CN108939154A - A kind of preparation method of collagen protein composite fiber bracket - Google Patents
A kind of preparation method of collagen protein composite fiber bracket Download PDFInfo
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- 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
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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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- 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
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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
- A61L27/58—Materials at least partially resorbable by the body
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Abstract
The present invention relates to a kind of preparation methods of collagen protein composite fiber bracket, belong to field of tissue engineering technology.The present invention prepares spinning solution as solute, by solvent of hexafluoroisopropanol using the good collagen of biocompatibility and polycaprolactone, spinning solution is placed in high-speed rotating spinning device, forms the composite fibrous scaffold containing collagen on collecting board through centrifugal action.The present invention prepares the composite fibrous scaffold containing collagen by the effect of centrifugal force, this method is easy to operate, efficient and safe, solve the problems, such as that processing efficiency is low and is difficult to direct processed collagen albumen existing for electrostatic spinning, can be made the tunica fibrosa of multiple diameter, thickness, size under the premise of taking into account spinnability.Composite fibrous scaffold even in fiber diameter distribution obtained, porosity be high, large specific surface area, can be used for filtering, the fields such as medical accessory, medicament slow release and organizational project.
Description
Technical field
The present invention relates to a kind of preparation methods of collagen protein composite fiber bracket, belong to field of tissue engineering technology.
Background technique
Organizational project, be it is a kind of combined with cell biology and material science, construct in vitro or in vivo tissue or
The new branch of science of organ.Wherein most basic thinking is to separate in vitro, cultivate cell inoculation to certain space structure
On bracket, by mutually sticking between cell, growing multiplication, extracellular matrix secretion, thus formed have certain structure and
The tissue or organ of function.And coarse, porous surface and the curvature of fiber size of fibrous material (including single fiber, multifilament)
There is certain influence to the oriented growth of cell, this fibrous material, can also be with while providing skeleton template for cell growth
For guiding, the histiocytic response of specification.Therefore, suitable timbering material is screened, is the key that building tissue engineering bracket.
In conjunction with the advantage of natural material and polymer material respectively, it is expected to filter out a kind of good biocompatibility, good mechanical performance
Composite fibrous scaffold.
Collagen (Collagen) is one in the most abundant protein of animal in-vivo content and extracellular matrix
Kind, it is the double-helix structure twisted by 3 α peptide chains, and 3 polypeptide chains are mutually twisted again with hydrogen bond, keeps its molecular structure non-
Often stablize, and there is low immunogenicity and good biocompatibility, is widely used in terms of organizational project.But individual glue
Former azelon material mechanical performance is poor, and it is impossible to meet the requirements of tissue engineering material mechanical property.Polycaprolactone
(Polycaprolactone, PCL) be it is a kind of make catalyst in metallo-organic compound (such as tetraphenyltin) by 6-caprolactone, two
Ring-opening polymerisation forms under the conditions of hydroxyl or trihydroxy make initiator, belongs to aggretion type polyester.Its with fabulous mechanical property,
Special carbon-chain structure makes it have good flexibility and machinability.And there is good biocompatibility, and degradable
For CO2And H2O, cell can on its substrate normal growth.Therefore it is widely used in fracture fixation material, sutures, medical auxiliary
The fields such as material, drug release material and tissue engineering bracket material.It the characteristics of using above-mentioned collagen and polycaprolactone, can make
The standby composite fibrous scaffold for having both good biocompatibility, mechanical property and degradability.
China Patent Publication No. is CN101156962A, and publication date is on April 9th, 2008, denomination of invention are as follows: contains collagen
The preparation method of the complex nanometer fibrous tissue renovation bracket of albumen, the document discloses a kind of prepare to repair containing collagen tissue
The method of multiple bracket, the advantage of incorporating collagen albumen and thermoplastic polyurethane respectively pass through electrospinning process composite
Obtain collagen/polyurethane composite fibre.China Patent Publication No. is CN107519524A, and publication date is 2017 12
Months 29 days, denomination of invention are as follows: a kind of polycaprolactone/collagen/quaternary ammonium salt electrospun composite fibers bracket and its preparation side
Method, the document discloses a kind of electrospun composite fibers with good flexibility, mechanical property and quick-acting haemostatic powder performance
The preparation method of bracket.
Because pure collagen is difficult to form, so above-mentioned two Wen Jun is using quiet usually with the compound preparation of other materials
Electrical spinning method prepares composite fibrous scaffold, but electrostatic spinning because of its low efficiency, need high-pressure electrostatic, solution to need the disadvantages of conductive
It is set to be difficult to large-scale use.
Summary of the invention
In view of the above-mentioned problems, the purpose of the present invention is to provide a kind of preparation method of collagen protein composite fiber bracket,
To achieve the goals above, technical solution of the invention are as follows:
A kind of preparation method of collagen protein composite fiber bracket, the preparation method sequentially include the following steps:
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 12:88~22:78, at room temperature with 300~700
Turn/speed of min stirs to being completely dissolved, obtain collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 12:88~22:78, at room temperature with 300~700
Turn/speed of min stirs to being completely dissolved, obtain polycaprolactone solution.
C, by the collagen solution prepared through step a and the polycaprolactone solution prepared through step b according to 3:7~6:4
Mass ratio mixing, stir evenly, above-mentioned solution be placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, is obtained
Spinning solution, the mass fraction of total soluble matters are 12~22%.
D, spinning solution being placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 3000~
10000rpm, symmetrical 2 spinneret orifices in spinning device side, spinning aperture 1mm, spinneret orifice connect conical spinneret, spray
Silk mouth end diameter is 0.2~0.64mm, and spinning device bottom is arranged flat collector sheets, collecting board and spinning device distance from bottom 5~
15cm, spinning solution form the composite fibrous scaffold containing collagen through centrifugal action on collecting board.
Due to using above technical scheme, a kind of preparation side of collagen protein composite fiber bracket proposed by the present invention
Method, low immunogenicity and good biocompatibility mainly in conjunction with collagen and the good mechanical property of polycaprolactone and
Degradability.Collagen and polycaprolactone are soluble in hexafluoroisopropanol, and polycaprolactone can provide certain glue for spinning solution
Degree, and hexafluoroisopropanol has high boiling characteristics, polycaprolactone jet stream is fast as the solvent evaporates during high speed spinning
Speed draws and attenuates and is solidified into fiber, while collagen is retained in composite fibrous scaffold material with the solidification of polycaprolactone.
The present invention overcomes conventional spinning methods can not be to collagen at the problem of silk, while also by natural material and conjunction
Fiber is formed after combining jointly at material.Fiber is hardly formed when collagen is using conventional spinning method, main cause is glue
Former molecular weight of albumen is lower, can not be orderly aligned in fiberizing, at the same apply compared with noticeable effort when be easily broken off,
Therefore fiber can not be formed continuously.Collagen itself is natural material, and polycaprolactone is a kind of synthetic material, material properties with
And solubility behavior has biggish difference, even if the property of material forming is entirely different both after dissolution.It is orderly in order to allow it to carry out
Combination, the present invention is the effect by centrifugal force, is regulated and controled under field conditions (factors), both obtains good combination, simultaneously
It is capable of forming the molecular structure of orientations, obtains the fiber of good mechanical properties.
The selected total soluble matters mass fraction of the present invention is 12~22%, the reason is that total mass fraction can be led lower than above-mentioned value
It causes solution viscosity too low, is difficult to form continuous fiber in spinning process and effective cured fiber content is too low；Total mass fraction is high
It is too high to will lead to solution viscosity in above-mentioned value, spinneret can be blocked during the spinning process and lead to not spin fiber.Institute of the present invention
The collagen and polycaprolactone mass ratio selected is 3:7~6:4, the reason is that mass ratio will lead to timbering material lower than above-mentioned value
Middle collagen content is too low, is difficult to play its effect in subsequent applications；Collagen is higher than upper with polycaprolactone mass ratio
Stating value will lead to the low continuous fiber for being difficult to form good mechanical properties of polycaprolactone content.
Method of the invention be throw away Polymer Solution to form jet stream through spinneret orifice by centrifugal force, and inertia force,
It under the collective effect of viscous force and air drag, is attenuated is formed by curing fiber rapidly, and quickly move to be formed again to collector
Condensating fiber bracket.This method is easy to operate, efficient and safe, solves the problems, such as the presence of electrostatic spinning, is taking into account spinnability
Under the premise of by adjust spinning parameter can prepare multiple diameter (1~100 μm), thickness, size tunica fibrosa or can be by changing
Become the shape of receiver to prepare fibrous framework of different shapes.Due to the preparation method more than using, in the work of centrifugal force
It is stretched with lower high molecular material and is arranged along specific direction, the fiber being collected simultaneously is oriented along spinning head direction of rotation and arranged
Column, thus it is entirely different with traditional obtained product of spinning process and electrostatic spinning.Composite fibrous scaffold fiber obtained
Diameter is evenly distributed, porosity is high, large specific surface area, can be used for filtering, the neck such as medical accessory, medicament slow release and organizational project
Domain.
Detailed description of the invention Fig. 1 is collagen protein composite fiber rack assumption diagram
Specific embodiment
Present invention is further described in detail in the following with reference to the drawings and specific embodiments.
A kind of preparation method of collagen protein composite fiber bracket, the preparation method sequentially include the following steps:
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 12:88~22:78, at room temperature with 300~700
Turn/speed of min stirs to being completely dissolved, obtain collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 12:88~22:78, at room temperature with 300~700
Turn/speed of min stirs to being completely dissolved, obtain polycaprolactone solution.
C, by the collagen solution prepared through step a and the polycaprolactone solution prepared through step b according to 3:7~6:4
Mass ratio mixing, stir evenly, above-mentioned solution be placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, is obtained
Spinning solution, the mass fraction of total soluble matters are 12~22%.
D, spinning solution being placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 3000~
10000rpm, symmetrical 2 spinneret orifices in spinning device side, spinning aperture 1mm, spinneret orifice connect conical spinneret, spray
Silk mouth end diameter is 0.2~0.64mm, and spinning device bottom is arranged flat collector sheets, collecting board and spinning device distance from bottom 5~
15cm, spinning solution form the composite fibrous scaffold containing collagen through centrifugal action on collecting board.
The present invention is due to the preparation method more than using, and high molecular material is stretched along spy under the influence of centrifugal force
Determine direction arrangement, the fiber being collected simultaneously is aligned along spinning head direction of rotation, prepared composite fibrous scaffold knot
Composition is shown in Fig. 1, and black arrow illustrates aligning for fiber in figure.
Specific embodiment
It is prepared as described above method.
Embodiment one
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 22:78, is stirred at room temperature with the speed of 300 turns/min
It mixes to being completely dissolved, obtains collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 22:78, is stirred at room temperature with the speed of 300 turns/min
It mixes to being completely dissolved, obtains polycaprolactone solution.
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the quality of 3:7
It than mixing, stirs evenly, above-mentioned solution is placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, obtain spinning original
Liquid, the mass fraction of total soluble matters are 22%.
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 10000rpm, spinneret
Symmetrical 2 spinneret orifices in device side, spinning aperture 1mm, spinneret orifice connect conical spinneret, and spinneret end diameter is
Flat collector sheets, collecting board and spinning device distance from bottom 5cm is arranged in 0.25mm, spinning device bottom, and spinning solution is through centrifugal action
The composite fibrous scaffold containing collagen is formed on collecting board.
Embodiment two
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 12:88, is stirred at room temperature with the speed of 400 turns/min
It mixes to being completely dissolved, obtains collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 12:88, is stirred at room temperature with the speed of 400 turns/min
It mixes to being completely dissolved, obtains polycaprolactone solution.
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the quality of 6:4
It than mixing, stirs evenly, above-mentioned solution is placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, obtain spinning original
Liquid, the mass fraction of total soluble matters are 12%.
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 3000rpm, spinning device
Symmetrical 2 spinneret orifices in side, spinning aperture 1mm, spinneret orifice connect conical spinneret, and spinneret end diameter is
Flat collector sheets, collecting board and spinning device distance from bottom 15cm is arranged in 0.25mm, spinning device bottom, and spinning solution is through centrifugal action
The composite fibrous scaffold containing collagen is formed on collecting board.
Embodiment three
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 16:84, is stirred at room temperature with the speed of 500 turns/min
It mixes to being completely dissolved, obtains collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 16:84, is stirred at room temperature with the speed of 500 turns/min
It mixes to being completely dissolved, obtains polycaprolactone solution.
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the quality of 4:6
Than mixing, above-mentioned solution is placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, is stirred evenly, obtains spinning original
Liquid, the mass fraction of total soluble matters are 16%.
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 7000rpm, spinning device
Symmetrical 2 spinneret orifices in side, spinning aperture 1mm, spinneret orifice connect conical spinneret, and spinneret end diameter is
Flat collector sheets, collecting board and spinning device distance from bottom 10cm is arranged in 0.41mm, spinning device bottom, and spinning solution is through centrifugal action
The composite fibrous scaffold containing collagen is formed on collecting board, and spinning solution is placed in a high-speed rotating cylindrical spinning device
In, spin out composite fibrous scaffold.
Example IV
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 20:80, is stirred at room temperature with the speed of 400 turns/min
It mixes to being completely dissolved, obtains collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 20:80, is stirred at room temperature with the speed of 400 turns/min
It mixes to being completely dissolved, obtains polycaprolactone solution.
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the quality of 5:5
Than mixing, above-mentioned solution is placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, is stirred evenly, obtains spinning original
Liquid, the mass fraction of total soluble matters are 20%.
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 6000rpm, spinning device
Symmetrical 2 spinneret orifices in side, spinning aperture 1mm, spinneret orifice connect conical spinneret, and spinneret end diameter is
Flat collector sheets, collecting board and spinning device distance from bottom 8cm is arranged in 0.41mm, spinning device bottom, and spinning solution is through centrifugal action
The composite fibrous scaffold containing collagen is formed on collecting board, and spinning solution is placed in a high-speed rotating cylindrical spinning device
In, spin out composite fibrous scaffold.
Embodiment five
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 12:88, is stirred at room temperature with the speed of 700 turns/min
It mixes to being completely dissolved, obtains collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 12:88, is stirred at room temperature with the speed of 700 turns/min
It mixes to being completely dissolved, obtains polycaprolactone solution.
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the quality of 3:7
It than mixing, stirs evenly, above-mentioned solution is placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, obtain spinning original
Liquid, the mass fraction of total soluble matters are 12%.
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 3500rpm, spinning device
Symmetrical 2 spinneret orifices in side, spinning aperture 1mm, spinneret orifice connect conical spinneret, and spinneret end diameter is
Flat collector sheets, collecting board and spinning device distance from bottom 12cm is arranged in 0.2mm, spinning device bottom, and spinning solution is through centrifugal action
The composite fibrous scaffold containing collagen is formed on collecting board, and spinning solution is placed in a high-speed rotating cylindrical spinning device
In, spin out composite fibrous scaffold.
Embodiment six
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 22:78, is stirred at room temperature with the speed of 600 turns/min
It mixes to being completely dissolved, obtains collagen solution.
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 22:78, is stirred at room temperature with the speed of 600 turns/min
It mixes to being completely dissolved, obtains polycaprolactone solution.
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the quality of 6:4
It than mixing, stirs evenly, above-mentioned solution is placed in 4~5h in the vacuum environment of -0.1~0MPa and removes bubble removing, obtain spinning original
Liquid, the mass fraction of total soluble matters are 22%.
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 8000rpm, spinning device
Symmetrical 2 spinneret orifices in side, spinning aperture 1mm, spinneret orifice connect conical spinneret, and spinneret end diameter is
Flat collector sheets, collecting board and spinning device distance from bottom 10cm is arranged in 0.41mm, spinning device bottom, and spinning solution is through centrifugal action
The composite fibrous scaffold containing collagen is formed on collecting board.
Claims (1)
1. a kind of preparation method of collagen protein composite fiber bracket, which is characterized in that the preparation method according to the following steps into
Row:
A, the preparation of collagen solution
Collagen is mixed with hexafluoroisopropanol according to the mass ratio of 12:88~22:78, at room temperature with 300~700 turns/min
Speed stir to being completely dissolved, obtain collagen solution；
B, the preparation of polycaprolactone solution
Polycaprolactone is mixed with hexafluoroisopropanol according to the mass ratio of 12:88~22:78, at room temperature with 300~700 turns/min
Speed stir to being completely dissolved, obtain polycaprolactone solution；
C, by the collagen solution prepared through step a with the polycaprolactone solution for preparing through step b according to the matter of 3:7~6:4
Amount stirs evenly, above-mentioned solution, which is placed in 4~5h in the vacuum environment of -0.1~0MPa, removes bubble removing, obtains spinning than mixing
Stoste, the mass fraction of total soluble matters are 12~22%；
D, spinning solution is placed in a high-speed rotating cylindrical spinning device, spinning device revolving speed is 3000~10000rpm, spray
Symmetrical 2 spinneret orifices in silk device side, spinning aperture 1mm, spinneret orifice connect conical spinneret, spinneret end diameter
For 0.2~0.64mm, flat collector sheets, collecting board and 5~15cm of spinning device distance from bottom, spinning solution is arranged in spinning device bottom
The composite fibrous scaffold containing collagen is formed on collecting board through centrifugal action.
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| CN111560709A (en) * | 2019-07-26 | 2020-08-21 | 上海交通大学医学院附属上海儿童医学中心 | Nanofiber electrospun membrane containing axitinib and preparation method and application thereof |
| CN111719244A (en) * | 2020-06-18 | 2020-09-29 | 山东理工大学 | Preparation of polycaprolactone/collagen composite nanofiber membrane with artemisinin slow-release function |
| CN112999403A (en) * | 2021-03-01 | 2021-06-22 | 中国科学院烟台海岸带研究所 | Preparation method of oral cavity repairing film |
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