CN106902098A - A kind of antitumor implantation film and preparation method thereof - Google Patents
A kind of antitumor implantation film and preparation method thereof Download PDFInfo
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- CN106902098A CN106902098A CN201710087762.3A CN201710087762A CN106902098A CN 106902098 A CN106902098 A CN 106902098A CN 201710087762 A CN201710087762 A CN 201710087762A CN 106902098 A CN106902098 A CN 106902098A
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- oncolytic virus
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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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
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- 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
- A61K35/66—Microorganisms or materials therefrom
- A61K35/76—Viruses; Subviral particles; Bacteriophages
- A61K35/761—Adenovirus
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- A61K35/76—Viruses; Subviral particles; Bacteriophages
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K35/66—Microorganisms or materials therefrom
- A61K35/76—Viruses; Subviral particles; Bacteriophages
- A61K35/768—Oncolytic viruses not provided for in groups A61K35/761 - A61K35/766
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- 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
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- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7007—Drug-containing films, membranes or sheets
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/042—Polysaccharides
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/148—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
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- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/425—Cellulose series
- D04H1/4258—Regenerated cellulose series
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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- 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
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- 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
- A61L2300/416—Anti-neoplastic or anti-proliferative or anti-restenosis or anti-angiogenic agents, e.g. paclitaxel, sirolimus
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Abstract
The present invention discloses a kind of antitumor implantation film and preparation method thereof, and the antitumor implantation film is the electro spinning nanometer fiber membrane for loading oncolytic virus, and preparation method is as follows：Step one, the biodegradable polymer with amino is prepared into the electrospinning film being made up of nanofiber with electrostatic spinning technique；Step 2, electro spinning nanometer fiber membrane described in step one is dipped into oncolytic virus solution, oncolytic virus is enriched on tunica fibrosa by electrostatic adsorption, obtain the antitumor implantation film.The problems such as local chemotherapy and drug resistance for facing below for tumor operation, and during oncolytic virus application intratumor injection technical limitation, the present invention is implanted directly into tumor resection site, local treatment carried out to tumors remaining, tumor recurrence is avoided, and has widened the range of application of oncolytic virus.The implantation film is biodegradable polymer, and good biocompatibility can be fully absorbed by human body, and can play the effect of preventing adhesion with physical isolation tumor resection site and peritonaeum.
Description
Technical field
The present invention relates to antitumor implantation film of one kind and preparation method thereof.
Background technology
Surgery excision is the maximally effective treatment method of cancer patient, but because tumor resection margin is difficult to define, it is easily residual
Invisible tumour cell is stayed, causes recurrence after operation rate higher, its 5 years recurrence rates may be up to 77%-100%.In order to prevent swelling
Knurl recurs, and Post operation generally requires to carry out chemotherapy.Traditional embolic chemotherapy is generally systemic vein administration, but because chemotherapeutics choosing
Selecting property is poor, has for normal cell and tumour cell almost same lethal, and systemic administration, toxicity is larger, side effect ratio
More, many patients are difficult to receive.In addition, long-term use chemotherapeutics can cause the drug resistance of tumour cell.Especially, liver
Dirty to wait removing toxic substances organ very insensitive for chemotherapeutics, this chemotherapeutic treatment to this kind of internal organs to serve greatly hinder to be made
With.
Oncolytic virus (oncolytic virus) is naturally occurring or by can selectively infect after artificial reconstructed
And crack a viroid of tumour cell.They by delete some replicated in normal cell institute it is required and in tumour cell
The unwanted viral key gene of institute is replicated so as to reach the effect of selectively targeted tumour cell, can selectivity infected tumor it is thin
Born of the same parents simultaneously replicate wherein, final cracking, kill tumour cell, and discharge the tumour that progeny virion further infects surrounding
Cell, and then without duplication or lethal effect in normal tissue cell, and it can be anti-with inducing specific antineoplastic immune
Should.Because oncolytic virus has the antitumor mechanism different from classic chemotherapy medicine, the tumour cell with chemoresistance is still
Sensitive so is treated to oncolytic virus.On October 27th, 2015, food and medicine surveillance authority of the U.S. (FDA) have been approved by pacifying into
(Amgen) company is used to treat focus in skin and the melanoma of lymph node by the herpes simplex virus of genetic modification.
2016 medical treatment cost regulators of Nian8Yue16 Britain --- United Kingdom National health is with clinical optimizing research institute (NICE) issue most
Whole guide, supports that oncolytic immunotherapy is used for into England and Wales national health services (NHS) system, for treating black
Plain knurl patient.But oncolytic virus cure needs virus concentration higher, therapeutic modality to be typically directly to note viral solution
It is mapped in the distinguishable tumour of naked eyes and with the whole tumour of viral solution saturation.Tumour of this administering mode for internal deep
For the invisible tumour cell of operation residual, it is difficult to realize.When therefore needing one kind badly for oncolytic virus application in knurl
The technology limitation of injection, and effective antitumor medicine and side the problems such as the local chemotherapy that faces below of tumor operation and drug resistance
Method.
The content of the invention
Based on above the deficiencies in the prior art, the problems such as the local chemotherapy faced below for tumor operation and drug resistance, with
And during oncolytic virus application intratumor injection technical limitation, the present invention is prepared for a kind of loading oncolytic with electrostatic spinning technique
The nanofiber implantation film of virus.Tumor resection site is implanted directly into tumor operation, oncolytic virus is sustained out by original position,
Selectively targeted tumour cell, carries out local treatment, it is to avoid tumor recurrence to tumors remaining, and has widened the application of oncolytic virus
Scope.
In order to solve the above-mentioned technical problem, the present invention provides a kind of antitumor implantation film, and the antitumor implantation film is negative
Carry the electro spinning nanometer fiber membrane of oncolytic virus.
Used as the preferred embodiment of above-mentioned technical proposal, the antitumor implantation film of one kind provided in an embodiment of the present invention enters one
Step includes the part or all of of following technical characteristic：
As the improvement of above-mentioned technical proposal, in one embodiment of the invention, the electro spinning nanometer fiber membrane be with
Biodegradable polymer with amino is electrospinning material, the high polymer nanometer fiber prepared using electrostatic spinning technique
Film.
Used as the improvement of above-mentioned technical proposal, in one embodiment of the invention, the oncolytic virus is that oncolytic is simple
One or more mixing in herpesviral, oncolytic adenovirus, oncolytic NDV.
A kind of preparation method of antitumor implantation film, it is characterised in that comprise the following steps：
The preparation of step one, electro spinning nanometer fiber membrane, with electrostatic spinning technique by the Biodegradable high-molecular with amino
Material is prepared into the electrospinning film being made up of nanofiber；
The preparation of step 2, the electro spinning nanometer fiber membrane of load oncolytic virus, by the electro spinning nano fiber described in step one
Film is dipped into oncolytic virus solution, and oncolytic virus is enriched to above electrospun fiber membrane by electrostatic adsorption, is born
The electro spinning nanometer fiber membrane for carrying oncolytic virus is antitumor implantation film.
As the preferred embodiment of above-mentioned technical proposal, a kind of system of antitumor implantation film provided in an embodiment of the present invention
Preparation Method further includes the part or all of of following technical characteristic：
It is in one embodiment of the invention, described with the biodegradable of amino as the improvement of above-mentioned technical proposal
Macromolecular material is preferably shitosan.
Used as the improvement of above-mentioned technical proposal, in one embodiment of the invention, the oncolytic virus is that oncolytic is simple
One or more mixing in herpesviral, oncolytic adenovirus, oncolytic NDV.
Used as the improvement of above-mentioned technical proposal, in one embodiment of the invention, the oncolytic virus solution is preferred
It is the oncolytic virus aqueous solution, the oncolytic virus solution concentration is 103~107pfu/mL。
As the improvement of above-mentioned technical proposal, in one embodiment of the invention, the antitumor implantation film is placed in-
100 DEG C~-5 DEG C preservations.
Used as the improvement of above-mentioned technical proposal, in one embodiment of the invention, the antitumor implantation film is placed in -80
Frozen under the conditions of DEG C.
Compared with prior art, technical scheme has the advantages that：
(1) antitumor implantation film of the invention can realize that local anti-tumor is treated, it is to avoid the poison of systemic chemotherapy is secondary anti-
Should；The drug concentration of tumor locus is improve, medicine usage amount is reduced, cost is reduced.(2) present invention is entered using oncolytic virus
Row antineoplaston, solves unwise to chemotherapeutics using the drug resistance problems and some tumour cells that are produced after chemotherapeutics
The problem of sense.(3) electro spinning nanometer fiber membrane that the present invention is used has larger specific surface area and porosity, is enrichment oncolytic disease
The good carrier of poison, and by being implanted into the easy tumour cell position that oncolytic virus is discharged into residual of the form of film, improve
The concentration of local oncolytic virus also breaches the technology limitation of oncolytic virus intratumor injection.(4) macromolecule that the present invention is used
Material is biodegradable polymer, and good biocompatibility can be fully absorbed by human body, is taken out without second operation.
(5) the Electrostatic Absorption method of the preparation method used by the present invention, electrostatic spinning technique and load oncolytic virus, goes for various
Macromolecular material and oncolytic virus, with universality.(6) the implantation film can with physical isolation tumor resection site and peritonaeum,
Play the effect of preventing adhesion.
Be prepared into shitosan Polymer material with high-specific surface area and hole using electrostatic spinning technique by the present invention
The nano fibrous membrane of rate, and oncolytic virus is loaded with it, tumor resection site is implanted directly into tumor operation, near killing
Potential tumour cell, it is to avoid tumor recurrence.
Described above is only the general introduction of technical solution of the present invention, in order to better understand technological means of the invention,
And can be practiced according to the content of specification, and in order to allow the above and other objects, features and advantages of the invention can
Become apparent, below in conjunction with preferred embodiment, describe in detail as follows.
Brief description of the drawings
Technical scheme in order to illustrate more clearly the embodiments of the present invention, will simply be situated between to the accompanying drawing of embodiment below
Continue.
Fig. 1：Antitumor implantation film action effect schematic diagram；
Fig. 2：Load the electron scanning micrograph of the electro spinning nanometer fiber membrane of oncolytic virus；
Fig. 3：Virus titer and the viral load amount loaded on electro spinning nanometer fiber membrane in oncolytic virus solution before vacuolar membrane
Graph of a relation；
Fig. 4：Load the antitumous effect for various human liver cancer cells of the electro spinning nanometer fiber membrane of oncolytic virus；
Fig. 5：The electro spinning nanometer fiber membrane of oncolytic virus is loaded for the lethality of Bel-7402 cells and the relation of MOI
Figure.
Specific embodiment
The following detailed description of specific embodiment of the invention, its as part of this specification, by embodiment come
Principle of the invention is illustrated, other aspects of the present invention, feature and its advantage will become apparent by the detailed description.
Macromolecular material is prepared into electro spinning nanometer fiber membrane using electrostatic spinning technique, nano fibrous membrane is immersed in molten
In tumor virus solution, oncolytic virus is enriched to above electrospun fiber membrane by electrostatic adsorption, obtains load oncolytic virus
Electro spinning nanometer fiber membrane.Tumor resection site is implanted directly into tumor operation, oncolytic virus is sustained out by original position, specifically
Property targets neoplastic cells, play local anti-tumor in situ effect.The effect of antitumor implantation film of the invention is as shown in Figure 1
Effect diagram.
Described macromolecular material is the biodegradable polymer with amino, such as shitosan.Electrostatic spinning skill
The Polymer Solution can be prepared into nano fibrous membrane by art, i.e., the three dimensions knot being made up of from 3nm -5 μm of fiber diameter
Structure.Because the specific surface area of nano-scale fiber is 1000 times of micron order fiber, the nano fibrous membrane being made up of nano-scale fiber
Specific surface area by several orders of magnitude bigger than the tunica fibrosa that is prepared by common spining technology.
Described oncolytic virus includes oncolytic herpes simplex virus, oncolytic adenovirus, oncolytic NDV etc..
The method of the load oncolytic virus is Electrostatic Absorption method.Because oncolytic virus is generally negatively charged, and by band amino
The electro spinning nanometer fiber membrane for preparing of macromolecular material in the solution can positively charged, can be rich by oncolytic virus by Electrostatic Absorption
Collect above electro spinning nanometer fiber membrane.And be enrichment oncolytic as described above, electro spinning nanometer fiber membrane has great specific surface area
The good carrier of virus, the implantation film can improve the concentration of local oncolytic virus after implanting.
Embodiment 1：Load the preparation method of the electro spinning nanometer fiber membrane of oncolytic virus
By shitosan, (viscosity average molecular weigh is about 850,000, and deacetylation is that 95%) (weight average molecular weight is with polyethylene glycol oxide
Ten thousand) 60 be dissolved in 90% acetic acid solution jointly, is configured to the Polymer Solution that mass fraction is 3.25%, wherein shitosan with
The mass ratio of polyethylene glycol oxide is 90:10.The Polymer Solution is placed in the sampling device of electrostatic spinning apparatus, voltage is adjusted
It is 20kv, shower nozzle is 15cm with the distance of collecting board, collects the nano fibrous membrane that thickness is 150~200 μm.Taken from collecting board
Lower nano fibrous membrane, is immersed in 1hr in the solution of potassium carbonate of 1mol/L, and clean and drying at room temperature is then washed with deionized,
To remove non-volatile clean acetum and polyethylene glycol oxide, the amino group of positively charged on release shitosan.Drying at room temperature
Preserved in device.The preparation method of the electro spinning nanometer fiber membrane of other macromolecular materials with amino is similar, simply gathers shell
Sugar changes other macromolecular materials into.
Dry chitosan nano fiber membrane is cut into 1 × 1cm2Fritter and be dipped into 0.5mL oncolytic virus solution
In, such as simple herpes virus (Wuhan Binhui Biological Technology Co., Ltd. gives), electrostatic adsorption is passed through at 4 DEG C will be with negative
The oncolytic virus of electricity is enriched on the nano fibrous membrane of surface positively charged, and the tunica fibrosa of absorption oncolytic virus, -80 are taken out after 2hr
DEG C freeze.
The electro spinning nanometer fiber membrane pattern of oncolytic virus is loaded with scanning electronic microscope observation.As shown in Fig. 2 oncolytic is sick
Malicious particle is attached to above electro spinning nanometer fiber membrane, and electrospinning film is made up of the nanofiber that diameter is about 150nm.
Embodiment 2：The control of oncolytic virus load capacity
It is 1 × 1cm by area2, weight is dipped into 0.5mL there are different virus to drip for the electro spinning nanometer fiber membrane of 1.5mg
In the oncolytic virus solution of degree, 2hr is stood at 4 DEG C.Before and after detection vacuolar membrane in oncolytic virus solution virus titer change, its
Difference is the viral load amount loaded on electro spinning nanometer fiber membrane.
As shown in the curve of Fig. 2, when concentration viral in oncolytic virus solution before vacuolar membrane from 7 × 103、1.11×105、
2.79×105Increase to 1.11 × 106Pfu/mL, load to oncolytic virus load capacity on electro spinning nanometer fiber membrane therewith from
1.29×103、3.35×104、8.4×104Increase to 3.35 × 105Virus in oncolytic virus solution before pfu/mg, i.e. vacuolar membrane
Concentration is bigger, and the oncolytic virus loaded on electro spinning nanometer fiber membrane is more.
Embodiment 3：Load the oncotherapy effect of the electro spinning nanometer fiber membrane of oncolytic virus
1) cell culture
Human hepatoma cell line Bel-7402 (being denoted as 2D-7402) is put with 10%FBS, 1% dual anti-DMEM complete mediums is contained
In 5%CO2, subculture in 37 DEG C of constant incubators.Its growing state is observed with inverted microscope, a second generation is passed within about 2 days,
The cell in exponential phase is taken for cytotoxicity experiment.
The drug-resistant cell strain 7402-5fu of the Bel-7402 5 FU 5 fluorouracils of resistance to chemotherapeutics (5-fu), its type of rearing with
2D-7402 is similar, and simply its nutrient solution is the DMEM complete mediums containing 20 μ g/mL 5-fu.
There is the property of stem cell with the Bel-7402 cells of 3D cell culture technology cultures, 3D-7402 is denoted as.
The type of rearing of human liver cancer cell HepG2 is identical with 2D-7402.
The drug-resistant cell strain HepG2-ADM of the resistance to chemotherapeutic drugs Doxorubicins of HepG2 (DOX), its type of rearing and HepG2 phases
Seemingly, simply its nutrient solution is the DMEM complete mediums containing 200ng/mL DOX.
2) cytotoxicity experiment
Above-mentioned various HCCs of the selection in exponential phase, are inoculated into 24 well culture plates, and every kind of cell is divided equally
It is 5 groups.Control group only adds culture medium；Blank film group, plus culture medium and electro spinning nanometer fiber membrane；Carry oncolytic virus film group, plus training
Support the electro spinning nanometer fiber membrane of base and load oncolytic virus；5-fu groups, plus the culture medium containing 20 μ g/mL 5-fu；Dox groups, plus contain
The culture medium of 200ng/mL DOX.
The each group cell of different cell lines is placed in 5%CO2, after cultivating 96hr in 37 DEG C of constant incubators, examined with srb assay
Survey its cell killing.Result is illustrated not as shown in figure 4, the cell survival rate of blank film group is similar with control group close to 100%
The electro spinning nanometer fiber membrane for loading oncolytic virus does not have cytotoxicity.It is resistance to from the point of view of the cell survival rate of 5-fu groups and Dox groups
Medicine cell 7402-fu and HepG2-ADM is insensitive for chemotherapeutics 5-fu and Dox, with chemotherapy resistance；Use 3D technology
The 3D-7402 cells with stem cell properties of culture also have certain drug resistance for 5-fu.But carry oncolytic virus film
Group, either for human liver cancer cell, the cell of the drug resistance HCC even 3D technology culture with stem cell properties
Very strong lethality (cell survival rate is less than 40%) is respectively provided with, illustrates that the electro spinning nanometer fiber membrane for loading oncolytic virus has very
Good antitumous effect.
Embodiment 4：Load the Execution of the electro spinning nanometer fiber membrane of oncolytic virus
By the electro spinning nanometer fiber membrane and Bel-7402 cells of the load oncolytic virus with different oncolytic virus load capacity
It is common to be incubated, detect its Execution such as above-mentioned method.MOI is oncolytic on the electro spinning nanometer fiber membrane for load oncolytic virus
The quantity of virus and the ratio of Bel-7402 cell numbers.Result is as shown in figure 5, when MOI increases from 0,0.01,0.3,0.8,1.3
When being added to 8.4, cell survival rate drops to 13.5% from 106%, 75.3%, 44%, 37.5%, 29.7%.Illustrate that load is molten
The electro spinning nanometer fiber membrane of tumor virus is relevant with the load capacity of oncolytic virus on film for the lethality of cell, with the increasing of load capacity
Plus and increase.
Using electrostatic spinning technique by the biodegradable polymer with amino, such as shitosan is prepared into the present invention
Nano fibrous membrane, then includes oncolytic herpes simplex virus, oncolytic adenovirus, oncolytic by electrostatic adsorption by oncolytic virus
NDV etc., is enriched to above electro spinning nanometer fiber membrane, obtains the electro spinning nanometer fiber membrane of load oncolytic virus.In tumour
Tumor resection site is implanted directly into operation, oncolytic virus is sustained out by original position, and selectively targeted tumour cell plays original position
Local anti-tumor is acted on.The concentration that improve local oncolytic virus also breaches the technology limitation of oncolytic virus intratumor injection.It is real
Checking is bright, and the electro spinning nanometer fiber membrane of load oncolytic virus of the invention is resistance to for various HCC Bel-7402 and HepG2
The property of medicine HCC 7402-5fu and HepG2-ADM, the 3D-7402 cells of the 3D technology culture even with stem cell properties
It is respectively provided with very strong lethality.Implantation film good biocompatibility of the invention, can be fully absorbed by human body, be taken without second operation
Go out, the effect of preventing adhesion can be played with physical isolation tumor resection site and peritonaeum.Preparation method used in the present invention, it is quiet
The Electrostatic Absorption method of Electrospinning and load oncolytic virus, goes for various macromolecular materials and oncolytic virus, has
Universality.
The above is the preferred embodiment of the present invention, can not limit the right model of the present invention with this certainly
Enclose, it is noted that for those skilled in the art, under the premise without departing from the principles of the invention, may be used also
To make some improvement and variation, these are improved and variation is also considered as protection scope of the present invention.
Claims (9)
1. a kind of antitumor implantation film, it is characterised in that：The antitumor implantation film is the electrospun nanofiber fibre for loading oncolytic virus
Dimension film.
2. a kind of antitumor implantation film as claimed in claim 1, it is characterised in that：The electro spinning nanometer fiber membrane is with band ammonia
The biodegradable polymer of base is electrospinning material, the high polymer nanometer fiber membrane prepared using electrostatic spinning technique.
3. a kind of antitumor implantation film as claimed in claim 1, it is characterised in that：The oncolytic virus is oncolytic herpe simplex
One or more mixing in virus, oncolytic adenovirus, oncolytic NDV.
4. it is a kind of it is antitumor implantation film preparation method, it is characterised in that comprise the following steps：
The preparation of step one, electro spinning nanometer fiber membrane, with electrostatic spinning technique by the biodegradable polymer with amino
It is prepared into the electrospinning film being made up of nanofiber；
The preparation of step 2, the electro spinning nanometer fiber membrane of load oncolytic virus, by the electro spinning nanometer fiber membrane leaching described in step one
Steep in oncolytic virus solution, oncolytic virus is enriched to above electrospun fiber membrane by electrostatic adsorption, obtain load molten
The electro spinning nanometer fiber membrane of tumor virus is antitumor implantation film.
5. the preparation method of a kind of antitumor implantation film as claimed in claim 4, it is characterised in that：The biodegradable height
Molecular material is preferably shitosan.
6. the preparation method of a kind of antitumor implantation film as claimed in claim 4, it is characterised in that：The oncolytic virus is molten
One or more mixing in knurl herpes simplex virus, oncolytic adenovirus, oncolytic NDV.
7. the preparation method of a kind of antitumor implantation film as claimed in claim 4, it is characterised in that：The oncolytic virus solution
The preferably oncolytic virus aqueous solution, the oncolytic virus solution concentration is 103~107pfu/mL。
8. the preparation method of a kind of antitumor implantation film as claimed in claim 4, it is characterised in that：The antitumor implantation film
It is placed in -100 DEG C~-5 DEG C preservations.
9. the preparation method of a kind of antitumor implantation film as claimed in claim 4, it is characterised in that：The antitumor implantation film
Frozen under the conditions of being placed in -80 DEG C.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020002413A1 (en) * | 2018-06-27 | 2020-01-02 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Multilayer patch for postoperative drug administration |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008013713A2 (en) * | 2006-07-24 | 2008-01-31 | Duke University | Coaxial electrospun fibers and structures and methods of forming same |
| US20090061496A1 (en) * | 2007-08-29 | 2009-03-05 | Dr. D. Graeser Ltd. | Encapsulation of bacteria and viruses in electrospun fibers |
| WO2009148488A2 (en) * | 2008-05-29 | 2009-12-10 | The General Hospital Corporation | Use of oncolytic herpes viruses for killing cancer stem cells |
| WO2014021954A2 (en) * | 2012-04-13 | 2014-02-06 | Trustees Of Tufts College | Magneto-sensitive silk fibroin-based materials |
| CN103933602A (en) * | 2014-04-22 | 2014-07-23 | 东华大学 | Preparation method of chitosan-based drug-loading composite antibacterial superfine fiber membrane |
| CN104822626A (en) * | 2012-09-28 | 2015-08-05 | 加州大学董事会 | Degradable silica nanoshells for ultrasonic imaging/therapy |
-
2017
- 2017-02-18 CN CN201710087762.3A patent/CN106902098B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008013713A2 (en) * | 2006-07-24 | 2008-01-31 | Duke University | Coaxial electrospun fibers and structures and methods of forming same |
| US20090061496A1 (en) * | 2007-08-29 | 2009-03-05 | Dr. D. Graeser Ltd. | Encapsulation of bacteria and viruses in electrospun fibers |
| WO2009148488A2 (en) * | 2008-05-29 | 2009-12-10 | The General Hospital Corporation | Use of oncolytic herpes viruses for killing cancer stem cells |
| WO2014021954A2 (en) * | 2012-04-13 | 2014-02-06 | Trustees Of Tufts College | Magneto-sensitive silk fibroin-based materials |
| CN104822626A (en) * | 2012-09-28 | 2015-08-05 | 加州大学董事会 | Degradable silica nanoshells for ultrasonic imaging/therapy |
| CN103933602A (en) * | 2014-04-22 | 2014-07-23 | 东华大学 | Preparation method of chitosan-based drug-loading composite antibacterial superfine fiber membrane |
Non-Patent Citations (7)
| Title |
|---|
| LAYING WU，等: "Electrospinning fabrication, structural and mechanical characterization of rod-like virus-based composite nanofibers", 《J. MATER. CHEM.》 * |
| SARA IBRAHIM，等: "Improved genistein loading and release on electrospun chitosan nanofiber blends", 《JOURNAL OF MOLECULAR LIQUIDS》 * |
| TAKAHARU OKADA，等: "Inactivated Sendai Virus (HVJ-E) Immobilized Electrospun Nanofiber for Cancer Therapy", 《MATERIALS》 * |
| WSALALHA，等: "Encapsulation of bacteria and viruses in electrospun nanofibres", 《NANOTECHNOLOGY》 * |
| 刘耕陶，等: "《当代药理学》", 31 May 2008, 中国协和医科大学出版社 * |
| 韩瑞发，等: "《浅表膀胱肿瘤的基础与临床》", 31 March 2004, 天津科学技术出版社 * |
| 马悦: "运用共载顺铂和姜黄素的静电纺丝纳米纤维防治宫颈癌术后局部复发", 《中国博士学位论文全文数据库医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020002413A1 (en) * | 2018-06-27 | 2020-01-02 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Multilayer patch for postoperative drug administration |
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