CN103599562A - Preparation method of micropatterned nanofiber - Google Patents
Preparation method of micropatterned nanofiber Download PDFInfo
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- CN103599562A CN103599562A CN201310586083.2A CN201310586083A CN103599562A CN 103599562 A CN103599562 A CN 103599562A CN 201310586083 A CN201310586083 A CN 201310586083A CN 103599562 A CN103599562 A CN 103599562A
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Abstract
The invention relates to a preparation method of a micropatterned nanofiber. A physical structure of a salivary gland acinar cell system is simulated within nanometer and micrometer scale ranges by combining a photoetching micropattern array and utilizing the advantage that an electrostatic spinning PLGA (Polylactide/Glycolide) nanofiber membrane serves as an engineering salivary gland tissue; due to the addition of the photoetching micropattern array, the surface area of a nanofiber base material can be remarkably increased. The nanofiber prepared by the invention has more practical application values in the field of preparation of salivary glands for tissue engineering.
Description
Technical field
The present invention relates to a kind of preparation method of PLGA nanofiber body, relate in particular to a kind of preparation method of the micro-patterning PLGA nanofiber body for glandula cell and histo-differentiation.
Background technology
Can not secrete enough salivas, namely salivary gland function is bad is a kind of important clinical problem, is the initial reason forming of xerostomia.Conventionally this imbalance being caused by autoimmune disease, radiation therapy or head injury can reduce a people's the ability of chewing, swallow, digest and speak, and more serious meeting causes oral cavity infection, gingival and decayed tooth.The Therapeutic Method that salivary gland function is bad, generally include and use some to have a lot of side effect medicines or artificial saliva, and these is all temporary transient and is inadequate.The possible bad method of salivary gland function that is used for the treatment of is set up an a kind of engineering salivary gland exactly, and this salivary gland can permanently replace forfeiture or the damage of acinar tissue.
Artificial saliva gland relates generally to the method for two kinds of organizational projects: a kind of method be by salivary gland epithelial cell line or initial cell self assembly in three-dimensional substrates, can be that artificial basement membrane or natural base material are as hyaluronic acid, but shortcoming is the airtight spheroid forming lacks interconnecting property, thereby be unfavorable for catching of saliva, and artificial basement membrane can not be realized human implantation; Another kind is to utilize flat synthetic or natural base material to organize salivary gland cell to be for monofilm, yet the salivary gland cell of growing in flat polymeric substrates often can not form tight johning knot, and this johning knot is necessary to the orientation secretion of saliva, and this method can not form the surface area that can provide enough large, the three-dimensional dendritic morphology that can produce enough salivas.
Electrostatic spinning nano fiber film is prepared the base material with high surface area with nano-scale structures that can analog cell epimatrix for a lot of research, these base materials are often used as timbering material for mesenchymal adhesion, extension and infiltration, in the fibrous nebulin matter in extracellular matrix, are assembled into three-dimensional net structure.With nano fibrous membrane, as base material, be used for salivary gland epithelial cell, polylactic acid copolymerization hydroxyacetic acid (PLGA) nanofiber has the ability to promote to produce the pattern of more three-dimensional epithelioid cell.
Summary of the invention
The present invention relates to a kind of preparation of PLGA nanofiber body of the micro-patterning for glandula cell differentiation and tissue, concrete preparation process is as follows:
(1) preparation of micro-patterning bar shaped array: the monocrystalline silicon piece of 200nm is cleaned to 2min with the sulphuric acid of volume ratio 3:1 and 30% hydrogenperoxide steam generator, then washing is dry, rotating speed backspin at 2500rpm is coated P20 adhesion promoter, the spin coating time is 50s, spin coating 5min under the condition that positive photoresist SPR2207.0 is 1000rpm at rotating speed, then cures 10min at 80 ℃; Then the 10min that exposes in contact photoetching machine, soft contact mode is for being less than 3mJ/cm
2after exposure, the resist 10min that develops under AZ300MIF developing agent under gentle stirring condition, improve temperature to 160 ℃ and cure 5min, the ratio that is 10:1 according to presoma and cross-linking agent ratio is thoroughly mixed polydimethylsiloxane, is spun on the silicon chip of patterning, and spin speed is 1000rpm, the spin coating time is 80s, then at 80 ℃ of heat treatment 6h.After heat treatment, polydimethylsiloxane is peeled off, be immersed in ethanol 1h to remove unnecessary photoresist.
(2) circular glass cover slide is first used Vectabond agent treated so that the adhesion of polymer and dry.Before carrying out electrostatic spinning, coverslip applies as tack coat with polydimethylsiloxane (～10 μ m) presoma/cross-linking agent of thin layer, the circular array of fixed size covers the surface of polydimethylsiloxane layer then at 80 ℃ of dry 10min after polydimethylsiloxane sheet cuts down, the coverslip that contains array is put into receive on paper tinsel and deposits electrostatic spinning nano fiber film, the PLGA that electrostatic spinning liquid contains 15wt% (solvent is perfluor isopropyl alcohol) and the NaCl of 2wt% and the SRB of 5 μ mol; Spinneret is the syringe of 19G, and spinning voltage is 20kV, and spinning distance is 20cm, and spinning liquid fltting speed is 2mL/h, and the spinning time is 1h.After electrostatic spinning finishes, sample is dry 24h at 37 ℃ immediately.
In the present invention, utilize Static Spinning PLGA nano fibrous membrane as the advantage of engineering salivary organization, in conjunction with the micro-pattern array of photoetching, within the scope of nanometer and micro-meter scale, simulate the physical arrangement of salivary gland acinous cell system, photoengraving pattern array add the surface area that can significantly strengthen nano fiber basis material, therefore at preparation engineering, with salivary gland field, there is actual using value.
The specific embodiment
In order to deepen the understanding of the present invention, below in conjunction with instantiation, the present invention is described in further detail:
(1) monocrystalline silicon piece of the preparation of micro-patterning bar shaped array: 200nm cleans 2min with the sulphuric acid of volume ratio 3:1 and 30% hydrogenperoxide steam generator, then washing is dry, rotating speed backspin at 2500rpm is coated P20 adhesion promoter, the spin coating time is 50s, spin coating 5min under the condition that positive photoresist SPR2207.0 is 1000rpm at rotating speed, then cures 10min at 80 ℃; Then the 10min that exposes in contact photoetching machine, soft contact mode is for being less than 3mJ/cm
2after exposure, the resist 10min that develops under AZ300MIF developing agent under gentle stirring condition, improve temperature to 160 ℃ and cure 5min, the ratio that is 10:1 according to presoma and cross-linking agent ratio is thoroughly mixed polydimethylsiloxane, is spun on the silicon chip of patterning, and spin speed is 1000rpm, the spin coating time is 80s, then at 80 ℃ of heat treatment 6h.After heat treatment, polydimethylsiloxane is peeled off, be immersed in ethanol 1h to remove unnecessary photoresist.
(2) circular glass cover slide is first used Vectabond agent treated so that the adhesion of polymer and dry.Before carrying out electrostatic spinning, coverslip applies as tack coat with the polydimethylsiloxane presoma/cross-linking agent of thin layer.The circular array of fixed size covers the surface of polydimethylsiloxane layer then at 80 ℃ of dry 10min after polydimethylsiloxane sheet cuts down, the coverslip that contains array is put into receive on paper tinsel and deposits electrostatic spinning nano fiber film, the PLGA that electrostatic spinning liquid contains 15wt% (solvent is perfluor isopropyl alcohol) and the NaCl of 2wt% and the SRB of 5 μ mol; Spinneret is the syringe of 19G, and spinning voltage is 20kV, and spinning distance is 20cm, and spinning liquid fltting speed is 2mL/h, and the spinning time is 1h; After electrostatic spinning finishes, sample is dry 24h at 37 ℃ immediately.
Claims (4)
1. a preparation method for the nanofiber body of micro-patterning, its step is as follows:
(1) preparation of micro-patterning bar shaped array:
The monocrystalline silicon piece of 200nm is cleaned to 2min with the hydrogenperoxide steam generator of sulphuric acid and 30%, then washing is dry, rotating speed backspin at 2500rpm is coated P20 adhesion promoter, the spin coating time is 50s, spin coating 5min under the condition that positive photoresist SPR2207.0 is 1000rpm at rotating speed, then at 80 ℃, cure 10min, the 10min that then exposes in contact photoetching machine, soft contact mode is for being less than 3mJ/cm
2after exposure, the resist 10min that develops under AZ300MIF developing agent under gentle stirring condition, improve temperature to 160 ℃ and cure 5min, the ratio that is 10:1 according to presoma and cross-linking agent ratio is thoroughly mixed polydimethylsiloxane, is spun on the silicon chip of patterning, and spin speed is 1000rpm, the spin coating time is 80s, then at 80 ℃ of heat treatment 6h; After heat treatment, polydimethylsiloxane is peeled off, be immersed in 1h in ethanol;
(2) circular glass cover slide is first used Vectabond agent treated so that the adhesion of polymer and dry; Before carrying out electrostatic spinning, coverslip applies as tack coat with one deck polydimethylsiloxane presoma/cross-linking agent, the circular array of fixed size covers the surface of polydimethylsiloxane layer then at 80 ℃ of dry 10min after polydimethylsiloxane sheet cuts down, the coverslip that contains array is put into receive on paper tinsel and deposits electrostatic spinning nano fiber film, the PLGA that electrostatic spinning liquid contains 15wt% (solvent is perfluor isopropyl alcohol) and the NaCl of 2wt% and the SRB of 5 μ mol, the spinning time is 1h; After electrostatic spinning finishes, sample is dry 24h at 37 ℃ immediately.
2. the preparation method of nanofiber body as claimed in claim 1, is characterized in that: the sulphuric acid of described volume ratio 3:1 and 30% hydrogenperoxide steam generator.
3. the preparation method of nanofiber body as claimed in claim 1, is characterized in that: described spinning condition is: the syringe that spinning head is 19G, and spinning voltage is 20kV, and spinning distance is 20cm, and spinning liquid fltting speed is 2mL/h.
4. the preparation method of nanofiber body as claimed in claim 1, is characterized in that: described spinning condition is: the thickness of described polydimethylsiloxane is less than 10 μ m.
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| CN201310586083.2A CN103599562A (en) | 2013-11-15 | 2013-11-15 | Preparation method of micropatterned nanofiber |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105251047A (en) * | 2015-09-30 | 2016-01-20 | 温州生物材料与工程研究所 | Electrospinning porous nanofiber matrix micropattern seal support material and preparation method and application thereof |
| CN108525013A (en) * | 2018-03-23 | 2018-09-14 | 南通大学 | A kind of preparation method of tissue-engineering graft constructed of the surface with micron-nano topological geometry |
| CN109414524A (en) * | 2016-05-12 | 2019-03-01 | 阿塞拉外科有限公司 | Tissue substitute material and method of tissue repair |
| US11173234B2 (en) | 2012-09-21 | 2021-11-16 | Washington University | Biomedical patches with spatially arranged fibers |
| US11224677B2 (en) | 2016-05-12 | 2022-01-18 | Acera Surgical, Inc. | Tissue substitute materials and methods for tissue repair |
| US11311366B2 (en) | 2010-06-17 | 2022-04-26 | Washington University | Biomedical patches with aligned fibers |
| CN114870095A (en) * | 2022-05-06 | 2022-08-09 | 南通大学 | Method for constructing tissue engineering graft with surface having anisotropic nano topological structure |
-
2013
- 2013-11-15 CN CN201310586083.2A patent/CN103599562A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| DAVID A. SOSCIA ET AL.: "Salivary gland cell differentiation and organization on micropatterned PLGA nanofiber craters", 《BIOMATERIALS》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11311366B2 (en) | 2010-06-17 | 2022-04-26 | Washington University | Biomedical patches with aligned fibers |
| US11471260B2 (en) | 2010-06-17 | 2022-10-18 | Washington University | Biomedical patches with aligned fibers |
| US11173234B2 (en) | 2012-09-21 | 2021-11-16 | Washington University | Biomedical patches with spatially arranged fibers |
| US11253635B2 (en) | 2012-09-21 | 2022-02-22 | Washington University | Three dimensional electrospun biomedical patch for facilitating tissue repair |
| US11596717B2 (en) | 2012-09-21 | 2023-03-07 | Washington University | Three dimensional electrospun biomedical patch for facilitating tissue repair |
| CN105251047A (en) * | 2015-09-30 | 2016-01-20 | 温州生物材料与工程研究所 | Electrospinning porous nanofiber matrix micropattern seal support material and preparation method and application thereof |
| CN109414524A (en) * | 2016-05-12 | 2019-03-01 | 阿塞拉外科有限公司 | Tissue substitute material and method of tissue repair |
| US11224677B2 (en) | 2016-05-12 | 2022-01-18 | Acera Surgical, Inc. | Tissue substitute materials and methods for tissue repair |
| US11826487B2 (en) | 2016-05-12 | 2023-11-28 | Acera Surgical, Inc. | Tissue substitute materials and methods for tissue repair |
| CN108525013A (en) * | 2018-03-23 | 2018-09-14 | 南通大学 | A kind of preparation method of tissue-engineering graft constructed of the surface with micron-nano topological geometry |
| CN114870095A (en) * | 2022-05-06 | 2022-08-09 | 南通大学 | Method for constructing tissue engineering graft with surface having anisotropic nano topological structure |
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Application publication date: 20140226 |