CN108478864A - Composite fibrous scaffold - Google Patents
Composite fibrous scaffold Download PDFInfo
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- CN108478864A CN108478864A CN201810206824.2A CN201810206824A CN108478864A CN 108478864 A CN108478864 A CN 108478864A CN 201810206824 A CN201810206824 A CN 201810206824A CN 108478864 A CN108478864 A CN 108478864A
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- 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
- A61L27/56—Porous materials, e.g. foams or sponges
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- 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
- A61L27/20—Polysaccharides
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- 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
- A61L27/225—Fibrin; Fibrinogen
-
- 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
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- 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
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Dermatology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dispersion Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Materials For Medical Uses (AREA)
Abstract
The present invention is proposed extracts a kind of composite fibrous scaffold from whole blood, and wherein preparation process includes being mixed with HPMC normal saline solutions before whole blood blood coagulation；The mixed liquor is centrifuged to obtain upper layer clear gel sample substance to be composite fibrous scaffold.The stickiness higher of the more traditional fibrin holder of composite fibrous scaffold of the present invention, and character is stablized.Various concentration and the HPMC of different stickiness can adjust the rheological property of this fibrous framework.The composite fibrous scaffold is rich in a variety of growth factors, and growth factor can not be secreted outside because combining closely with web, be released again when being stimulated (such as wound healing).
Description
The application is application number 2017106649584, title composite fibrous scaffold and preparation method thereof, the applying date 2017
On August 7, divisional application.
Technical field
The present invention relates to Medical rack fields, particularly relate to a kind of composite fibrous scaffold.
Background technology
Regeneration and restoration is based on stem cell, growth factor and holder.Wherein holder is a kind of three dimensional matrix,
It can play a role under growth factor-induced to stem cell and provide environment.Good holder needs to meet claimed below：1 mould
Quasi- natural extracellular matrix；2 adhere to convenient for cell, migration, proliferation and differentiation；3 promote angiogenesis；4 good biology drops
Solution rate；5 physical properties appropriate are to support the structure of regenerating tissues；6 apertures are about 15-20um, are grown into convenient for new vessels.
Existing holder is divided into biological support and artificial synthesized holder, wherein biological support (such as rich platelet according to source
It is blood plasma platelet rich plasma, PRP, rich platelet fiber platelet rich fibrin, PRF, collagen, deacetylated
Chitin, hyaluronic acid etc.) due to being better than artificial synthesized holder with good biocompatibility.Wherein PRP and PRF be from
The fibrin containing Platelet Concentrate extracted in blood is used in regeneration field relatively broad in recent years.PRP preparation process
It is to centrifuge the whole blood of anti-freezing by two steps, adds calcium ion and fibrin ferment is made；PRF preparation process is directly to pass through whole blood
It is made after crossing primary centrifugation.Since PRP and PRF had not only had fibrous reticular structure but also a variety of growth factors are contained, are dry
Cell proliferation and differentiation and promotion regeneration provide good medium, are current better biological supports.
But there is also defects by PRP and PRF：They all do not have stable viscoelasticity, cannot be according to biological tissue again
It is raw to require progress moulding.And the apertures PRP are smaller (about 1-5 μm), although the apertures PRF are slightly larger (about 15-20 μm), PRF is being held
It is contracted to be less than 10 μm by the back aperture of external force deformation, therefore no matter the aperture of PRP or PRF is all not in full conformity with stem cell branch
The demand of frame.
In view of this, change is traditional to extract fibrinous preparation method from whole blood, to overcome drawbacks described above and obtain
It is very necessary to a kind of new ideal stem cell holder.
Bibliography：
1 Lee is wise and farsighted, Zhang Xiuming, Yin Yonglei, and the external free radical cracking of simple army political affairs Piao's hydroxypropyl methyl celluloses is ground
Study carefully chemistry and bioengineering .2013, vol.30No.6.
2.Dohan,D.M.,J.Choukroun,A.Diss,S.L.Dohan,et al.,Platelet-rich fibrin
(PRF):a second-generation platelet concentrate.Part II:platelet-related
biologic features.Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006.101
(3):p.e45-50.
Invention content
The present invention proposes a kind of composite fibrous scaffold, solve Character instability, aperture in the prior art it is small, can not basis
Regeneration requirements carry out moulding problem.
The present invention polymerize to form addition high molecular material hydroxypropyl first in the netted configuration process of fibrin in fibrinogen
Cellulose (hydroxypropyl methylcellulose, HPMC) and fibrin are compounded to form a kind of new fiber branch
Frame.
The technical proposal of the invention is realized in this way：
A kind of preparation method of composite fibrous scaffold, including,
The normal saline solution and whole blood for mixing HPMC, obtain mixed liquor；
The mixed liquor is centrifuged before the mixed liquor blood coagulation, it is composite fibre branch to obtain upper layer clear gel sample substance
Frame.
The HPMC uses collagen, hyaluronic acid, cellulose, chitin or hyaluronic acid as a preferred technical solution,
It is replaced with the synthetic product of cellulose.
Wherein, the chemical constitution of hyaluronic acid is as follows：
The chemical structure of general formula of cellulose is：
The chemical mechanical general formula of chitin is：
A concentration of 6-10% of the normal saline solution of the HPMC as a preferred technical solution,.
A concentration of 3-5%s of the HPMC in mixed liquor as a preferred technical solution,.
The centrifugation time is 8-15 minutes as a preferred technical solution,.
The process for preparation of the normal saline solution of HPMC is as follows as a preferred technical solution,：HPMC pulvis is first used final
After the 70-80 degree physiological saline solutions of the 1/3 of volume, then pour into 2/3 room temperature physiological saline stirring and dissolving.
A kind of composite fibrous scaffold is the above-mentioned clear gel sample substance being prepared.
Advantageous effect
(1) stickiness of composite fibrous scaffold of the invention is compared with PRP and PRF highers.This is because various concentration and difference
The HPMC of stickiness can adjust the rheological property of this fibrous framework.
(2) composite fibrous scaffold of the invention compares PRP and PRF, without apparent fibrous structure, but homogeneous has
The spawn of viscosity, character are stablized, and the various clinicals application such as wound surface smearing and skin-grafting is met.
(3) composite fibrous scaffold of the invention is rich in a variety of growth factors, and growth factor and web combine closely and
It is preserved, is released again when being stimulated (such as wound healing).The concentration mensuration result of ELISA also demonstrates the present invention
The outer secretory volume of growth factor is less than PRP (p<0.05), with PRF no significant differences.
(3) compared with PRP and PRF, the fiber aperture of composite fibrous scaffold of the invention is more big (to be shone referring to attached drawing Electronic Speculum
Piece 1-3).The apertures PRP are about 1-5um, and the apertures PRF are about 5-10um, and aperture of the present invention is about 15-20um, is more advantageous to new life
Blood vessel is grown into.Reason is：HPMC is high molecular material, its presence affects the fiber egg in fibrin fiber forming process
The polymerization of Bai Danti.On the other hand, the carboxyl in the hydroxyl and fibrinogen in HPMC forms hydrogen bond, changes fibrin
Secondary structure.
(5) the high molecular material HPMC content very littles used in preparation method of the invention, and HPMC is as implantation material
Expect without rejection, and can degradation in vivo [1].
Description of the drawings
It, below will be to embodiment in order to illustrate more clearly of embodiment of the present invention or technical solution in the prior art
Or attached drawing needed to be used in the description of the prior art is briefly described, it should be apparent that, the accompanying drawings in the following description is only
It is some embodiments of the present invention, for those of ordinary skill in the art, in the premise of not making the creative labor property
Under, other drawings may also be obtained based on these drawings.
Fig. 1 is electromicroscopic photograph of the present invention.
Fig. 2 is PRP electromicroscopic photographs.
Fig. 3 is PRF electromicroscopic photographs.
Fig. 4 fat stem cells cultivated on culture medium after figure；
A is that live/dead is dyed after 5%HPMC is co-cultured 7 with fat stem cell；
B is fat stem cell after ordinary culture medium culture 7 days.
Fig. 5 is the present invention, the ELISA testing results of PRF and PRP secreted in vitro cell factors.
Specific implementation mode
The technical scheme in the embodiments of the invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is common
The every other embodiment that technical staff is obtained without making creative work belongs to the model that the present invention protects
It encloses.
The reagent used in following embodiments is commercially available.
Embodiment 1
(1) a kind of preparation method of composite fibrous scaffold, including,
(1) HPMC normal saline solutions are prepared；
By HPMC pulvis first use final volume 1/3 70-80 degree physiological saline solutions after, then pour into 2/3 room temperature life
Saline dissolving is managed, the HPMC normal saline solutions of a concentration of 6-10% are configured to.
Wherein, the model E50 (medical type) of HPMC selections.
(2) 5-20mL animals or the whole blood of patient are collected with vacuum blood collection tube.
(3) whole blood for collecting HPMC normal saline solutions made from step (1) and step (2) is 1 in centrifuge tube：1 is mixed
It closes, obtains mixed liquor, HPMC final concentrations are diluted to 3-5%.
Wherein, mixing need to be completed before blood coagulation, within about 1 minute.
(4) (300-500g) is centrifuged before the mixed liquor blood coagulation of step (3) 8-15 minutes, obtain upper layer clear gel
Sample substance, lower layer are red blood cell layer, levels volume about 1：1.
(5) disposable plastic dropper is used to collect upper layer gel-like substance to get composite fibrous scaffold.
Compared with traditional blood platelet concentration of fibre protein product PRP and PRF, its stickiness higher and dimensionally stable.
HPMC is clinical common thickener, and the HPMC of the concentration and the different stickiness of selection that adjust HPMC can adjust this fibrous framework
Rheological property.
The composite fibrous scaffold is rich in a variety of growth factors.Document records PRP secretions growth factor concentration and is significantly higher than blood
Clearly, the growth factor concentration of opposite PRF secretions is significantly lower than serum；Reason is the close life of the fiber forming process of PRF
Situation is managed, structure is uniform and orderly, and growth factor is locked in fibrous reticular structure well；And the fiber of PRP is suddenly
Formed, structure is then stiff and unordered, growth factor can not with web in conjunction with and secreting outside [2].The compound fibre of the present invention
Dimensional scaffold forming process is similar to PRF, growth factor and web in conjunction with and can not secrete outside, in (such as surface of a wound that is stimulated
Healing) when release again.The concentration mensuration result of ELISA also demonstrates this point.
Compared with simple PRP and PRF, the fiber aperture of composite fibrous scaffold of the invention is bigger, and about 15um more has
It is grown into (referring to attached drawing 1-3) conducive to new vessels.Reason is：HPMC is high molecular material, its presence is affected in fiber egg
The polymerization of fibrin monomer in white fiber forming process.On the other hand, the carboxyl in the hydroxyl and fibrinogen in HPMC
Hydrogen bond is formed, fibrinous secondary structure is changed.
Traditional PRP and PRF is made of the fiber of solid phase and the serum of liquid phase, if such as squeezing or stirring by outer force effect
Mix, after serum and web separate, the rapid collapse of web and actual clinical can not be suitable for.The present invention is without apparent fiber
Shape structure, but the homogeneous spawn with viscosity meet the various clinicals application such as wound surface smearing and skin-grafting.
(2) experiment in vitro：
The HPMC of 5% concentration and fat stem cell are co-cultured 7.
Live/dead dyes the cytoplasm that Green fluorescence is living cells, and red is the nucleus of dead cell.Referring to Fig. 4
Live/dead is dyed after shown 5%HPMC is co-cultured 7 with fat stem cell, the results showed that 5%HPMC makees cytotoxic
With.Therefore stem cell stent applications be can be used as in regenerative medicine.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
With within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention god.
Claims (3)
1. a kind of composite fibrous scaffold, which is characterized in that the composite fibrous scaffold is clear gel sample substance；
Its preparation includes：
The normal saline solution and whole blood for mixing HPMC, obtain mixed liquor；
The mixed liquor is centrifuged before the mixed liquor blood coagulation, it is composite fibrous scaffold to obtain clear gel sample substance.
2. a kind of composite fibrous scaffold according to claim 1, which is characterized in that the model E50 of the HPMC selections.
3. a kind of composite fibrous scaffold according to claim 1, which is characterized in that the preparation of the normal saline solution of HPMC
Process is as follows：After HPMC pulvis is first 70-80 degree physiological saline solutions with 1/3 temperature of final volume, then pour into 2/3 it is normal
Warm saline stirring and dissolving.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201810206824.2A CN108478864B (en) | 2017-08-07 | 2017-08-07 | Composite fiber stent |
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| CN201810206824.2A CN108478864B (en) | 2017-08-07 | 2017-08-07 | Composite fiber stent |
| CN201710664958.4A CN107412878B (en) | 2017-08-07 | 2017-08-07 | Composite fibrous scaffold and preparation method thereof |
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| CN201710664958.4A Active CN107412878B (en) | 2017-08-07 | 2017-08-07 | Composite fibrous scaffold and preparation method thereof |
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| IL300830B1 (en) * | 2023-02-21 | 2024-04-01 | Reddress Ltd | An implantable object and a method of preparing it |
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| CN109432501A (en) * | 2018-10-30 | 2019-03-08 | 上海交通大学医学院附属第九人民医院 | Purposes of the composite fibrous scaffold in the composite material for preparing nasal plastic |
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| IL300830B1 (en) * | 2023-02-21 | 2024-04-01 | Reddress Ltd | An implantable object and a method of preparing it |
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| CN108478864B (en) | 2020-10-23 |
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