolymer A and curing agent the B mixing of dimethyl silicone polymer described in the step one of present embodiment one can be prepared
Dimethyl silicone polymer (PDMS), is liquid, buys from the U.S. (USA) Dow Corning companies, model
Sylgard184。
Phospholipid capsule bubble ruptures in the presence of the trishydroxymethylaminomethane aqueous solution in present embodiment step 5, in elasticity
Assembling is sprawled in substrate and forms immobilized artificial membrane.
Reduce in present embodiment step 6 or remove the external force to elastic substrates, elastic substrates surface is spread over originally
Immobilized artificial membrane can reconfigure to form phosphatide pipe.
Phospholipid capsule bubble, immobilized artificial membrane and phosphatide Guan Jun described in present embodiment exist in aqueous.
The advantage of present embodiment：
First, present embodiment realizes the simple and quick preparation of phosphatide pipe using elastic substrates, and the tubular state of phosphatide is good, yield
It is higher；It is widely used in the associated biomolecule membrane property in the fields such as cell biology, membrane biophysics, biochemistry, bionics,
Cell is simulated, the research such as pharmaceutical carrier and target administration and application；
2nd, present embodiment utilizes the recoverable resilient characteristic of substrate, realizes the simple and quick preparation of phosphatide pipe, fills out
The blank on phosphatide tube preparation method is mended, with simple to operate, low energy consumption, required mild condition, generation phosphatide pipe controllability
The advantages of good；
3rd, the length of phosphatide pipe prepared by present embodiment is 2 μm~100 μm, a diameter of 0.3 μm~4 μm；
4th, the method that a kind of utilization elastic substrates of present embodiment prepare phosphatide pipe is with low cost, and technique is simple, is adapted to work
Industry metaplasia produces phosphatide pipe.
Present embodiment is applied to prepare phosphatide pipe.
Embodiment two：Present embodiment is with the difference of embodiment one：Elasticity described in step one
The elastic modelling quantity of substrate is 1MPa~10MPa.Other steps are identical with embodiment one.
Embodiment three：One of present embodiment and embodiment one or two difference is：Institute in step one
The prepolymer A and curing agent B of the dimethyl silicone polymer stated mass ratio are (10~10.5):1.Other steps and specific implementation
Mode one or two is identical.
Embodiment four：One of present embodiment and embodiment one to three difference is：Institute in step one
The prepolymer A and curing agent B of the dimethyl silicone polymer stated mass ratio are (10.5~11):1.Other steps and specific implementation
Mode one to three is identical.
Embodiment five：One of present embodiment and embodiment one to four difference is：Institute in step 2
The power for the ultrasonic cleaning stated is 300W, and the temperature of ultrasonic cleaning is 23~30 DEG C.Other steps and embodiment one to
Four is identical.
Embodiment six：One of present embodiment and embodiment one to five difference is：Institute in step 2
State plasma cleaner cleaning when technological parameter be：Vacuum is less than 100Pa, and rf frequency is 13.56MHz, radio frequency work(
Rate is 150W.Other steps are identical with embodiment one to five.
Embodiment seven：One of present embodiment and embodiment one to six difference is：Institute in step 4
Phospholipid capsule bubble is DOPC vesica, phosphatidyl glycerol vesica or oleoylphosphatidyl in the phospholipid capsule bubble aqueous solution stated
Choline vesica；A diameter of 1 μm~100 μm of described phospholipid capsule bubble.Other steps are identical with embodiment one to six.
Embodiment eight：One of present embodiment and embodiment one to seven difference is：Phosphorus in step 4
The concentration of phospholipid capsule bubble is 1000/μ L~1400/μ L in the lipid vesicle aqueous solution.Other steps and embodiment one to
Seven is identical.
Embodiment nine：One of present embodiment and embodiment one to eight difference is：Institute in step 6
The length for the phosphatide pipe stated is 2 μm~100 μm, a diameter of 0.3 μm~4 μm.Other steps and the phase of embodiment one to eight
Together.
Embodiment ten：One of present embodiment and embodiment one to nine difference is：Institute in step 4
The volume ratio for the trishydroxymethylaminomethane aqueous solution that the phospholipid capsule bubble aqueous solution stated is 0.5mol/L with concentration is 1:(2~6).
Other steps are identical with embodiment one to nine.
Beneficial effects of the present invention are verified using following examples：
Embodiment one：A kind of method that utilization elastic substrates prepare phosphatide pipe, is specifically realized by the following steps：
First, elastic substrates are prepared：
The prepolymer A of dimethyl silicone polymer and curing agent B is mixed, then carries out spin coating, the bullet that thickness is 0.1mm is obtained
Property substrate；
The prepolymer A and curing agent B of dimethyl silicone polymer described in step one mass ratio are 10:1；
The elastic modelling quantity of elastic substrates described in step one is 6MPa；
2nd, clean：
It is clear for 0.1mm elastic substrates ultrasound to the thickness obtained in step one using absolute ethyl alcohol and deionized water respectively
15min is washed, plasma cleaner cleaning 30s is reused, obtains the elastic substrates of cleaning；
The power of ultrasonic cleaning described in step 2 is 300W, and the temperature of ultrasonic cleaning is 25 DEG C；
Described in step 2 plasma cleaner cleaning when technological parameter be：Vacuum is less than 100Pa, rf frequency
For 13.56MHz, radio-frequency power is 150W；
3rd, clean elastic substrates are in extended state using external force, thickness is changed into the 2/3 of original thickness, is in
The elastic substrates of extended state；
4th, the phospholipid capsule bubble aqueous solution and concentration are mixed for the 0.5mol/L trishydroxymethylaminomethane aqueous solution, obtained
Mixed liquor；
Phospholipid capsule bubble is DOPC vesica, dioleoyl phosphorus in the phospholipid capsule bubble aqueous solution described in step 4
The concentration of phosphatidylcholine vesica is 1200/μ L；A diameter of 10 μm~50 μm of described phospholipid capsule bubble；
The phospholipid capsule bubble aqueous solution and the trishydroxymethylaminomethane aqueous solution of the concentration for 0.5mol/L described in step 4
Volume ratio be 1:4；
5th, the mixed liquor obtained in step 4 is placed in the elastic substrates in extended state obtained in step 3,
50min is stood in the case where temperature is 40 DEG C again, the DOPC immobilized artificial membranes sprawled in elastic substrates；
6th, the external force to elastic substrates is removed, the thickness of elastic substrates is changed into original thickness, is obtained in elastic substrates
DOPC phosphatide pipes.
Clean elastic substrates are to utilize two phases in extended state by utilization external force described in the step 3 of embodiment one
Same cylindrical drum docking, the joint that clean elastic substrates are fixed between two identical cylindrical drums recycles inflation
To clean elastic substrates inflating pressure at the circular hole that pump passes through cylindrical drum so that clean elastic substrates are in extended state,
Thickness is changed into the 2/3 of original thickness, obtains the elastic substrates in extended state.
Fig. 1 is the fluorescent microscopy images of the DOPC immobilized artificial membranes sprawled obtained in the step 5 of embodiment one；
From fig. 1, it can be seen that a diameter of 10 μm~50 μm of DOPC phospholipid capsule bubbles.
Fig. 2 is the fluorescent microscopy images for the DOPC phosphatide pipes that the step 6 of embodiment one is obtained；
As can be seen from Figure 2, the length of DOPC phosphatide pipe is 2~100 μm, and diameter range is 0.3~3 μm.
Fig. 3 is the process chart of embodiment one, and A is clean elastic substrates in Fig. 3, and B is first cylindrical drum, and C is
Second cylindrical drum, D is inflating pressure, and E is the DOPC phospholipid capsule bubble aqueous solution, and F is DOPC immobilized artificial membranes, and G is DOPC phosphatide pipes, 1
For the joint that clean elastic substrates are fixed between two identical cylindrical drums, 2 drawing is in for clean elastic substrates
State is stretched, 3 is add the phospholipid capsule bubble aqueous solution, and 4 is form the DOPC immobilized artificial membranes sprawled, and 5 be the DOPC phosphatide pipes formed.
Claims (10)
1. a kind of method that utilization elastic substrates prepare phosphatide pipe, it is characterised in that a kind of utilization elastic substrates prepare phosphatide pipe
What method was specifically realized by the following steps：
First, elastic substrates are prepared：
The prepolymer A of dimethyl silicone polymer and curing agent B is mixed, then carries out spin coating, thickness is obtained for 0.01mm~0.3mm
Elastic substrates；
The prepolymer A and curing agent B of dimethyl silicone polymer described in step one mass ratio are (10~11):1；
2nd, clean：
The thickness obtained in step one is surpassed for 0.01mm~0.3mm elastic substrates using absolute ethyl alcohol and deionized water respectively
Sound cleans 10min~20min, reuses plasma cleaner cleaning 30s, obtains the elastic substrates of cleaning；
3rd, clean elastic substrates are in extended state using external force, thickness is changed into the 1/3~2/3 of original thickness, is in
The elastic substrates of extended state；
4th, the phospholipid capsule bubble aqueous solution and concentration are mixed for the 0.5mol/L trishydroxymethylaminomethane aqueous solution, mixed
Liquid；
The phospholipid capsule bubble aqueous solution and body of the concentration for the 0.5mol/L trishydroxymethylaminomethane aqueous solution described in step 4
Product is than being 1:(2~10)；
5th, the mixed liquor obtained in step 4 is placed in the elastic substrates in extended state obtained in step 3, then
Temperature is standing 40min~60min, the immobilized artificial membrane sprawled in elastic substrates at 30 DEG C~50 DEG C；
6th, reduce or remove the external force to elastic substrates, the thickness of elastic substrates is changed into the 85%~100% of original thickness,
Phosphatide pipe is obtained in elastic substrates.
2. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step one
The elastic modelling quantity of described elastic substrates is 1MPa~10MPa.
3. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step one
The prepolymer A and curing agent B of described dimethyl silicone polymer mass ratio are (10~10.5):1.
4. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step one
The prepolymer A and curing agent B of described dimethyl silicone polymer mass ratio are (10.5~11):1.
5. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step 2
The power of described ultrasonic cleaning is 300W, and the temperature of ultrasonic cleaning is 23~30 DEG C.
6. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step 2
Technological parameter during described plasma cleaner cleaning is：Vacuum is less than 100Pa, and rf frequency is 13.56MHz, radio frequency
Power is 150W.
7. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step 4
Phospholipid capsule bubble is DOPC vesica, phosphatidyl glycerol vesica or oleoyl phospholipid in the described phospholipid capsule bubble aqueous solution
Phatidylcholine vesica；A diameter of 1 μm~100 μm of described phospholipid capsule bubble.
8. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step 4
The concentration of phospholipid capsule bubble is 1000/μ L~1400/μ L in the phospholipid capsule bubble aqueous solution.
9. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step 6
The length of described phosphatide pipe is 2 μm~100 μm, a diameter of 0.3 μm~4 μm.
10. the method that a kind of utilization elastic substrates according to claim 1 prepare phosphatide pipe, it is characterised in that in step 4
The volume ratio for the trishydroxymethylaminomethane aqueous solution that the described phospholipid capsule bubble aqueous solution is 0.5mol/L with concentration is 1:(2~
6)。
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| CN201710167659.XA CN106955358B (en) | 2017-03-21 | 2017-03-21 | Method for preparing phospholipid tube by using elastic substrate |
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| CN201710167659.XA CN106955358B (en) | 2017-03-21 | 2017-03-21 | Method for preparing phospholipid tube by using elastic substrate |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080299086A1 (en) * | 2006-09-21 | 2008-12-04 | Tohoku University | Cultured muscle cells with high metabolic activity and method for production of the cultured muscle cells |
| WO2010034487A2 (en) * | 2008-09-23 | 2010-04-01 | Silence Therapeutics Ag | Means for inhibiting the expression of orc-1 |
| CN103173353A (en) * | 2011-12-23 | 2013-06-26 | 国家纳米科学中心 | Multilayer tubular structural cell culture bracket as well as preparation method and use thereof |
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2017
- 2017-03-21 CN CN201710167659.XA patent/CN106955358B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080299086A1 (en) * | 2006-09-21 | 2008-12-04 | Tohoku University | Cultured muscle cells with high metabolic activity and method for production of the cultured muscle cells |
| WO2010034487A2 (en) * | 2008-09-23 | 2010-04-01 | Silence Therapeutics Ag | Means for inhibiting the expression of orc-1 |
| CN103173353A (en) * | 2011-12-23 | 2013-06-26 | 国家纳米科学中心 | Multilayer tubular structural cell culture bracket as well as preparation method and use thereof |
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