CN105268027A - Bone tissue engineering stent and preparation method thereof - Google Patents
Bone tissue engineering stent and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a bone tissue engineering stent and a preparation method thereof. The bone tissue engineering stent is prepared from PLGA, isoniazide, rifampicin, allogeneic bone power and bone morphogenetic protein at the weight ratio of (87-51) to (1-6) to (1-6) to (10-30) to (1-7); according to the bone tissue engineering stent, polylactic acid-hydroxyacetic acid material, allogeneic bone power, antituberculosis drugs and bone morphogenetic protein are combined by adopting a 3D printing technology, a bioactive stent is manufactured by adopting the 3D printing technology, the physical, chemical and biological properties of the stent are detected, and the bone tissue engineering stent has the good biocompatibility and the good bone-formation promotion capacity and is biodegradable.
Description
Technical field
The present invention relates to a kind of medical material and preparation method thereof, especially a kind of bone tissue engineering scaffold and preparation method thereof.
Background technology
Bone tuberculosis is the outer tuberculosis of common Secondary cases lung, and account for all lungy about 3%, wherein about half involves spinal column, can cause patient's lifelong disability as delayed the diagnosis and treatment, even dead.Spinal tuberculosis as the most common form of bone tuberculosis, in recent years also along with increase lungy is in rising trend.Tubercule bacillus is to medicaments insensitives such as rifampicin (RFI) isoniazids (INH).The medicines such as Rimactazid can penetrate in host cell, all have kill and (or) inhibitory action the tubercule bacillus of intraor extracellular.But long-term orally in a large number often cause multiple untoward reaction, even cause the infringement of the organ such as liver, kidney.How to solve the recurrence of the Cranial defect of the postoperative bulk of focal cleaning, local tuberculose focus abscess, and how to shorten the course for the treatment of of antituberculotics, under the prerequisite not damaging other organs, the drug level that tuberculose focus position can also be made to remain valid is a problem demanding prompt solution.The alternative of administering mode is the breach solved the problem.Bone tissue engineer is the new way of clinical treatment Cranial defect and bone does not connect.Bone tissue engineer relates to timbering material and somatomedin, and finding a kind ofly has good biocompatibility, has and promote osteogenic ability preferably and can biodegradable timbering material be focus and the difficult point of research, is also one of key element of bone tissue engineer.Bone morphogenetic protein (BMP) is the most important skeletal growth factor of generally acknowledged osteogenic activity, and BMP can break up to one-tenth cartilage, osteoblast directional proliferation by inducing bone mesenchymal stem cell, thus promotes Bone Defect Repari.
The new material forming preparation process being representative with 3D printing technique makes the 3D support preparing various form more convenient.Compare other 3D printing techniques, in 3D biodegradable polymer deposition technique, forming materials process does not need the environmental condition such as high temperature, laser, less to the damage of contained medicine and somatomedin, thus can be used to the biodegradation high molecular support preparing combination drug and somatomedin.
Against the background of the prior art, adopt 3D Method of printing to make the resistive connection nuclearity bone tissue engineering scaffold with slow release pastille and osteoinductive, be expected to for the operative treatment of bone tuberculosis provides new thinking.
Summary of the invention
Technical problem to be solved by this invention is to provide bone tissue engineering scaffold.
Another technical problem to be solved by this invention is the preparation method providing above-mentioned bone tissue engineering scaffold, adopts 3D solution printing technique to build bone tissue engineering scaffold.
For solving the problems of the technologies described above, technical scheme of the present invention is:
A kind of bone tissue engineering scaffold, be made up of PLGA (polyglycolic-polylactic acid), isoniazid (INH), rifampicin (RFP), allogeneic bone meal and bone morphogenetic protein (BMP-2), wherein, the weight ratio of described PLGA, isoniazid, rifampicin, allogeneic bone meal and bone morphogenetic protein is followed successively by 87-51:1-6:1-6:10-30:1-7.
Preferably, above-mentioned bone tissue engineering scaffold, in described PLGA, the percentage by weight of lactic acid and hydroxyacetic acid is 15%-85%:85%-15%, molecular mass 50,000-12 ten thousand D.
Preferably, above-mentioned bone tissue engineering scaffold, the granularity of described allogeneic bone meal is 1-10 μm.
Preferably, above-mentioned bone tissue engineering scaffold, the granularity of described allogeneic bone meal is 5 μm.
The preparation method of above-mentioned bone tissue engineering scaffold, according to three-dimensional computed tomography technology (computedtomography, CT) to the three-dimensional reconstruction of Cranial defect position data, by the accurately successively 3D accumulation of material, prepare slow release degraded pastille and skeletal growth factor support, concrete steps are as follows:
(1) be dissolved in dichloromethane (DCM) and acetone by formula ratio by PLGA, wherein the volume ratio of dichloromethane and acetone is 1:1, is made into 10%-25%PLGA solution;
(2) by formula ratio allogeneic bone meal, under 20 DEG C of conditions, progressively join on a small quantity in step (1) in PLGA solution, sealing magnetic stirs 3 hours, until form homogeneous mixture solotion;
(3) join in above-mentioned mixed solution respectively by formula ratio isoniazid, rifampicin, bone morphogenetic protein, sealing ultrasonic agitation makes spray solution in 1 hour;
(4) spray solution is transferred in injection needle, and be placed in 4 DEG C of pre-coolings 30 minutes, be convenient to print to obtain suitable viscosity;
(5) pneumatic 3D printer is placed in super-clean bench also with ultra violet lamp sterilizing in 30 minutes, the injection needle that printing liquid is housed of pre-cooling is arranged on Support for impression mechanism and runs print routine, wherein, print needle diameter 100 microns, print speed 5 mm/second is set, material silk spacing is 120 microns, and the air pressure for extruded material silk is 0.3MPa; Print in the culture dish of receiving platform by material silk, by the accurately successively 3D accumulation of material, make slow release degraded pastille and skeletal growth factor support, after lyophilization, 4 DEG C save backup.
Prepare the resistive connection nuclearity bone tissue engineer bioactive bracket with slow release pastille and osteoinductive thus.
The invention has the beneficial effects as follows:
Described bone tissue engineering scaffold, adopt 3D printing technique, poly lactic-co-glycolic acid material, allogeneic bone meal, antituberculotics and bone morphogenetic protein (BMP) etc. are combined, utilize 3D printing technique to manufacture bioactive bracket, and detection carried out to the physical chemistry of this support and biological property there is good biocompatibility, have and promote osteogenic ability preferably and can biodegradation.
Confirm by measuring, in bone tissue engineering scaffold of the present invention, PLGA has good biocompatibility, degradable absorbability, PLGA as Biodegradable material in biomedical applications in occupation of critical role, through being elected to be bone alternate material in a large number; Do not reach orthopaedics internal fixtion requirement problem to solve PLGA mechanics of materials intensity, the allogeneic bone meal of specified quantitative be filled in PLGA material, with strengthen PLGA material mechanical strength, more easily absorb by freshman bone tissue and good biocompatibility.
Described bone tissue engineering scaffold contains the BMP of bone inductive effect and has steady release, and osteocyte inducing action is obvious, has and well promotes knitting effect.In bone support, add antituberculotics isoniazid, rifampicin makes lesions position long-term sustained release high concentration medicine, systemic drug consumption can be reduced, shorten medication times, suppress the generation of tulase.The advantage of allogeneic bone meal is used to be, homogeneous allogenic bone bone meal is through physics and chemistry process, defat deproteinization, its antigenicity is eliminated, do not cause obvious immunological rejection after implant into body, and residual collagen component contributes to the combination with BMP, have and good organize affinity and adhesion, can bone conduction effect be played, direct bone can be formed with osseous tissue and be connected; Meanwhile, be rich in the necessary aminoacid sequence of host material, and have the structural arrangement with host's skeleton homology, host tissue cells is more accessible and give absorbing and eliminating.This kind of material source enriches, easy to make, and at its function adaptability, organize and melt mutually, be better than synthetic material in physicochemical property, biological degradability and cost; Along with the degraded of PLGA and disengaging of medicine define more cavity on three-dimensional porous structure support after in stent implant, the expression of osteoblast differentiation and promotion osteoblast under BMP-2 induction, accelerate knitting, the degradable tubercular drugs of final PLGA all discharges, and the area of new bone cell bone meal adsorbed completely in support forms new bone.
Preparation method utilizes three-dimensional computed tomography technology (computedtomography, CT) to the three-dimensional reconstruction of Cranial defect position data, before surgery bone support is printed, avoid the misery of second operation, bone tissue engineering scaffold prepared by this 3D printing technique has unique advantage in support personalization, accuracy, mechanical strength, hole adjustment, space structure complexity.
Accompanying drawing explanation
Fig. 1 is bone tissue engineering scaffold sample drawing;
Fig. 2 is PLGABMP-2INHRFP homogeneous allogenic bone support bone support scanning electron microscopic picture;
Fig. 3 is isoniazid (INH), rifampicin (RFP) releasing curve diagram;
Fig. 4 is BMP-2 cumulative release curve chart.
Detailed description of the invention
In order to make those skilled in the art better understand technical scheme of the present invention, below in conjunction with detailed description of the invention, technical scheme of the present invention is described in further detail.
Embodiment 1
(1) PLGA10g (25:75, w/w) PLGA is dissolved in 100ml dichloromethane and acetone mixed solution, wherein dichloromethane: acetone (v/v)=1:1, is made into 10%PLGA solution.
(2) by 15g5 μm of allogeneic bone meal, under 20 DEG C of conditions, progressively join on a small quantity in 10%PLGA solution, sealing magnetic stirs 3 hours, until form homogeneous mixture solotion.
(3) isoniazid (INH) 3g, rifampicin (RFP) 1g, bone morphogenetic protein (BMP-2) 1g join in above-mentioned mixed solution respectively, and sealing ultrasonic agitation makes spray solution in 1 hour.
(4) in order to obtain suitable viscosity to print, this solution is transferred in injection needle, and be placed in 4 DEG C of pre-coolings 30 minutes.
(5) pneumatic 3D printer is placed in super-clean bench also with ultra violet lamp sterilizing in 30 minutes.The injection needle printing liquid that is equipped with of pre-cooling is arranged on Support for impression mechanism, runs print routine according to code compiled in advance.Print needle diameter 100 microns, arrange print speed 4 mm/second, material silk spacing is 150 microns, and the air pressure for extruded material silk is 0.3MPa.Material silk is printed in the culture dish of receiving platform, by the accurately successively 3D accumulation of material, makes slow release degraded pastille and skeletal growth factor support.After lyophilization, 4 DEG C save backup.Prepare the resistive connection nuclearity bone tissue engineering scaffold with slow release pastille and osteoinductive thus.
Embodiment 2
(1) PLGA is dissolved in dichloromethane (DCM) and acetone, is made into 10%-25% solution.Such as, 30g (85:15, w/w) PLGA is dissolved in 200ml dichloromethane and acetone mixed solution, wherein dichloromethane: acetone (v/v)=1:1, is made into 15%PLGA solution.
(2) by 15g7 μm of allogeneic bone meal, under 20 DEG C of conditions, progressively join on a small quantity in 15%PLGA solution, sealing magnetic stirs 3 hours, until form homogeneous mixture solotion.
(3) isoniazid (INH) 4g, rifampicin (RFP) 3g, bone morphogenetic protein (BMP-2) 2g join in above-mentioned mixed solution respectively, and sealing ultrasonic agitation makes spray solution in 1 hour.
(4) in order to obtain suitable viscosity to print, this solution is transferred in injection needle, and be placed in 4 DEG C of pre-coolings 30 minutes.
(5) pneumatic 3D printer is placed in super-clean bench also with ultra violet lamp sterilizing in 30 minutes.The injection needle printing liquid that is equipped with of pre-cooling is arranged on Support for impression mechanism, runs print routine according to code compiled in advance.Print needle diameter 100 microns, arrange print speed 5 mm/second, material silk spacing is 120 microns, and the air pressure for extruded material silk is 0.3MPa.Material silk is printed in the culture dish of receiving platform, by the accurately successively 3D accumulation of material, makes slow release degraded pastille and skeletal growth factor support.After lyophilization, 4 DEG C save backup.Prepare the resistive connection nuclearity bone tissue engineering scaffold with slow release pastille and osteoinductive thus.
Embodiment 3
(1) PLGA is dissolved in dichloromethane (DCM) and acetone, is made into 10%-25% solution.Such as, 30g (50:50, w/w) PLGA is dissolved in 300ml dichloromethane and acetone mixed solution, wherein dichloromethane: acetone (v/v)=1:1, is made into 10%PLGA solution.
(2) by 30g7 μm of allogeneic bone meal, under 20 DEG C of conditions, progressively join on a small quantity in 10%PLGA solution, sealing magnetic stirs 3 hours, until form homogeneous mixture solotion.
(3) isoniazid (INH) 2g, rifampicin (RFP) 4g, bone morphogenetic protein (BMP-2) 2g join in above-mentioned mixed solution respectively, and sealing ultrasonic agitation makes spray solution in 1 hour.
(4) in order to obtain suitable viscosity to print, this solution is transferred in injection needle, and be placed in 4 DEG C of pre-coolings 30 minutes.
(5) pneumatic 3D printer is placed in super-clean bench also with ultra violet lamp sterilizing in 30 minutes.The injection needle printing liquid that is equipped with of pre-cooling is arranged on Support for impression mechanism, runs print routine according to code compiled in advance.Print needle diameter 100 microns, arrange print speed 5 mm/second, material silk spacing is 120 microns, and the air pressure for extruded material silk is 0.3MPa.Material silk is printed in the culture dish of receiving platform, by the accurately successively 3D accumulation of material, makes slow release degraded pastille and skeletal growth factor support.After lyophilization, 4 DEG C save backup.Prepare the resistive connection nuclearity bone tissue engineering scaffold with slow release pastille and osteoinductive thus.
Embodiment 4
(1) PLGA is dissolved in dichloromethane (DCM) and acetone, is made into 10%-25% solution.Such as, 25g (50:50, w/w) PLGA is dissolved in 100ml dichloromethane and acetone mixed solution, wherein dichloromethane: acetone (v/v)=1:1, is made into 25%PLGA solution.
(2) by 18g5 μm of allogeneic bone meal, under 20 DEG C of conditions, progressively join on a small quantity in 25%PLGA solution, sealing magnetic stirs 3 hours, until form homogeneous mixture solotion.
(3) isoniazid (INH) 4g, rifampicin (RFP) 5g, bone morphogenetic protein (BMP-2) 3g join in above-mentioned mixed solution respectively, and sealing ultrasonic agitation makes spray solution in 1 hour.
(4) in order to obtain suitable viscosity to print, this solution is transferred in injection needle, and be placed in 4 DEG C of pre-coolings 30 minutes.
(5) pneumatic 3D printer is placed in super-clean bench also with ultra violet lamp sterilizing in 30 minutes.The injection needle printing liquid that is equipped with of pre-cooling is arranged on Support for impression mechanism, runs print routine according to code compiled in advance.Print needle diameter 100 microns, arrange print speed 5 mm/second, material silk spacing is 120 microns, and the air pressure for extruded material silk is 0.3MPa.Material silk is printed in the culture dish of receiving platform, by the accurately successively 3D accumulation of material, makes slow release degraded pastille and skeletal growth factor support.After lyophilization, 4 DEG C save backup.Prepare the resistive connection nuclearity bone tissue engineering scaffold with slow release pastille and osteoinductive thus.
Test case:
Carry out analysis to embodiment 1 gained bone tissue engineering scaffold to measure:
(1) surface observation and scanning electron microscopic observation are carried out to prepared bone tissue engineering scaffold.Support is cut open, sightingpiston carries out electron-microscope scanning after metal spraying, under the accelerating potential of 15kV, observe surface and cross-section structure.The results are shown in Figure 1, Fig. 2.
(2) high performance liquid chromatograph is adopted to carry out isoniazid (INH), the research of rifampicin (RFP) release mensuration to prepared bone tissue engineering scaffold.Measure isoniazid (INH) standard curve and rifampicin (RFP) standard curve.
Be soaked in by bone tissue engineering scaffold in 100mlPBS solution, sealing is placed in 37 DEG C of shaking baths.Get 100 μ l leachates at 1d, 5d, 10d, 15d, 20d, 25d, 30d, 35d, 40d, 45d, 50d, 55d, 60d, 65d, 70d, 75d, 80d, 85d, 90d time point respectively, supplement the fresh PBS solution of lOO μ l to keep soak constancy of volume simultaneously.Drug release concentration determination is carried out according to isoniazid (INH) standard curve determination method and rifampicin (RFP) standard curve determination method.Its drug release patterns is shown in Fig. 3.
(3) carry out bone morphogenetic protein (BMP-2) release profiles to prepared bone tissue engineering scaffold to measure
Get one piece of bone tissue engineering scaffold, invaded in the PBS solution of 50ml, 37 DEG C of incubations, respectively at 1d, 5d, 10d, 15d, 20d, 25d, 30d, 35d, 40d, 45d, 50d, 55d, 60d, 65d, 70d, 75d, 80d, 85d, 90d sample lml, after taking sample, supplement the PBS solution of same volume, application ELISA method detects the concentration of BMP-2 at every turn, calculates the percent of accumulative release BMP-2.Get standard substance dilution be 31.25,62.5,125,250,500,1000,2000Pg/ml, measure OD value, with On value for abscissa, C is vertical coordinate, carry out standard straight-line matching with CurveExpertl.3 software, try to achieve BMP-2 concentration, be multiplied by the concentration obtaining actual BMP-2 after 10.BMP-2 release profiles is shown in Fig. 4.
(4) biocompatibility mensuration is carried out to prepared bone tissue engineering scaffold
Tetrazolium salts (MTT) colorimetry is adopted to carry out evaluation of its biocompatibility to PLGA bioactive bracket extract.The object of this test is carried out in vitro contacting cultivating at support sample lixiviating solution and cell, by the observation to cellular morphology, propagation and inhibitory effect, and the vitro cytotoxicity effect of evaluation test sample.Cell strain: NCTCclone929 (l cell L-929), derives from National Institute for Food and Drugs Control.
Blank: through the MEM cell culture fluid (lixiviate medium) of 0.22m frit.
Negative control: surface area is that the high density polyethylene (HDPE) (production of Jiangmen city Xinhui District Hao Tian Trade Co., Ltd.) of 30cm2 adds lixiviate medium 5.0ml, puts 37 DEG C, lixiviate 24h.
Positive control: containing the lixiviate medium of 5% dimethyl sulfoxide (the special Chemical Company in Rui Jin, Tianjin, lot number: on October 26th, 2013).
Mammalian cell In vitro culture is used for the existing very long history of Cytotoxic evaluation of biomaterial and medical apparatus and instruments, and in ISO10993-5:2009, the cell of recommendation is l cell (NCTCclone929).This kind of cell injuring model technology is selected in this test, support sample lixiviating solution and cell are carried out contacting cultivating in vitro, using cell culture fluid (lixiviate medium) as blank, the high density polyethylene (HDPE) lixiviating solution of known non-toxic as negative control, containing the cell culture fluid of 5% dimethyl sulfoxide as positive control.By to the observation of cellular morphology, propagation and inhibitory effect and mensuration, the toxic action after detecting test sample exposing cell, cell in vitro produced.The method can be predicted there is reasonability.
Prepared by cell suspension: by the L-929 cell of normal Secondary Culture, through 0.25% pancreatin (SCIENTIFICRESEARCHSPECIAL, lot number: 27250018) cell suspension that concentration is 1 × 104/ml is made in digestion, be inoculated in 96 orifice plates, every hole 200l, often organize 6 holes, put 37 DEG C of cultivation 24h in 5%CO2 incubator.
Lixiviating solution exchanges: after cell culture 24h, discard original fluid, blank group uses lixiviate Medium Exchange, negative control group lixiviate ratio is that 6cm2/ml high density polyethylene (HDPE) lixiviating solution exchanges, the lixiviate Medium Exchange of positive controls containing 5% dimethyl sulfoxide, test specimen group lixiviating solution exchanges.To put in 5%CO2 incubator 37 DEG C, cultivate 72h.
The mensuration of absorbance: after changing culture fluid, get the orifice plate cultivating 72h, every hole adds 40l5g/L tetrazolium salts (MTT) (SCIENTIFICRESEARCHSPECIAL, effect duration: 062014) solution discards liquid in hole after continuing to cultivate 4h, add 150L dimethyl sulfoxide, put 10min that agitator vibrates, measure each hole absorbance at microplate reader (Lei Bo company of Finland, MK3) 570nm and 630nm place.Be calculated as follows the relative rate of increase (RGR).
A-sample sets (negative, positive group) absorbance, A0-blank group absorbance
Cell-cytotoxic reaction stage division is in table 1.
Table 1
| Rank | Relative rate of increase % |
| 0 grade | ≥100 |
| 1 grade | 80～99 |
| 2 grades | 50～79 |
| 3 grades | 30～49 |
| 4 grades | 0～29 |
Judge according to cell-cytotoxic reaction grade scale, the reaction of negative control group should be not more than 1 grade, and positive controls is at least 3 grades.
The biocompatibility of bone tissue engineering scaffold is evaluated by the cell toxicity test of support.72h under this experimental condition: negative control group RGR is 99.1%, and cell-cytotoxic reaction is 1 grade; Positive controls RGR is 4.0%, and cell-cytotoxic reaction is 4 grades.Pilot system reaction is set up.The relative rate of increase RGR of lixiviating solution cell is 90.3%, and cell-cytotoxic reaction is 1 grade.
Above-mentioned detailed description of this bone tissue engineering scaffold and preparation method thereof being carried out with reference to detailed description of the invention; illustrative instead of determinate; several embodiments can be listed according to institute's limited range; therefore in the change do not departed under general plotting of the present invention and amendment, should belong within protection scope of the present invention.
Claims (5)
1. a bone tissue engineering scaffold, it is characterized in that: be made up of PLGA, isoniazid, rifampicin, allogeneic bone meal and bone morphogenetic protein, wherein, the weight ratio of described PLGA, isoniazid, rifampicin, allogeneic bone meal and bone morphogenetic protein is followed successively by 87-51:1-6:1-6:10-30:1-7.
2. bone tissue engineering scaffold according to claim 1, is characterized in that: in described PLGA, the percentage by weight of lactic acid and hydroxyacetic acid is 15%-85%:85%-15%, molecular mass 50,000-12 ten thousand D.
3. bone tissue engineering scaffold according to claim 1, is characterized in that: the granularity of described allogeneic bone meal is 1-10 μm.
4. bone tissue engineering scaffold according to claim 3, is characterized in that: the granularity of described allogeneic bone meal is 5 μm.
5. the preparation method of bone tissue engineering scaffold described in claim 1, it is characterized in that: according to the three-dimensional reconstruction of three-dimensional computed tomography technology to Cranial defect position data, by the accurately successively 3D accumulation of material, prepare slow release degraded pastille and skeletal growth factor support, concrete steps are as follows:
(1) be dissolved in dichloromethane and acetone by formula ratio by PLGA, wherein the volume ratio of dichloromethane and acetone is 1:1, is made into 10%-25%PLGA solution;
(2) by formula ratio allogeneic bone meal, under 20 DEG C of conditions, progressively join on a small quantity in step (1) in PLGA solution, sealing magnetic stirs 3 hours, until form homogeneous mixture solotion;
(3) join in above-mentioned mixed solution respectively by formula ratio isoniazid, rifampicin, bone morphogenetic protein, sealing ultrasonic agitation makes spray solution in 1 hour;
(4) spray solution is transferred in injection needle, and be placed in 4 DEG C of pre-coolings 30 minutes, be convenient to print to obtain suitable viscosity;
(5) pneumatic 3D printer is placed in super-clean bench also with ultra violet lamp sterilizing in 30 minutes, the injection needle that printing liquid is housed of pre-cooling is arranged on Support for impression mechanism and runs print routine, wherein, print needle diameter 100 microns, print speed 5 mm/second is set, material silk spacing is 120 microns, and the air pressure for extruded material silk is 0.3MPa; Print in the culture dish of receiving platform by material silk, by the accurately successively 3D accumulation of material, make slow release degraded pastille and skeletal growth factor support, after lyophilization, 4 DEG C save backup.
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| CN106474555A (en) * | 2016-11-04 | 2017-03-08 | 暨南大学 | A kind of polycaprolactone of 3D printing/oyster shell powder composite and preparation and application and the bone tissue engineering scaffold based on which |
| CN106474555B (en) * | 2016-11-04 | 2019-09-17 | 暨南大学 | A kind of polycaprolactone of 3D printing/oyster shell powder composite material and preparation and application and the bone tissue engineering scaffold based on it |
| CN107469153A (en) * | 2017-09-20 | 2017-12-15 | 华南农业大学 | Degradable composite bone repairing material and preparation method thereof and the application in 3D printing |
| CN109010911A (en) * | 2018-09-03 | 2018-12-18 | 南京市第医院 | A kind of anti-bone tuberculosis bone tissue engineering scaffold of 3D printing PCL- rifampicin isoniazid streptomysin and preparation method and application |
| CN110180033A (en) * | 2019-05-16 | 2019-08-30 | 深圳市第二人民医院 | Carry the three-dimensional porous rack and preparation method thereof of neferine |
| CN114569539A (en) * | 2022-01-24 | 2022-06-03 | 赵振群 | Novel antibiotic release system, preparation method and application thereof |
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