CN108836354A - A kind of preparation method of microneedle array biosensor - Google Patents
A kind of preparation method of microneedle array biosensor Download PDFInfo
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- CN108836354A CN108836354A CN201810370629.3A CN201810370629A CN108836354A CN 108836354 A CN108836354 A CN 108836354A CN 201810370629 A CN201810370629 A CN 201810370629A CN 108836354 A CN108836354 A CN 108836354A
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- microneedle array
- preparation
- array
- rough surface
- molecule
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
- A61B5/14503—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
- A61B5/14532—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue for measuring glucose, e.g. by tissue impedance measurement
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
- A61B5/6846—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive
- A61B5/6847—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive mounted on an invasive device
- A61B5/685—Microneedles
Abstract
The invention discloses a kind of preparation methods of microneedle array biosensor, including：Make the conductive microneedle array with rough surface；Molecule is identified in the rough surface modified biological of the microneedle array；Degradable high polymer material is added on the microneedle array surface for having modified the biological identification molecule.Microneedle array in the microneedle array biosensor has rough surface; facilitate in puncture process to modification in the protection of the biological identification molecule on microneedle array surface, and the degradable high polymer material of microneedle array outer layer addition can also realize the second protection to biological identification molecule.
Description
Technical field
The present invention relates to biomedical engineering fields, concretely relate to a kind of preparation of microneedle array biosensor
Method.
Background technique
Microneedle array biosensor be in microneedle array surface modification biological identification molecule, in physical examination is surveyed, micropin
Array biosensor, which punctures, to be entered after skin, and biological identification molecule is directly contacted with body fluid, and biological identification molecule can be special
Property is specifically bound with the biomolecule in tissue fluid, while generating electronics, by the concentration of biomolecule with electric signal
Form show.
Have in physical examination measuring tool using microneedle array biosensor and small, painless or hypodynia, skin are injured to human skin
Skin can restore to the original state in a short time, microneedle array is inserted into human body skin corium not and will cause the advantages such as biofouling.For example,
Microneedle array biosensor is used for blood sugar test and does not need that blood was collected by the common blood sugar test in physical examination survey, can be straight
The glucose content in measurement tissue fluid is connect, the blood-sugar content in glucose content and blood in tissue fluid is essentially identical, only
It is to have delay in 5-10 minutes, and carry out blood sugar test according to blood glucose meter common in the market, patient needs to take by finger tip
The method of blood measures blood glucose anywhere or anytime, but finger tip, which takes, can generate pain when blood, and finger tip takes the wound of blood cannot be
Restore in several minutes, be easy to cause infection.
Although microneedle array biosensor can be used for surveying in physical examination, it is conventionally used to the material master of production microneedle array
If metal, monocrystalline silicon and polymer, monocrystalline silicon microneedle array brittleness is larger, and micropin is caused to be easily broken off.Polymer micro needle battle array
Big powers' degree is not big enough, is not easy to puncture cuticula, and metal micro-needle array develops to obtain comparative maturity, and safety also has certain guarantee,
But these types of microneedle array all have the shortcomings that one it is common, that is, the microneedle array prepared is all smooth surface, is being punctured through
Biological identification molecule of the modification in microneedle array surface can fall off because of frictional force in journey, cause entry into intracorporal microneedle array
It cannot achieve the purpose that survey in physical examination.
Summary of the invention
It is micro- in the microneedle array biosensor it is an object of the invention to prepare a kind of microneedle array biosensor
Needle array has rough surface, and the biological identification molecule that rough surface can modify microneedle array surface plays protection and makees
With, in puncture process will not because of frictional force effect and large area falls off, facilitate in puncture process to modification in micropin
The protection of the biological identification molecule of array surface, and the degradable high polymer material of microneedle array outer layer addition can also be realized
To the second protection of biological identification molecule.
In order to solve the above technical problems, the technical solution adopted by the present invention is：
A kind of preparation method of microneedle array biosensor, including：Make the conductive micropin with rough surface
Array；Molecule is identified in the rough surface modified biological of the microneedle array；In the micropin for having modified the biological identification molecule
Array surface adds degradable high polymer material.
The microneedle array biosensor being prepared using the above method, for physical examination survey, outer layer it is degradable
High molecular material can puncture in several minutes that enter dermal layer of the skin in micropin to be dissolved, and exposes biological identification molecule, biology is known
Other molecule can be specifically bound with the biomolecule in tissue fluid, while generate electronics, by the concentration of biomolecule with electricity
The form of signal shows.
Further, it includes 3D printing, chemical wet etching that production, which has the method for the conductive microneedle array of rough surface,.
3D printing is to be mapped as needed using software, then printed on 3D printer, and the surface printed is even roughness degree,
Chemical wet etching is that photoresist solidifies under the action of uv light, forms coarse microneedle array surface.Preferably, since laser is beaten
The surface roughness that hole generates is obvious, microneedle array mold is prepared using laser boring, in the process for preparing microneedle array mold
In in the microneedle array mold inner-cavity formed gradient rough surface；Conductive polymer material is cast in the microneedle array
In mold, demoulding forms the conductive microneedle array with rough surface.
Specifically, using laser boring to prepare the material of the microneedle array mold referred in microneedle array mold can be aluminium
Piece, during preparing microneedle array mold using laser boring, trepanning velocity is 100~1000mm/s, and punching number is
At 100~2000 times, the disk diameter of laser boring is 0.2mm, a length of 6.8mm of array of discs, width 6.8mm, disk battle array
The diameter of column is 0.2mm, and array horizontal spacing is 0.6mm, longitudinal pitch 0.6mm, 12 × 12 arrays.
Certainly, the mold materials of microneedle array are also possible to copper sheet, iron plate, stainless steel material etc., trepanning velocity and number
Changed according to specific requirement, the needle height that different trepanning velocities is generated with number and roughness difference, disk is laser boring
Path, the disk diameter of laser boring is that the setting of 0.2mm is the preferable aperture measured according to experiment, and the size in aperture can shadow
The length and diameter of micropin is rung, and will affect the distribution number of aperture on pore membrane so that influencing the distribution of microneedle array.
Further, the rough surface modified biological using chemical crosslink technique in microneedle array identifies molecule；
Further, it under the action of high voltage electric field, is sprayed on the microneedle array surface for having modified the biological identification molecule
Apply one layer of degradable high polymer material with biocompatibility.Further, the degradable high polymer material of spraying
For PVP or PVA, further, the intensity of high voltage electric field can be 10~20kV, and the high voltage electric field intensity of 10~20kV can
To guarantee to form one layer more uniformly in the PVP or PVA of the microneedle array surface spraying for having modified the biological identification molecule
Nano fibrous membrane.
In the above-mentioned methods, further, the microneedle array material can be glassy metal, the microneedle array material
It can also be doping carbon black and/or the conductive polymer material of graphene and/or carbon nanotube and/or carbon fiber.
Compared with prior art, the beneficial effects of the invention are as follows：
The conductive microneedle array with rough surface is made, the rough surface of microneedle array can protect the life of modification
Object identify molecule, in puncture process will not because of frictional force effect and large area falls off, can be in puncture process
Biological identification molecule is protected.And degradable high molecular material is sprayed to microneedle array surface and may be implemented to know biology
The second protection of other molecule.This layer of high molecular material be it is degradable, can enter several minutes of tissue fluid in dissolve, exposure birth
Object identifies molecule, will not have an impact to the sensitivity of biosensor, and has good biocompatibility, will not be to people
Body generates adverse reaction.
Therefore, the invention enables the microneedle array biosensors of preparation can realize in puncture process to bio-identification
The protection of molecule, and can be realized the second protection to biological identification molecule, to realize in physical examination survey to biological identification molecule
Peak use rate, while making the preparation process of microneedle array biosensor simpler.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of microneedle array biosensor in the embodiment of the present invention；
Fig. 2 is the schematic device of spray on polymer material in the embodiment of the present invention.
Specific embodiment
The present invention is further illustrated With reference to embodiment.
Embodiment 1
As shown in Figure 1, microneedle array biosensor includes microneedle array 1, the biology knowledge in 1 surface modification of microneedle array
Other molecule 2 and the degradable high polymer material 3 of spraying, the microneedle array biosensor in embodiment 1 exist for glucose
Physical examination is surveyed.Specific preparation method is as follows：
A. microneedle array 1 is prepared：Microneedle array mold is prepared using laser boring, in the process for preparing microneedle array mold
In in the microneedle array mold inner-cavity formed gradient rough surface；It will be in liquefaction state, conductive microneedle array material
It is cast in the microneedle array mold, demoulding forms conductive microneedle array, and the material of microneedle array mold is aluminium flake, swashs
Light trepanning velocity is set as 200mm/s, when punching number is set as 1200 times, under the conditions of above-mentioned setting, by the aluminium of laser boring
Piece can produce the coarse surface of gradient, so as to form gradient rough surface in the inner cavity of microneedle array mold.Laser is beaten
The disk diameter in hole is 0.2mm, and disk diameter is that the setting of 0.2mm is the preferable aperture measured according to experiment, array of discs
Number can be designed according to the actual situation, a length of 6.8mm for the array of discs drawn in the present embodiment, and width is
6.8mm, the diameter of array of discs are 0.2mm, and array horizontal spacing is 0.6mm, longitudinal pitch 0.6mm, 12 × 12 arrays；This
Microneedle array material used in embodiment is glassy metal, and glassy metal is heated to its glass transition temperature, is poured
Microneedle array mold inner-cavity after laser boring demoulds after cooling, obtains conductive microneedle array.
B. in 1 surface modification biological identification molecule 2 of microneedle array.In the present embodiment, biological identification molecule 2 is specially enzyme
Molecule, using chemical crosslink technique in microneedle array surface modification biological identification molecule.The specific steps are：On 1 surface of microneedle array
Instillation enzyme solutions are placed in 4 DEG C of environment until microneedle array 1, is immersed in crosslinking agent after dry and carries out chemical friendship by drying
Connection, makes enzyme molecule be fixed on 1 surface of microneedle array by chemical bond.Selected enzyme molecule is the Portugal of 8mg/ml in the present embodiment
Grape carbohydrate oxidase 50uL, selected chemical cross-linking agent are glutaraldehyde.The glucose oxidase for selecting 8mg/ml is to its concentration
The optium concentration obtained after parameter optimization is carried out, under the concentration, microneedle array biosensor response is maximum.Selected penta
The concentration of dialdehyde is 0.2%, and 4h is impregnated in 4 DEG C of environment, and selecting crosslinking 4h is the Best Times of optimum experimental, is crosslinked 4h
When, glucose oxidase can be fixed on microneedle array biosensor surface with maximal efficiency.
C. degradable high polymer material 3 is sprayed.Referring to fig. 2, under the action of high voltage electric field, in the micropin that step b is obtained
1 surface of array sprays one layer of preferable degradable high polymer material of biocompatibility, to realize to biological identification molecule into one
Step protection.Specially：The intensity of high voltage electric field used in the present embodiment is 15kV, and the high molecular material of spraying is PVP, at this
Under voltage, in 1 surface of microneedle array that step b is obtained, the PVP of spraying forms one layer of relatively uniform nano fibrous membrane, Ke Yishi
Now to the further protection of glucose oxidase.
The microneedle array biosensor obtained according to above-mentioned steps is used to survey in physical examination, the degradable high score on surface
Sub- material can puncture in several minutes that enter dermal layer of the skin in micropin to be dissolved, and is exposed biological identification molecule, is being touched life
After object molecule, electric signal can be generated.
Embodiment 2
The present embodiment 2 and the difference of above-described embodiment 1 are that the microneedle array biosensor of the present embodiment 2 is used for gallbladder
Sterol is surveyed in physical examination.Selected enzyme molecule is the cholesterol oxidase 50uL of 10mg/ml in step b, in the same manner as in Example 1,
Enzyme molecule is also to be modified by chemical crosslink technique in the surface of microneedle array 1, and selected chemical cross-linking agent is still glutaraldehyde.Choosing
It is the optium concentration for obtain after parameter optimization to its concentration with the cholesterol oxidase of 10mg/ml, under the concentration, micropin
Array biosensor response is maximum.The concentration of selected glutaraldehyde is 0.2%, and 4h is impregnated in 4 DEG C of environment, selects and hands over
Connection 4h is the Best Times of optimum experimental, and when being crosslinked 4h, cholesterol oxidase can be fixed on microneedle array biology with maximal efficiency
Sensor surface.
Remaining step can refer to embodiment 1.
In preparation method provided by the above embodiment, the microneedle array mold prepared by the method for laser boring can
To generate gradient rough surface, surface roughness is obvious, and the rough surface of the microneedle array prepared by mold can be preferably
Protect biological identification molecule, in puncture process will not because of frictional force effect and large area falls off, can puncture
Biological identification molecule is protected in the process.And degradable high molecular material is sprayed to microneedle array surface and may be implemented pair
The second protection of biological identification molecule.This layer of high molecular material be it is degradable, can enter several minutes of tissue fluid in dissolve, cruelly
Expose biological identification molecule, the sensitivity of biosensor will not be had an impact, and there is good biocompatibility, no
Adverse reaction can be generated to human body.
Therefore, the present embodiment is realized the microneedle array biosensor of preparation in puncture process and is known to biology
The protection of other molecule, and can be realized the second protection to biological identification molecule and surveyed with realizing in physical examination, biological knowledge may be implemented
The peak use rate of other molecule, while making the preparation process of microneedle array biosensor simpler.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair
The restriction of embodiments of the present invention.For those of ordinary skill in the art, may be used also on the basis of the above description
To make other variations or changes in different ways.There is no necessity and possibility to exhaust all the enbodiments.It is all this
Made any modifications, equivalent replacements, and improvements etc., should be included in the claims in the present invention within the spirit and principle of invention
Protection scope within.
Claims (9)
1. a kind of preparation method of microneedle array biosensor, which is characterized in that including：
Make the conductive microneedle array with rough surface；
Molecule is identified in the rough surface modified biological of the microneedle array；
Degradable high polymer material is added on the microneedle array surface for having modified the biological identification molecule.
2. a kind of preparation method of microneedle array biosensor according to claim 1, which is characterized in that the production
There is the step of conductive microneedle array of rough surface, specially：
Microneedle array mold is prepared using laser boring, in the microneedle array mold during preparing microneedle array mold
Gradient rough surface is formed on inner cavity；Conductive polymer material is cast in the microneedle array mold, demoulding, which is formed, to be had
The conductive microneedle array of rough surface.
3. a kind of preparation method of microneedle array biosensor according to claim 2, which is characterized in that
The material of the microneedle array mold is aluminium flake, during preparing microneedle array mold using laser boring, punching
Speed is 100~1000mm/s, when punching number is 100~2000 times；
The disk diameter of laser boring is 0.2mm, a length of 6.8mm of array of discs, and the diameter of width 6.8mm, array of discs is
0.2mm, array horizontal spacing are 0.6mm, longitudinal pitch 0.6mm, 12 × 12 arrays.
4. a kind of preparation method of microneedle array biosensor according to any one of claim 1 to 3, feature exist
In,
Rough surface modified biological using chemical crosslink technique in the microneedle array identifies molecule.
5. a kind of preparation method of microneedle array biosensor according to claim 4, which is characterized in that
Under the action of high voltage electric field, spraying one layer on the microneedle array surface for having modified the biological identification molecule has biology
The degradable high polymer material of compatibility.
6. a kind of preparation method of microneedle array biosensor according to claim 5, which is characterized in that
The high molecular material is PVA or PVP.
7. a kind of preparation method of microneedle array biosensor according to claim 6, which is characterized in that
The intensity of the high voltage electric field is 10~20kV.
8. a kind of preparation method of microneedle array biosensor according to claim 7, which is characterized in that
The microneedle array material is glassy metal.
9. a kind of preparation method of microneedle array biosensor according to claim 7, which is characterized in that
The microneedle array material is the conducting polymer of doping carbon black and/or graphene and/or carbon nanotube and/or carbon fiber
Material.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115184421A (en) * | 2022-06-21 | 2022-10-14 | 中山大学 | Microneedle blood glucose sensor, preparation method thereof and differential pulse voltammetry test method |
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