CN115970069A - Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating - Google Patents
Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating Download PDFInfo
- Publication number
- CN115970069A CN115970069A CN202211596392.3A CN202211596392A CN115970069A CN 115970069 A CN115970069 A CN 115970069A CN 202211596392 A CN202211596392 A CN 202211596392A CN 115970069 A CN115970069 A CN 115970069A
- Authority
- CN
- China
- Prior art keywords
- pirfenidone
- ureteral stent
- preparation
- stent tube
- nanoparticles
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 98
- ISWRGOKTTBVCFA-UHFFFAOYSA-N pirfenidone Chemical compound C1=C(C)C=CC(=O)N1C1=CC=CC=C1 ISWRGOKTTBVCFA-UHFFFAOYSA-N 0.000 title claims abstract description 58
- 229960003073 pirfenidone Drugs 0.000 title claims abstract description 56
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 239000011248 coating agent Substances 0.000 title claims abstract description 27
- 238000000576 coating method Methods 0.000 title claims abstract description 27
- 239000002131 composite material Substances 0.000 title claims abstract description 24
- 239000000725 suspension Substances 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000008367 deionised water Substances 0.000 claims abstract description 19
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 19
- 238000003756 stirring Methods 0.000 claims abstract description 16
- 238000005406 washing Methods 0.000 claims abstract description 13
- CTENFNNZBMHDDG-UHFFFAOYSA-N Dopamine hydrochloride Chemical compound Cl.NCCC1=CC=C(O)C(O)=C1 CTENFNNZBMHDDG-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229960001149 dopamine hydrochloride Drugs 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 9
- 238000002791 soaking Methods 0.000 claims abstract description 8
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 7
- 238000011534 incubation Methods 0.000 claims abstract description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 24
- 229940043267 rhodamine b Drugs 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 14
- 239000000839 emulsion Substances 0.000 claims description 11
- 239000002953 phosphate buffered saline Substances 0.000 claims description 8
- 230000001804 emulsifying effect Effects 0.000 claims description 7
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 6
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 6
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 6
- 238000009210 therapy by ultrasound Methods 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 5
- 238000002372 labelling Methods 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 2
- 238000004945 emulsification Methods 0.000 claims 1
- 238000001727 in vivo Methods 0.000 claims 1
- 229920001690 polydopamine Polymers 0.000 claims 1
- 208000031481 Pathologic Constriction Diseases 0.000 abstract description 19
- 230000036262 stenosis Effects 0.000 abstract description 19
- 208000037804 stenosis Diseases 0.000 abstract description 19
- 102000008186 Collagen Human genes 0.000 abstract description 6
- 108010035532 Collagen Proteins 0.000 abstract description 6
- 229920001436 collagen Polymers 0.000 abstract description 6
- 230000008021 deposition Effects 0.000 abstract description 5
- 230000000642 iatrogenic effect Effects 0.000 abstract description 4
- 229940099456 transforming growth factor beta 1 Drugs 0.000 abstract description 4
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 abstract description 2
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 abstract description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 11
- 210000000626 ureter Anatomy 0.000 description 10
- 206010046405 Ureteric injury Diseases 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000009297 electrocoagulation Methods 0.000 description 5
- 230000006378 damage Effects 0.000 description 4
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 210000004379 membrane Anatomy 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- 239000006180 TBST buffer Substances 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000001356 surgical procedure Methods 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 2
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 206010016654 Fibrosis Diseases 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000002033 PVDF binder Substances 0.000 description 2
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 2
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- 239000012670 alkaline solution Substances 0.000 description 2
- 230000003367 anti-collagen effect Effects 0.000 description 2
- 230000000417 anti-transforming effect Effects 0.000 description 2
- 238000006065 biodegradation reaction Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000013270 controlled release Methods 0.000 description 2
- 229960003638 dopamine Drugs 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 230000004761 fibrosis Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 210000000713 mesentery Anatomy 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- 208000009911 Urinary Calculi Diseases 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000002399 angioplasty Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 229940096422 collagen type i Drugs 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 210000001731 descending colon Anatomy 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000916 dilatatory effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000010102 embolization Effects 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 102000034240 fibrous proteins Human genes 0.000 description 1
- 108091005899 fibrous proteins Proteins 0.000 description 1
- 238000002695 general anesthesia Methods 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 238000012151 immunohistochemical method Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000003331 infrared imaging Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 238000011587 new zealand white rabbit Methods 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 230000036573 scar formation Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000004304 subcutaneous tissue Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000011870 unpaired t-test Methods 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method of a ureteral stent tube carrying a pirfenidone nanoparticle composite coating, which comprises the following steps: s1, firstly, preparing a suspension loaded with pirfenidone nanoparticles in advance; s2, then, putting the ureteral stent tube into a phosphate buffer solution containing dopamine hydrochloride for incubation; s3, stirring for a certain time at room temperature to form a polydopamine-modified ureteral stent; and S4, washing the polydopamine-modified ureteral stent tube obtained in the step S3 with deionized water twice, and then soaking the stent in the suspension of the pirfenidone nanoparticles. The invention can inhibit ureteral stenosis by reducing the expression of transforming growth factor beta 1 and the deposition of collagen by coating the pirfenidone nano particle composite coating on the ureteral stent tube, therefore, the ureteral stent using the nano particle/PFD composite coating is considered to be an effective method for preventing ureteral stenosis caused by iatrogenic operation.
Description
Technical Field
The invention relates to the technical field of preparation of ureteral stent tubes, in particular to a preparation method of a ureteral stent tube carrying a pirfenidone nanoparticle composite coating.
Background
In recent years, with the continuous application of ureteroscopy technology, the ureteral injury is increased. The reported probability of ureter drug damage at home and abroad is 60% -94%. In particular, the incidence of ureteral stenosis due to iatrogenic procedures is increasing as ureteroscopy is performed on upper urinary tract stones. Therefore, how to prevent ureteral stenosis caused by iatrogenic operation, especially ureteral stenosis after ureteroscopy operation, is a problem which needs to be solved urgently by urinary surgeons today.
Ureteral injury repair is generally fibrous scar repair, and is pathologically characterized by massive proliferation of fibroblasts and excessive deposition of proteoglycans in collagen and extracellular matrix, resulting in disorganization of collagen fibers. The research shows that TGF-beta 1 is highly expressed in fibroblasts and is a cytokine which is most closely related to scar formation. In this study, we attempted to further explore the mechanism of ureteral stenosis by using animal ureteral injury models to simulate ureteral injury from ureteroscopy procedures. At present, main ureteral injury animal models at home and abroad comprise a unilateral ureteral complete obstruction model and an electrocoagulation injury model. Therefore, we tried to further investigate the pathogenesis of ureteral stenosis by using a rabbit ureteral injury model closer to ureteral stenosis by clinical ureteroscopic electrocoagulation.
There are many methods for treating ureteral stenosis, including endoscopic, balloon-dilation, laparoscopic or open surgical angioplasty, etc. After surgery, ureteral stent tubes are typically placed within the ureter. Although the ureteral stent tube has a certain effect in dilating the ureter and preventing stenosis, there are some patients who have a mild stenosis again after removing the ureteral stent tube. Therefore, there is an urgent clinical need to develop more effective stent tubes to prevent ureteral stenosis. In view of the above defects, it is actually necessary to design a preparation method of the pirfenidone-loaded nanoparticle composite coating ureteral stent tube.
Disclosure of Invention
The invention aims to provide a preparation method of a ureteral stent tube carrying a pirfenidone nanoparticle composite coating, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: the preparation method of the ureteral stent tube carrying the pirfenidone nano particle composite coating comprises the following steps:
s1, firstly, preparing a suspension loaded with pirfenidone nanoparticles in advance;
s2, then, putting the ureteral stent tube into a phosphate buffer solution containing dopamine hydrochloride for incubation;
s3, stirring for a certain time at room temperature to form a polydopamine-modified ureteral stent;
s4, washing the polydopamine-modified ureteral stent tube obtained in the step S3 with deionized water twice, and then soaking the stent in a suspension of pirfenidone nanoparticles to form a ureteral stent tube loaded with a pirfenidone nanoparticle coating;
s5, observing the surface morphology of the ureteral stent tube wrapped with the nanoparticles by using a scanning electron microscope;
and S6, finally, labeling the nanoparticle complex by rhodamine B, and detecting the distribution of the pirfenidone nanoparticles on the ureteral stent tube.
Preferably, in step S1, the preparation of the pirfenidone nanoparticle-loaded suspension includes the following steps:
s11, firstly, emulsifying phosphate buffered saline, pirfenidone and polylactic acid-glycolic acid in dichloromethane, and performing ultrasonic treatment for 1min by using an ultrasonic homogenizer to obtain a first mixture;
s12, adding a polyvinyl alcohol solution into the mixture I obtained in the step S11, and performing ultrasonic treatment to form an emulsion;
s13, stirring the emulsion obtained in the step S12 at room temperature for 24 hours to completely evaporate dichloromethane to obtain nano particles;
s14, centrifuging the nanoparticles obtained in the step S13 at the speed of 12,000 revolutions per minute of less than 4 ℃ for 5min;
and S15, finally, washing the nanoparticles for multiple times by using deionized water, and suspending the nanoparticles in the deionized water to obtain a suspension loaded with the pirfenidone nanoparticles.
Preferably, in step S6, the preparation of the nanoparticle complex labeled with rhodamine B comprises the following steps:
s21, firstly, preparing a suspension containing rhodamine B labeled nanoparticles in advance;
s22, then, incubating the ureteral stent tube obtained in the step S4 in a phosphate buffer solution containing 0.5mg/ml dopamine hydrochloride, and then stirring for 3 hours at room temperature to form a polydopamine-modified ureteral stent tube;
s23, washing the polydopamine-modified ureteral stent tube twice with deionized water, and then soaking the polydopamine-modified ureteral stent tube in the rhodamine B-labeled nanoparticle suspension prepared in step S21 to form the rhodamine B nanoparticle/pirfenidone-coated ureteral stent tube.
Preferably, in step S21, the preparation of the rhodamine B labeled nanoparticle suspension comprises the following steps:
s211, firstly, emulsifying 100ul of phosphate buffered saline and rhodamine B in 2ml of dichloromethane containing 20mg of PLGA, adding the mixture, and carrying out ultrasonic treatment in an ice bath for 0.5min to obtain a second mixture;
s212, then, 4.5ml of 1.5% PVA was added to the second mixture and sonicated to form a multiple emulsion;
s213, next, the emulsion is stirred at room temperature for 24h to completely evaporate the dichloromethane, and the nanoparticles are centrifuged at a speed of 12,000 rpm, which is lower than 4 ℃, for 5min;
and S214, finally, washing the nano particles with deionized water for three times, and finally suspending in the deionized water to obtain the rhodamine B marked nano particle suspension.
Preferably, in step S2, the concentration of dopamine hydrochloride is 0.5mg/ml, and the pH value of the phosphate buffer is 8.5.
Preferably, in step S3, the stirring time period for stirring is 3 hours.
Compared with the prior art, the preparation method of the ureteral stent tube loaded with the pirfenidone nanoparticle composite coating can be used for soaking a clinically common ureteral stent tube in an alkaline solution containing dopamine to form an adhesive coating on the surface of the stent, and then incubating the dopamine-modified stent tube and the nanoparticle/pirfenidone composite to form the ureteral stent tube loaded with the pirfenidone nanoparticle coating. In the treatment of ureteral stenosis, there have been few reports of the use of nanoparticles as drug delivery platforms. In the present study, we innovatively loaded nanoparticle/pirfenidone complexes onto ureteral stent tubes, and examined the biocompatibility, release characteristics, and tissue distribution of the nanoparticle/pirfenidone complex-coated ureteral stent tubes through biodegradation and controlled release of the nanoparticles. Finally, we evaluated the efficacy of the nanoparticle/pirfenidone composite coated ureteral stent to inhibit ureteral fibrosis by pathological section staining and Western Blot to detect gross anatomical changes, collagen deposition and expression of various fibrous proteins, to achieve the effect of preventing ureteral stenosis, and applied the pirfenidone nanoparticle composite coating on the ureteral stent tube can inhibit ureteral stenosis by reducing the expression of transforming growth factor β 1 and the deposition of collagen, and therefore, we considered that the use of the nanoparticle/PFD composite coated ureteral stent is an effective method for preventing ureteral stenosis caused by iatrogenic manipulations.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a technical scheme that: the preparation method of the ureteral stent tube carrying the pirfenidone nano particle composite coating comprises the following steps:
s1, firstly, preparing a suspension loaded with pirfenidone nanoparticles in advance;
s2, then, putting the ureteral stent tube into a phosphate buffer solution containing dopamine hydrochloride for incubation;
s3, stirring for a certain time at room temperature to form a polydopamine-modified ureteral stent;
s4, washing the polydopamine-modified ureteral stent tube obtained in the step S3 with deionized water twice, and then soaking the stent in a suspension of pirfenidone nanoparticles to form a ureteral stent tube loaded with a pirfenidone nanoparticle coating;
s5, observing the surface morphology of the ureteral stent tube wrapped with the nanoparticles by using a scanning electron microscope;
and S6, finally, labeling the nanoparticle compound by rhodamine B, and detecting the distribution of the pirfenidone nanoparticles on the ureteral stent.
Wherein, in step S1, the preparation of the pirfenidone nanoparticle-loaded suspension includes the following steps:
s11, firstly, emulsifying phosphate buffered saline, pirfenidone and polylactic acid-glycolic acid in dichloromethane, and carrying out ice bath for 1min by using an ultrasonic homogenizer to obtain a first mixture;
s12, adding polyvinyl alcohol into the mixture I obtained in the step S11 to form an emulsion;
s13, emulsifying the emulsion obtained in the step S12 again for 3min by using ultrasonic waves in an ice bath, and stirring for 24h at room temperature to completely evaporate dichloromethane to obtain nano particles;
s14, centrifuging the nanoparticles obtained in the step S13 at a speed of less than 4 ℃ and 12,000 revolutions per minute for 5min;
and S15, finally, washing the nanoparticles for multiple times by using deionized water, and suspending the nanoparticles in the deionized water to obtain a suspension loaded with the pirfenidone nanoparticles.
In step S6, the preparation of the nanoparticle complex labeled with rhodamine B comprises the following steps:
s21, firstly, preparing a suspension containing rhodamine B labeled nano particles in advance;
s22, then, incubating the ureteral stent tube obtained in the step S4 in a phosphate buffer solution containing 0.5mg/ml dopamine hydrochloride, and stirring for 3 hours at room temperature to form a polydopamine-modified ureteral stent tube;
s23, washing the polydopamine-modified ureteral stent tube twice with deionized water, and then soaking the polydopamine-modified ureteral stent tube in the rhodamine B-labeled nanoparticle suspension prepared in step S21 to form the rhodamine B nanoparticle/pirfenidone-coated ureteral stent tube.
In step S21, the preparation of the rhodamine B labeled nanoparticle suspension includes the following steps:
s211, firstly, emulsifying 100ul of phosphate buffered saline and rhodamine B in 2ml of dichloromethane containing 20mg of PLGA, adding the mixture, and carrying out ultrasonic treatment in an ice bath for 0.5min to obtain a second mixture;
s212, then, adding 4.5ml of 1.5% PVA to the second mixture and sonicating to form a multiple emulsion;
s213, stirring the emulsion at room temperature for 24h to completely evaporate dichloromethane, and centrifuging the nanoparticles at a speed of 12,000 rpm lower than 4 ℃ for 5min;
and S214, finally, washing the nano particles with deionized water for three times, and finally suspending in the deionized water to obtain the rhodamine B marked nano particle suspension.
In step S2, the concentration of dopamine hydrochloride is 0.5mg/ml, and the pH value of the phosphate buffer is 8.5.
Wherein, in the step S3, the stirring time is 3h.
The cytotoxicity of the nanoparticle/pirfenidone complex coated ureteral stent tubes was then determined by the CCK8 method. The determination method comprises the following steps: sterile ureteral stent tubes of 20mm length were cut to 2mm length and placed in 96-well plates. Next, the cell suspension was added to 96 wells. After 24h and 48h incubation, 10 mg/ml CCK8 dye solution was added to each well and incubation continued at 37 ℃ under 5% carbon dioxide for 4h. The absorbance of each well was measured at 450nm using an enzyme-linked immunosorbent assay. The cell viability was 100% with untreated cells as control. Experiments were performed in triplicate.
Animal experiment model for ureter damage
The experimental animals were male New Zealand white rabbits at 20 weeks of age. Animal experiments were approved by the university of south Tong laboratory animal ethics Committee (approval No.: S20210301-991). The experimental rabbits were randomly divided into a scald-making module, an unmodified ureteral stent treatment group, and an NP/pirfenidone ureteral stent treatment group, each group containing 3 rabbits. All rabbits were intramuscularly injected with fast-sleeping novi II (1 ml/kg) + Shutai 50 (0.4 ml/kg). After general anesthesia, the rabbit was fixed on the operating table in supine position. Taking the middle incision of the abdomen, incising the skin, subcutaneous tissue and abdominal rectus muscle layer by layer, and opening the peritoneum. After entering the abdominal cavity, the descending colon and the mesentery are pushed away, the adipose tissue is separated bluntly, and a section of ureter with the length of about 1cm is dissociated. Inserting the LK-3 type electrocoagulation guide wire into the ureter, electrocauterizing the ureter for 3 seconds by using 10W electrocoagulation, and removing the guide wire. The colon mesentery is replaced and the abdominal cavity is closed. Three days after surgery, daily intramuscular injections of cephalosporin prevented infection.
In the treatment group, as in the above method, a section of ureter tissue having a length of 1cm was isolated, a longitudinal incision was made in the middle of the ureter, an unmodified ureteral stent and an NP/pirfenidone ureteral stent were placed, respectively, and then thermal injury was performed using an electrocoagulation guidewire. Animals were sacrificed 2 weeks post-surgery and the ureteral stenosis and treatment (about 1 cm) were removed, comparing changes in the gross specimens and expression of the relevant cytokines.
HE staining and immunohistochemical staining analysis
After 2 weeks of ureteral injury and stent placement, each rabbit was sacrificed by air embolization. The bilateral ureters and kidneys of the rabbits were then removed in their entirety. Tissues were preserved in 10% formalin solution. The specimens were dehydrated, embedded in paraffin, and cut into 5-micron-thick sections. Changes in ureteral endothelial cells and luminal area were observed by staining with hematoxylin-eosin (H & E).
Expression levels of TGF-beta 1 in ureteral tissue were detected using immunohistochemical methods. Incubate with 3% hydrogen peroxide at room temperature for 5-10min to block endogenous peroxidase activity. The water bath is set to 100 ℃; the sections were placed in citrate buffer (daceae, glosteux, denmark) for 5min for antigen retrieval. Slides were washed 3 times with PBS for 5min each and blocked with 5% BSA for 2h. Sections were then incubated with rabbit anti-transforming growth factor β 1 antibody (1, 21898-1-AP, proteintech) overnight at 4 ℃. Next, the slides were washed with PBS and incubated with secondary antibody for 1h. Sections were stained with DAB for color development and then counterstained with hematoxylin. The sections were sealed with neutral gel and the tissue images were observed and evaluated with a microscope (Leka DMR3000,234 Leica Microsystem, bensiemm, germany).
Western Blot analysis
Total proteins of ureteral tissues of the US group, the US + ureteral stent tube group and the US + NP/pirfenidone ureteral stent tube group were extracted and subjected to Western blot analysis. Protein samples were separated on SDS-PAGE gels and transferred to polyvinylidene fluoride (PVDF) membranes. Then washed with TBST buffer (50 mM Tris-HCl,100mM NaC I, 0.1% Tween-20, pH 7.6) and blocked with 5% skim milk powder in TBST for 2h. Then, the cells were incubated with rabbit anti-transforming growth factor β 1 antibody (1, 21898-1-AP, protetech), rabbit anti-collagen type I antibody (1, gb114197, servicebio) and rabbit anti-collagen type III antibody (1, 1000, gb111323, servicebio) overnight at 4 ℃. All experiments were repeated three times. The following day, primary antibody was taken, membranes were washed 3 times with TBST at room temperature for 10min, and membranes were incubated with secondary antibody overnight at 4 ℃. The membrane was washed as above, and the protein bands were visualized using an Odyssey Infrared imaging System ((LICOR, lincoln, NE, USA.) the expression of the protein bands was determined using ImageJ software and normalized to GAPDH.
Statistical analysis
All values are expressed as means ± Standard Deviation (SD). The difference between groups was measured by unpaired t-test. One-way analysis of variance (ANOVA) was performed using GraphPad Prism9 software. P values <0.05 were statistically significant.
In summary, according to the preparation method of the ureteral stent tube loaded with the pirfenidone nanoparticle composite coating, the ureteral stent tube which is commonly used in clinic is soaked in an alkaline solution containing dopamine to form an adhesive coating on the surface of the stent, and then the dopamine-modified stent tube is incubated with the nanoparticle/pirfenidone composite to form the ureteral stent tube loaded with the pirfenidone nanoparticle coating. In the treatment of ureteral stenosis, there have been few reports of the use of nanoparticles as drug delivery platforms. In the present study, we innovatively loaded nanoparticle/pirfenidone complexes onto ureteral stent tubes, and examined the biocompatibility, release characteristics, and tissue distribution of the nanoparticle/pirfenidone complex-coated ureteral stent tubes through biodegradation and controlled release of the nanoparticles. Finally, the effect of inhibiting ureteral fibrosis by the nanoparticle/pirfenidone composite coating ureteral stent tube is evaluated by pathological section staining and Western Blot to detect gross anatomical change, collagen deposition and expression of various fibrin, so as to achieve the effect of preventing ureteral stenosis.
In the description of the present invention, unless otherwise expressly specified or limited, the terms "disposed," "mounted," "connected," and "secured" are to be construed broadly, e.g., as meaning fixedly connected, detachably connected, or integral to; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. The preparation method of the ureteral stent tube carrying the pirfenidone nano particle composite coating is characterized by comprising the following steps: the method comprises the following steps:
s1, firstly, preparing a suspension carrying pirfenidone nanoparticles in advance;
s2, then, putting the ureteral stent tube into a phosphate buffer solution containing dopamine hydrochloride for incubation;
s3, stirring for a certain time at room temperature to form a polydopamine modified ureteral stent;
s4, washing the polydopamine-modified ureteral stent tube obtained in the step S3 with deionized water twice, and then soaking the stent in a suspension of pirfenidone nanoparticles to form a ureteral stent tube loaded with a pirfenidone nanoparticle coating;
s5, observing the surface morphology of the ureteral stent tube wrapped with the nanoparticles by using a scanning electron microscope;
and S6, finally, labeling the nanoparticle compound by rhodamine B, and detecting the distribution condition of the pirfenidone nanoparticles in tissues after the application of the pirfenidone nanoparticles in the ureteral stent in vivo.
2. The preparation method of the ureteral stent tube with the pirfenidone nanoparticle composite coating according to claim 1, wherein the preparation method comprises the following steps: in step S1, preparation of a pirfenidone nanoparticle-loaded suspension is prepared, including the steps of:
s11, firstly, emulsifying phosphate buffered saline, pirfenidone and polylactic acid-glycolic acid in dichloromethane, and performing ultrasonic treatment for 1min by using an ultrasonic homogenizer to obtain a first mixture;
s12, adding a polyvinyl alcohol solution into the mixture I obtained in the step S11, and performing ultrasonic emulsification for 3min;
s13, stirring the emulsion obtained in the step S12 at room temperature for 24 hours to completely evaporate dichloromethane to obtain nano particles;
s14, centrifuging the nanoparticles obtained in the step S13 at the speed of 12,000 revolutions per minute of less than 4 ℃ for 5min;
and S15, finally, washing the nanoparticles for multiple times by using deionized water, and suspending the nanoparticles in the deionized water to obtain a suspension loaded with the pirfenidone nanoparticles.
3. The preparation method of the ureteral stent tube with the pirfenidone-loaded nanoparticle composite coating according to claim 2, wherein the preparation method comprises the following steps: in step S6, preparation of a nanoparticle complex labeled with rhodamine B, comprising the steps of:
s21, firstly, preparing a suspension containing rhodamine B labeled nanoparticles in advance;
s22, then, incubating the ureteral stent tube obtained in the step S4 in a phosphate buffer solution containing 0.5mg/ml dopamine hydrochloride, and then stirring for 3 hours at room temperature to form a polydopamine-modified ureteral stent tube;
s23, washing the polydopamine-modified ureteral stent tube twice with deionized water, and then soaking the polydopamine-modified ureteral stent tube in the rhodamine B-labeled nanoparticle suspension prepared in step S21 to form the rhodamine B nanoparticle/pirfenidone-coated ureteral stent tube.
4. The preparation method of the pirfenidone-nanoparticle-loaded composite coated ureteral stent tube according to claim 3, which comprises the following steps: in step S21, preparation of a suspension of rhodamine B labeled nanoparticles, comprising the steps of:
s211, firstly, emulsifying 100ul of phosphate buffered saline and rhodamine B in 2ml of dichloromethane containing 20mg of PLGA, and carrying out ultrasonic treatment for 0.5min in an ice bath to obtain a second mixture;
s212, then, 4.5ml of 1.5% PVA was added to the second mixture and sonicated to form a multiple emulsion;
s213, next, the emulsion is stirred at room temperature for 24h to completely evaporate the dichloromethane, and the nanoparticles are centrifuged at a speed of 12,000 rpm, which is lower than 4 ℃, for 5min;
and S214, finally, washing the nano particles with deionized water for three times, and finally suspending in the deionized water to obtain the rhodamine B marked nano particle suspension.
5. The preparation method of the ureteral stent tube with the pirfenidone-loaded nanoparticle composite coating according to claim 1, wherein the preparation method comprises the following steps: in step S2, the concentration of dopamine hydrochloride is 0.5mg/ml, and the pH value of the phosphate buffer is 8.5.
6. The preparation method of the ureteral stent tube with the pirfenidone nanoparticle composite coating according to claim 1, wherein the preparation method comprises the following steps: in step S3, the stirring time period of the stirring was 3 hours.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211596392.3A CN115970069A (en) | 2022-12-13 | 2022-12-13 | Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating |
| PCT/CN2023/126065 WO2024125091A1 (en) | 2022-12-13 | 2023-10-24 | Method for preparing ureteral stent tube coated with pirfenidone-carrying nanoparticle composite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211596392.3A CN115970069A (en) | 2022-12-13 | 2022-12-13 | Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115970069A true CN115970069A (en) | 2023-04-18 |
Family
ID=85971486
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211596392.3A Pending CN115970069A (en) | 2022-12-13 | 2022-12-13 | Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN115970069A (en) |
| WO (1) | WO2024125091A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024125091A1 (en) * | 2022-12-13 | 2024-06-20 | 南通大学附属医院 | Method for preparing ureteral stent tube coated with pirfenidone-carrying nanoparticle composite |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110996931A (en) * | 2017-06-16 | 2020-04-10 | 埃维克辛公司 | Compositions and methods for treating fibrotic diseases |
| CN211911903U (en) * | 2019-11-26 | 2020-11-13 | 中国科学院金属研究所 | Ureteral stent for inhibiting recurrent stricture stimulation symptoms and calculus |
| WO2021142086A1 (en) * | 2020-01-08 | 2021-07-15 | Synthis Therapeutics, Inc. | Alk5 inhibitor conjugates and uses thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2877164B1 (en) * | 2012-07-24 | 2023-07-05 | Avalyn Pharma Inc. | Aerosol pirfenidone and pyridone analog compounds |
| CN109893682A (en) * | 2019-04-16 | 2019-06-18 | 东北大学 | A kind of degradable metal ureter bracket and preparation method with composite construction |
| CN110464882A (en) * | 2019-07-29 | 2019-11-19 | 东华大学 | A kind of ureter rack tube and preparation method thereof that hydrophilic antimicrobial is degradable |
| KR102331879B1 (en) * | 2019-09-20 | 2021-11-29 | 의료법인 성광의료재단 | Nanoparticles including improved coatings and their uses |
| CN113663142A (en) * | 2021-09-03 | 2021-11-19 | 中国科学院长春应用化学研究所 | Multifunctional coating suitable for urinary medical instruments, and preparation method and application thereof |
| CN115970069A (en) * | 2022-12-13 | 2023-04-18 | 南通大学附属医院 | Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating |
-
2022
- 2022-12-13 CN CN202211596392.3A patent/CN115970069A/en active Pending
-
2023
- 2023-10-24 WO PCT/CN2023/126065 patent/WO2024125091A1/en unknown
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110996931A (en) * | 2017-06-16 | 2020-04-10 | 埃维克辛公司 | Compositions and methods for treating fibrotic diseases |
| CN211911903U (en) * | 2019-11-26 | 2020-11-13 | 中国科学院金属研究所 | Ureteral stent for inhibiting recurrent stricture stimulation symptoms and calculus |
| WO2021142086A1 (en) * | 2020-01-08 | 2021-07-15 | Synthis Therapeutics, Inc. | Alk5 inhibitor conjugates and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| UESHIMA, EISUKE, 等: "Macrophage-secreted TGF-β1 contributes to fibroblast activation and ureteral stricture after ablation injury", AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, vol. 317, no. 1, 31 July 2019 (2019-07-31), pages 52 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024125091A1 (en) * | 2022-12-13 | 2024-06-20 | 南通大学附属医院 | Method for preparing ureteral stent tube coated with pirfenidone-carrying nanoparticle composite |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2024125091A1 (en) | 2024-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Braided thin‐walled biodegradable ureteral stent: Preliminary evaluation in a canine model | |
| CN113557040B (en) | Acoustic extracellular matrix hydrogels and uses thereof | |
| Dias et al. | Neovaginoplasty using Nile tilapia fish skin as a new biologic graft in patients with Mayer-Rokitansky-Küster-Hauser syndrome | |
| CN115970069A (en) | Preparation method of ureteral stent tube carrying pirfenidone nanoparticle composite coating | |
| Eggink et al. | In utero repair of an experimental neural tube defect in a chronic sheep model using biomatrices | |
| Yan et al. | Ligustrazine nanoparticles nano spray’s activation on Nrf2/ARE pathway in oxidative stress injury in rats with postoperative abdominal adhesion | |
| WO2022228356A1 (en) | Ptmc-based intestinal anastomosis stent of bioabsorbable flexible elastomer and preparation method therefor | |
| US20230364301A1 (en) | Methods for preparation of a terminally sterilized hydrogel or colloidal suspension derived from extracellular matrix, and uses thereof | |
| Wei et al. | A novel tetra-PEG based hydrogel for prevention of esophageal stricture after ESD in a porcine model | |
| CN109846864A (en) | Isoliquiritigenin and its application in the acute lungs liver for the treatment of and brain tissue impairment | |
| CN102599995A (en) | Pipe stent with double-layered structure and preparation method of pipe stent | |
| Chen et al. | Study of poly-ɛ-caprolactone membranes for pleurodesis | |
| CN114377131A (en) | Use of 5-hydroxytryptamine receptor subtypes 3 and 4 and antagonists thereof for treating post-cholecystectomy diarrhea | |
| Chen et al. | Protective effect of sulodexide on podocyte injury in adriamycin nephropathy rats | |
| Lampus et al. | The influence of topical mitomycin-C on total fibroblasts, epithelialization, and collagenization in anoplasty wound healing in Wistar rats | |
| Inagaki et al. | Transplantation of autologous oral mucosal epithelial cell sheets inhibits the development of acquired external auditory canal atresia in a rabbit model | |
| CN118050510A (en) | Application of lysine hydroxylase serving as action target in preparation of medicament for treating urethral fibrosis | |
| Gentileschi et al. | Simultaneous correction of breast hypertrophy and vaginal agenesis: Aesthetic surgery to the aid of reconstructive surgery | |
| CN116270713B (en) | Application of miR-1246 in preparation of medicament for promoting tendon bone healing | |
| RU2738163C1 (en) | Method of treating uretero-vaginal fistulas following complete hysterectomy | |
| CN112121149B (en) | Medicine for treating postoperative abdominal adhesion and new application thereof | |
| US11944650B2 (en) | Method of treating or preventing hernia formation | |
| Geng et al. | Clinical effects of laparoscopic treatment of unilateral inguinal indirect hernia in children with occult patent processus vaginalis and nano-silver antibacterial dressing in postoperative wound recovery | |
| CN116270555A (en) | Application of composite nanofiber membrane in preparation of medicine for promoting diabetic wound healing | |
| KR101927419B1 (en) | Antiadhesive reagent containing silk fibroin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |