CN107050512A - A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material - Google Patents
A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material Download PDFInfo
- Publication number
- CN107050512A CN107050512A CN201710330358.4A CN201710330358A CN107050512A CN 107050512 A CN107050512 A CN 107050512A CN 201710330358 A CN201710330358 A CN 201710330358A CN 107050512 A CN107050512 A CN 107050512A
- Authority
- CN
- China
- Prior art keywords
- solution
- dopa
- preparation
- structural modification
- tissue renovation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Materials For Medical Uses (AREA)
Abstract
A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material, belongs to composite biological material preparation field, and transformation of the way Preparation Method mainly includes：Prepare the amine-modified solution of DOPA, prepare silk fibroin nano-fiber film, the spraying amine-modified solution of DOPA.By DOPA structural modification fibroin albumen tissue renovation material made from this method, the cell adhesion and bioactivity of tissue renovation material are further increased.
Description
Technical field
The invention belongs to composite biological material preparation field, and in particular to a kind of DOPA structural modification fibroin albumen tissue is repaiied
The preparation method of multiple material.
Background technology
Biomaterial, which is developed so far, has gone through three Main Stages.Since 1980s, with medical treatment, health care
And the bio-medical material promoted for the purpose of quality of life etc. achieves quick development, has had more than 50 kinds of implantation instrument quilts
Applied to clinic, the engineering material of technology maturation is typically derived from, and with biocompatibility and defective tissue alternative functions,
Such as heart artificial blood vessel, joint prosthesis and the artificial kidney for being widely used to clinic.Above-mentioned bio-medical material, with one
Universal general character:Biologically inert.I.e. bio-medical material develops followed principle and is to try to acceptor to the different of implantation instrument
Thing reaction is preferably minimized.From last century eighties the to the nineties, the emphasis of biomedical materials field is gradually turned by biologically inert
To bioactivity and controlled degradation, second generation bio-medical material and product are developed.
Later stage 1990s, third generation biomaterial is accompanied by the foundation of organizational engineering, develops and send out rapidly
What exhibition was got up.Using Tissue Engineering Biomaterials support as representative, it combines engineering science and life science principle, careful to plant
Born of the same parents, which provide, is adapted to its growth, matrix synthesis and the biology space for playing other functions, overcomes in the past single cell and moves
The shortcomings of cell is difficult to survive in plant, matrix synthesis capability is low, the structure for engineered tissue provides cell carrier
And structure stand.By bioactive materials and degradation material, the two independent concepts have been combined this kind of bio-medical material
Come, molecular modification is carried out in degradation material, the interaction for causing cellular integration plain, induced cell proliferation, differentiation, and
The synthesis and assembling of extracellular matrix.But the performance for the biomaterial for being applied to organizational project at present need further to carry
It is high, it is impossible to meet the demand of modern medicine.
The content of the invention
In order to solve problems of the prior art, the invention provides a kind of DOPA structural modification fibroin albumen tissue
The preparation method of repair materials, further increases the cell adhesion and bioactivity of tissue renovation material.
The present invention uses following technical scheme：
A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material, comprises the following steps：
Step one：By 350mg Gelatins in 60mL water, heating water bath is to 50 DEG C of stirrings to being completely dissolved obtained gelatin
Solution；
Step 2：By gelatin solution and 150mgN- hydroxysuccinimides, 191mg1- ethyls-(3- dimethylaminos third
Base) phosphinylidyne diimine and 103mg dopamines mixing, in electric heating constant temperature tank 40 DEG C stirring 48 hours after, solution is placed in
Analyse in bag, be put into the beaker for filling ultra-pure water, be placed in electric heating constant temperature tank and stir 2-3day under the conditions of 40 DEG C, and every
8h changes a ultra-pure water；
Step 3：The solution remained in bag filter is taken out, is placed in -20 DEG C of household freezers and freezes 8h, then be placed on freeze dryer
48h is dried, adhesiveness gelatin is obtained；
Step 4：Adhesiveness gelatin is made to the 3mg/ml aqueous solution, pH value of water solution is adjusted to 7 with HCl and NaOH, i.e.,
Obtain the amine-modified solution of DOPA；
Step 5：2L deionized waters are heated to boiling, 4g sodium carbonate and 5g silkworm raw silks is added, boils after 30min, are used
Deionized water is cleaned up and dried, then dissolves silk with 9.3mol/L LiBr solution, and concentration of ordinary dissolution 20wt% treats completely molten
Solution is loaded into bag filter after solution, deionized water dialysis 72h, 9000r/min centrifugation 20min is repeated twice, obtains fibroin albumen molten
Liquid；
Step 6：Silk fibroin protein solution is injected in polyethylene ware, fibroin protein film is obtained after natural air drying, by fibroin egg
Tunica albuginea is dissolved in the formic acid of mass concentration 98%, is then charged into syringe and is carried out electrostatic spinning, obtains silk fibroin nano-fiber film；
Step 7：By the amine-modified solution even application of DOPA and silk fibroin nano-fiber film surface, and in 37 DEG C of dryings
Case is dried after 12h, is rinsed 3 times with phosphate buffer (PBS), is produced tissue renovation material.
It is preferred that, the bag filter remaining molecules amount described in step 2 is 10000Da.
It is preferred that, ultrapure water consumption is to cover bag filter in described step two.
It is preferred that, the spinning parameter of electrostatic spinning is in step 5：Spinning syringe needle internal diameter 0.9mm, spinning voltage is in 10-
15KV, distance is in 8-15cm, and speed is 0.1-O.5m/h.
It is preferred that, the thickness sprayed in step 6 is 5-30nm.
The beneficial effects of the present invention are：
1) dopamine is a kind of compound in neurohormone, containing abundant ethylamino and catechol active function groups,
The surface of any body can be almost sticked to, particularly with organic surface effect more preferably.Gelatin after DOPA is amine-modified sticks
Property is higher, and forms on material surface dopamine-gelatin layer of polymerization.The modification of dopamine, is not only increased fixed
The content of gelatin, while promoting the adhesion of cell, strengthens the cell compatibility of material surface.
2) substantial amounts of hydroxyl and a small amount of carboxyl and amino are contained in gelatin molecule structure, with extremely strong hydrophily.Cause
This, with the amino of dopamine condensation reaction can occur for the carboxyl in gelatin molecule.There are gelatin other synthetic materials can not compare
Biocompatibility, degradability and the bioactivity of plan.
3) fibroin albumen biocompatibility is good, and degradation speed is slow and catabolite has low immunogenicity to tissue,
Nutritious and repair ability is organized to skin, periodontal etc., fibroin albumen provides field as timbering material for cell growth, metabolism
Institute, suitable environment is provided to form new organization, the differentiation that can also regulate and control and induce in addition, forms new with functional group
Knit and organ.
Embodiment
The present invention is described in further detail with reference to embodiments.
Embodiment 1
A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material, comprises the following steps：
Step one：By 350mg Gelatins in 60mL water, heating water bath is to 50 DEG C of stirrings to being completely dissolved obtained gelatin
Solution；
Step 2：By gelatin solution and 150mgN- hydroxysuccinimides, 191mg1- ethyls-(3- dimethylaminos third
Base) phosphinylidyne diimine and 103mg dopamines mixing, in electric heating constant temperature tank 40 DEG C stirring 48 hours after, solution is placed in
Analyse in bag, described bag filter remaining molecules amount is 10000Da, bag filter is put into the beaker for filling ultra-pure water, ultra-pure water
Consumption stirs 2-3day to cover bag filter position, and beaker being placed in into electric heating constant temperature tank under the conditions of 40 DEG C, and every 8h
Change a ultra-pure water；
Step 3：The solution remained in bag filter is taken out, is placed in -20 DEG C of household freezers and freezes 8h, then be placed on freeze dryer
48h is dried, adhesiveness gelatin is obtained；
Step 4：Adhesiveness gelatin is made to the 3mg/ml aqueous solution, pH value of water solution is adjusted to 7 with HCl and NaOH, i.e.,
Obtain the amine-modified solution of DOPA；
Step 5：2L deionized waters are heated to boiling, 4g sodium carbonate and 5g silkworm raw silks is added, boils after 30min, are used
Deionized water is cleaned up and dried, then dissolves silk with 9.3mol/L LiBr solution, and concentration of ordinary dissolution 20wt% treats completely molten
Solution is loaded into bag filter after solution, deionized water dialysis 72h, 9000r/min centrifugation 20min is repeated twice, obtains fibroin albumen molten
Liquid；
Step 6：Silk fibroin protein solution is injected in polyethylene ware, fibroin protein film is obtained after natural air drying, by fibroin egg
Tunica albuginea is dissolved in the formic acid of mass concentration 98%, is then charged into syringe and is carried out electrostatic spinning, obtains silk fibroin nano-fiber film；
Step 7：By the amine-modified solution even application of DOPA and silk fibroin nano-fiber film surface, and in 37 DEG C of dryings
Case is dried after 12h, is rinsed 3 times with phosphate buffer (PBS), is produced tissue renovation material.
Cell adhesion experiments：The HUVEC cells of culture are taken, are digested, cell count, with 5 × 103Cell/mL density is inoculated with
On tissue renovation material made from embodiment 1 and blank control group fibroin material, per hole 1mL, 0.5h, 1h, 2h are inoculated with
Afterwards, rinsed with sterile PBS solution, as a result Microscopic observation and counting of taking a picture see the table below 1.It can be seen that amine-modified by DOPA
The cell quantity that fibroin albumen tissue renovation material afterwards is adhered to does not pass through the thin of the blank control group of modification significantly more than
Born of the same parents' quantity.
Table 1
Cell proliferation experiment：The HUVEC cells of culture are taken, are digested, cell count, with 5 × 103Cell/mL density is inoculated with
On tissue renovation material made from embodiment 1 and blank control group fibroin material, per hole 1mL, liquid was changed every two days, is inoculated with
After 5day, Cell counting Kit WST-8 detects the cytoactive of material surface, the results are shown in Table 2, it can be deduced that pass through dopamine
The cytoactive of fibroin albumen tissue renovation material after modification is slightly above the cytoactive of blank assay group.
Table 2
Claims (5)
1. a kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material, it is characterised in that comprise the following steps：
Step one：By 350mg Gelatins in 60mL water, heating water bath is molten to obtained gelatin is completely dissolved to 50 DEG C of stirrings
Liquid；
Step 2：By gelatin solution and 150mg N- hydroxysuccinimides, 191mg 1- ethyls-(3- dimethylaminopropyls)
Phosphinylidyne diimine and 103mg dopamines mixing, in electric heating constant temperature tank 40 DEG C stirring 48 hours after, solution is placed in bag filter
In, it is put into the beaker for filling ultra-pure water, is placed in electric heating constant temperature tank and 2-3day is stirred under the conditions of 40 DEG C, and changed every 8h
Ultra-pure water；
Step 3：The solution remained in bag filter is taken out, is placed in -20 DEG C of household freezers and freezes 8h, then is placed on freeze dryer dry
48h, obtains adhesiveness gelatin；
Step 4：Adhesiveness gelatin is made to the 3mg/ml aqueous solution, pH value of water solution is adjusted to 7 with HCl and NaOH, i.e., much
The amine-modified solution of bar；
Step 5：2L deionized waters are heated to boiling, 4g sodium carbonate and 5g silkworm raw silks is added, boils after 30min, spend from
Sub- water is cleaned up and dried, then dissolves silk with 9.3mol/L LiBr solution, concentration of ordinary dissolution 20wt%, until completely dissolved
Solution is loaded into bag filter, deionized water dialysis 72h, 9000r/min centrifugation 20min is repeated twice, obtains silk fibroin protein solution；
Step 6：Silk fibroin protein solution is injected in polyethylene ware, fibroin protein film is obtained after natural air drying, by fibroin protein film
It is dissolved in the formic acid of mass concentration 98%, is then charged into syringe and carries out electrostatic spinning, obtain silk fibroin nano-fiber film；
Step 7：By the amine-modified solution even application of DOPA and silk fibroin nano-fiber film surface, and it is dry in 37 DEG C of drying boxes
After dry 12h, rinsed 3 times with phosphate buffer (PBS), produce tissue renovation material.
2. a kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material according to claim 1, it is special
Levy and be：Bag filter remaining molecules amount described in step 2 is 10000Da.
3. a kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material according to claim 1, it is special
Levy and be：Ultrapure water consumption is to cover bag filter in described step two.
4. a kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material according to claim 1, it is special
Levy and be, the spinning parameter of electrostatic spinning is in step 5：Spinning syringe needle internal diameter 0.9mm, spinning voltage is in 10-15KV, distance
In 8-15cm, speed is 0.1-O.5m/h.
5. a kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material according to claim 1, it is special
Levy and be：The thickness sprayed in step 6 is 5-30nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710330358.4A CN107050512A (en) | 2017-05-11 | 2017-05-11 | A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710330358.4A CN107050512A (en) | 2017-05-11 | 2017-05-11 | A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107050512A true CN107050512A (en) | 2017-08-18 |
Family
ID=59597090
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710330358.4A Pending CN107050512A (en) | 2017-05-11 | 2017-05-11 | A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107050512A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109364295A (en) * | 2018-09-30 | 2019-02-22 | 上海交通大学医学院附属第九人民医院 | Fibroin albumen-dopamine-E7 small peptide compound rest and its preparation method and application |
| CN109403046A (en) * | 2018-10-16 | 2019-03-01 | 西南交通大学 | A kind of preparation method of the natural fibroin microfibre of dopamine regulation |
| CN111437441A (en) * | 2020-05-19 | 2020-07-24 | 重庆大学 | Drug-loaded KGN (KGN) nano-fiber scaffold and preparation method and application thereof |
| CN112640952A (en) * | 2020-12-16 | 2021-04-13 | 重庆市林业科学研究院 | Bamboo shoot preservative and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101579246A (en) * | 2009-05-31 | 2009-11-18 | 苏州大学 | Artificial silk fibroin nano-fiber nerve repair conduit and preparation method thereof |
| CN103721295A (en) * | 2013-12-19 | 2014-04-16 | 浙江大学 | Method for increasing biocompatibility and mechanical property of fibroin biomaterials |
-
2017
- 2017-05-11 CN CN201710330358.4A patent/CN107050512A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101579246A (en) * | 2009-05-31 | 2009-11-18 | 苏州大学 | Artificial silk fibroin nano-fiber nerve repair conduit and preparation method thereof |
| CN103721295A (en) * | 2013-12-19 | 2014-04-16 | 浙江大学 | Method for increasing biocompatibility and mechanical property of fibroin biomaterials |
Non-Patent Citations (1)
| Title |
|---|
| 杨茜: ""多巴结构用于材料表面的仿生修饰及其生物学基础研究"", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109364295A (en) * | 2018-09-30 | 2019-02-22 | 上海交通大学医学院附属第九人民医院 | Fibroin albumen-dopamine-E7 small peptide compound rest and its preparation method and application |
| CN109364295B (en) * | 2018-09-30 | 2021-08-24 | 上海交通大学医学院附属第九人民医院 | Silk fibroin-dopamine-E7 short peptide composite scaffold and preparation method and application thereof |
| CN109403046A (en) * | 2018-10-16 | 2019-03-01 | 西南交通大学 | A kind of preparation method of the natural fibroin microfibre of dopamine regulation |
| CN111437441A (en) * | 2020-05-19 | 2020-07-24 | 重庆大学 | Drug-loaded KGN (KGN) nano-fiber scaffold and preparation method and application thereof |
| CN112640952A (en) * | 2020-12-16 | 2021-04-13 | 重庆市林业科学研究院 | Bamboo shoot preservative and preparation method thereof |
| CN112640952B (en) * | 2020-12-16 | 2023-08-29 | 重庆市林业科学研究院 | Bamboo shoot preservative and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | Marine collagen scaffolds in tissue engineering | |
| Aigner et al. | Biomedical applications of recombinant silk‐based materials | |
| CN107050512A (en) | A kind of preparation method of DOPA structural modification fibroin albumen tissue renovation material | |
| CN101736430B (en) | Method for preparing silk fibroin nano-fibre with skin-care effect | |
| Zheng et al. | A review of recent progress on collagen‐based biomaterials | |
| CN100493626C (en) | Cell culturing rack material and its preparation | |
| Fang et al. | A biocompatible vascularized graphene oxide (GO)-collagen chamber with osteoinductive and anti-fibrosis effects promotes bone regeneration in vivo | |
| CN102220017B (en) | Injectable moulded silk fibroin hydrogel and preparation method thereof | |
| CN106729984A (en) | A kind of Isin glue collagen repairs sponge and preparation method thereof | |
| CN101856510B (en) | Preparation method of composite nano-fiber support material of silk fibroin and calcium silicate | |
| Bakhshandeh et al. | A review on advances in the applications of spider silk in biomedical issues | |
| Chen et al. | Silk fibroin combined with electrospinning as a promising strategy for tissue regeneration | |
| CN103083720B (en) | Silk fibroin tube and preparation method thereof | |
| CN107137764A (en) | A kind of preparation method of DOPA structural modification PLA tissue renovation material | |
| Liu et al. | Flexible water-absorbing silk-fibroin biomaterial sponges with unique pore structure for tissue engineering | |
| CN107789666A (en) | A kind of inwall micro-patterning small-caliber artificial blood vessel | |
| CN107149702A (en) | A kind of preparation of poly-dopamine modified lithium porous support | |
| Guo et al. | Microfluidic 3D printing polyhydroxyalkanoates-based bionic skin for wound healing | |
| Yan et al. | Preparation and applications of keratin biomaterials from natural keratin wastes | |
| CN109395162B (en) | Preparation method of natural protein-based bionic structure bone scaffold | |
| CN102133432A (en) | Preparation method of silk fibroin micropore bracket | |
| Dong et al. | Electrospun nanofibrous membranes of recombinant human collagen type III promote cutaneous wound healing | |
| CN108452366B (en) | Cod skin gelatin composite hemostatic dressing and preparation method thereof | |
| CN106822992A (en) | The preparation method of the implantation human body degradable biomaterial based on silk fibroin | |
| CN1210335C (en) | Process for preparing dual-layer combined chitosan-gelatin-mucinase scaffold material |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170818 |
|
| RJ01 | Rejection of invention patent application after publication |