CN105919694A - Multi-layer electrospun membrane and use thereof - Google Patents
Multi-layer electrospun membrane and use thereof Download PDFInfo
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- CN105919694A CN105919694A CN201610200737.7A CN201610200737A CN105919694A CN 105919694 A CN105919694 A CN 105919694A CN 201610200737 A CN201610200737 A CN 201610200737A CN 105919694 A CN105919694 A CN 105919694A
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- electrospinning
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/0063—Implantable repair or support meshes, e.g. hernia meshes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2210/00—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2210/0076—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof multilayered, e.g. laminated structures
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- Cardiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Engineering & Computer Science (AREA)
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Abstract
The invention relates to the technical field of nano-biomaterials, in particular to a multi-layer electrospun membrane and the use thereof. The multi-layer electrospun membrane includes an inner layer material, an intermediate layer material and an outer layer material. The multi-layer electrospun membrane is used as a dura mater substitute material, and has significantly improved performances as compared with a collagen matrix through comparison tests on mechanical properties, porosity, water absorption and water repellency.
Description
Technical field
The present invention relates to nano meter biomaterial technical field, be specifically related to a kind of cladding electrospinning film and use thereof
On the way.
Background technology
1. dural substitutes present Research
Cerebral dura mater is wrapped around the membrane structure outermost layer outside brain and spinal cord, is the firmest a kind of connective
Tissue thin film, the inner surface of natural cerebral dura mater structure is the monolayer fibroblast that a Rotating fields is fine and close,
Thin and smooth；More fibroblast and a small amount of collagen are contained in intermediate layer；Outer layer is thicker, containing relatively
Many collagen component and a small amount of fibroblast.Such structure inner face is smooth and entirety ten
That divides is tough and tensile, it is possible to well plays protection brain and prevents the effect of cerebrospinal fluid seepage.Outside due to brain
Cerebral dura mater must be cut when wound, the cerebral tumor and cerebrovascular carry out operation of opening cranium；Meningorrhagia needs coagulation
Hemostasis；Tumor invades cerebral dura mater, needs to make meninges excision；Cerebral dura mater damage often results in defect, these
Factor causes dura defect, damage, cerebrospinal leak, intracranial infection of common occurrence, therefore outside nerve
Section's operation it is frequently necessary to carry out duramater reparation.
Dural substitutes is mainly used in brain convex surface and skull base surgery, first performs the operation at brain convex surface
In, because of the open decompression of meninges, Yi Jiyi in wound coup injury, the infiltration of tumor, operation process
Need during hard brain (ridge) the film defect that a little geneogenous factors cause to repair, substitution material in art
Application percentage about about 30%；Secondly in basis cranii surgical operation, due to basis cranii cerebral dura mater relatively
Thin, and fit closely with basis cranii skeleton so that it is once occur that defect easily causes cerebrospinal leak,
As fracture of skull base, especially anterior cranial fossa and fracture of middle fossa will easily cause cerebrospinal rhinorrhea and otorrhea.
The tumors such as the meningioma of basis cranii, chordoma usually corrode infiltration and arrive skull and the cerebral dura mater of surrounding, are cutting
Except generally also attaching part meninges and skull while tumor, this is the root of postoperative generation cerebrospinal leak
Source, so cerebral dura mater needs the probability repaired higher in skull base surgery, accounts for 50% left side of operation ratio
Right [1].
The character that preferably dura mater substitution material should possess includes: 1. close with normal cerebral dura mater life
Thing mechanical characteristic, has certain elasticity, toughness and stands stitching, can effectively recover Subdural space
The integrity in chamber.2. there is stable biocompatibility, do not cause host to reject and inflammatory reaction, nothing
Potential toxic and side effects.3. can provide support for the autogenous repairing of dura mater, be beneficial to fibroblast raw
Length and migration, thus promote dura mater to regenerate；New dura mater forms rear graft and can be gradually absorbed, in case
Longer-term persistence brings the harmful effects such as chronic inflammatory reaction in internal foreign body to health.The most do not increase art
Rear intracranial infection rate and the incidence rate of intracranial hematoma.5. nervous tissue and surrounding tissue are not caused secondary
Infringement, without potential pathophorous risk.6. it is prone to sterilization store, economical and easily available, easy to use.
The material that at present dura defect reparation uses the most clinically mainly has a few class: autologous fascia tissue,
Allohisto compatibility, xenogenic biological materials, synthetic material etc..
Autologous tissue such as Temporal fascia, cranial periosteum, hat shape key film, autologous fascia lata etc. are traditionally
The duramater reparation substitution material used the earliest, the most still has clinician to select, because these materials
Deriving from autologous, histocompatibility is good, will not produce rejection, and existing enchashment with, be not required to two
Secondary sterilization, disease will not be infected also do not result in any immunoreation [2-4], but source of drawing material, take
Material size and shape is limited, increases the operating procedure in operation, brings new iatrogenic wound to patient
Wound and misery, time especially when open craniocerebral injury, large area dura defect, be more difficult to look for
Repair to suitably self fascia tissue, do not meet the needs of modern neuro surgery development.
Allohisto compatibility such as lyophilizing human cerebral dura mater, at thickness, tension stress intensity, fibre architecture etc.
Aspect has the feature identical with self cerebral dura mater, implants conveniently, can preserve for a long time.But because there being report
Road can be propagated Creutzfeldt-Jakob disease (CJD) and have potential viral infection risk [5,6],
Clinical practice [7] is forbidden by some countries.Once had the allogeneic acellular dermis of harmless treatment for
Duramater reparation is such as[8], due to identical risk, present this material is seldom faced
Bed is used.
Heteroplasm's peplos such as pig peritoneum, cattle and sheep pericardium,Deng the most once for cerebral dura mater
Repairing [9,10], these materials are similar to mankind's fascia in composition quality, tougher resistance to stitching and cutting
Cutting out, convenient to operate in operation, material source the most relatively human body fascia has a clear superiority in.But it is because it
Degradable can not absorb, and these animal tissue's materials yet suffer from potential CJD and infect possible,
And have the risk producing rejection.
Along with the progress of biomaterial science, the such as collagen protein of the active skull cap components in heteroplasm,
Fibrins etc. are extracted and carry out restructuring and harmless treatment is made thin-film material and repaiied for cerebral dura mater
Mend and become a kind of new trend.Cerebral dura mater based on collagen stroma is such as in recent years Become to be becoming increasingly popular.Collagen protein is a kind of extracellular protein, for extracellular base
The main component of matter, is twisted into spiral fibrous proteins by 3 peptide chains, and collagen protein is human body
Interior content rich in protein, accounts for more than the 30% of whole body gross protein.Have now been found that more than 16 kinds
Collagen molecules, wherein based on type i collagen, be distributed widely in skin, skeleton, blood vessel etc. many
Number connective tissue, its function is to maintain skin and the form of histoorgan and structure, is also to repair each damage
The important source material material of injured tissue.Collagen protein can make blood coagulation, has coagulation function, can conduct
Wound hemostasis dressing, for first aid and treatment wound.Collagen protein is possible not only to be identified by cell,
Cell is also had chemotactic characteristic, has and have good biocompatibility, promote cell adhesion, breed, accelerate
Wound healing, no antigen, can the advantage such as natural degradation, catabolite avirulence.Collagen protein
Biocompatibility is derived from interaction good between host cell and tissue, is either being inhaled
Receiving previous crops is neoblastic skeleton, or being entered host by absorption and assimilation becomes a part for tissue, all
There is good interaction with pericellular substrate, show interactional harmony, and become
For cell and tissue normal physiological function part of the whole.It can separate brain as meninges substitution material
Organize with on brain, promote that healing up of traumatic tissues and self cerebral dura mater are newborn, and without obvious inflammatory reaction,
So general effect is satisfied [11-16] in clinical practice.But collagen stroma also has substantially
Shortcoming, as big in self fragility, mechanical strength is poor, for cerebral tissue provide protection limited, collagen-based
Matter fragile structure sew up difficulty, be not easy to fix, cerebral dura mater is closed and the best, postoperative be there is cerebrospinal leak
Risk etc..Some other kinds of natural high molecular substance such as chitosan, alginate water-settings in recent years
Glue etc. are also used for the research of dural repairment material, are not the most the most ripe in application aspect, the widest
General used [17-19] by clinical operation.
Synthetic inert material is also commonly used for dural repairing, and these materials mostly are polyphosphazene polymer
Compound, once conventional have expanded PTFE (ePTFE),Polyurethane,
Deng, these biomaterials have extraordinary mechanical strength and reasonable resistance to blocking, can be effective
Preventing the adhesion [20-25] of cerebral tissue, material is easy to manufacture, it is easy to sterilization stores, and does not has transmitted virus
Risk.Shortcoming be these inert materials can not natural degradation, have and cause granulation tissue to be formed and induction is slow
Sexual stimulus causes long-term foreign body reaction possible.The degradation material of synthetic gets more and more in recent years
For dural substitutes study such as polylactic acid (PLA), polycaprolactone (PCL), poly butyric
Ester (PHB), polypropylene glycol (PPG) etc., these materials can natural degradation in vivo, thus avoid
The infection risk caused because of longer-term persistence and immunoreation [26-33], these degradation materials have continued people
Mechanical property that work inert material is good and histocompatibility, it is possible to the intensity remaining enough is easy to operation
Operate and prevent cerebrospinal leak, but be to lack biological activity as high molecular polymerization product deficiency, right
Wound healing and self cerebral dura mater new life are not significantly promoted effect.And these high molecular polymer water
Solve afterproduct and can produce local slightly acidic environment, unfavorable to cell and tissue growth.
2. electrostatic spinning technique
Along with progress and the development of nanotechnology, electrostatic spinning obtains attention and the utilization of height in recent years,
But before its basic ideas may trace back to more than 100 year.People are from research to the understanding of Electrospun
Liquid electrojet behavior under the electric field starts.Raleigh in 1882 etc. have studied drop at electric field
The wild effect of middle appearance, when electric field force exceedes surface tension, balance originally is broken, outstanding
Drop at spinning head has just cleaved into a series of charged droplets, and this wild effect was claimed later
For " Raleigh Instability " [34].Since 1915, Taylor etc. have studied drop and charging fibre
The problem that dimension bundle crushes under the electric field, along with electric field is strengthened, drop is gradually elongated, and works as electric field
When power and surface tension numerical value are equal, being the formation of the circular cone that drift angle is 49.3 °, this circular cone was later
It is referred to as Taylor cone.On the basis of liquid electric spray process has been had basic understandings, electrostatic spinning by
Gradually it is applied to the preparation of fiber, thus develops into the electrostatic spinning technique obtaining polymer nanofiber.
Within 1934, Formlals has delivered first patent of electrostatic spinning technique, illustrates and uses electrostatic field
Prepare technique and the device [35] of macromolecule filament.Simons diameter in 1966 is ultra-fine, weight is extremely light,
And the nanofiber with different pattern is made for corresponding non-woven fabrics [36].Last century the nineties
Reneker place research group improves electrostatic spinning technique and development is made that significant contribution, not only
Including series macromolecular being carried out Electrospun, also fiber formation mechenism in spinning process is done detailed explaining
State.The basic and applied research to electrostatic spinning technique has been started from these global research workers.
Electrostatic spinning is completely different with traditional fibre manufacture method, is spinning of polymeric spray electrostatic stretch
Silk method.First by volt high-pressure electrostatics thousand of to up to ten thousand on polymer solution or melt band, charged is poly-
Compound drop under the effect of electric field force capillary tube Taylor conical point be accelerated；When electric field force foot
Time the biggest, polymer drop overcomes surface tension to form injection thread；Thread is solvent in course of injection
Evaporation or solidification, finally fall on the reception device, forms the fiber felt of similar nonwoven cloth-like, thus obtains
To micron or the processing technique of nano level superfine fiber.Electrostatic spinning process was broadly divided into for three stages: the
One stage, liquid stream produce and linearly stablize preliminary drawing-off；Second stage, unstable
Non-axis symmetry whip is dynamic makes fluid be further stretched；Phase III, thread is dried and is frozen into diameter several
The fiber of nanometer extremely hundreds of nanometer.How liquid stream diameter is by 1 millimeter (capillary outlet diameter)
The mechanism dropping to less than 1 micron is not currently fully understood, but generally believes except one, three stages made
Outside fiber attenuates, second stage has played decisive role [37].
Electrostatic spinning technique may be used for production and is difficult to by the acquisition of traditional non-woven measuring fiber manufacturing technology
Fiber, fiber diameter range has higher surface from tens nanometers to a few micrometers, electrospinning material
Volume ratio, porosity and adjustability, can form all size and shape by parameter adjustment, in addition
Change electrospinning film component proportioning can control to reach desired material character and function.Electrostatic spinning is with it
Manufacture the advantages such as device is simple, spinning is with low cost, it is various to spin substance classes, technique is controlled,
Become effectively one of main path preparing nano-fiber material.The nanofiber that electrostatic spinning manufactures
Diameter is suitable less than the microfilament in cell, with extracellular matrix, the micro-pore knot in addition communicated with each other
Structure, these features enable electrospun fibers felt to simulate natural extracellular matrix (ECM) knot well
Structure, provides preferable microenvironment for cell growth, promotes cell and substrate and cell and intercellular phase
Interaction, guide tissue regeneration and reparation.Some electrospinning raw materials have good biocompatibility and can
Degradability, can enter human body as carrier, and be readily absorbed by；In addition electrostatic spinning nano fiber has very
The good characteristics such as big specific surface area, porosity, therefore it causes researcher at biomedical sector
Give more sustained attention, and obtain in many-sides such as medicine controlled releasing, repair in trauma, bioengineered tissues
Well application [38-43].Electrospinning process currently mainly has following a few class.
Mixed solution electrostatic spinning, will be dissolved in same solvent by two or more polymer, or will
Two or more polymer solution mixes, and carries out electrostatic spinning again, obtain many after forming homogeneous solution
The superfine fibre electrospinning film of component.Mixed solution electrospinning has both sides advantage: on the one hand some gathers
Compound is difficult to electrostatic spinning, add another kind of the most compatible can the polymer of electrospinning, it is possible to
Preparation is containing the electrospinning film of these polymer, thus expand can the scope [44-46] of electrospinning material.Separately
On the one hand the fiber obtained by electrospinning can get both the advantage of multiple polymers, makes obtained electrospinning film
There is wider application [47,48].
Altogether electrostatic spinning, refer to two or more polymer solution under the effect of electric field force independently
Electrospinning, uses rotary drum or rotating disk for receiving device, realizes multiple polymers fiber by the rotation of rotary drum
Compound.Its advantage compared with mixing electrospinning is, when two kinds of polymer, can not to be dissolved in same molten
During agent, common-battery spins and still can realize the compound of superfine fibre and be interweaved together, so that
The performance of the electrospinning film arrived is more sophisticated [49,50].
Multilamellar electrostatic spinning, i.e. first electric spinning polymer A, then by another kind polymer B electrospinning to
On the polymer A electrospinning film completed, then continue to electric spinning polymer C... by that analogy, it is possible to obtain many
The fiber membrane structure of layer heterogeneity, so that electrospinning film possesses different functional layers, extends electrospinning
The application of film.
Coaxial electrostatic spinning, is to be improved device on the basis of conventional electrostatic spinning, prepare core/
A kind of method of shell knot fine fibre.The range of application of coaxial electrostatic spinning is more extensive, can use polymer
Template, use is provided can not individually to carry out the conducts such as the material such as medicine dispersant of electrospinning fibroblast as shell
Core, the nano level superfine fiber of preparation knockout/shell structure.This method creates a kind of nano level thing
Matter slow-releasing system, advantage is on the premise of ensureing high surface and porosity, and sheathing material is internal
Active medicine provide protection so that it is can material in sustained-release core and preferably play biological activity
Effect, it is to avoid the burst drug release phenomenon [51-55] that common medicine carrying electrospun fibers felt the most easily occurs.
Orientation electrostatic spinning, is the arrangement ordering by the improvement of electro-spinning equipment makes electrospinning fibre,
The trend of electrospinning fibre can be rearranged according to artificial design, thus obtained Nanowire can be made
Dimension felt possesses the ability of more preferable mechanical property and induced cell growth.This structure is suitable for manufacturing
Artificial-muscle and blood vessel etc. are organized, because can simulate these tissues by orderly fiber alignment
Natural structure, builds for organizational project and they provides probability [56-60].
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Summary of the invention
It is an object of the invention to solve the problems referred to above, it is provided that a kind of cladding electrospinning film and application thereof, energy
Enough solve the problems referred to above.
The present invention solves the problems referred to above and be the technical scheme is that
A kind of cladding electrospinning film, including inner layer material, middle layer material, cladding material, it presses following step
Suddenly it is prepared from:
1) preparation of inner layer material: inner layer material composition is PLA, concentration of dope 5%wt, electrospinning is joined
Number for flow velocity be 1.0ml/h, voltage 12kV, receiving range 15cm, electrostatic spinning 2h；
2) preparation of layer material in: middle level material selection PLA/PCL (1/1) group, composition is
50%PLA50%PCL, concentration of dope 5%wt, electrospinning parameter be flow velocity be 1.0ml/h, voltage 12kV,
Receiving range 15cm, electrostatic spinning 4h；
3) preparation of cladding material: cladding material chooses collagen/PLA (4/1) group, composition is 80% collagen
20%PLA, concentration of dope 6%wt, electrospinning parameter is flow velocity 1.2ml/h, voltage 14kV, receive away from
From 15cm, electrostatic spinning 6h.
Further, described electrospinning conditions is indoor temperature 25 DEG C, relative humidity 40%.
Further, the thickness after described cladding electrospinning film electro static spinning completes is
212.34±42.24μm。
The purposes of a kind of cladding electrospinning film, applies to dural substitutes by cladding electrospinning film.
The invention has the beneficial effects as follows:
1. electrospinning film and collagen stroma contrast, enterprising in mechanical property, porosity, water absorption rate, water proofing property
Row contrast test, electrospinning film properties is significantly increased.
2.HSF (fibroblast) can grow on electrospinning film layer materials and normally rise in value, acridine
Orange dyeing is better than internal layer and culture plate with MTT detection display cell proliferation of growth in material outer layer
Comparison；Adhesion experiment display cladding material is conducive to HSF cell adhesion；Quantitative PCR and Western blot
Result show that on cladding material the HSF cell of growth is in extracellular matrix secretion is functionally better than
Layer and culture plate compare.
3. after subcutaneous rat implants electrospinning film, histiocyte can attach on material ectonexine and grow,
HE dyeing display cladding material is raw compared with having more fibroblast and epidermal-like cell to migrate in inner layer material
Grow into inside.Build new zealand rabbit dura defect animal model, implant cladding electrospinning film substitution material
Postoperative animal recovers good, infection of incisional wound and intracranial infection sign does not occurs.When electrospinning membrane material takes out
Inner surface and cerebral tissue then fit tightly with surrounding tissue without obvious adhesion, material outer layer, have during stripping
Certain resistance；The cerebral tissue surface color of transplantation site and periphery is normal, without substantially damage, edema,
Inflammatory reaction.Electrospinning film HE dyeing visible material outer layer has more fibroblast peacekeeping epidermal-like cell compared with internal layer
Grow into wherein；Implant site cerebral tissue HE dyeing display electrospinning film is with self fascia matched group the most substantially
, all there is not the vestige of obvious inflammatory reaction and granulation tissue hyperplasia in difference.
Accompanying drawing explanation
Fig. 1 is containing collagen, PLA, PCL cladding electrospinning membrane structure ideograph；
Fig. 2 is that cladding electrospinning film contrasts mechanical test figure with collagen stroma；
Fig. 3 is cladding electrospinning film external degradation weight-loss ratio curve chart；
Fig. 4 is the proliferative conditions that HSF cell cultivates 1d, 3d, 5d on layer materials and culture plate
(* is P ＜ 0.05) figure；
Fig. 5 is HSF cell cell adhesion quantity (* is P ＜ 0.05) on layer materials and culture plate
Figure；
Fig. 6 is the mRNA of HSF cell Colla1A2, Eln on electrospinning film layer materials and culture plate
Express (* is P ＜ 0.05) figure；
Fig. 7 is the albumen of HSF cell Colla1A2, Eln on electrospinning film layer materials and culture plate
Express (* is P ＜ 0.05).
Detailed description of the invention
As it is shown in figure 1, a kind of cladding electrospinning film of the present invention, including inner layer material, middle layer material,
Cladding material, it is prepared from according to the following steps:
1) preparation of inner layer material: inner layer material composition is PLA, concentration of dope 5%wt, electrospinning is joined
Number for flow velocity be 1.0ml/h, voltage 12kV, receiving range 15cm, electrostatic spinning 2h；
2) preparation of layer material in: middle level material selection PLA/PCL (1/1) group, composition is
50%PLA50%PCL, concentration of dope 5%wt, electrospinning parameter be flow velocity be 1.0ml/h, voltage 12kV,
Receiving range 15cm, electrostatic spinning 4h；
3) preparation of cladding material: cladding material chooses collagen/PLA (4/1) group, composition is 80% collagen
20%PLA, concentration of dope 6%wt, electrospinning parameter is flow velocity 1.2ml/h, voltage 14kV, receive away from
From 15cm, electrostatic spinning 6h.
Further, described electrospinning conditions is indoor temperature 25 DEG C, relative humidity 40%.
Further, the thickness after described cladding electrospinning film electro static spinning completes is
212.34±42.24μm。
The purposes of a kind of cladding electrospinning film, applies to dural substitutes by cladding electrospinning film.
The mechanical property of electrospinning film: cladding electrospinning film dry state and hygrometric state and commercially available collagen stroma mechanical test
Result is as shown in Figure 2.Electrospinning film is peak load, tensile strength, elongation at break under dry and wet state
The most relatively collagen stroma dry state increases significantly.37 DEG C of normal saline of electrospinning film soak mechanics after 48h
Being decreased obviously does not occurs in performance, and collagen stroma soak mechanical property after 48h occur in that notable under
Fall.
Electrospinning film water absorption rate: collagen stroma water absorption rate % is 91.4 ± 2.3, cladding electrospinning film contains PLA
With hydrophobic polymer polymer such as PCL, water absorption rate % is 56.2 ± 2.1, substantially less than collagen stroma.
Electrospinning membrane porosity: the three-decker of cladding electrospinning film is all manufactured by electrostatic spinning technique, fiber
Very thin, and have larger aperture diameter ratio, therefore porosity % more a height of 82.2 ± 3.7, collagen stroma
Porosity % is 61.3 ± 4.9, substantially less than electrospinning film.
The leakage resistance of electrospinning film: in the composition of cladding electrospinning film, PLA Yu PCL is waterproof material, electricity
Spinning film resistance to hydrostatic pressure power is 5.84 ± 0.72KPa, and the resistance to hydrostatic pressure power of collagen stroma is 0.88 ± 0.24
KPa is substantially less than electrospinning film.
The external degradation of electrospinning film: cladding electrospinning film, collagen stroma and PLA electrospinning film are through 37 DEG C
All occur in that the situation of loss of weight after artificial cerebrospinal fluid immersion in vitro, extend three kinds of materials with soak time and subtract
Heavily rate all continues to increase.After wherein PLA soaks 16d, weight-loss ratio % is 12.88 ± 0.34, loss of weight speed
Slower；And collagen stroma immersion 16d weight-loss ratio % is 25.67 ± 0.32, loss of weight speed is very fast；Cladding
Electrospinning film weight-loss ratio is therebetween.Weight-loss ratio curve is as shown in Figure 3.
HSF cell MTT detection on ectonexine electrospinning membrane material: inside and outside MTT detection display electrospinning film
HSF cell on layer material and on (matched group) at the bottom of cultivation plate hole is thin with each group of the prolongation of incubation time
Born of the same parents all present and gradually breed trend, and after cultivating 5d, in material outer layer, the cell OD value of growth is significantly
Higher than internal layer group and matched group, as shown in Figure 4.
HSF cell cell adhesion experiments on electrospinning film layer materials: HSF cell is respectively inside and outside
Cell adhesion experiments on layer electrospinning membrane material and culture plate shows, extends each group of cell in time at material
Adhesion quantity on material all has increase, the HSF cell quantity adhered on electrospinning film cladding material after 7h
It is significantly higher than internal layer group and matched group, as shown in Figure 5.
HSF cell quantitative PCR on electrospinning film layer materials: Primer Premier 5.0 is soft
The parameter of part design quantification PCR primer is, amplified production length 80～150bp, primer length 21 ± 4
Bp, other parameter acquiescence, design principle is to select amplified production to cross over the primer pair of intron as far as possible,
5 ' ends of editor's primer are to avoid the occurrence of hair clip or dimeric structure and to have the mispairing of amplified production, electrospinning
On film layer materials and culture plate, HSF cell is after cultivating 5d, the cell of growth on cladding material
The mrna expression level of Colla1A2 and Eln is significantly higher than internal layer group and matched group, as shown in Figure 6.
HSF cell Western blot on electrospinning film layer materials: electrospinning film layer materials
And HSF cell is after cultivating 5d on culture plate, on cladding material, the cell of growth expresses Colla1A2
With the protein band optical density of Eln apparently higher than remaining two groups, show that its protein expression level is significantly high
In internal layer group and matched group, as shown in Figure 7.
Embodiment 1
Electrospinning film layer materials SD subcutaneous rat implants experiment
Male SD rat, body weight is 200～about 300g, and quantity is 12, separately raises in cages, and feeds
The feedstuff of Experimental Animal Center supply, freely drinks water.Observe 3 days without exception after proceed by experiment.
All experimental rats are all anaesthetized with the chloral hydrate solution of lumbar injection 300mg/kg, and back is shaved
Hair, normal sterile processes, and dorsal midline lateral symmetry position is respectively made one and is parallel to center line, length
Reach subcutaneous otch for the 3cm degree of depth, about every rat back, implant inner layer material and outer layer material respectively
Material.
Postoperative external use sterile gauze is wrapped up, the outer dressing of replacing every day, individually raise in cages conventional feeds and drinking-water,
Observe otch and ambient conditions, normally take out stitches after 7d.
Being subcutaneously implanted Post operation skin gradually to heal, otch is without inflammatory exudate, without incrustation, otch week
Enclosing without obvious red swelling of the skin, it is the poorest to implant between the rat cutting part of different two groups electrospinning film
Not, all equal healing states of laboratory animal otch are good, and 7d disruption of wound does not occurs after taking out stitches.Implant
After 10d, all rats all survive, and operating scissors starts face again, it is seen that surrounding tissue cells is on electrospinning film
Growth, when peeling off electrospinning film, electrospinning film inner layer material and surrounding tissue are without obvious adhesion, electrospinning film
Cladding material then has adhesion with surrounding tissue, has certain resistance during stripping；6 are remained after implanting 20d
Survival of rats, electrospinning film taking-up process is similar with group after implantation 10d, electrospinning film inner layer material and surrounding
Organizing without obvious adhesion, electrospinning film cladding material is the tightst with surrounding tissue growth, has resistance during stripping
Power.It is similar that all experimental rat electrospinning films take out situation, has no that electrospinning film surrounding tissue has obvious inflammation
Disease reflection also occurs without local hydrops and infection conditions.
Take out electrospinning film to observe under scanning microscope, cladding material grows the cellular layer of 10d, 20d
Completely covers material surface；A small amount of cell attachment and life is only had after inner layer material surface 10d, 20d
Long, but do not merge in flakes, it is clear that the fibre structure that material surface is blank.
Examine under a microscope after the electrospinning film section HE dyeing taken off, cladding material has relatively many cells
Migration is grown into inside it, and cell growth is intensive, cellular layer is thicker, the cell grown into fusiformis, mostly
The fibroblast of dihedral and flat star is main, is additionally also shown in a small amount of epithelioid cell, and inside is not
See significantly other inflammatory cells；On inner layer material, only a small amount of cell grows and migrates in material
Portion, cell is sparse, cellular layer is relatively thin, and cell type of growing into is based on fibroblast and a small amount of epithelium
Like cell.
Embodiment 2
Cladding electrospinning film implants the experiment of dura defect new zealand rabbit
New zealand rabbit 6, male and female are random, and 2～3kg, quarantine and individually raise in cages for 7 days, maintain ambient temperature
Feedstuff with Experimental Animal Center supply of regularly throwing something and feeding.
The pentobarbital sodium 70mg/kg body weight using 3% is anaesthetized in rabbit ear edge intravenous injection, animal surgery
Platform fixes head, binder fixing head, infuses and intubates permission and breathe in art, 2% iodine tincture and 75% ethanol
Sterilization.The long straight cut of center line 4cm is cut scalp and is reached skull, exposes skull.The rear side of crown line, cranium
Bore the bone window of mill diameter about 12mm, expose cerebral dura mater.Wipe out the circular hard of 1 about 8mm × 8mm
Meninges and arachnoidea defect.
Cladding electrospinning film is cut into the circular implant of about 1.0cm diameter, and internal layer PLA implants on surface down
At defect, insert a small amount of medical gelatin sponge and prevent material from loosening, the amount inserted about with the skull of defect
Edge is concordant, conventional each layer fascia and the scalp sewed up, and conventional dressing covers wrapping.Matched group uses animal
Self fascia tissue is inserted at dura defect, and operation process is identical.
Postoperative reinforcement of individually raising in cages is observed, and more change dressings after 2d, after 7d, otch is taken out stitches.Extensive according to animal
Multiple situation is given and the forage feed such as fresh vegetables, and (department of cerebral surgery is postoperative for ease of seeing not to give analgesic drug product
Examine neurosigns situation of change, be typically cautious use of analgesic drug product) postoperative 1～3 day, penicillin 200,000 U/
Secondary, 1 time/d, intramuscular injection.
The all laboratory animals of Post operation recover good, and wound healing is good without infecting sign, and local is without long-pending
Liquid, oozes out without secretions, and otch cerebrospinal leak does not occurs after taking out stitches, and occurs without Deviant Behavioies such as epilepsies；
Operative site is without cyst, without scleroma and locally bulging or hardening.Postoperative feed water inlet is normal, and animal lives
Dynamic normal, do not find the obvious dyskinesia.
After postoperative 30d, laboratory animal all survives, and the material that defect of skull position is implanted is observed in operation again
Situation, there is the growth of obvious cell on material outer layer surface together with being grown in surrounding tissue, peels off electrospinning
Resistance is had during film；Electrospinning film inner surface and cerebral tissue are easy without obvious adhesion, stripping, implant site brain
Tissue surface is relatively smooth, and color is normal, have no obvious inflammatory reaction, and cerebral tissue is without obvious edema mark
As.
The cladding electrospinning film HE dyeing taken out after implanting 30d, cladding electrospinning film inner layer has a small amount of cell to move
Moving up into, cell is sparse, cellular layer is relatively thin；Electrospinning film outer layer then has greater number of cell migration to enter
Entering material internal, cellular layer is thicker.
Cladding electrospinning film implants dura defect New Zealand with using animal self fascia tissue (matched group)
The implant site cerebral tissue HE dyeing taken out after rabbit 30d, cerebral tissue all presents normal dyeing, in having no
Property the inflammatory cell such as granulocyte, lymphocyte reaction.
Skilled person will appreciate that of the industry, the present invention is not restricted to the described embodiments, above-mentioned reality
Execute the principle that the present invention is simply described described in example and description, without departing from present invention spirit and model
On the premise of enclosing, the present invention also has various changes and modifications, and these changes and improvements both fall within requirement
In the scope of the invention of protection.Claimed scope is by appending claims and equivalence thereof
Thing defines.
Claims (4)
1. a cladding electrospinning film, it is characterised in that include inner layer material, middle layer material, outer layer material
Expecting, it is prepared from according to the following steps:
1) preparation of inner layer material: inner layer material composition is PLA, concentration of dope 5%wt, electrospinning is joined
Number for flow velocity be 1.0ml/h, voltage 12kV, receiving range 15cm, electrostatic spinning 2h；
2) preparation of layer material in: middle level material selection PLA/PCL (1/1) group, composition is
50%PLA50%PCL, concentration of dope 5%wt, electrospinning parameter be flow velocity be 1.0ml/h, voltage 12kV,
Receiving range 15cm, electrostatic spinning 4h；
3) preparation of cladding material: cladding material chooses collagen/PLA (4/1) group, composition is 80% collagen
20%PLA, concentration of dope 6%wt, electrospinning parameter is flow velocity 1.2ml/h, voltage 14kV, receive away from
From 15cm, electrostatic spinning 6h.
A kind of cladding electrospinning film the most according to claim 1, it is characterised in that: described Static Spinning
Strand part is indoor temperature 25 DEG C, relative humidity 40%.
A kind of cladding electrospinning film the most according to claim 1, it is characterised in that: described cladding electricity
Spinning the thickness after film electro static spinning completes is 212.34 ± 42.24 μm.
4. the purposes of a cladding electrospinning film, it is characterised in that: cladding electrospinning film is applied to cerebral dura mater
Substitution material.
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| CN108066043A (en) * | 2017-12-22 | 2018-05-25 | 山东赛克赛斯生物科技有限公司 | A kind of medical embedded sticking patch and preparation method and application |
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| CN115364284A (en) * | 2022-09-01 | 2022-11-22 | 东南大学泰州生物医药与医疗器械研究院 | Absorbable composite anti-adhesion membrane and preparation method thereof |
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