CN113234711A - Biomineralization thrombin, preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and preparation method thereof - Google Patents
Biomineralization thrombin, preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and preparation method thereof Download PDFInfo
- Publication number
- CN113234711A CN113234711A CN202110528638.2A CN202110528638A CN113234711A CN 113234711 A CN113234711 A CN 113234711A CN 202110528638 A CN202110528638 A CN 202110528638A CN 113234711 A CN113234711 A CN 113234711A
- Authority
- CN
- China
- Prior art keywords
- thrombin
- gauze
- biomineralized
- biomineralization
- preparation
- 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
- 108090000190 Thrombin Proteins 0.000 title claims abstract description 120
- 229960004072 thrombin Drugs 0.000 title claims abstract description 120
- 230000002439 hemostatic effect Effects 0.000 title claims abstract description 74
- 230000033558 biomineral tissue development Effects 0.000 title claims abstract description 55
- 238000002360 preparation method Methods 0.000 title claims abstract description 36
- 239000000463 material Substances 0.000 claims abstract description 70
- 229910021645 metal ion Inorganic materials 0.000 claims abstract description 38
- 239000012621 metal-organic framework Substances 0.000 claims abstract description 28
- 239000002994 raw material Substances 0.000 claims abstract description 21
- 239000010452 phosphate Substances 0.000 claims abstract description 14
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims abstract description 14
- 229910001463 metal phosphate Inorganic materials 0.000 claims abstract description 8
- 239000005995 Aluminium silicate Substances 0.000 claims description 44
- 235000012211 aluminium silicate Nutrition 0.000 claims description 44
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims description 44
- 239000004927 clay Substances 0.000 claims description 35
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 claims description 32
- 239000000758 substrate Substances 0.000 claims description 29
- 229910052751 metal Inorganic materials 0.000 claims description 21
- 239000002184 metal Substances 0.000 claims description 21
- 238000002791 soaking Methods 0.000 claims description 20
- 239000007788 liquid Substances 0.000 claims description 17
- 229960003638 dopamine Drugs 0.000 claims description 16
- 238000001035 drying Methods 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 claims description 8
- 239000011230 binding agent Substances 0.000 claims description 8
- 150000003839 salts Chemical class 0.000 claims description 8
- 239000013154 zeolitic imidazolate framework-8 Substances 0.000 claims description 8
- MFLKDEMTKSVIBK-UHFFFAOYSA-N zinc;2-methylimidazol-3-ide Chemical compound [Zn+2].CC1=NC=C[N-]1.CC1=NC=C[N-]1 MFLKDEMTKSVIBK-UHFFFAOYSA-N 0.000 claims description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 7
- 239000001110 calcium chloride Substances 0.000 claims description 7
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 7
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 claims description 6
- 239000001263 FEMA 3042 Substances 0.000 claims description 6
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 6
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 claims description 6
- FXNFHKRTJBSTCS-UHFFFAOYSA-N baicalein Chemical compound C=1C(=O)C=2C(O)=C(O)C(O)=CC=2OC=1C1=CC=CC=C1 FXNFHKRTJBSTCS-UHFFFAOYSA-N 0.000 claims description 6
- KTVIXTQDYHMGHF-UHFFFAOYSA-L cobalt(2+) sulfate Chemical compound [Co+2].[O-]S([O-])(=O)=O KTVIXTQDYHMGHF-UHFFFAOYSA-L 0.000 claims description 6
- 229910000366 copper(II) sulfate Inorganic materials 0.000 claims description 6
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 6
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 6
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 6
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 6
- 235000015523 tannic acid Nutrition 0.000 claims description 6
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 claims description 6
- 229940033123 tannic acid Drugs 0.000 claims description 6
- 229920002258 tannic acid Polymers 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 5
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 claims description 4
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 4
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 claims description 3
- PFTAWBLQPZVEMU-ZFWWWQNUSA-N (+)-epicatechin Natural products C1([C@@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-ZFWWWQNUSA-N 0.000 claims description 3
- PFTAWBLQPZVEMU-UKRRQHHQSA-N (-)-epicatechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-UKRRQHHQSA-N 0.000 claims description 3
- IPMYMEWFZKHGAX-UHFFFAOYSA-N Isotheaflavin Natural products OC1CC2=C(O)C=C(O)C=C2OC1C(C1=C2)=CC(O)=C(O)C1=C(O)C(=O)C=C2C1C(O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-UHFFFAOYSA-N 0.000 claims description 3
- IKMDFBPHZNJCSN-UHFFFAOYSA-N Myricetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC(O)=C(O)C(O)=C1 IKMDFBPHZNJCSN-UHFFFAOYSA-N 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- UXRMWRBWCAGDQB-UHFFFAOYSA-N Theaflavin Natural products C1=CC(C2C(CC3=C(O)C=C(O)C=C3O2)O)=C(O)C(=O)C2=C1C(C1OC3=CC(O)=CC(O)=C3CC1O)=CC(O)=C2O UXRMWRBWCAGDQB-UHFFFAOYSA-N 0.000 claims description 3
- 239000000440 bentonite Substances 0.000 claims description 3
- 229910000278 bentonite Inorganic materials 0.000 claims description 3
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 claims description 3
- 239000005313 bioactive glass Substances 0.000 claims description 3
- 235000005487 catechin Nutrition 0.000 claims description 3
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 claims description 3
- 229950001002 cianidanol Drugs 0.000 claims description 3
- LPTRNLNOHUVQMS-UHFFFAOYSA-N epicatechin Natural products Cc1cc(O)cc2OC(C(O)Cc12)c1ccc(O)c(O)c1 LPTRNLNOHUVQMS-UHFFFAOYSA-N 0.000 claims description 3
- 235000012734 epicatechin Nutrition 0.000 claims description 3
- 235000004515 gallic acid Nutrition 0.000 claims description 3
- 229940074391 gallic acid Drugs 0.000 claims description 3
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 3
- PCOBUQBNVYZTBU-UHFFFAOYSA-N myricetin Natural products OC1=C(O)C(O)=CC(C=2OC3=CC(O)=C(O)C(O)=C3C(=O)C=2)=C1 PCOBUQBNVYZTBU-UHFFFAOYSA-N 0.000 claims description 3
- 235000007743 myricetin Nutrition 0.000 claims description 3
- 229940116852 myricetin Drugs 0.000 claims description 3
- IPMYMEWFZKHGAX-ZKSIBHASSA-N theaflavin Chemical compound C1=C2C([C@H]3OC4=CC(O)=CC(O)=C4C[C@H]3O)=CC(O)=C(O)C2=C(O)C(=O)C=C1[C@@H]1[C@H](O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-ZKSIBHASSA-N 0.000 claims description 3
- 235000014620 theaflavin Nutrition 0.000 claims description 3
- 229940026509 theaflavin Drugs 0.000 claims description 3
- 239000005909 Kieselgur Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 230000000740 bleeding effect Effects 0.000 abstract description 9
- 102000009123 Fibrin Human genes 0.000 abstract description 5
- 108010073385 Fibrin Proteins 0.000 abstract description 5
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 abstract description 5
- 108010049003 Fibrinogen Proteins 0.000 abstract description 5
- 102000008946 Fibrinogen Human genes 0.000 abstract description 5
- 239000008280 blood Substances 0.000 abstract description 5
- 210000004369 blood Anatomy 0.000 abstract description 5
- 229950003499 fibrin Drugs 0.000 abstract description 5
- 229940012952 fibrinogen Drugs 0.000 abstract description 5
- 239000000243 solution Substances 0.000 description 47
- 239000011701 zinc Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 239000008055 phosphate buffer solution Substances 0.000 description 11
- 239000007853 buffer solution Substances 0.000 description 10
- 238000004108 freeze drying Methods 0.000 description 10
- 230000023597 hemostasis Effects 0.000 description 10
- 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 9
- 208000032843 Hemorrhage Diseases 0.000 description 9
- 239000000835 fiber Substances 0.000 description 7
- 230000007246 mechanism Effects 0.000 description 6
- 229910001868 water Inorganic materials 0.000 description 6
- 239000000853 adhesive Substances 0.000 description 5
- 230000001070 adhesive effect Effects 0.000 description 5
- 239000002105 nanoparticle Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- LXBGSDVWAMZHDD-UHFFFAOYSA-N 2-methyl-1h-imidazole Chemical compound CC1=NC=CN1 LXBGSDVWAMZHDD-UHFFFAOYSA-N 0.000 description 4
- PZKFSRWSQOQYNR-UHFFFAOYSA-N 5-methyl-1h-1,2,4-triazole Chemical compound CC1=NC=NN1 PZKFSRWSQOQYNR-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 208000027418 Wounds and injury Diseases 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 229920001690 polydopamine Polymers 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000007983 Tris buffer Substances 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- YDMBNDUHUNWWRP-VJBWXMMDSA-N (2s)-1-[(2r)-2-amino-3-phenylpropanoyl]-n-[(2s)-5-(diaminomethylideneamino)-1-(4-nitroanilino)-1-oxopentan-2-yl]piperidine-2-carboxamide Chemical compound C([C@@H](N)C(=O)N1[C@@H](CCCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NC=1C=CC(=CC=1)[N+]([O-])=O)C1=CC=CC=C1 YDMBNDUHUNWWRP-VJBWXMMDSA-N 0.000 description 2
- MOMKYJPSVWEWPM-UHFFFAOYSA-N 4-(chloromethyl)-2-(4-methylphenyl)-1,3-thiazole Chemical compound C1=CC(C)=CC=C1C1=NC(CCl)=CS1 MOMKYJPSVWEWPM-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 108010094028 Prothrombin Proteins 0.000 description 2
- 102100027378 Prothrombin Human genes 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 108010039286 S 2238 Proteins 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 229910001431 copper ion Inorganic materials 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 238000001000 micrograph Methods 0.000 description 2
- 230000001089 mineralizing effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 238000011056 performance test Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- 229940039716 prothrombin Drugs 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 235000019983 sodium metaphosphate Nutrition 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- WMFHUUKYIUOHRA-UHFFFAOYSA-N (3-phenoxyphenyl)methanamine;hydrochloride Chemical compound Cl.NCC1=CC=CC(OC=2C=CC=CC=2)=C1 WMFHUUKYIUOHRA-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 229920000297 Rayon Polymers 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- -1 antifungals Substances 0.000 description 1
- 239000000739 antihistaminic agent Substances 0.000 description 1
- 229940125715 antihistaminic agent Drugs 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 229940019700 blood coagulation factors Drugs 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- JIJSGQYJSRWCLG-UHFFFAOYSA-L disodium;2-[hydroxy(2-sulfonatoethyl)amino]ethanesulfonate Chemical compound [Na+].[Na+].[O-]S(=O)(=O)CCN(O)CCS([O-])(=O)=O JIJSGQYJSRWCLG-UHFFFAOYSA-L 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 1
- 239000002657 fibrous material Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000010954 inorganic particle Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 description 1
- 210000002741 palatine tonsil Anatomy 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
- 239000002964 rayon Substances 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- GYDJEQRTZSCIOI-LJGSYFOKSA-N tranexamic acid Chemical compound NC[C@H]1CC[C@H](C(O)=O)CC1 GYDJEQRTZSCIOI-LJGSYFOKSA-N 0.000 description 1
- 229960000401 tranexamic acid Drugs 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 210000002268 wool Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/18—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing inorganic materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/38—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/44—Medicaments
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6429—Thrombin (3.4.21.5)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21005—Thrombin (3.4.21.5)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
Abstract
The invention provides biomineralization thrombin, a preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and a preparation method thereof, belonging to the technical field of surgical medical supplies; the preparation method comprises the following steps: biomineralization of thrombin by metal ions and phosphate to obtain biomineralized thrombin; or the thrombin is mineralized by the preparation raw material of the metal organic framework material to obtain the biomineralized thrombin. The thrombin can stabilize the structure of the thrombin after biomineralization, so that the thrombin can be stored at room temperature for a long time and can be directly loaded on gauze. When the gauze loaded with the biomineralized thrombin contacts with blood, the biological mineralized thrombin can quickly catalyze fibrinogen to form network fibrin so as to achieve the aim of stopping bleeding.
Description
Technical Field
The invention relates to the technical field of surgical medical supplies, in particular to biomineralization thrombin, a preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and a preparation method thereof.
Background
The rescue time can be prolonged by preventing excessive blood loss in emergency treatment and sudden accidents, and the survival rate of patients is improved. The tourniquet and bandage for first aid is mainly a tourniquet and a bandage at present, but the tourniquet and bandage can not meet the requirements of wound first aid development from the aspects of first aid mode and first aid effect. The prior art discloses a hemostatic product for acute major hemorrhage, namely 'Xudun' quick-acting hemostatic powder, but the biggest problems of the 'Xudun' quick-acting hemostatic powder are as follows: in the treatment process, part of the hemostatic powder possibly enters blood vessels to form thrombus, so that the risk of serious hemostatic sequelae is caused; in addition, the main components of the hemostatic powder are inorganic particles which cannot be absorbed and metabolized in the body and can be accumulated in the body, and the long-term safety of the hemostatic powder cannot be guaranteed.
The prior art also discloses the use of kaolin for the preparation of hemostatic gauze. Kaolin is an inert mineral substance without any animal or vegetable protein, and has no allergy when contacting with wound and high safety. Currently marketed kaolin hemostatic materials are mainly from us Z-medical company, which utilizes kaolin to contact and activate FXII, activate the characteristics of internal blood coagulation pathway, and develops a series of kaolin hemostatic materials, including kaolin battle wound gauze, kaolin interventional hemostatic bandage, kaolin tonsil hemostatic sponge, kaolin hemostatic pad, and the like, aiming at different bleeding amounts and bleeding parts. However, the kaolin hemostatic gauze still has certain defects, because the kaolin activates an endogenous coagulation mechanism to further activate prothrombin to generate thrombin, and the thrombin catalyzes fibrinogen to form network fibrin to achieve the purpose of hemostasis, and the hemostasis mechanism is single. The existing kaolin hemostatic gauze mainly contains kaolin, has single hemostatic component and single hemostatic mechanism, so that the hemostatic speed needs to be further improved. The defect that the high hemostasis speed needs to be further improved can be overcome by adding thrombin into the kaolin hemostatic gauze, but the thrombin is directly loaded on the gauze and then needs to be stored in a refrigerator below 4 ℃, the thrombin is inactivated within a short time at room temperature, and the requirement of conventional first aid cannot be met.
Disclosure of Invention
The invention aims to provide biomineralization thrombin, a preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and a preparation method thereof. The hemostatic time of the rapid hemostatic gauze prepared based on the biomineralized thrombin and the clay material is obviously shortened.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of biomineralized thrombin, which comprises the following steps:
biomineralization of thrombin by metal ions and phosphate to obtain biomineralized thrombin; or the thrombin is mineralized by the preparation raw material of the metal organic framework material to obtain the biomineralized thrombin.
Preferably, the metal ion is selected from Cu2+、Ca2+、Mg2+、Co2+、Fe2+、Mn2+And Zn2+One or more of them.
Preferably, the metal ion is derived from a metal salt; the metal salt is preferably selected from CuSO4、CaCl2、MgCl2、CoSO4、FeSO4、MnSO4And Zn (CH)3COO)2One or more of them.
Preferably, the metal organic framework material is selected from ZIF-8 and/or MAF-7.
The invention also provides the biomineralization thrombin prepared by the preparation method in the scheme, and the particle size of the biomineralization thrombin is 5 nm-50 μm.
The invention also provides application of the biomineralization thrombin in the scheme in preparation of a hemostatic product.
The invention also provides a rapid hemostatic gauze prepared from the biomineralization thrombin based on the scheme, which comprises the following components:
a gauze substrate;
a clay material and biomineralized thrombin distributed on the gauze substrate.
Preferably, the clay material is selected from one or more of kaolin, activated clay, bentonite, bioactive glass and diatomite.
Preferably, the clay material is adhered to the gauze substrate by a binder, and the binder is selected from one or more of dopamine, tannic acid, gallic acid, 5,6, 7-trihydroxyflavone, tannic acid, myricetin, catechin, epicatechin and theaflavin.
The invention also provides a preparation method of the rapid hemostatic gauze in the scheme, which comprises the following steps:
after the gauze substrate distributed with the clay material is subjected to first soaking in a pretreatment solution, the gauze substrate is placed in a mineralization solution to be subjected to second soaking, and drying is carried out, so that the rapid hemostatic gauze is obtained;
the solute in the pretreatment solution comprises metal ion phosphate or metal element donor raw material of metal organic framework material;
the mineralized liquid comprises thrombin and a mineralizer, the mineralizer comprises a first mineralizer and/or a second mineralizer, the first mineralizer comprises metal ions and phosphate, and the second mineralizer comprises raw materials for preparing metal organic framework materials.
The invention provides a preparation method of biomineralized thrombin, which comprises the following steps: biomineralization of thrombin by metal ions and phosphate to obtain biomineralized thrombin; or the thrombin is mineralized by the preparation raw material of the metal organic framework material to obtain the biomineralized thrombin. The thrombin can stabilize the structure of the thrombin through biomineralization, so that the thrombin can be stored at room temperature for a long time and can be directly loaded on a hemostatic product carrier. When the hemostatic product loaded with biomineralized thrombin contacts with blood, the fibrinogen can be rapidly catalyzed to form network fibrin so as to achieve the purpose of hemostasis.
The invention also provides a rapid hemostatic gauze, which comprises: a gauze substrate; a clay material distributed on the gauze substrate; and biomineralized thrombin distributed on the gauze substrate; the biomineralization thrombin is obtained by biomineralization of thrombin by metal ion phosphate or metal organic framework materials. In the invention, the clay material can activate prothrombin to generate thrombin by activating an endogenous coagulation mechanism, and the thrombin catalyzes fibrinogen to form network fibrin so as to achieve the aim of stopping bleeding. When the biomineralization thrombin contacts with blood, the biomineralization thrombin can quickly catalyze the fibrinogen to form network fibrin so as to achieve the purpose of hemostasis. Therefore, the hemostatic gauze has a mechanism of activating endogenous hemostasis by the clay material and a quick hemostasis mechanism by thrombin, and can realize the purpose of quick hemostasis compared with the traditional hemostatic gauze with single kaolin.
Drawings
FIG. 1 is an electron microscope image of micro-nano particles of phosphate biomineralization thrombin of different metal ions in example 1;
FIG. 2 is SEM results corresponding to different kaolin-based hemostatic gauze of example 3;
FIG. 3 shows Cu in example 32+SEM results corresponding to the phosphate biomineralization thrombin/kaolin rapid hemostatic gauze;
FIG. 4 is a drawing of a QuikClot Combat Gauze entity;
FIG. 5 is a drawing of a QuikClot Combat Gauze-X Ray Gauze;
FIG. 6 is QuikClot EMS Rolled Gauze;
figure 7 results of hemostatic performance tests of different groups of gauze.
Detailed Description
The invention provides a preparation method of biomineralized thrombin, which comprises the following steps:
biomineralization of thrombin by metal ions and phosphate to obtain biomineralized thrombin; or the thrombin is mineralized by the preparation raw material of the metal organic framework material to obtain the biomineralized thrombin.
In the present invention, the metal ion is preferably selected from Cu2+、Ca2+、Mg2+、Co2+、Fe2+、Mn2+And Zn2+More preferably selected from Cu2+And/or Zn2+。
In the inventionThe metal ion is preferably derived from a metal salt; the metal salt is preferably selected from CuSO4、CaCl2、MgCl2、CoSO4、FeSO4、MnSO4And Zn (CH)3COO)2One or more of them.
In the present invention, the phosphate is preferably derived from a phosphate buffer solution; the pH value of the phosphate buffer solution is preferably 7.4; the molar concentration of the phosphate buffer solution is preferably 0.1M.
In the present invention, the metal organic framework material is preferably selected from ZIF-8 and/or MAF-7. In the present invention, the ZIF-8 is preferably prepared from a starting material comprising Zn (OAc)2·2H2O and 2-methylimidazole; said Zn (OAc)2·2H2The molar ratio of O and 2-methylimidazole is preferably 1: (5-30), more preferably 1: 16; the raw material for preparing the MAF-7 preferably comprises Zn (NO)3)2·6H2O, 3-methyl-1, 2, 4-triazole and NH3·H2O; the Zn (NO)3)2·6H2O, 3-methyl-1, 2, 4-triazole and NH3·H2The proportion of O is preferably 0.04 mM: 0.64 mM: 6 μ L.
In the invention, the preparation method specifically comprises the following steps:
mixing metal salt, phosphate buffer solution and thrombin, and standing and mineralizing to obtain biomineralized thrombin; alternatively, the first and second electrodes may be,
mixing the preparation raw materials of the metal organic framework material with thrombin, standing and mineralizing to obtain the biomineralized thrombin.
In the invention, the temperature of the standing mineralization is preferably 0-40 ℃, and more preferably 20 ℃; the standing mineralization time is 1-72 hours.
The invention also provides biomineralization thrombin prepared by the preparation method in the scheme.
In the present invention, the particle size of the biomineralized thrombin is preferably 1nm to 50 μm, more preferably 1nm to 10 μm, and still more preferably 5nm to 3 μm.
The invention also provides application of the biomineralization thrombin in the scheme in preparation of a hemostatic product.
The invention also provides a rapid hemostatic gauze prepared from the biomineralization thrombin based on the scheme, which comprises the following components:
a gauze substrate;
a clay material and biomineralized thrombin distributed on the gauze substrate.
In the present invention, the gauze substrate comprises a woven or non-woven fibrous material including, but not limited to, cotton, silk, wool, plastic, cellulose, rayon, polyester, or combinations thereof.
In the invention, the clay material is selected from one or more of kaolin, activated clay, bentonite, bioactive glass and diatomite.
In the present invention, the clay material is preferably adhered to the gauze substrate by a binder, the binder is preferably one or more selected from dopamine, tannic acid, gallic acid, 5,6, 7-trihydroxyflavone, tannic acid, myricetin, catechin, epicatechin, and theaflavin, and more preferably dopamine.
The invention takes dopamine as an adhesive, phenolic hydroxyl in dopamine molecules has strong interaction with silicon hydroxyl in clay materials and cellulose hydroxyl in gauze, and the dopamine molecules also can generate chemical reaction to form covalent bonds. Therefore, the dopamine in the invention loads kaolin on the surface of the gauze fiber with strong acting force, compared with the conventional kaolin-based hemostatic gauze, the dopamine is more firm in loading of kaolin, so that the kaolin is greatly reduced in falling off when contacting blood.
The invention takes dopamine molecules as the adhesive and has the following advantages: the polydopamine formed on the surface of the gauze fiber has strong adsorption performance on metal ions when the polydopamine is adhered to the clay material, so that the assembly of biomineralized thrombin on the surface of the gauze is promoted; polydopamine formed on the surface of gauze fiber when clay material is adhered can adsorb the enriched part of blood coagulation factors due to the existence of phenolic hydroxyl groups of the polydopamine, so that blood coagulation is promoted; the dopamine has good biocompatibility, and the hemostatic gauze prepared by using the dopamine as the binder is softer and convenient to use.
In the present invention, the rapid hemostatic gauze preferably further comprises: a hemostatic material disposed on the gauze substrate in combination with the clay material; the hemostatic material is selected from one or more of magnesium sulfate, sodium metaphosphate, calcium chloride, dextrin, hydrated magnesium sulfate, sodium metaphosphate hydrate, calcium chloride hydrate, dextrin hydrate, ascorbic acid, tranexamic acid, rutin and thrombin.
In the present invention, the rapid hemostatic gauze preferably further comprises: at least one of the following materials in combination with the clay material distributed on the gauze substrate: antibiotics, antifungals, antimicrobials, anti-inflammatory agents, analgesics, and antihistamines.
The invention also provides a preparation method of the rapid hemostatic gauze in the scheme, which comprises the following steps:
after the gauze substrate distributed with the clay material is subjected to first soaking in a pretreatment solution, the gauze substrate is placed in a mineralization solution to be subjected to second soaking, and drying is carried out, so that the rapid hemostatic gauze is obtained;
the solute in the pretreatment solution comprises metal ions or metal element donor raw materials of metal organic framework materials;
the mineralized liquid comprises thrombin and a mineralizer, the mineralizer comprises a first mineralizer and/or a second mineralizer, the first mineralizer comprises metal ions and phosphate, and the second mineralizer comprises raw materials for preparing metal organic framework materials.
In the invention, the gauze substrate distributed with the clay material is preferably prepared by the following method: and soaking the gauze substrate in the clay material dispersion liquid, and drying to obtain the gauze substrate distributed with the clay material.
In the present invention, the clay material dispersion preferably uses a Tris-HCl buffer solution as a solvent, and includes the following components in concentration: 1-1250 mg/mL of adhesive and 1-100 mg/mL of clay material; the concentration of the clay material is preferably 30 mg/mL; the concentration of the adhesive is preferably 250-500 mg/mL.
In the invention, the molar concentration of the Tris-HCl buffer solution is preferably 0.05 mol/L; the pH value of the Tris-HCl buffer solution is 8.5; in the specific implementation process of the invention, the preparation method of the Tris-HCl buffer solution comprises the following steps: adding 0.2mol/L HCl aqueous solution into 0.2mol/L Tris buffer solution, and then adding deionized water for dilution; the volume ratio of the Tris buffer solution to the HCl aqueous solution to the deionized water is 5: 3: 8.
in the invention, the soaking process is preferably accompanied with shaking treatment; the oscillation frequency is preferably 250-300 rpm; the soaking temperature is preferably 20-25 ℃; the soaking time is preferably 5-6 h. In the present invention, the drying mode is preferably drying; the drying temperature is preferably 45-50 ℃; the drying time is preferably 8-10 h.
After obtaining the gauze substrate distributed with the clay material, the invention preferably re-soaks the obtained gauze substrate distributed with the clay material in Tris-HCl buffer solution to make the adhesive more effectively adhere the clay material to the gauze; the concentration of the binding agent in the Tris-HCl buffer solution of the binding agent is preferably 125-1250 mg/mL, and more preferably 250-500 mg/mL.
After obtaining the gauze substrate distributed with the clay material, the gauze substrate distributed with the clay material is soaked in a pretreatment solution for the first time, then is soaked in a mineralization solution for the second time, and is dried to obtain the rapid hemostatic gauze;
the solute of the pretreatment solution comprises metal ions or metal element donor raw materials of metal organic framework materials;
the mineralized liquid comprises thrombin; further comprises one of the following components:
1) a metal ion;
2) preparing the raw material of the metal organic framework material.
In the present invention, when the solute of the pretreatment liquid includes metal ions, the mineralization liquid preferably further includes metal ions; when the solute of the pretreatment solution comprises a metal element donor material of a metal organic framework material, the mineralization solution further comprises a material for preparing the metal organic framework material.
In the present invention, the kind of the metal element in the pretreatment liquid is preferably the same as the kind of the metal ion in the mineralization liquid; alternatively, the kind of the metal element donor material of the metal organic framework material in the pretreatment liquid is preferably the same as that of the metal element donor material of the metal organic framework material in the mineralized liquid.
In the invention, the molar concentration of the metal ions in the pretreatment solution is preferably 100-120 mM; the molar concentration of the metal element donor raw material of the metal organic framework material is preferably 0.04-0.05 mM.
In the invention, the first soaking process is preferably performed with shaking; the oscillation frequency is preferably 250-300 rpm; the first soaking temperature is preferably 20-30 ℃, and further preferably 25 ℃; the first soaking time is preferably 5-6 h. In the present invention, the first soaking is used for making the gauze substrate adsorb corresponding metal ions or metal element donor raw materials of metal organic framework materials until the adsorption is saturated.
In the invention, the mineralized liquid preferably takes phosphate buffer solution as a solvent; the pH value of the phosphate buffer solution is 7.4; the concentration of the phosphate buffer solution is preferably 0.1M.
In the invention, the concentration of thrombin in the mineralized liquid is preferably 0.1-2.5 KU/mL.
In the invention, when the mineralized liquid contains metal ions, the molar concentration of the metal ions in the mineralized liquid is 0.01-5 mM, preferably 0.8-1.5 mM.
In the invention, when the mineralization liquid comprises raw materials for preparing the metal organic framework material; the molar concentration of the metal element donor raw material in the raw material for preparing the metal organic framework material is 0.001-2 mM, and preferably 0.02-0.05 mM.
In the present invention, the mineralized liquid preferably comprises the following components: phosphate buffer solution and thrombin; also comprises metal salt or raw materials for preparing metal organic framework materials; the volume ratio of the phosphate buffer solution to the metal salt solution to the thrombin solution is 98: (1-2): (2-3); the molar concentration of the metal salt solution is 0.8-1.5 mM; the concentration of the thrombin solution is 0.1-2.5 KU/mL.
In the invention, the second soaking temperature is preferably 0-35 ℃, and more preferably 20 ℃; and the second soaking time is 20-24 h. In the present invention, the second soaking is used to biomineralize the metal ion or metal organic framework material donor material with thrombin.
In the present invention, the drying is preferably freeze-drying; the temperature of the freeze drying is preferably-20 to-50 ℃; the freeze-drying time is preferably 24 h.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. 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.
Example 1
1mL of 120mM CuSO was added4、CaCl2、CoSO4、FeSO4、MnSO4、Zn(CH3COO)2To the solution, 98mL of phosphate buffer solution (PBS pH 7.4, 0.1M) was added, followed by 2mL of thrombin solution (1 KU/mL). And standing the final solution at room temperature for 24h, and performing centrifugal precipitation and freeze drying to obtain the biomineralized thrombin mineralized by different metal ions.
Preparation of ZIF-8 biomineralized thrombin: at room temperature, 2mL of thrombin solution (1KU/mL) was added to 28mL of a solution containing Zn (OAc)2·2H2O (0.04mM), 2-methylimidazole (0.64mM) and distilled water. And standing the final solution at room temperature for 24h, and performing centrifugal precipitation and freeze drying to obtain the ZIF-8 biomineralization thrombin.
Preparation of MAF-7 biomineralized thrombin: at room temperature, 2mL of thrombin solution (1KU/mL) was added to 28mL of a solution containing Zn (NO)3)2·6H2O (0.04mM), 3-methyl-1, 2, 4-triazole (0.12mM), 10% NH3·H2O (60. mu.L) in distilled water. And standing the final solution at room temperature for 24h, performing centrifugal precipitation, and freeze-drying to obtain the MAF-7 biomineralization thrombin.
An electron microscope image of the different metal ion biomineralization thrombin micro-nano particles is shown in figure 1. As can be seen from FIG. 1, the forms of nanoparticles obtained by biomineralization of different metal ions are different, and the particle size is 1-3000 nm.
Thermal stability of biomineralization thrombin micro-nano particles mineralized by different metal ions
Measurement of the thermal stability of the biomineralized thrombin mineralized by the different metal ions obtained: placing the sample tube filled with a certain amount of metal ion biomineralization thrombin in an oven at 50 ℃/70 ℃ for 2h, taking out and cooling to room temperature, and adding 1ml of PBS buffer solution preheated at 37 ℃ to uniformly disperse the sample in the solution. Then, 2ml of a solution (8X 10) of a thrombin chromogenic substrate S2238 (preheated at 37 ℃ C.) was added to the above solution-4M) and the mixture was placed in a 37 ℃ water bath for 5 min. The supernatant was collected by centrifugation and the absorbance was immediately measured at 405 nm. For the untreated sample, the sample tube was placed in a 37 ℃ water bath for 10min, and 1ml of a pre-warmed PBS buffer and a thrombin chromogenic substrate S2238 solution (8X 10)-4M). The mixture was placed in a 37 ℃ water bath for 10 min. The supernatant was collected by centrifugation and the absorbance measured at 405nm, the results of which are shown in Table 1, the thermal stability of the zinc/copper ion biomineralized thrombin was the best.
TABLE 1 thermal stability of different metal ion biomineralization thrombin micro-nano particles
EXAMPLE 2 preparation of different biomineralized Thrombin hemostatic gauze
First, a square of dopamine modified gauze with a side length of about 20cm was soaked in 30mL of 120mM CuSO4、CaCl2、CoSO4、FeSO4、MnSO4、Zn(CH3COO)2The solution was shaken at 30 ℃ for 5h, then immersed in 98mL of phosphate buffer (PBS pH 7.4, 0.1M), and 1mL of 120mM CuSO was added thereto4、CaCl2、CoSO4、FeSO4、MnSO4、Zn(CH3COO)2The solution was then added 2mL of thrombin solution (1 KU/mL). And standing the final solution at room temperature for 24h, taking out the gauze, and freeze-drying to obtain the biomineralization thrombin rapid hemostatic gauze mineralized by different metal ions.
Preparing ZIF-8 biomineralization thrombin rapid hemostatic gauze: at room temperature, dopamine-modified square gauze approximately 20cm on a side was soaked in 30mL of 0.04mM Zn (OAc)2·2H2O solution was shaken at 30 ℃ for 5h and then immersed in 28mL of a solution containing Zn (OAc)2·2H2O (0.04mM), 2-methylimidazole (0.64mM), and 2mL of a thrombin solution (1KU/mL) in distilled water. And standing the final solution at room temperature for 24h, taking out the gauze, and freeze-drying to obtain the ZIF-8 biomineralization thrombin rapid hemostatic gauze.
Preparing MAF-7 biomineralization thrombin rapid hemostatic gauze: square dopamine-modified gauze with a side length of about 20cm is soaked in 30mL of 0.04mM Zn (NO) respectively3)2·6H2Shaking in O solution at 30 deg.C for 5h, and soaking in 28mL solution containing Zn (NO)3)2·6H2O (0.04mM), 3-methyl-1, 2, 4-triazole (0.12mM), 10% NH3·H2O (60. mu.L) and 2mL of thrombin solution (1KU/mL) in distilled water. And standing the final solution at room temperature for 24h, taking out the gauze, and freeze-drying to obtain the MAF-7 biomineralization thrombin rapid hemostatic gauze.
Example 3
(1) Preparing kaolin-based hemostatic gauze: preparing 100ml of dopamine Tris-HCl buffer solutions with different concentrations according to the proportion shown in the table 2, adding kaolin, performing ultrasonic treatment until the kaolin is uniformly dispersed in the dopamine solution for about 30min, adding square gauze with the side length of about 20cm based on the condition that no kaolin exists at the bottom of a bottle, shaking at the temperature of 25 ℃ for 6h in a shaking table at the rpm of 250rpm, and drying in a drying oven at the temperature of 50 ℃ for 10h to obtain the kaolin-based hemostatic gauze. And (3) soaking the obtained dried kaolin-based hemostatic gauze in a dopamine Tris-HCl buffer solution again, and repeating the steps without adding new kaolin.
Preparation of Tris-HCl buffer solution (0.05mol/LpH ═ 8.5): 6mL of 0.2mol/L HCl solution was added to 10mL of 0.2mol/L Tris buffer solution, followed by 24mL of deionized water.
TABLE 2 Kaolin-based hemostatic gauze
SEM results corresponding to different groups of kaolin-based hemostatic gauze are shown in fig. 2, where the gauze fibers without kaolin adhered thereto have smooth surfaces, and the fibers with kaolin adhered thereto have rough surfaces and kaolin particles, and the results indicate that kaolin can be adhered to the surfaces of the gauze fibers in all the preparation methods.
(2) Preparing the rapid hemostatic gauze:
the No. 3 kaolin based hemostatic gauze obtained in the step (1) is respectively soaked in 30mL of 120mM CuSO4Shaking the mixture in an aqueous solution at 30 deg.C for 5h, soaking the mixture in 98mL phosphate buffer (PBS pH 7.4, 0.1M), and adding 1mL 120mM CuSO4After the aqueous solution, 2mL of thrombin solution (1KU/mL, solvent pH 7.4, phosphate buffer solution at a concentration of 0.1M) was added. Standing the final solution at 20 ℃ for 24h, taking out gauze, and freeze-drying to obtain Cu2+Biomineralization thrombin/kaolin rapid hemostatic gauze. Cu2+The SEM results corresponding to the biomineralized thrombin/kaolin fast hemostatic gauze are shown in fig. 3. As can be seen from FIG. 3, the gauze fibers had both the kaolin flakes (1 in FIG. 3) and the Cu flakes2+Biomineralized thrombin (2 in fig. 3).
(3) Setting a control group 1, a control group 2, a control group 3 and an experimental group 1; kaolin is adopted for control group 1, control group 2 and control group 3 for hemostasisGauze, QuikClot Combat Gauze (see FIG. 4), QuikClot Combat Gauze-X Ray Gauze (see FIG. 5) and QuikClot EMS Rolled Gauze Gauze (see FIG. 6), respectively. The experimental group adopts the Cu prepared in the step (2)2+Biomineralization thrombin/kaolin rapid hemostatic gauze.
Performing hemostasis performance tests on the different groups of gauze: rat strain and liver resection model: the strain: SD rats, which are widely used in pharmacology, toxicology, drug efficacy, and GLP experiments; ② age of mice: 7 weeks; ③ weight: about 250 g; fourthly, anesthesia: intraperitoneal injection of chloral hydrate; trauma: approximately 2cm of liver was excised. A hemostasis mode: firstly, placing gauze under the liver, cutting off 2cm of liver, and freely bleeding for 30 s; secondly, firstly pressing the gauze on the bleeding part of the wound for 1 min; and thirdly, opening the gauze every 30 seconds, and observing the bleeding condition until the bleeding is completely stopped.
The results of the experiment are shown in FIG. 7. As can be seen from fig. 7, the copper ion biomineralization thrombin/kaolin rapid hemostatic gauze of the experimental group 1 has shorter hemostatic time, lower bleeding amount and better hemostatic performance than those of the control group 1, the control group 2 and the control group 3.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A method for preparing biomineralized thrombin comprises the following steps:
biomineralization of thrombin by metal ions and phosphate to obtain biomineralized thrombin; or the thrombin is mineralized by the preparation raw material of the metal organic framework material to obtain the biomineralized thrombin.
2. The method according to claim 1, wherein the metal ion is selected from Cu2+、Ca2+、Mg2+、Co2 +、Fe2+、Mn2+And Zn2+One or more ofSeveral kinds of them.
3. The production method according to claim 1 or 2, wherein the metal ion is derived from a metal salt; the metal salt is preferably selected from CuSO4、CaCl2、MgCl2、CoSO4、FeSO4、MnSO4And Zn (CH)3COO)2One or more of them.
4. The method for preparing according to claim 1, wherein the metal organic framework material is selected from ZIF-8 and/or MAF-7.
5. The biomineralized thrombin prepared by the preparation method of any one of claims 1 to 4, wherein the biomineralized thrombin has a particle size of 5nm to 50 μm.
6. Use of biomineralized thrombin according to claim 5 in the preparation of a hemostatic product.
7. A rapid hemostatic gauze prepared based on the biomineralized thrombin of claim 5, comprising:
a gauze substrate;
a clay material and biomineralized thrombin distributed on the gauze substrate.
8. The rapid hemostatic gauze of claim 7, wherein the clay material is selected from one or more of kaolin, activated clay, bentonite, bioactive glass, and diatomaceous earth.
9. The rapid hemostatic gauze according to claim 7 or 8, wherein the clay material is adhered to the gauze substrate by a binder selected from one or more of dopamine, tannic acid, gallic acid, 5,6, 7-trihydroxyflavone, tannic acid, myricetin, catechin, epicatechin, and theaflavin.
10. The method for preparing the rapid hemostatic gauze of any one of claims 7 to 9, comprising the following steps:
after the gauze substrate distributed with the clay material is subjected to first soaking in a pretreatment solution, the gauze substrate is placed in a mineralization solution to be subjected to second soaking, and drying is carried out, so that the rapid hemostatic gauze is obtained;
the solute in the pretreatment solution comprises metal ion phosphate or metal element donor raw material of metal organic framework material;
the mineralized liquid comprises thrombin and a mineralizer, the mineralizer comprises a first mineralizer and/or a second mineralizer, the first mineralizer comprises metal ions and phosphate, and the second mineralizer comprises raw materials for preparing metal organic framework materials.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110528638.2A CN113234711A (en) | 2021-05-14 | 2021-05-14 | Biomineralization thrombin, preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110528638.2A CN113234711A (en) | 2021-05-14 | 2021-05-14 | Biomineralization thrombin, preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113234711A true CN113234711A (en) | 2021-08-10 |
Family
ID=77134422
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110528638.2A Pending CN113234711A (en) | 2021-05-14 | 2021-05-14 | Biomineralization thrombin, preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113234711A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108379644A (en) * | 2018-03-03 | 2018-08-10 | 青岛大学 | A kind of preparation method for the efficient hemostatic material being suitable for open drowned wound |
| CN111450252A (en) * | 2020-03-25 | 2020-07-28 | 暨南大学 | Medicine for targeted blocking of tumor blood vessels and preparation method and application thereof |
-
2021
- 2021-05-14 CN CN202110528638.2A patent/CN113234711A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108379644A (en) * | 2018-03-03 | 2018-08-10 | 青岛大学 | A kind of preparation method for the efficient hemostatic material being suitable for open drowned wound |
| CN111450252A (en) * | 2020-03-25 | 2020-07-28 | 暨南大学 | Medicine for targeted blocking of tumor blood vessels and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 吴忠隐等: "可溶性鸡蛋壳膜蛋白-聚丁二酸丁二醇酯共同静电纺丝负载凝血酶制备新型止血敷料的可行性研究", 《中国血管外科杂志（电子版）》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Calcium-modified microporous starch with potent hemostatic efficiency and excellent degradability for hemorrhage control | |
| Dai et al. | Molecular imprinted macroporous chitosan coated mesoporous silica xerogels for hemorrhage control | |
| Hu et al. | Antibacterial hemostatic dressings with nanoporous bioglass containing silver | |
| Fan et al. | Morphology-controllable cellulose/chitosan sponge for deep wound hemostasis with surfactant and pore-foaming agent | |
| KR20070117589A (en) | Mineral technologies (mt) for acute hemostasis and for the treatment of acute wounds and chronic ulcers | |
| WO2020233547A1 (en) | Sponge for traumatic hemostasis, preparation method therefor and use thereof | |
| KR101693696B1 (en) | Hemostatic porous structure and process for preparing the same | |
| CN106806931A (en) | A kind of starch compound hemostatic dressing of mesoporous silicon dioxide micro-sphere | |
| CN109731121A (en) | A kind of preparation method of the cellulose containing mesoporous silicon oxide and chitosan combine dressing | |
| CN109172857A (en) | A kind of outer wound compound hemostatic material and preparation method thereof | |
| Wang et al. | Calcium alginate/silk fibroin peptide/Bletilla striata polysaccharide blended microspheres loaded with tannic acid for rapid wound healing | |
| CN105903062B (en) | A kind of aperture and the controllable mesoporous silicon rapid hemostasis powder and preparation method of the equal monodisperse of partial size | |
| Shi et al. | Preparation and application of quick hemostatic gauze based on biomimetic mineralized thrombin | |
| Jia et al. | Hydrophobic aerogel-modified hemostatic gauze with thermal management performance | |
| Zhang et al. | Fabrication of effective mesoporous silica materials for emergency hemostasis application | |
| KR102645889B1 (en) | Hemostatic fabric containing trypsin and method for manufacturing the same | |
| CN113234711A (en) | Biomineralization thrombin, preparation method and application thereof, and rapid hemostatic gauze based on biomineralization thrombin and preparation method thereof | |
| Ding et al. | Ca-Ga double doping strategy to fabricate hemostatic mesoporous silica nanoparticles (MSN) with antibacterial activity | |
| CN106540310B (en) | Absorbable rapid hemostatic material and preparation method thereof | |
| Szymonowicz et al. | The evaluation of resorbable haemostatic wound dressings in contact with blood in vitro | |
| US8389498B2 (en) | Spinning solution and method for manufacturing biomaterial fibers | |
| CN113694247B (en) | Preparation method of multifunctional composite hemostatic sponge | |
| CN108815560B (en) | Porous tissue plugging material, preparation method thereof and plugging product | |
| CN111249518B (en) | Hemostatic composition and preparation method thereof | |
| CN111150877B (en) | Mesoporous silicon nano-particles, preparation method and application |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210810 |