CN105944143B - Based on bionical tissue of acellular matrix-cell-three-dimensional fiber network and preparation method thereof - Google Patents
Based on bionical tissue of acellular matrix-cell-three-dimensional fiber network and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of based on bionical tissue of acellular matrix-cell-three-dimensional fiber network and preparation method thereof, includes at least compound acellular matrix hydrogel and epithelial cell based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network；The compound acellular matrix hydrogel is using the good three-dimensional fiber network material of biocompatibility as the hydrogel of skeleton；The good three-dimensional fiber network material of biocompatibility is the network structure material being wound by the good fiber of biocompatibility.Of the invention can be simulated based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network according to histoorgan structure, had a good application prospect, can be widely applied in field of biomedicine in organizational project.
Description
Technical field
The invention belongs to compound acellular matrix hydrogel preparation technical fields, are related to a kind of based on acellular matrix-cell-
Bionical tissue of three-dimensional fiber network and preparation method thereof.
Background technique
Hydrogel (Hydrogel) is the gel using water as decentralized medium, and a kind of macromolecule network system, property is soft,
It is similar to extracellular matrix in nature, shape, intensity is controllable and the characteristic of loose and porous inner surface is more next because it can arbitrarily change
It is applied in bioengineered tissue more.
All water-soluble or hydrophilic macromolecules can form water by certain chemical crosslinking or physical crosslinking
Gel.These macromolecules can be divided into natural and synthesis two major classes by its source.Natural hydrophilic macromolecule includes that polysaccharide (forms sediment
Powder, cellulose, alginic acid, hyaluronic acid, chitosan etc.) and polypeptide (collagen, polylysine, poly- L- Glutamic Acid etc.).It closes
At hydrophilic high mol include alcohol, acrylic acid and its derivative species (polyacrylic acid, polymethylacrylic acid, polyacrylamide, poly- N-
Gather for acrylamide etc.).Natural hydrogel has good biocompatibility and biological degradability, extensively by the pass of people
Note.
The difference of natural polymer hydrogel network bonding, can be divided into physical gel and chemical gel.Physical gel is logical
Cross the formation such as the physical force such as winding of electrostatic interaction, hydrogen bond, chain, this gel be it is non-permanent, it is solidifying by heating
Glue can be changed into solution, so also referred to as pseudo gel or heat-convertible gel.Such as gelatin by reduce temperature can be formed it is solidifying
Glue, and collagen forms gel by increasing temperature, silk fibroin protein solution can form hydrogel under high pressure.Chemical gel is
The three-dimensional network polymer being cross-linked to form by chemical bond is permanent, also known as true gel.Such as first in hyaluronic acid carboxyl
Upper introducing furan group is chemically crosslinked plastic using the PEG and furan nucleus of two terminal modified maleimides.But these substances are all
It is one-component, can not simulates completely and mix the natural tissues constituted by various ingredients, the biological function provided also extremely has
Limit.Acellular matrix (ECM) is to obtain complete tissue or organ removal cell and some antigenic component residues, is remained
The natural skeleton of extracellular matrix, containing collagenous fibres, glycoprotein, vascular endothelial growth factor etc., good biocompatibility
And biological degradability, it is ideal water-setting collagen-based materials.However, cannot be born big between hydrogel for isotropic material
Load, mechanical performance is poor, therefore constrains application range.
Enhance its maximum benefit of hydrogel competence exertion.In order to improve the mechanical property of hydrogel, many research workers
It is made that improvement, 104487103 A of patent publication No. discloses a kind of method of fiber reinforcement hydrogel composite material i.e. in water
The mode of fiber dispersion is added in gel bodies.105085791 A of patent publication No. discloses a kind of hydrophobic microballoon toughening increasing
The preparation method of strong physical crosslinking double-network hydrogel.But it is more likely to change the structure of gel bodies all to reach enhancing mechanics
The effect of performance changes original attribute of gel, and if transparency reduces, internal structure changes.Hydrogel is influenced simultaneously
Internal existing migration ingredient such as drug, vascular endothelial growth factor etc., to make the disabler of entire gel.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of based on acellular matrix-cell-three-dimensional fiber network
Bionical tissue and preparation method thereof, to solve, structural strength is insufficient in the prior art and the migration speed of the effective adding ingredient in inside
Spend fast technical problem.It is compound de- thin that the present invention uses the good three-dimensional fiber network material of biocompatibility to prepare for skeleton
Cytoplasmic matrix hydrogel can achieve the effect of enhancing.Meanwhile it proposing imitative based on acellular matrix-cell-three-dimensional fiber network
Raw tissue, forms sandwich structure, is then simulated according to histoorgan structure, has application well in organizational project
Prospect.
In the present invention, term " hydrogel precursor " refers to a kind of existence form before can be used to form hydrogel.
It is of the invention based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, it is described based on de- cell base
The bionical tissue of matter-cell-three-dimensional fiber network includes at least compound acellular matrix hydrogel and epithelial cell；It is described compound
Acellular matrix hydrogel is using the good three-dimensional fiber network material of biocompatibility as the hydrogel of skeleton；The biofacies
The good three-dimensional fiber network material of capacitive is the network structure material being wound by the good fiber of biocompatibility.Implantation
After organism, inflammation, rejection etc. will not be generated.The good three-dimensional fiber network material of biocompatibility refers to implantation
In organism, inflammation and rejection etc. are not generated, and there is good biological degradability.
As a preferred technical scheme:
As described above based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, the biocompatibility is good
Good three-dimensional fiber network material is natural or synthesis the good three-dimensional fiber network material of biocompatibility；The biology
Fibre diameter in the good three-dimensional fiber network material of compatibility is 500nm~3 μm, and pore diameter range is 1.0 μm -9 μm, hole
Rate is 65%-90%, and micropore is evenly distributed；The main component of the acellular matrix hydrogel is water, porosity 50%-
90%, pore diameter range is 10 μm -450 μm；The acellular matrix of the acellular matrix hydrogel be derived from urethra, ureter,
Bladder, blood vessel, small intestine, alimentary canal, vagina or fallopian tubal acellular matrix；The good fiber of biocompatibility is fibroin
Azelon, poly (glycolide-co-lactide) fiber or polycaprolactone fiber.
It is described including at least compound as described above based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network
Acellular matrix hydrogel and epithelial cell are one of following scenario described:
Compound acellular matrix hydrogel situation in conjunction with epithelial cell:
Epithelial cell is planted on compound acellular matrix hydrogel；
Compound acellular matrix hydrogel upper surface covers hydrogel, and in compound acellular matrix hydrogel and hydrogel
Between plant epithelial cell；
Compound acellular matrix hydrogel upper surface covers hydrogel, and epithelial cell is planted in hydrogel upper surface；
Compound acellular matrix hydrogel upper surface cover two layers of hydrogel, epithelial cell be planted in two layers of hydrogel it
Between；
Alternatively, further including one of following scenario described further:
Compound acellular matrix hydrogel situation in conjunction with smooth muscle cell:
Plant smooth muscle cell in compound acellular matrix hydrogel lower surface；
Compound acellular matrix hydrogel lower surface covers hydrogel, and in compound acellular matrix hydrogel and hydrogel
Between plant smooth muscle cell；
Compound acellular matrix hydrogel lower surface covers hydrogel, and smooth muscle cell is planted in hydrogel lower surface；
Compound acellular matrix hydrogel lower surface cover two layers of hydrogel, smooth muscle cell be planted in two layers of hydrogel it
Between；
The compound acellular matrix hydrogel is acellular matrix hydrogel and the good three-dimensional of the biocompatibility
Network of fibers material is whole or part is compound；The entirety is compound to be referred in acellular matrix hydrogel integrally comprising bio-compatible
The good three-dimensional fiber network material of property；The part is compound to refer to the upper of the good three-dimensional fiber network material of biocompatibility
Surface and lower surface and its superficial layer and acellular matrix hydrogel are compound；
The hydrogel is the hydrogel that acellular matrix is prepared, specially bladder acellular matrix hydrogel, blood
Pipe acellular matrix hydrogel, urethra acellular matrix hydrogel or mucous membrane of small intestine acellular matrix hydrogel；
As described above based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, the compound de- cell base
Also containing migration ingredient in matter hydrogel, the migration ingredient dispersed encapsulated is in the hydrogel.
As described above based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, the migration ingredient refers to
Migration ingredient is dispersed in acellular matrix hydrogel, but does not generate any chemical reaction；The migration ingredient is blood vessel endothelium
Growth factor, insulin, aspirin, Ketoprofen, Shu Tian be logical or one or more of Finasteride.
As described above based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, the acellular matrix water
The storage temperature of gel is at 4 DEG C~40 DEG C.
The present invention also provides the preparation method based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, institutes
Stating preparation method is infusion method, cladding process or spray coating method.
Preparation method as described above prepares acellular matrix hydrogel precursor solution first, then impregnate, coat or
Spraying, then stand several minutes at 37 DEG C and form compound acellular matrix hydrogel；Usually stand 3~30min.
The immersion are as follows:
Before gel is not formed, the good three-dimensional fiber network material of biocompatibility is soaked in the de- cell base
Matter precursor solution；
The coating are as follows:
Before gel is not formed, it is good that the acellular matrix hydrogel precursor solution is coated in biocompatibility
Three-dimensional fiber network material upper surface, lower surface or upper and lower surfaces, form one layer of good three-dimensional fiber of biocompatibility
The stepped construction that network structure and acellular matrix hydrogel precursor solution are constituted；Or it is further that biocompatibility is good
Three-dimensional fiber network material fits in coated acellular matrix hydrogel precursor solution layer, forms multi-layer biological compatibility
The stepped construction that good three-dimensional fiber network structure and acellular matrix hydrogel precursor solution are constituted；
The spraying are as follows:
It is before gel is not formed, the acellular matrix hydrogel precursor solution spraying is good in biocompatibility
Three-dimensional fiber network material upper surface, lower surface or upper and lower surfaces, form one layer of good three-dimensional fiber of biocompatibility
The stepped construction that network structure and acellular matrix hydrogel precursor solution are constituted；Or it is further that biocompatibility is good
Three-dimensional fiber network material fits in sprayed acellular matrix hydrogel precursor solution layer, forms multi-layer biological compatibility
The stepped construction that good three-dimensional fiber network structure and acellular matrix hydrogel precursor solution are constituted.
The preparation of acellular matrix hydrogel precursor solution is made in acellular matrix；
Freeze grinding again after the acellular matrix material made is freeze-dried, is broken into powder；In hydrochloric acid solution
It is middle to use pepsin digestion acellular matrix powder about 48h, acellular matrix pre-gel solution is formed, it is de- thin to be added about 1/10
The NaOH solution of cytoplasmic matrix pre-gel solution volume, wherein the concentration of NaOH solution is 0.01mol/L, wherein adding about 1/
10 × PBS of 9 acellular matrix pre-gel solution volumes, wherein the concentration of PBS is 0.0.1mol/L, and it is solidifying to form acellular matrix
Glue precursor solution.
Preparation method as described above, the good three-dimensional fiber network material of biocompatibility are prepared as Static Spinning
Silk method or 3 D weaving method.
Preparation method as described above, be additionally added in the preparation process of acellular matrix hydrogel precursor solution migration at
Point.
The mechanism of action of the invention are as follows:
Hydrogel be strand crosslink and the three-dimensional network of molecular scale that is formed, migration ingredient is from wherein sustained release master
It is related with diffusion velocity of the migration ingredient in gel.The diffusion velocity first aspect is related with hydrogel structure, i.e. water-setting
The three-dimensional network density of glue, network pore size and distribution, cross-link intensity, distribution of charges and intensity；Second aspect and drug knot
Structure is related with content, such as drug size, drugloading rate, pharmaceutical molecular weight, load position, drug distribution of charges and intensity；Third party
Face is related with environmental response condition, i.e. ambient temperature, pH value, electric field, magnetic field, light etc..
Three-dimensional fiber network material is that fiber is intertwined the three-dimensional network shape bulk material being staggered to form, and is belonged to micro-nano
The three-dimensional net structure of scale, excellent in mechanical performance, can be used for enhancing hydrogel and obtain has isotropism mechanical behavior special
The composite structure of sign, while the fibre diameter in three-dimensional fiber network is 500nm~3 μm, pore diameter range is 1.0 μm of -9 μ
M, porosity 65%-90%, and micropore is evenly distributed.
In view of above-mentioned, the present invention provides a kind of preparations of the bionical tissue of acellular matrix-cell-three-dimensional fiber network
Method is the preparation of compound acellular matrix hydrogel first, i.e., acellular matrix hydrogel and biocompatibility is good
Three-dimensional fiber network material is compound by spray coating method, infusion method, cladding process progress physics.Main component in hydrogel is water,
Porosity is 50%-90%, and pore diameter range is 10 μm -450 μm, therefore its aperture is greater than the aperture of three-dimensional fiber network.The two
After compound, the macroporous structure in hydrogel can completely or partially be filled by three-dimensional fiber network structure.On the one hand, three-dimensional network
Network plays the role of three-dimensional framework in hydrogel, to enhance the mechanical property of hydrogel.On the other hand, three-dimensional network
Network can fill the portion void that reason water occupies in hydrogel, also can construct mixing portion fiber in polymer in hydrogel.
Since the density of three-dimensional fiber network is greater than water, diffusion velocity of the drug in composite hydrogel is slower than in no three-dimensional fiber network
Diffusion velocity in hydrogel, therefore drug can be delayed in hydrogel to the sustained release of material outer.In addition, when hydrogel construct it is poly-
When closing mixing portion fiber in object, due to unsupported drug in three-dimensional fiber network polymer, it is carried in hydrogel
Drug diffusion rate can be also delayed.It is more lasting that above-mentioned two reason will lead to the medicament slow release time in composite hydrogel.
By the three-dimensional fiber network material for selecting that there is different pore size, fibre diameter, porosity, or by changing three-dimensional fiber network
Polymer and hydrogel construct the complex method of polymer, composite denseness, and the intensity and drug diffusion of composite hydrogel is adjusted
Behavior.
Since three-dimensional fiber network and hydrogel network belong to the three-dimensional network of different scale, three-dimensional fiber network and water
The compound main body three-dimensional net structure that can't change hydrogel of gel, such as the phase of crosslink density, hydrogel material and drug
The environment-responsive of interaction and hydrogel.
Acellular matrix is mainly derived from the tissue or organ of people or mammal, is mainly derived from urethra, ureter, wing
The tissue such as Guang, blood vessel, small intestine, alimentary canal, vagina or fallopian tubal is by complete tissue or organ removal cell and antigenic component
Obtained complete extracellular matrix, is made of collagen, polysaccharide, glycoprotein and vascular endothelial growth factor, there is good life
Object compatibility and biological degradability.Therefore, using the good three-dimensional fiber network material of biocompatibility as the de- cell base of skeleton
Matter hydrogel compound system has good biocompatibility, can apply to bioengineered tissue, can be used for tissue repair, and full
The reparation of sufficient different parts irregular shape needs.
Histoorgan has general character, is the official jargon structure of mucous membrane (also known as epithelium) and smooth muscle composition, and different tissues need
Cell be broadly divided into two major classes, epithelium (mucous membrane) cell and smooth muscle cell, as nerve and vascular tissue, content it is opposite
In epithelium and smooth muscle much less, therefore we only need to introduce the factor, then grow into after implanting by surrounding tissue.Cause
This, is prepared into sandwich structure for acellular matrix/cell/three-dimensional fiber network material, then carries out mould according to histoorgan
It is quasi-, sandwich structure is laid out and is arranged, is formed based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network,
It is had a good application prospect in organizational project.
Beneficial effect
(1) acellular matrix hydrogel overcomes that traditional natural hydrogel component is single, biological function is single in the present invention
The shortcomings that.
(2) direct combination of three-dimensional fiber network of the invention and gel, will not change gel this body structure can reach
Enhance the purpose of hydrogel.
(3) structure relative to hydrogel loose and porous inner surface, three-dimensional fiber network compact structure, for hydrogel Chinese medicine
Object, the package of active factors are fine and close, can play the effect of sustained release, reduce the rate of release of drug, to improve its utilization rate.
(4) preparation is simple by the present invention, and the compound acellular matrix hydrogel of preparation has good three-dimensional space
Between structure, good mechanical properties, good biocompatibility can be widely applied in field of biomedicine.
(5) can be flexible and changeable according to institutional framework based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network,
Simulated tissue organ structure well has a good application prospect in organizational project.
Detailed description of the invention
Fig. 1 is the scanning electron microscope diagram of composite hydrogel；
Fig. 2 is the schematic diagram that embodiment 1 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 3 is the schematic diagram that embodiment 2 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 4 is the schematic diagram that embodiment 3 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 5 is the schematic diagram that embodiment 4 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 6 is the schematic diagram that embodiment 5 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 7 is the schematic diagram that embodiment 6 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 8 is the schematic diagram that embodiment 7 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Fig. 9 is the schematic diagram that embodiment 8 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Figure 10 is the schematic diagram that embodiment 9 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Figure 11 is the schematic diagram that embodiment 10 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Figure 12 is the schematic diagram that embodiment 11 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Figure 13 is the schematic diagram that embodiment 12 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Figure 14 is the schematic diagram that embodiment 13 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network；
Figure 15 is the schematic diagram that embodiment 14 prepares the gained bionical tissue of acellular matrix-cell-three-dimensional fiber network.
In figure: 1 is epithelial cell, and 2 be hydrogel, and 3 be compound acellular matrix hydrogel, and 4 be smooth muscle cell；
3-1 is the upper surface of the good three-dimensional fiber network material of biocompatibility and its superficial layer and acellular matrix water
Gel is compound；
3-2 is the good three-dimensional fiber network material of biocompatibility of the compound compound acellular matrix hydrogel in part
Material.
Specific embodiment
The invention will be further elucidated with reference to specific embodiments.It should be understood that these embodiments are merely to illustrate this hair
It is bright rather than limit the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, art technology
Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Fixed range.
It is of the invention based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, for including at least compound de- thin
Cytoplasmic matrix hydrogel and epithelial cell, the compound acellular matrix hydrogel, for the good three-dimensional fiber of biocompatibility
Network material is the hydrogel of skeleton, i.e., includes the good three-dimensional fiber network material of biocompatibility in acellular matrix hydrogel
Material；The good three-dimensional fiber network material of biocompatibility is the network being wound by the good fiber of biocompatibility
Structural material will not generate inflammation, rejection etc. after being implanted into organism.Fig. 1 is the scanning electron microscope of composite hydrogel
Figure, it can be seen from the figure that hydrogel is closely connect as weaker zone, with three-dimensional fiber network structure as compacted zone, and
It is mutually communicated.The good three-dimensional fiber network of biocompatibility is natural or synthesis the good three-dimensional of biocompatibility
Network of fibers；Fibre diameter in the good three-dimensional fiber network of biocompatibility is 500nm~3 μm, and pore diameter range is
1.0 μm -9 μm (the fiber aperture of the good three-dimensional fiber network material of biocompatibility is no more than 20 μm), porosity is
65%-90%, and micropore is evenly distributed；Main component in the acellular matrix hydrogel is water, porosity 50%-
90%, pore diameter range is 10 μm -450 μm.
The compound acellular matrix hydrogel and epithelial cell of including at least is one of following scenario described:
Compound acellular matrix hydrogel situation in conjunction with epithelial cell:
Epithelial cell is planted on compound acellular matrix hydrogel；
Compound acellular matrix hydrogel upper surface covers hydrogel, and in compound acellular matrix hydrogel and hydrogel
Between plant epithelial cell；
Compound acellular matrix hydrogel upper surface covers hydrogel, and epithelial cell is planted in hydrogel upper surface；
Compound acellular matrix hydrogel upper surface cover two layers of hydrogel, epithelial cell be planted in two layers of hydrogel it
Between；
Alternatively, further including one of following scenario described further:
Compound acellular matrix hydrogel situation in conjunction with smooth muscle cell:
Plant smooth muscle cell in compound acellular matrix hydrogel lower surface；
Compound acellular matrix hydrogel lower surface covers hydrogel, and in compound acellular matrix hydrogel and hydrogel
Between plant smooth muscle cell；
Compound acellular matrix hydrogel lower surface covers hydrogel, and smooth muscle cell is planted in hydrogel lower surface；
Compound acellular matrix hydrogel lower surface cover two layers of hydrogel, smooth muscle cell be planted in two layers of hydrogel it
Between；
The good fiber of biocompatibility is that fibroin fiber, poly (glycolide-co-lactide) fiber or polycaprolactone are fine
Dimension etc.；
The acellular matrix of the acellular matrix hydrogel is to be derived from nerve, spinal cord, tendon, muscle, mucous membrane of urinary bladder, small
Intestinal mucosa, heart, liver, kidney or cartilaginous tissue acellular matrix.
Also containing migration ingredient in the compound acellular matrix hydrogel, the migration ingredient dispersed encapsulated is described de-
In cellular matrix hydrogel.
The migration ingredient refers to that migration ingredient is dispersed in acellular matrix hydrogel, but it is anti-not generate any chemistry
It answers；The migration ingredient be that vascular endothelial growth factor, insulin, aspirin, Ketoprofen, Shu Tian be logical or Finasteride in
One or more.
The storage temperature of the compound acellular matrix hydrogel is at 4 DEG C~40 DEG C.
Embodiment 1
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from urethra, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds insulin, and wherein the mass ratio of insulin and acellular matrix is 0.05:100, forms de- cell base
Matter gel precursor solution.Before gel is not formed, above-mentioned acellular matrix precursor solution is coated in and is compiled by three-dimensional
The fibre diameter of weave preparation is 3 μm, and pore diameter range is 8-9 μm, porosity 90%, and micropore is evenly distributed biocompatibility
The upper surface of good poly (glycolide-co-lactide) network of fibers material forms one layer of three-dimensional fiber network structure and acellular matrix
The stepped construction that hydrogel precursor solution is constituted, then stands 30min at 37 DEG C again, forms compound acellular matrix water-setting
Glue；One layer of hydrogel precursor solution, specially bladder acellular matrix are coated in compound acellular matrix hydrogel surface again
Precursor solution stands 5min at 37 DEG C and continues gel, forms bionical group of acellular matrix-cell-three-dimensional fiber network
It knits, as shown in Figure 2；Main component in compound acellular matrix hydrogel is water, porosity 90%, pore diameter range 400-
450 μm, storage temperature is 40 DEG C.
Embodiment 2
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from ureter, and freeze grinding again after being freeze-dried is beaten
It is broken into powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds aspirin, and wherein the mass ratio of aspirin and acellular matrix is 1:20, forms de- cell base
Matter gel precursor solution.Before gel is not formed, above-mentioned acellular matrix precursor solution is sprayed on and is compiled by three-dimensional
The fibre diameter of weave preparation is 3 μm, and pore diameter range is 8-9 μm, porosity 90%, and micropore is evenly distributed biocompatibility
The upper surface of good poly (glycolide-co-lactide) network of fibers material forms one layer of three-dimensional fiber network structure and acellular matrix
The stepped construction that hydrogel precursor solution is constituted；3min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；?
Compound acellular matrix hydrogel surface plants epithelial cell, then the compound acellular matrix hydrogel table in plantation epithelial cell
Face sprays one layer of hydrogel precursor solution, specially blood vessel acellular matrix hydrogel precursor solution, then at 37 DEG C it is quiet
9min is set, gel is continued, forms compound acellular matrix hydrogel upper surface covering hydrogel, and in compound acellular matrix water
The bionical tissue of acellular matrix-cell-three-dimensional fiber network that epithelial cell is planted between gel and hydrogel, such as Fig. 3 institute
Show；Main component in compound acellular matrix hydrogel is water, and porosity 70%, pore diameter range is 15-60 μm, storage temperature
Degree is 8 DEG C.
Embodiment 3
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from bladder, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds Ketoprofen, and wherein the mass ratio of Ketoprofen and acellular matrix is 1:10, and it is solidifying to form acellular matrix
Above-mentioned acellular matrix precursor solution is coated in front of gel is not formed and passes through 3 D weaving method by glue precursor solution
The fibre diameter of preparation is 1.8 μm, and pore diameter range is 3-5 μm, porosity 80%, and the micropore biocompatibility that is evenly distributed is good
The upper surface of good poly (glycolide-co-lactide) network of fibers material, forms one layer of three-dimensional fiber network structure and acellular matrix water
The stepped construction that gel precursor solution is constituted；9min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；Exist again
Compound acellular matrix hydrogel surface coats one layer of hydrogel precursor solution, specially before urethra acellular matrix hydrogel
Liquid solution is driven, 5min is stood at 37 DEG C and continues gel, so that compound acellular matrix hydrogel upper surface is covered hydrogel, upper
Chrotoplast is planted in hydrogel upper surface, then coats one layer in the compound acellular matrix hydrogel surface of plantation epithelial cell
Hydrogel precursor solution, specially urethra acellular matrix hydrogel precursor solution, then 9min is stood at 37 DEG C, continue
Gel, formed compound acellular matrix hydrogel upper surface cover two layers of hydrogel, epithelial cell be planted in two layers of hydrogel it
Between the bionical tissue of acellular matrix-cell-three-dimensional fiber network, as shown in Figure 4；In compound acellular matrix hydrogel
Main component is water, and porosity 78%, pore diameter range is 150-200 μm, and storage temperature is 10 DEG C.
Embodiment 4
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from blood vessel, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L, adds vascular endothelial growth factor, and the mass ratio of medium vascular endothelial growth factor and acellular matrix is
0.025:100, forming acellular matrix gel precursor solution will be prepared before gel is not formed by method of electrostatic spinning
Fibre diameter be 500nm, pore diameter range is 1-2 μm, porosity 65%, and the micropore biocompatibility that is evenly distributed is good
Fibroin fiber network material be cut into diameter be 14mm disk, it is molten to be soaked in above-mentioned 2mL acellular matrix presoma
In liquid, is formed in fibroin fiber network material hole and construct polymer, acellular matrix containing acellular matrix hydrogel
It is also mutual comprising structure containing three-dimensional fiber network material in hydrogel hole；3min is stood at 37 DEG C again, is formed compound
Acellular matrix hydrogel；Then epithelial cell is planted in compound acellular matrix upper surface, forms compound acellular matrix water
The bionical tissue of acellular matrix-cell-three-dimensional fiber network of epithelial cell is planted in gel upper surface, as shown in Figure 5；It is compound
Main component in acellular matrix hydrogel is water, and porosity 50%, pore diameter range is 10-50 μm, storage temperature 4
℃。
Embodiment 5
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares and is derived from gastral acellular matrix material, and freeze grinding again after being freeze-dried is beaten
It is broken into powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds Shu Tian and leads to, wherein the mass ratio for the body An Tongyu acellular matrix that relaxes is 1:18, forms de- cell base
Matter gel precursor solution, before gel is not formed, the micropore biocompatibility that is evenly distributed prepared by method of electrostatic spinning is good
Good polycaprolactone network of fibers material is soaked in above-mentioned 2mL acellular matrix precursor solution, forms polycaprolactone fiber
Polymer is constructed containing acellular matrix hydrogel in network material hole, also contains three-dimensional in acellular matrix hydrogel hole
Network of fibers material mutually includes structure；Fibre diameter is 1.8 μm, and pore diameter range is 6.5-7.0 μm, porosity 79%；
15min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；Then in compound acellular matrix hydrogel material table
Epithelial cell is planted in face, then sprays one layer of hydrogel precursor solution, specially mucous membrane of small intestine acellular matrix hydrogel forerunner
Liquid solution stands 15min at 37 DEG C and continues gel, formed between compound acellular matrix hydrogel and hydrogel and plant epithelium
The bionical tissue of acellular matrix-cell-three-dimensional fiber network of cell, as shown in Figure 6；In compound acellular matrix hydrogel
Main component be water, porosity 80%, pore diameter range be 150-250 μm, storage temperature be 20 DEG C.
Embodiment 6
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from vagina, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds Finasteride, and wherein the mass ratio of Finasteride and acellular matrix is 1:5, forms acellular matrix
The fibre diameter prepared by 3 D weaving method is 2.5 μm before gel is not formed by gel precursor solution, aperture model
Enclosing is 5-6 μm, porosity 85%, and the micropore good fibroin fiber network material of biocompatibility that is evenly distributed impregnates
In above-mentioned 2mL acellular matrix precursor solution, is formed in fibroin fiber network material hole and contain acellular matrix
Hydrogel constructs polymer, also containing three-dimensional fiber network material mutually includes structure in acellular matrix hydrogel hole；
18min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；In the upper and lower table of compound acellular matrix hydrogel
Epithelial cell and smooth muscle cell are planted in face respectively, form the bionical tissue of acellular matrix-cell-three-dimensional fiber network, such as
Shown in Fig. 7；Main component in compound acellular matrix hydrogel is water, and porosity 70%, pore diameter range is 250-300 μ
M, storage temperature are 30 DEG C.
Embodiment 7
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from fallopian tubal, and freeze grinding again after being freeze-dried is beaten
It is broken into powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the mixture of insulin and vascular endothelial growth factor, wherein insulin and vascular endothelial growth factor
Mass ratio be the mass ratio of 2:5, insulin and vascular endothelial growth factor and acellular matrix be 0.1:100, formed de- thin
Above-mentioned acellular matrix precursor solution is sprayed on by quiet by cytoplasmic matrix gel precursor solution before gel is not formed
The fibre diameter of electrical spinning method preparation is 1.9 μm, and pore diameter range is 6.9-8.5 μm, porosity 78%, and micropore is evenly distributed
The upper and lower surfaces of the good poly (glycolide-co-lactide) network of fibers material of biocompatibility form one layer of three-dimensional fiber network knot
The stepped construction that structure and acellular matrix hydrogel precursor solution are constituted；25min is stood at 37 DEG C again, is formed compound de- thin
Cytoplasmic matrix hydrogel；One layer of hydrogel precursor solution is respectively coated in compound acellular matrix hydrogel upper and lower surfaces again, has
Body is blood vessel acellular matrix hydrogel precursor solution, and 15min is stood at 37 DEG C and continues gel, then by epithelial cell
It is planted in hydrogel upper surface, smooth muscle cell is planted in hydrogel lower surface, forms acellular matrix-cell-three-dimensional fiber
The bionical tissue of network, as shown in Figure 8；Main component in compound acellular matrix hydrogel is water, porosity 66%, hole
Diameter range is 300-360 μm, and storage temperature is 25 DEG C.
Embodiment 8
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from urethra, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the mixture of aspirin and Ketoprofen, and wherein the quality of aspirin and Ketoprofen is 3:2, A Si
The mass ratio of woods and Ketoprofen and acellular matrix is 1:9, acellular matrix gel precursor solution is formed, in the non-shape of gel
At before, it is 1.6 μm that acellular matrix precursor solution, which is coated in through fibre diameter prepared by 3 D weaving method, aperture model
Enclosing is 5.5-6.6 μm, porosity 81%, and micropore is evenly distributed the good fibroin fiber network material of biocompatibility
Upper and lower surfaces；Form the stacking knot of one layer of three-dimensional fiber network structure and acellular matrix hydrogel precursor solution composition
Structure；20min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；It is upper and lower in compound acellular matrix hydrogel again
Epithelial cell and smooth muscle cell are planted in surface respectively, and one layer of water then is respectively coated in epithelial cell and smooth muscle cells
Gel precursor solution, specially mucous membrane of small intestine acellular matrix hydrogel precursor solution stand 24min at 37 DEG C and continue
Gel forms the bionical tissue of acellular matrix-cell-three-dimensional fiber network, as shown in Figure 9；Compound acellular matrix water-setting
Main component in glue is water, and porosity 70%, pore diameter range is 300-350 μm, and storage temperature is 30 DEG C.
Embodiment 9
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from ureter, and freeze grinding again after being freeze-dried is beaten
It is broken into powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the mixture that Ketoprofen and Shu Tian lead to, and the mass ratio that wherein Ketoprofen and Shu Tian lead to is 3:4, ketone
The logical mass ratio with acellular matrix of ibuprofen and Shu Tian is 2:7, forms acellular matrix gel precursor solution, gel not
Before formation, it is 2.5 μm that above-mentioned acellular matrix precursor solution, which is sprayed on through fibre diameter prepared by 3 D weaving method,
Pore diameter range is 7-7.8 μm, porosity 70%, and the micropore good poly (glycolide-co-lactide) of biocompatibility that is evenly distributed is fine
The upper and lower surfaces for tieing up network material, form the good three-dimensional fiber network structure of one layer of biocompatibility and acellular matrix water
The stepped construction that gel precursor solution is constituted；18min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；Again
One layer of bladder acellular matrix hydrogel precursor solution is respectively coated in compound acellular matrix hydrogel upper and lower surfaces,
15min is stood at 37 DEG C and continues gel, epithelial cell is then being planted in hydrogel upper surface, smooth muscle cell is planted in water
Then one layer of hydrogel precursor solution is respectively coated in epithelial cell and smooth muscle cells, specially in gel lower surface
Blood vessel acellular matrix hydrogel precursor solution stands 15min at 37 DEG C and continues gel, forms acellular matrix-cell-
The bionical tissue of three-dimensional fiber network, as shown in Figure 10；Main component in acellular matrix hydrogel is water, and porosity is
65%, pore diameter range is 320-400 μm, and storage temperature is 35 DEG C.
Embodiment 10
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from small intestine, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the logical mixture of aspirin, Ketoprofen and Shu Tian, wherein aspirin, Ketoprofen and Shu Tian
Logical mass ratio is 2:1:3, and the logical mass ratio with acellular matrix of aspirin, Ketoprofen and Shu Tian is 1:10, is formed de-
The fibre diameter prepared by 3 D weaving method is by cellular matrix gel precursor solution before gel is not formed
900nm, pore diameter range are 1-3 μm, porosity 65%, and micropore is evenly distributed the good fibroin fiber of biocompatibility
Network material is soaked in above-mentioned acellular matrix precursor solution, is formed in fibroin fiber network material hole containing de- thin
Cytoplasmic matrix hydrogel constructs polymer, also mutually includes containing three-dimensional fiber network material in acellular matrix hydrogel hole
Structure；Epithelial cell and smooth muscle cell are planted respectively in compound acellular matrix hydrogel material upper and lower surfaces again, are then existed
One layer of hydrogel precursor solution, specially urethra acellular matrix water-setting is respectively coated in epithelial cell and smooth muscle cells
Glue precursor solution continues gel, forms the bionical tissue of acellular matrix-cell-three-dimensional fiber network, as shown in figure 11；
Main component in compound acellular matrix hydrogel is water, and porosity 70%, pore diameter range is 380-430 μm, storage temperature
Degree is 5 DEG C.
Embodiment 11
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from blood vessel, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the mixture of insulin and vascular endothelial growth factor, wherein insulin and vascular endothelial growth factor
Mass ratio be the mass ratio of 2:5, insulin and vascular endothelial growth factor and acellular matrix be 0.1:100, formed de- thin
Above-mentioned acellular matrix precursor solution is sprayed on by quiet by cytoplasmic matrix gel precursor solution before gel is not formed
The fibre diameter of electrical spinning method preparation is 1.9 μm, and pore diameter range is 6.9-8.5 μm, porosity 78%, and micropore is evenly distributed
The upper and lower surfaces of the good poly (glycolide-co-lactide) network of fibers material of biocompatibility form one layer of three-dimensional fiber network knot
The stepped construction that structure and acellular matrix hydrogel precursor solution are constituted；25min is stood at 37 DEG C again, is formed compound de- thin
Cytoplasmic matrix hydrogel；One layer of hydrogel precursor solution is respectively coated in compound acellular matrix hydrogel upper and lower surfaces again, has
Body is bladder acellular matrix hydrogel precursor solution, and 15min is stood at 37 DEG C and continues gel, then by epithelial cell
It is planted in hydrogel upper surface, smooth muscle cell is planted in hydrogel lower surface, then coats one layer in smooth muscle cells
Hydrogel precursor solution, specially urethra acellular matrix hydrogel precursor solution stand 18min at 37 DEG C and continue to coagulate
Glue forms the bionical tissue of acellular matrix-cell-three-dimensional fiber network, as shown in figure 12；Compound acellular matrix hydrogel
In main component be water, porosity 66%, pore diameter range be 300-360 μm, storage temperature be 25 DEG C.
Embodiment 12
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares and is derived from gastral acellular matrix material, and freeze grinding again after being freeze-dried is beaten
It is broken into powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the mixture of insulin and vascular endothelial growth factor, wherein insulin and vascular endothelial growth factor
Mass ratio be the mass ratio of 2:5, insulin and vascular endothelial growth factor and acellular matrix be 0.1:100, formed de- thin
Above-mentioned acellular matrix precursor solution is coated in by quiet by cytoplasmic matrix gel precursor solution before gel is not formed
The fibre diameter of electrical spinning method preparation is 1.9 μm, and pore diameter range is 6.9-8.5 μm, porosity 78%, and micropore is evenly distributed
The upper and lower surfaces of the good poly (glycolide-co-lactide) network of fibers material of biocompatibility form one layer of three-dimensional fiber network knot
The stepped construction that structure and acellular matrix hydrogel precursor solution are constituted；28min, the compound de- cell of shape are stood at 37 DEG C again
Matrix hydrogel；One layer of hydrogel precursor solution, specially blood vessel are coated in compound acellular matrix hydrogel upper surface again
Acellular matrix hydrogel precursor solution stands 15min at 37 DEG C and continues gel, epithelial cell is then being planted in water
Smooth muscle cell is planted in compound acellular matrix hydrogel lower surface, then is sprayed in smooth muscle cells in gel upper surface
One layer of hydrogel precursor solution, specially bladder acellular matrix hydrogel precursor solution, at 37 DEG C stand 10min after
Continuous gel, forms the bionical tissue of acellular matrix-cell-three-dimensional fiber network, as shown in figure 13；Acellular matrix hydrogel
In main component be water, porosity 66%, pore diameter range be 300-360 μm, storage temperature be 25 DEG C.
Embodiment 13
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from vagina, and freeze grinding again after being freeze-dried is smashed
At powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L adds the mixture that Ketoprofen and Shu Tian lead to, and the mass ratio that wherein Ketoprofen and Shu Tian lead to is 3:4, ketone
The logical mass ratio with acellular matrix of ibuprofen and Shu Tian is 2:7, forms acellular matrix gel precursor solution, gel not
Before formation, it is 2.5 μm that above-mentioned acellular matrix precursor solution, which is sprayed on through fibre diameter prepared by 3 D weaving method,
Pore diameter range is 7-7.8 μm, porosity 70%, and the micropore good poly (glycolide-co-lactide) of biocompatibility that is evenly distributed is fine
The upper surface for tieing up network material, forms the good three-dimensional fiber network structure of one layer of biocompatibility and acellular matrix hydrogel
The stepped construction that precursor solution is constituted；18min is stood at 37 DEG C again, forms compound acellular matrix hydrogel；Again multiple
It closes acellular matrix hydrogel upper surface and sprays one layer of urethra acellular matrix hydrogel precursor solution, stood at 37 DEG C
15min continues gel, epithelial cell is then being planted in hydrogel upper surface, in compound acellular matrix hydrogel lower surface
Smooth muscle cell is planted, then one layer of hydrogel precursor solution is respectively coated in epithelial cell and smooth muscle cells, specifically
For bladder acellular matrix hydrogel precursor solution, 8min is stood at 37 DEG C and continues gel, it is thin to form acellular matrix-
The bionical tissue of born of the same parents-three-dimensional fiber network, as shown in figure 13；Main component in acellular matrix hydrogel is water, porosity
It is 65%, pore diameter range is 320-400 μm, and storage temperature is 35 DEG C.
Embodiment 14
A kind of preparation method of the bionical tissue of acellular matrix-cell-three-dimensional fiber network, is compound de- cell first
The preparation of matrix hydrogel prepares the acellular matrix material for being derived from fallopian tubal, and freeze grinding again after being freeze-dried is beaten
It is broken into powder；Pepsin digestion acellular matrix powder about 48h is used in hydrochloric acid solution, and it is molten to form acellular matrix pregel
The NaOH solution of about 1/10 acellular matrix pre-gel solution volume is added in liquid, and wherein the concentration of NaOH solution is
0.01mol/L adds 10 × PBS of about 1/9 acellular matrix pre-gel solution volume, and wherein the concentration of PBS is
0.01mol/L, adds vascular endothelial growth factor, and the mass ratio of medium vascular endothelial growth factor and acellular matrix is
0.025:100, forming acellular matrix gel precursor solution will be prepared before gel is not formed by method of electrostatic spinning
Fibre diameter be 500nm, pore diameter range is 1-2 μm, porosity 65%, and the micropore biocompatibility that is evenly distributed is good
Fibroin fiber network material be cut into diameter be 14mm disk, it is molten to be soaked in above-mentioned 2mL acellular matrix presoma
In liquid, is formed in fibroin fiber network material hole and construct polymer, acellular matrix containing acellular matrix hydrogel
It is also mutual comprising structure containing three-dimensional fiber network material in hydrogel hole；3min is stood at 37 DEG C again, is formed compound
Acellular matrix hydrogel；Epithelial cell smooth muscle cell is planted respectively in compound acellular matrix hydrogel upper and lower surfaces,
Smooth muscle cells spray one layer of hydrogel precursor solution, and specially mucous membrane of small intestine acellular matrix hydrogel precursor is molten
Liquid stands 9min at 37 DEG C and continues gel, forms the bionical tissue of acellular matrix-cell-three-dimensional fiber network, such as Figure 15
It is shown；Main component in compound acellular matrix hydrogel is water, and porosity 50%, pore diameter range is 10-50 μm, storage
Temperature is 4 DEG C.
Claims (9)
1. a kind of preparation method based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network, it is characterized in that: described
Preparation method is infusion method；It is described that compound take off is included at least based on the bionical tissue of acellular matrix-cell-three-dimensional fiber network
Cellular matrix hydrogel and epithelial cell, alternatively, also including smooth muscle cell further；The compound acellular matrix water
Gel is using the good three-dimensional fiber network material of biocompatibility as the hydrogel of skeleton；The biocompatibility good three
Tieing up network of fibers material is the network structure material being wound by the good fiber of biocompatibility；The compound de- cell base
Matter hydrogel is that acellular matrix hydrogel and the good three-dimensional fiber network material of the biocompatibility are integrally compound；It is described
Entirety is compound to be referred in acellular matrix hydrogel integrally comprising the good three-dimensional fiber network material of biocompatibility.
2. preparation method according to claim 1, which is characterized in that the preparation method specifically: prepare first de- thin
Cytoplasmic matrix hydrogel precursor solution, then impregnates, then stands 3~30min at 37 DEG C and form compound acellular matrix water
Gel；
The immersion are as follows:
Before gel is not formed, before the good three-dimensional fiber network material of biocompatibility is soaked in the acellular matrix
Drive liquid solution.
3. preparation method according to claim 2, which is characterized in that the good three-dimensional fiber network of biocompatibility
The preparation method of material is method of electrostatic spinning or 3 D weaving method.
4. preparation method according to claim 2, which is characterized in that the preparation of acellular matrix hydrogel precursor solution
It is additionally added migration ingredient in the process.
5. preparation method according to claim 1, which is characterized in that the good three-dimensional fiber network of biocompatibility
Material is natural or synthesis the good three-dimensional fiber network material of biocompatibility；The good three-dimensional of biocompatibility
Fibre diameter in network of fibers material is 500nm~3 μm, and pore diameter range is 1 μm -9 μm, porosity 65%-90%, and micro-
Pore size distribution is uniform；The main component of the compound acellular matrix hydrogel is water, porosity 50%-90%, pore diameter range
It is 10 μm -450 μm；The acellular matrix of the compound acellular matrix hydrogel be derived from urethra, ureter, blood vessel, small intestine,
The acellular matrix of alimentary canal, vagina or fallopian tubal；The good fiber of biocompatibility is fibroin fiber, the friendship of poly- second
Ester lactide fiber or polycaprolactone fiber.
6. preparation method according to claim 1, which is characterized in that described to include at least compound acellular matrix hydrogel
It is one of following scenario described with epithelial cell:
Compound acellular matrix hydrogel situation in conjunction with epithelial cell:
Epithelial cell is planted on compound acellular matrix hydrogel；
Compound acellular matrix hydrogel upper surface covers hydrogel, and between compound acellular matrix hydrogel and hydrogel
Plant epithelial cell；
Compound acellular matrix hydrogel upper surface covers hydrogel, and epithelial cell is planted in hydrogel upper surface；
Compound acellular matrix hydrogel upper surface covers two layers of hydrogel, and epithelial cell is planted between two layers of hydrogel；
Alternatively, further including one of following scenario described further:
Compound acellular matrix hydrogel situation in conjunction with smooth muscle cell:
Plant smooth muscle cell in compound acellular matrix hydrogel lower surface；
Compound acellular matrix hydrogel lower surface covers hydrogel, and between compound acellular matrix hydrogel and hydrogel
Plant smooth muscle cell；
Compound acellular matrix hydrogel lower surface covers hydrogel, and smooth muscle cell is planted in hydrogel lower surface；
Compound acellular matrix hydrogel lower surface covers two layers of hydrogel, and smooth muscle cell is planted between two layers of hydrogel；
The hydrogel is the hydrogel that acellular matrix is prepared.
7. preparation method according to claim 1, which is characterized in that also contain in the compound acellular matrix hydrogel
Ingredient is migrated, the migration ingredient dispersed encapsulated is in the acellular matrix hydrogel.
8. preparation method according to claim 7, which is characterized in that it is multiple that the migration ingredient refers to that migration ingredient is dispersed in
It closes in acellular matrix hydrogel, but does not generate any chemical reaction；The migration ingredient is vascular endothelial growth factor, pancreas islet
Element, aspirin, Ketoprofen, Shu Tian be logical or one or more of Finasteride.
9. preparation method according to claim 1, which is characterized in that the storage temperature of the compound acellular matrix hydrogel
Degree is at 4 DEG C~40 DEG C.
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| CN109157305B (en) * | 2018-09-25 | 2021-05-25 | 广州锐澄医疗技术有限公司 | Composite artificial cornea and preparation method thereof |
| CN110106148B (en) * | 2019-05-16 | 2020-10-13 | 中国人民解放军军事科学院军事医学研究院 | Tissue engineering nerve tissue and construction method thereof |
| CN110124086B (en) * | 2019-05-23 | 2020-06-30 | 中山大学 | Composite nanofiber pad, hydrogel/sponge dressing, preparation method and application |
| CN113230461B (en) * | 2021-05-06 | 2022-02-15 | 东华大学 | Micro-nanofiber-hydrogel tissue engineering ureter and preparation method thereof |
| CN113790958A (en) * | 2021-09-01 | 2021-12-14 | 沈阳恒生医用科技有限公司 | High-molecular bionic tissue and application thereof |
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| CN104274864A (en) * | 2013-07-11 | 2015-01-14 | 上海市第六人民医院 | Method for establishing three-dimensional tissue in vitro |
| CN104971380A (en) * | 2014-04-11 | 2015-10-14 | 烟台隽秀生物科技有限公司 | Acellular matrix repairing gel and new method for preparing the same |
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