CN107213140A - A kind of preparation method of medullotherapy duplicature - Google Patents

A kind of preparation method of medullotherapy duplicature Download PDF

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Publication number
CN107213140A
CN107213140A CN201710455029.2A CN201710455029A CN107213140A CN 107213140 A CN107213140 A CN 107213140A CN 201710455029 A CN201710455029 A CN 201710455029A CN 107213140 A CN107213140 A CN 107213140A
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China
Prior art keywords
stem cell
duplicature
medullotherapy
preparation
chitosan
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CN201710455029.2A
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Chinese (zh)
Inventor
顾军
冯晓军
范健
纪立军
宋晓丽
徐林
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Xishan People's Hospital Of Wuxi
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Xishan People's Hospital Of Wuxi
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Priority to CN201710455029.2A priority Critical patent/CN107213140A/en
Publication of CN107213140A publication Critical patent/CN107213140A/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
    • A61K9/7023—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms
    • A61K9/703—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms characterised by shape or structure; Details concerning release liner or backing; Refillable patches; User-activated patches
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/51—Umbilical cord; Umbilical cord blood; Umbilical stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1825—Fibroblast growth factor [FGF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/185—Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
    • 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
    • A61K47/34—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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
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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
    • A61L27/14—Macromolecular materials
    • A61L27/20—Polysaccharides
    • 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
    • A61L27/36—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
    • A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A61L2300/414—Growth factors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
    • A61L2300/62—Encapsulated active agents, e.g. emulsified droplets
    • A61L2300/624—Nanocapsules
    • A—HUMAN NECESSITIES
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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
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction

Abstract

The present invention provides a kind of preparation method of medullotherapy duplicature, 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 stem 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 stem 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

A kind of preparation method of medullotherapy duplicature
Technical field
Present invention relates to a kind of preparation method of medullotherapy duplicature.
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：
A kind of preparation method of medullotherapy duplicature, comprises the following steps：
1）Polylactic acid film is made as outer side seal layer；
2）Bioactie agent is loaded with bioactivity glass and mixed with stem cell suspension, chitosan solution, using spin coating, drop Mixed solution is covered in polylactic acid membrane by coating method.
Further, 2）In loaded and bioactie agent and mixed with chitosan solution with bioactivity glass, using rotation Apply, mixed solution is covered in polylactic acid membrane by drop coating method.
3）After chitosan film is molded, by stem cell suspension, in the chitosan film using uniform immersion previous step Portion carries out the culture of stem cell.
Further, 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 stem cell is Mesenchymal Stem Cells from Umbilical Cord, i.e. HUMSCs.
Further, the stem cell is selected from：Schwann cell, medulla mesenchyma cell, fat mesenchymal stem cell, umbilical cord 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 stem cell HUMSCs of the neural activity factor, it is desirable to more effectively repair spinal cord injury.The support Using the good histocompatbility of chitosan and stem cell easily in the characteristic for adhering to and breeding thereon, bioactivity glass pair is utilized Bioactie agent（bFGF、BD-NF、NT-3、IGF）Pay(useful) load continues with finite concentration release characteristics with control, by making It is provided simultaneously with following effect：
1, duplicature 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. 1.
4, duplicature periphery is closed with Medical Living Creature Gum, to ensure that active factors and stem cell are overflow not around duplicature 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 duplicature 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 duplicature of the present invention, the nerve tract for blocking or being partially damaged applied to reparation, it is not necessary to suture directly will be double Tunic is attached to the damaged part of nerve tract.Its dense outer layer is used for the active factors for stopping internal layer and stem cell to the party Face is permeated and outwardly oozed out by the tunic；On the one hand internal layer is used within a certain period of time, form continuous concentration medicine Separate out, on the other hand to offer stem cell at spinal cord injury.The technical problems to be solved by the invention, each layer is each in duplicature From effect and the technique effect that plays, it is completely different with CN201510716279.8, it is creative.
Brief description of the drawings
Fig. 1 is that structure of the present invention and medicine separate out direction of motion schematic diagram.
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）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.
3）By the chitosan film of certain molecular weight（Matter average molecular weight is 5000-1000000, preferably 5000-100000）, it is dry Bioactie agent bullet bioactivity glass is loaded in cell suspension and previous step to be dissolved into acidic aqueous solution（The acid bag of use Include hydrochloric acid, acetic acid, nitric acid）, concentration is 0.1-10 wt%, and mixed solution is covered in into PLA using methods such as spin coating, drop coatings Film.
Embodiment 2
Compare, be in place of the difference of embodiment 2,3 with embodiment 1）In with bioactivity glass load bioactie agent simultaneously Mixed with chitosan solution, mixed solution is covered in by polylactic acid membrane using spin coating, drop coating method.
4）After chitosan film is molded, by stem cell suspension, in the chitosan film using uniform immersion previous step Portion carries out the culture of stem cell.

Claims (8)

1. a kind of preparation method of medullotherapy duplicature, it is characterized in that：Comprise the following steps：
1）Polylactic acid film is made as outer side seal layer；
2）Bioactie agent is loaded with bioactivity glass and mixed with stem cell suspension, chitosan solution, using spin coating, drop Mixed solution is covered in polylactic acid membrane by coating method.
2. a kind of preparation method of medullotherapy duplicature as described in the appended claim 1, it is characterised in that：
2）In loaded and bioactie agent and mix with chitosan solution with bioactivity glass, using spin coating, drop coating method general Mixed solution is covered in polylactic acid membrane；
3）After chitosan film is molded, stem cell suspension enters inside the chitosan film using uniform immersion previous step The culture of row stem cell.
3. a kind of preparation method of medullotherapy duplicature as described in claims 1 or 2, it is characterised in that：Biology is living Sex factor is the factor that stimulates neuronal growth.
4. a kind of preparation method of medullotherapy duplicature as claimed in claim 3, it is characterised in that：The nerve growth Stimulating factor is one or several kinds of combinations in bFGF, BD-NF, NT-3, IGF.
5. a kind of preparation method of medullotherapy duplicature as claimed in claim 4, it is characterised in that：Compound membrane support week While being closed with the Medical Living Creature Gum of impermeable the stimulate neuronal growth factor and stem cell.
6. a kind of preparation method of medullotherapy duplicature as claimed in claim 5, it is characterised in that：The mesoporous biological Activity glass bead replaces with ethylene glycol lactic acid copolymer（PEG-PLA）Nanoparticle.
7. a kind of preparation method of medullotherapy duplicature as recited in claim 6, it is characterised in that：The stem cell is Mesenchymal Stem Cells from Umbilical Cord, i.e. HUMSCs.
8. a kind of preparation method of medullotherapy duplicature as recited in claim 7, it is characterised in that：The stem cell choosing From：Schwann cell, medulla mesenchyma cell, fat mesenchymal stem cell, umbilical cord mesenchymal stem cells, NSC or induction One or more combination in multipotential stem cell.
CN201710455029.2A 2017-06-16 2017-06-16 A kind of preparation method of medullotherapy duplicature Pending CN107213140A (en)

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Application publication date: 20170929