CN107252500A - The preparation method that a kind of sandwich style is combined membrane support - Google Patents
The preparation method that a kind of sandwich style is combined membrane support Download PDFInfo
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- CN107252500A CN107252500A CN201710461345.0A CN201710461345A CN107252500A CN 107252500 A CN107252500 A CN 107252500A CN 201710461345 A CN201710461345 A CN 201710461345A CN 107252500 A CN107252500 A CN 107252500A
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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Abstract
The present invention provides the preparation method that a kind of sandwich style is combined membrane support, and the PLA/bioactivity glass/chitosan sandwich style for building load bioactie agent is combined membrane carrier；Comprise the following steps：1）Polylactic acid film is made respectively, is loaded the mesopore bioactive glass of bioactie agent, is loaded the chitosan film of seed cell as outer side seal layer, sandwich of layers and inner side controlled release and stem cell shelf layer；2）The sandwich of layers for the bioactivity glass that bioactie agent will be loaded is covered on polylactic acid film, then the chitosan film for loading seed cell is covered in into the compound membrane carrier of structure sandwich style on bioactivity glass layer.To solve to have sustained damage（But it is dead）Single nerve tract, the problem of prior art can not be repaired and treated；Existing drug-loading system can not realize the targeted release of medicine, it is impossible to the problem of ensureing the high concentration of damage local factors.
Description
Technical field
Present invention relates to the preparation method that a kind of sandwich style is combined membrane support.
Background technology
The most common method of administration for promoting nerve growth or nerve regneration medicine at present is vein or intrathecal injection.Also have with solidifying
Cementing conjunction active factors pad pasting spreads on spinal cord surface, using the L- poly-D-lysine binding activity factors as stenter to implant spinal cord, with
Solve the problems, such as that the factor passes through blood-brain barrier.
Drug delivery system and stem cell biological support can be used in treating spinal cord injury, but have no that both make in combination at present
With.The method of the stem cell three-dimensional rack Bridging nerve united application similar with having had in material as（One kind is used to bridge
The composite repairing material and its support CN201510716279.8 of defect nerve）.But three-dimensional rack Bridging nerve is controlled at present
Treatment method, has the following disadvantages：
On the one hand to having sustained damage（But it is dead）Single nerve tract, it is impossible to reach repair and treatment purpose；It is another
Aspect during bridge joint because the spinal cord of injury region is cut out one it is similar with three-dimensional rack shape --- be typically
Tubulose --- breach, therefore to unmarred nerve cell at this and can sustain damage（But it is dead）Nerve cell enter
Row is removed, and as a result causes the further loss of cord cell at this.
2nd, existing drug-loading system can not realize the targeted release of medicine, it is impossible to ensure the high concentration of damage local factors；
3, most drug-loading systems are easy to degraded, and drugloading rate is low, and insoluble drug release can not be through spinal cord injury repair process all the time；
4, drug-loading system volume is excessive, may oppress spinal cord, while not being conducive to suturing endorchis, causes leakage of cerebrospinal to be formed.
The content of the invention
The object of the invention：
1 there is provided a kind of preparation method of the drug-loading system of new medullotherapy, it is to avoid using load medicine traditional in background technology
, it is necessary to unmarred nerve cell at this and sustain damage during system（But it is dead）Nerve cell be purged,
To cause the further loss of cord cell at this.
2 there is provided a kind of preparation method of new drug-loading system, solves the targeted release of medicine, the problem of drugloading rate is low.
3 there is provided a kind of preparation method of new drug-loading system, and solving insoluble drug release can not repair through spinal cord injury
Journey all the time the problem of.
4 there is provided a kind of preparation method of new drug-loading system, and the existing drug-loading system volume of solution is excessive, may oppress ridge
Marrow, while the problem of not being conducive to suture endorchis.
The technical solution adopted by the present invention is：
The preparation method that a kind of sandwich style is combined membrane support, builds PLA/bioactivity glass of load bioactie agent
Glass/chitosan sandwich style is combined membrane carrier；Comprise the following steps：
1）Polylactic acid film is made respectively, is loaded the mesopore bioactive glass of bioactie agent, is loaded the shell of seed cell
Glycan film is used as outer side seal layer, sandwich of layers and inner side controlled release and stem cell shelf layer；
2）The sandwich of layers that the bioactivity glass of bioactie agent will be loaded is covered on polylactic acid film, then will load seed
The chitosan film of cell, which is covered on bioactivity glass layer, builds the compound membrane carrier of sandwich style.
Further, 1）It is middle to make the chitosan film without load seed cell；
2）It is middle that the chitosan film without load seed cell is covered on bioactivity glass layer；
3）By stem cell suspension, the culture of seed cell is carried out inside the chitosan film using uniform immersion previous step.
Further, 1）In do not make chitosan film；2）In not by chitosan film be covered in bioactivity glass layer on；3）
Stem cell and the outstanding solution of chitosan mixing are made, the mixed solution is covered in by bioactivity glass using spin coating, drop coating method
On layer, build class sandwich style and be combined membrane carrier.
Further, each layer is glued with the Medical Living Creature Gum that can permeate the stimulate neuronal growth factor and stem cell
Even.
Further, the sandwich of layers load bioactie agent is the factor that stimulates neuronal growth.
Further, the factor that stimulates neuronal growth is one or several kinds of in bFGF, BD-NF, NT-3, IGF
Combination.
Further, composite membrane support perimeter is entered with the Medical Living Creature Gum of impermeable the stimulate neuronal growth factor and stem cell
Row closing.
Further, the mesopore bioactive glass bead replaces with ethylene glycol lactic acid copolymer（PEG-PLA）Nanometer
Microballoon.
Further, the seed cell is Mesenchymal Stem Cells from Umbilical Cord, i.e. HUMSCs.
Further, the seed cell is selected from：Schwann cell, medulla mesenchyma cell, fat mesenchymal stem cell, navel
With the one or more combination in mescenchymal stem cell, NSC or induced multi-potent stem cell.
The technical scheme factor that will stimulate neuronal growth is combined membrane carrier and organizational project Stem Cell Science and organically combined, with
Load the compound membrane support combination seed cell HUMSCs of the neural activity factor, it is desirable to more effectively repair spinal cord injury.The branch
Frame, easily in the characteristic for adhering to and breeding thereon, utilizes bioactivity glass using the good histocompatbility of chitosan and stem cell
To bioactie agent（bFGF、BD-NF、NT-3、IGF）Pay(useful) load continues with finite concentration release characteristics with control, passes through
It is set to be provided simultaneously with following effect：
1, composite membrane can directly be pasted on damage spinal cord position, and on the one hand overcoming bioactie agent can not be by blood brain screen
The problem of barrier, on the other hand the spinal cord on damage location periphery need not be purged, it is to avoid cord cell enters one at this
Step loss.
2, the time limit is acted on according to bioactie agent, the valid density that periphery cord cell needs is repaired, scientific design is situated between
Load medicine total amount and rate of release, the concentration of hole bioactive glass bead, improve stress efficacy；
3, dense outer layer can not pass through active factors and stem cell so that active factors and stem cell can only inner layer directions（I.e.
Damage location）Arrow in targeted release, such as Fig. 3.
4, composite membrane support perimeter is closed with Medical Living Creature Gum, to ensure that active factors and stem cell are not overflow from branch frame peripheral
Go out, it is ensured that drug-loading system one side targeted release, the time of the release on the other hand ensured and concentration；
5, the characteristics of composite membrane has ultra-thin（Thickness is no more than 2mm）, will not simultaneously be conducive to because of the excessive compressing spinal cord of volume
Endorchis is sutured, prevents leakage of cerebrospinal from being formed.
Need to do a special instruction, and immediate prior art of the invention, be：One kind is used to bridge defect god
The composite repairing material and its support CN201510716279.8 of warp.It discloses a kind of compound repairing for being used to bridge defect nerve
Multiple material and its support, including：The composite is arranged in order by fibroin layer-collagen layer-high polymer layer and constituted,
The T-shaped or rectangle design of high molecular polymerization layer, using when by folding or convolution formation fibroin layer is during internal layer, collagen layer are
Layer, high polymer layer are the bridge grafting nerves support of outer layer.But its technical problem to be solved, and the technology effect played
Really, and the present invention be completely it is different.
The technical problem that CN201510716279.8 is solved is to realize the quick bridge joint of defect nerve, and it is substantially background skill
" nerve tract that bridge joint is blocked " introduced in art.Itself in use, first by support by fold or convolution formation fibroin layer for internal layer,
The bridge grafting nerves support that collagen layer is middle level, high polymer layer is outer layer, secondly by the support two after folding or certificate folding
The nerve tract two ends held and blocked are sutured by surgical method.Wherein fibroin layer not only guides CO2 laser weld, and separates
Different nerves, prevent from forming neuroma.Collagen layer provides the various nutrients of neural axon growth/required.High polymer layer,
With good mechanical property there is provided operation stitching or the required mechanical strength that coincide, and perineural intrusion is prevented, prevented
The only formation of fibrous scar.
The compound membrane support of the present invention, the nerve tract for blocking or being partially damaged applied to reparation, it is not necessary to suture, directly
Composite membrane is attached to the damaged part of nerve tract（Such as Fig. 2）.Its dense outer layer is used for the work for stopping sandwich of layers and internal layer
Sex factor and stem cell are permeated to this aspect and outwardly oozed out by the tunic（Such as Fig. 3）；Sandwich of layers is used in a timing
In, form the precipitation of continuous concentration medicine；Internal layer provides stem cell.The technical problems to be solved by the invention, composite membrane is each
Layer respective effect and the technique effect played in composite membrane, it is completely different with CN201510716279.8, with creation
Property.
Brief description of the drawings
Fig. 1 is the sandwich style composite membrane support schematic diagram of the present invention.
The compound membrane support of sandwich style that Fig. 2 is the present invention is attached at spinal cord schematic diagram.
Fig. 3 is composite membrane stent drug release direction（Path）.
Embodiment
Embodiment 1
1）Make the PLA of certain molecular weight（Matter average molecular weight is 5000-1000000, preferably 5000-100000）, dissolving
Into organic solvent（Organic solvent includes Isosorbide-5-Nitrae-dioxane, acetone, chloroform/alcohol mixed solvent, dichloromethane, four
Hydrogen furans, and above-mentioned solvent mixed solvent）, concentration is 0.1-10 wt%, prepares PLA using methods such as spin coating, drop coatings thin
Film, is used as outer side seal layer；
2）By the chitosan film of certain molecular weight（Matter average molecular weight is 5000-1000000, preferably 5000-100000）, it is dissolved into
In acidic aqueous solution（The acid of use includes hydrochloric acid, acetic acid, nitric acid）, concentration is 0.1-10 wt%, using methods such as spin coating, drop coatings
Chitosan film is prepared, inner side controlled release layer and stem cell support is used as；
3）By stem cell suspension, it is uniformly immersed using the method successively injected and trained inside the chitosan film of previous step
Support；
4）The preparation process of mesopore bioactive glass is made, it is described in detail below.By ethyl orthosilicate, calcium nitrate, F127,2M
Nitric acid by certain mass than add absolute ethyl alcohol mixed liquor in stir（Typical proportions are 2.7 g TEOS, 1.18 g Ca
(NO3) 2 4H2O, 4.2 g F127 and 0.08 g 2 M HNO3 add 8 g absolute ethyl alcohols）, 3 h are stirred at room temperature obtains molten
Glue.Gained colloidal sol is put into vacuum drying chamber and carries out sol-gel process at 50 DEG C, after after collosol and gel drying, in Muffle
Sintering removes template at 600 DEG C in stove, produces mesopore bioactive glass.Then gained mesopore bioactive glass is ground
Into uniform powder, this powder is MBGs.The saturated solution for the factor that stimulates neuronal growth is prepared, is loaded to using perfusion
Mesopore bioactive glass（Hereinafter referred to as MBGs）In, specific implementation method is as follows：Infiltrated with certain density drug solution
The MBGs powder of known quality, is placed in vacuum drying chamber, and its rapid draing is made under vacuum condition, completes once to irrigate week
Phase.It is repeated several times after the completion of its adjustable drugloading rate of perfusion, most rear bearing medicine, is placed in drying box, it is standby.Record perfusion number of times,
Calculate theoretical drugloading rate.Sandwich of layers is used as using mesopore bioactive glass load bioactie agent
5）It is as follows that stacked system builds sandwich style composite membrane carrier method：PLA is prepared using methods such as spin coating, drop coatings thin
Film loads bioactie agent with bioactivity glass and is covered in polylactic acid film as outer side seal layer, then by by shell
Glycan film or chitosan solution, which are covered on bioactivity glass layer, builds the compound membrane carrier of sandwich style.
Embodiment 2
Compare, be in place of the difference of embodiment 2,5 with embodiment 1）By each layer with can permeate stimulate neuronal growth the factor and
The Medical Living Creature Gum of stem cell carries out adhesion.
Claims (10)
1. the preparation method that a kind of sandwich style is combined membrane support, it is characterized in that：Comprise the following steps：
1）Polylactic acid film is made respectively, is loaded the mesopore bioactive glass of bioactie agent, is loaded the shell of seed cell
Glycan film is used as outer side seal layer, sandwich of layers and inner side controlled release and stem cell shelf layer；
2）The sandwich of layers that the bioactivity glass of bioactie agent will be loaded is covered on polylactic acid film, then will load seed
The chitosan film of cell, which is covered on bioactivity glass layer, builds the compound membrane carrier of sandwich style.
2. the preparation method that a kind of sandwich style is combined membrane support as described in the appended claim 1, it is characterised in that：
1）It is middle to make the chitosan film without load seed cell；
2）It is middle that the chitosan film without load seed cell is covered on bioactivity glass layer；
3）By stem cell suspension, the culture of seed cell is carried out inside the chitosan film using uniform immersion previous step.
3. the preparation method that a kind of sandwich style is combined membrane support as described in the appended claim 1, it is characterised in that：
1）In do not make chitosan film；2）In not by chitosan film be covered in bioactivity glass layer on；3）Make stem cell and
The mixed solution, is covered on bioactivity glass layer, builds class three by the outstanding solution of chitosan mixing using spin coating, drop coating method
Mingzhi's formula is combined membrane carrier.
4. the preparation method that a kind of sandwich style as described in one of claim 1-3 is combined membrane support, it is characterised in that：Will be each
Layer carries out adhesion with the Medical Living Creature Gum that can permeate the stimulate neuronal growth factor and stem cell.
5. the preparation method that a kind of sandwich style is combined membrane support as claimed in claim 4, it is characterised in that：Sandwich of layers is stated to bear
It is the factor that stimulates neuronal growth to carry bioactie agent.
6. the preparation method that a kind of sandwich style is combined membrane support as claimed in claim 5, it is characterised in that：The nerve is raw
Long stimulating factor is one or several kinds of combinations in bFGF, BD-NF, NT-3, IGF.
7. the preparation method that a kind of sandwich style is combined membrane support as recited in claim 6, it is characterised in that：Compound membrane support
Closed with the Medical Living Creature Gum of impermeable the stimulate neuronal growth factor and stem cell on periphery.
8. the preparation method that a kind of sandwich style is combined membrane support as recited in claim 7, it is characterised in that：The mesoporous life
Thing activity glass bead replaces with ethylene glycol lactic acid copolymer（PEG-PLA）Nanoparticle.
9. the preparation method that a kind of sandwich style is combined membrane support as claimed in claim 8., it is characterised in that：The kind is careful
Born of the same parents are Mesenchymal Stem Cells from Umbilical Cord, i.e. HUMSCs.
10. the preparation method that a kind of sandwich style is combined membrane support as claimed in claim 9, it is characterised in that：The seed
Cell is selected from：Schwann cell, medulla mesenchyma cell, fat mesenchymal stem cell, umbilical cord mesenchymal stem cells, NSC
Or the one or more combination in induced multi-potent stem cell.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710461345.0A CN107252500A (en) | 2017-06-16 | 2017-06-16 | The preparation method that a kind of sandwich style is combined membrane support |
| CN201810564926.1A CN108714248A (en) | 2017-06-16 | 2018-06-04 | A kind of production method of the compound membrane support of sandwich style |
| CN201810563658.1A CN108744046A (en) | 2017-06-16 | 2018-06-04 | A kind of compound membrane support of sandwich style |
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| CN201710461345.0A CN107252500A (en) | 2017-06-16 | 2017-06-16 | The preparation method that a kind of sandwich style is combined membrane support |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108714248A (en) * | 2017-06-16 | 2018-10-30 | 无锡市锡山人民医院 | A kind of production method of the compound membrane support of sandwich style |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108714248A (en) * | 2017-06-16 | 2018-10-30 | 无锡市锡山人民医院 | A kind of production method of the compound membrane support of sandwich style |
| CN108744046A (en) * | 2017-06-16 | 2018-11-06 | 无锡市锡山人民医院 | A kind of compound membrane support of sandwich style |
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