CN101347403A - Method for preparing organic metal compound anti-cancer medicine with electrostatic spinning technique - Google Patents

Method for preparing organic metal compound anti-cancer medicine with electrostatic spinning technique Download PDF

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CN101347403A
CN101347403A CNA2008100418981A CN200810041898A CN101347403A CN 101347403 A CN101347403 A CN 101347403A CN A2008100418981 A CNA2008100418981 A CN A2008100418981A CN 200810041898 A CN200810041898 A CN 200810041898A CN 101347403 A CN101347403 A CN 101347403A
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solution
dichloromethane
nozzle needle
product
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吴庆生
陈平
丁亚平
张琨
朱子春
李升�
陈义军
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Tongji University
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Abstract

The invention relates to the fields of nano-biomedicines and nano-materials, in particular to a method for preparing an organic metal complex anticancer drug by electrostatic spinning technology. The specific steps of the method are as follows: solution A and solution B are respectively prepared; the solution A and the solution B are evenly mixed and then ejected by a nozzle needle; the ambient temperature is controlled at 18-20 DEG C and the ambient humidity is 45%-50%; the space between collecting shields is 14-15cm, the voltage is 10-20KV, and the mass velocity of an injection pump is 2-8mL/h; and aluminum foil collecting product fiber is arranged at collecting shield. The method of the invention can control the drug content, thickness of the shell layer, fineness of the micro/nanometer fiber, and the like, of the product only by controlling the conditions such as drug content, voltage, flow rate of the nozzle needle, etc., by which the position and time specific release of the drugs is expected to realize, and a new carrier for oral administration of some medicines is expected to be provided. By adopting the method, micro/nanometer drug fiber membrane with large area and great thickness can be spun in a short time. The method has the advantages of high drug recovery rate, large single-time output of the product, easy storage of the product, and the like, thus being valuable for industrial application and expected to be expanded to a large-scale application.

Description

Method with preparing organic metal compound anti-cancer medicine with electrostatic spinning technique
Technical field
The invention belongs to nano biological medicine and material technology field, be specifically related to a kind of method with preparing organic metal compound anti-cancer medicine with electrostatic spinning technique.
Background technology
Cancer is one of principal disease of serious harm human health, and cancer has become the second largest killer who is only second to the cardiovascular diseases.Capture cancer is the research topic of attracting attention in the world always.The investigation of World Health Organization (WHO) shows that the cancer patient increases just year by year.At present people have carried out extensive studies to cancer, particularly to the research of cancer therapy drug.
1979, the Koepf reported first of Berlin, Germany polytechnical university Cp 2TiCl 2Active anticancer, and it has been carried out systematic research.Find that cyclopentadienyl titanium dichloride has the broad spectrum anticancer activity, and present the crossing drug resistant activity more weak with cisplatin.But also there are two big difficulties in the application of cyclopentadienyl titanium dichloride, the one, and low at the aqueous phase dissolubility, the 2nd, shortcomings such as its acting duration weak point.These drawbacks limit the use of this medicine.Therefore research improves cyclopentadienyl titanium dichloride antineoplaston effect very significance.Cisplatin [cis dichloro two ammino platinum (II), cisplatin], be that first generation platinum-containing anticancer drug has tangible active anticancer, and by clinical verification, it has significant curative effect to carcinoma of testis and uterus carcinoma, be the choice drug of these two kinds of cancers of treatment, it also can be used for treating bronchogenic carcinoma, laryngocarcinoma, cervical cancer, lymphatic cancer, bladder cancer and osteosarcoma etc.Cisplatin has become countries in the world one of the most effective cancer therapy drug clinically at present.Though the antitumaous effect of cisplatin class medicine is strong, it has toxic and side effects such as nausea and vomiting when having serious nephrotoxicity, neurotoxicity, gastrointestinal toxicity and ototoxicity and use, and these toxic and side effects are restricted cisplatin in process of clinical application.Studies show that cisplatin concentration height in tissue is consistent with its antitumaous effect and toxic and side effects.Therefore research does not reduce the oncotherapy effect again when reducing the normal structure toxic and side effects very significance.
Electrostatic spinning technique is directly to prepare one of most important, effective method of nanofiber.The electrostatic spinning process device is simple, can influence the structure of material by changing preparation condition, reaches the purpose of control drug release amount and speed.Utilize this technology a lot of conventional medicine molecules can be joined in the spinning liquid, rely on the effect of electric field force that medicine is evenly spread in the polymer fiber, thereby avoided the degeneration of active component to lose efficacy effectively.Because solvent volatilization fast in the electrostatic spinning process, medicine can exist in the nanofiber with very little granule.And superfine fibre has very high specific surface area, and it can be controlled drug slow as drug release carrier and discharge, and improves the dissolution velocity of medicine in water.Particularly for the lower medicine of dissolubility in the water, superfine fibre can improve bioavailability of medicament greatly as its release vehicle.At present also not about the organic metal cancer therapy drug being carried out the research report of micro-nanoization with the anti-cancer properties that improves them with electrostatic spinning technique.
Summary of the invention
The objective of the invention is to prepare the method for the organic compound anti-cancer medicine of metal,, improve the bioavailability of medicine and reduce its toxic and side effects to reach the release of control cancer therapy drug with electrostatic spinning technique.
What the present invention proposed prepares the method for organic metal cancer therapy drug with electrostatic spinning technique, and concrete steps are as follows:
(1) solution preparation: preparation is that the quality of solvent is the solution A of 0.2-2.0wt% with the dichloromethane, places in the glass dish and stir 1.5-2.5h under the magnetic stirring apparatus effect, to dissolving fully; Preparation is that the quality of solvent is the solution B of 3-10wt% with the dichloromethane; Place in the glass dish and under the magnetic stirring apparatus effect, stir 1.5-2.5h, to dissolving fully;
(2) spinning process: adopt electrostatic spinning apparatus to carry out spinning, Teflon pipe (Teflon pipe) 1 one ends link to each other with liquid storage pipe 7 by stainless pin 2, and the other end links to each other with nozzle needle 6.Collect screen 5 ground connection, place under the nozzle needle 6.HV generator 4 is connected on the stainless pin 2.Is to be added in the liquid storage pipe 7 behind the 1/5-1/1 mix homogeneously solution A and solution B according to volume ratio, under HV generator applies voltage condition, by nozzle needle 6 ejections, obtains required product from collecting screen 5, and wherein, the temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; Nozzle needle is 14-15 centimetre to the distance of collecting screen; The voltage of HV generator is 15-20KV; The mass velocity of syringe pump is 5-8mL/h.
Wherein, described solution A and solution B are respectively following solution:
1. solution A is the dichloromethane solution of cyclopentadienyl titanium dichloride, and solution B is the dichloromethane solution of Poly-L-lactic acid (PLLA);
Perhaps
2. solution A is the dichloromethane solution of cisplatin, and cisplatin can be used N earlier, and dinethylformamide dissolves fully, solution is joined in the dichloromethane to get final product then), solution B is the dichloromethane solution of Poly-L-lactic acid (PLLA).
Among the present invention, described in the step (1) in the solution A, at dichloromethane is that the mass ratio of cyclopentadienyl titanium dichloride in the solvent is 0.2-2.0wt%, and the electrospinning effect was better when the mass ratio of PLLA was 5wt%, is easy to spin in the short time that area is big, the higher nano fibrous membrane of thickness.
Among the present invention, products obtained therefrom can select for use means such as XRD, SEM, TG that its structure and pattern are characterized.The medicine release in vitro situation of products obtained therefrom can be measured by the following method: cyclopentadienyl titanium dichloride uv-visible absorption spectra method, cisplatin is measured with the ICP method.
Among the present invention, the anti-cancer properties of products obtained therefrom is measured by mtt assay.
Mechanism of the present invention is: after at first solution A and solution B being mixed, several thousand to volt high-pressure electrostatics up to ten thousand on the mixed liquor band, and charged polymer liquid drops under the effect of electric field force and is accelerated at Taylor conical point capillaceous.When electric field force is enough big, mixes drop and overcome surface tension formation injection thread.Thread is solvent evaporation or curing in course of injection, finally drop on and collect on the screen, be transformed into diameter because the quick volatilization of solvent, the liquid of ejection can form the also rapid curing of superfine spraying and reach the following or nano level superfine fibre of micron, shield collection by the collection of ground connection.In this process, drop has certain electrostatic pressure usually and is in the middle of the electric field, therefore, when jet from capillary end when collecting the screen motion, hastening phenomenon appears in the capital, thereby has caused the stretching of jet in electric field, forms the fiber felt of similar non-weaving cloth shape.
The present invention has the following advantages:
1, preparation process is simple, and production cost is lower.
2, because the present invention only need be by conditions such as control medicament contg, voltage, inside and outside nozzle needle the flow velocity just medicament contg, shell thickness, micro/nano-fibre thickness etc. of may command product, therefore further research be expected to realize medicine the location, regularly discharge.Be expected to simultaneously carry enough a kind of new carrier for the oral drug delivery of some drugs.
3, utilize the inventive method can spin that area is big, the higher micro-/ nano iatric fiber film of thickness in the short time.Have that medicine response rate height, product single output are big, product is easy to advantages such as preservations, very valuable concerning commercial Application, and expect to be generalized to large-scale application.
Description of drawings
Fig. 1 is used electric spinning equipment sketch map in the experiment.
Fig. 2 is four kinds of micrometer/nanometer fibers obtaining among the embodiment 1-4 and the SEM photo of pure PLLA.Wherein, (a) being the SEM photo of embodiment 1, (b) is the SEM photo of embodiment 2, (c) is the SEM photo of embodiment 3, (d) for being the SEM photo of embodiment 4.
The micrometer/nanometer fiber stereoscan photograph of Fig. 3 for obtaining among the embodiment.Wherein, (a) being the SEM photo of embodiment 1, (b) is the SEM photo of embodiment 2, (c) is the SEM photo of embodiment 3, (e) is the SEM photo of embodiment 4.
Fig. 4 is raw material cyclopentadienyl titanium dichloride (A), PLLA fiber (B) and be loaded with the XRD figure of the PLLA fiber (C) of cyclopentadienyl titanium dichloride.
Fig. 5 for the vitro drug release curve of the fibrous membrane of electrospinning preparation (37 ℃, pH7.4PBS).Wherein, curve a discharges the performance of cyclopentadienyl titanium dichloride for fibrous membrane under the situation that does not add E.C. 3.4.21.64, curve b discharges the performance of cyclopentadienyl titanium dichloride for the performance of the release of fibrous membrane under the situation that adds 10mg/L E.C. 3.4.21.64 cyclopentadienyl titanium dichloride, curve c for fibrous membrane under the situation that adds the 50mg/L E.C. 3.4.21.64.
Fig. 6 is the experimental result of mtt assay (the cyclopentadienyl titanium dichloride concentration that fibrous membrane discharges is 5.2,10.4 and 15.6mg/L, is expressed as PLLA5.2 respectively, PLLA10.4 and PLLA15.6).
Number in the figure: 1 is the Teflon pipe, and 2 is stainless pin, and 3 is syringe pump, and 4 is HV generator, and 5 for collecting screen, and 6 is nozzle needle, and 7 is liquid storage pipe.
The specific embodiment
The invention is further illustrated by the following examples.
Embodiment 1:
1) accurately measure PLLA (mass ratio of PLLA is 5wt% in dichloromethane solvent) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.After accurately measuring cyclopentadienyl titanium dichloride (the cyclopentadienyl titanium dichloride mass ratio is 0.4wt%) solute and dichloromethane solvent with analytical balance, place and under the magnetic stirring apparatus effect, stir 2.5h in the clean glass dish, obtain solution B to dissolving fully.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 14.0KV; The mass velocity of syringe pump is 6.0mL/h; Collect screen and collect the product fiber.The product that obtains is a micron order, and size range is 1200-1500nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation, result such as Fig. 5 by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties, result such as Fig. 6 with mtt assay.Fig. 5 shows that this system has the release action of controlling cyclopentadienyl titanium dichloride preferably.Fig. 6 shows that the cyclopentadienyl titanium dichloride that this system discharges still has the good anticancer performance.
Embodiment 2:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.After accurately measuring cyclopentadienyl titanium dichloride (the cyclopentadienyl titanium dichloride mass ratio is 0.6wt%) solute and dichloromethane solvent with analytical balance, place in the clean glass dish that stir about 2h obtains solution B to dissolving fully under the magnetic stirring apparatus effect.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 16.0KV; The mass velocity of syringe pump is 6.0mL/h; Collect the screen place and collect the product fiber.The product that obtains is a micron order, and size range is 1300-1750nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.
Embodiment 3:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.After accurately measuring cyclopentadienyl titanium dichloride (the cyclopentadienyl titanium dichloride mass ratio is 0.8wt%) solute and dichloromethane solvent with analytical balance, place in the clean glass dish that stir about 2h obtains solution B to dissolving fully under the magnetic stirring apparatus effect.
2) solution A and solution B are sprayed by nozzle needle after mixing.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 16.0KV; The mass velocity of syringe pump is 6.0mL/h; Collect the screen place and collect the product fiber.The product that obtains is a micron order, and size range is 1800-2250nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.
Embodiment 4:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.After accurately measuring cyclopentadienyl titanium dichloride (the cyclopentadienyl titanium dichloride mass ratio is 1.0wt%) solute and dichloromethane solvent with analytical balance, place in the clean glass dish that stir about 2h obtains solution B to dissolving fully under the magnetic stirring apparatus effect.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 20.0KV; The mass velocity of syringe pump is 6.0mL/h; Collect the screen place and place aluminium foil collection product fiber.The product that obtains is a micron order, and size range is 700-2120nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.
Embodiment 5:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.Accurately measure cisplatin (the cisplatin mass ratio is 0.2wt%) solute with analytical balance, use earlier N, dinethylformamide dissolves fully, then solution is joined in the dichloromethane of certain volume, place stir about 2h dissolving extremely fully under the magnetic stirring apparatus effect in the clean glass dish, obtain solution B.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 16KV; The mass velocity of syringe pump is 6.0mL/h; Collect the screen place and collect the product fiber.The product that obtains is a nanoscale, and size range is 150-400nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.
Embodiment 6:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.Accurately measure cisplatin (the cisplatin mass ratio is 0.2wt%) solute with analytical balance, use earlier N, dinethylformamide dissolves fully, then solution is joined in the dichloromethane of certain volume, place stir about 2h dissolving extremely fully under the magnetic stirring apparatus effect in the clean glass dish, obtain solution B.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 14.0KV, and the mass velocity of syringe pump is 6.0mL/h; Collect the screen place and collect the product fiber.The product that obtains is a nanoscale, and size range is 120-460nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.
Embodiment 7:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.Accurately measure cisplatin (the cisplatin mass ratio is 0.2wt%) solute with analytical balance, use earlier N, dinethylformamide dissolves fully, then solution is joined in the dichloromethane of certain volume, place stir about 2h dissolving extremely fully under the magnetic stirring apparatus effect in the clean glass dish, obtain solution B.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 16KV; The mass velocity of syringe pump is 4.0mL/h; Collect the screen place and collect the product fiber.The product that obtains is a micro/nano level, and size range is 360-2200nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.
Embodiment 8:
1) accurately measure PLLA (mass ratio of PLLA is 5wt%) solute and dichloromethane solvent with analytical balance after, place in the clean glass dish that stir about 2h obtains solution A to dissolving fully under the magnetic stirring apparatus effect.Accurately measure cisplatin (the cisplatin mass ratio is 0.2wt%) solute with analytical balance, use earlier N, dinethylformamide dissolves fully, then solution is joined in the dichloromethane of certain volume, place stir about 2h dissolving extremely fully under the magnetic stirring apparatus effect in the clean glass dish, obtain solution B.
2) will be added in the liquid storage pipe behind solution A and the solution B mix homogeneously, under HV generator applies voltage condition, spray by nozzle needle.The temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; The spacing of collecting screen is 14-15 centimetre.Voltage is 18.4KV; The mass velocity of syringe pump is 6.0mL/h; Collect the screen place and collect the product fiber.The product that obtains is a micro/nano level, and size range is 200-1000nm.
3) products obtained therefrom characterizes its structure and pattern with means such as XRD, TEM, SEM; Measure medicine release in vitro situation by the uv-visible absorption spectra of cyclopentadienyl titanium dichloride; And measure its anti-cancer properties with mtt assay.

Claims (1)

1, a kind ofly prepare the method for organic metal cancer therapy drug, it is characterized in that concrete steps are as follows with electrostatic spinning technique:
(1) solution preparation: preparation is that the quality of solvent is the solution A of 0.2-2.0wt% with the dichloromethane, places in the glass dish and stir 1.5-2.5h under the magnetic stirring apparatus effect, to dissolving fully; Preparation is that the quality of solvent is the solution B of 3-10wt% with the dichloromethane; Place in the glass dish and under the magnetic stirring apparatus effect, stir 1.5-2.5h, to dissolving fully;
(2) spinning process: adopt electrostatic spinning apparatus to carry out spinning, Teflon pipe (1) one end links to each other with liquid storage pipe (7) by stainless pin (2), the other end links to each other with nozzle needle (6), collect screen (5) ground connection, place under the nozzle needle (6), HV generator (4) is connected on the stainless pin (2); Is to be added in the liquid storage pipe (7) behind the 1/5-1/1 mix homogeneously solution A and solution B according to volume ratio, apply under the voltage condition at HV generator (4),, obtain required product from collecting screen (5) by nozzle needle (6) ejection, wherein, the temperature that controls environment is 18-20 ℃; Ambient humidity 45%-50%; Nozzle needle is 14-15 centimetre to the distance of collecting screen; The voltage of HV generator is 15-20KV; The mass velocity of syringe pump is 5-8mL/h;
Wherein, described solution A and solution B are respectively following solution:
3. solution A is the dichloromethane solution of cyclopentadienyl titanium dichloride, and solution B is the dichloromethane solution of Poly-L-lactic acid;
Perhaps
4. solution A is the dichloromethane solution of cisplatin, and solution B is the dichloromethane solution of Poly-L-lactic acid.
CNA2008100418981A 2008-08-20 2008-08-20 Method for preparing organic metal compound anti-cancer medicine with electrostatic spinning technique Pending CN101347403A (en)

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Cited By (2)

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Publication number Priority date Publication date Assignee Title
CN102660889A (en) * 2012-04-13 2012-09-12 东华大学 Fiber-floc suspension and preparation method thereof
RU2491961C2 (en) * 2009-03-10 2013-09-10 Медприн Редженератив Медикал Текнолоджис Ко., Лтд. Artificial dura mater and method of its production

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2491961C2 (en) * 2009-03-10 2013-09-10 Медприн Редженератив Медикал Текнолоджис Ко., Лтд. Artificial dura mater and method of its production
US8795708B2 (en) 2009-03-10 2014-08-05 Medprin Regenerative Medical Technologies Co., Ltd. Treating defective dura with synthetic artificial dura substitute
US9211180B2 (en) 2009-03-10 2015-12-15 Medprin Regenerative Medical Technologies Co., Ltd. Method for treating defective dura mater
US9271822B2 (en) 2009-03-10 2016-03-01 Medprin Regenerative Medical Technologies Co., Ltd. Artificial dura mater and manufacturing method thereof
CN102660889A (en) * 2012-04-13 2012-09-12 东华大学 Fiber-floc suspension and preparation method thereof

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