CN115814144A - Silk fibroin adhesive hemostatic powder and preparation method and application thereof - Google Patents
Silk fibroin adhesive hemostatic powder and preparation method and application thereof Download PDFInfo
- Publication number
- CN115814144A CN115814144A CN202211481795.3A CN202211481795A CN115814144A CN 115814144 A CN115814144 A CN 115814144A CN 202211481795 A CN202211481795 A CN 202211481795A CN 115814144 A CN115814144 A CN 115814144A
- Authority
- CN
- China
- Prior art keywords
- silk fibroin
- hemostatic powder
- powder
- adhesive
- solution
- 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
- 108010022355 Fibroins Proteins 0.000 title claims abstract description 176
- 239000000843 powder Substances 0.000 title claims abstract description 131
- 230000002439 hemostatic effect Effects 0.000 title claims abstract description 126
- 239000000853 adhesive Substances 0.000 title claims abstract description 83
- 230000001070 adhesive effect Effects 0.000 title claims abstract description 83
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 230000000740 bleeding effect Effects 0.000 claims abstract description 22
- 238000007789 sealing Methods 0.000 claims abstract description 8
- 230000035876 healing Effects 0.000 claims abstract description 7
- 230000008439 repair process Effects 0.000 claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 75
- 238000006722 reduction reaction Methods 0.000 claims description 50
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 37
- -1 polyphenol compounds Chemical class 0.000 claims description 26
- 235000013824 polyphenols Nutrition 0.000 claims description 26
- 239000008367 deionised water Substances 0.000 claims description 24
- 229910021641 deionized water Inorganic materials 0.000 claims description 24
- PBVAJRFEEOIAGW-UHFFFAOYSA-N 3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochloride Chemical compound Cl.OC(=O)CCP(CCC(O)=O)CCC(O)=O PBVAJRFEEOIAGW-UHFFFAOYSA-N 0.000 claims description 15
- 239000003638 chemical reducing agent Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 15
- 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 14
- 239000001263 FEMA 3042 Substances 0.000 claims description 14
- 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 14
- 238000002156 mixing Methods 0.000 claims description 14
- 229920002258 tannic acid Polymers 0.000 claims description 14
- 235000015523 tannic acid Nutrition 0.000 claims description 14
- 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 14
- 229940033123 tannic acid Drugs 0.000 claims description 14
- 150000003573 thiols Chemical class 0.000 claims description 14
- 230000000975 bioactive effect Effects 0.000 claims description 13
- AMXOYNBUYSYVKV-UHFFFAOYSA-M lithium bromide Chemical compound [Li+].[Br-] AMXOYNBUYSYVKV-UHFFFAOYSA-M 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 11
- 238000007873 sieving Methods 0.000 claims description 10
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 8
- WMBWREPUVVBILR-WIYYLYMNSA-N (-)-Epigallocatechin-3-o-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=C(O)C=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-WIYYLYMNSA-N 0.000 claims description 7
- WMBWREPUVVBILR-UHFFFAOYSA-N GCG Natural products C=1C(O)=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-UHFFFAOYSA-N 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 7
- 229940030275 epigallocatechin gallate Drugs 0.000 claims description 7
- 238000000227 grinding Methods 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- XMOCLSLCDHWDHP-IUODEOHRSA-N epi-Gallocatechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC(O)=C(O)C(O)=C1 XMOCLSLCDHWDHP-IUODEOHRSA-N 0.000 claims description 6
- 238000001914 filtration Methods 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
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 239000003102 growth factor Substances 0.000 claims description 4
- 229920002545 silicone oil Polymers 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
- LSHVYAFMTMFKBA-PZJWPPBQSA-N (+)-catechin-3-O-gallate Chemical compound O([C@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=CC=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 LSHVYAFMTMFKBA-PZJWPPBQSA-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
- LSHVYAFMTMFKBA-TZIWHRDSSA-N (-)-epicatechin-3-O-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=CC=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 LSHVYAFMTMFKBA-TZIWHRDSSA-N 0.000 claims description 3
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 3
- 229920001661 Chitosan Polymers 0.000 claims description 3
- 102000008186 Collagen Human genes 0.000 claims description 3
- 108010035532 Collagen Proteins 0.000 claims description 3
- LSHVYAFMTMFKBA-UHFFFAOYSA-N ECG Natural products C=1C=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 LSHVYAFMTMFKBA-UHFFFAOYSA-N 0.000 claims description 3
- 108010010803 Gelatin Proteins 0.000 claims description 3
- XMOCLSLCDHWDHP-UHFFFAOYSA-N L-Epigallocatechin Natural products OC1CC2=C(O)C=C(O)C=C2OC1C1=CC(O)=C(O)C(O)=C1 XMOCLSLCDHWDHP-UHFFFAOYSA-N 0.000 claims description 3
- 239000002202 Polyethylene glycol Substances 0.000 claims description 3
- 229920000615 alginic acid Polymers 0.000 claims description 3
- 239000000783 alginic acid Substances 0.000 claims description 3
- 235000010443 alginic acid Nutrition 0.000 claims description 3
- 229960001126 alginic acid Drugs 0.000 claims description 3
- 150000004781 alginic acids Chemical class 0.000 claims description 3
- 239000005312 bioglass Substances 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
- 235000005487 catechin Nutrition 0.000 claims description 3
- 229940045110 chitosan Drugs 0.000 claims description 3
- 229950001002 cianidanol Drugs 0.000 claims description 3
- 229920001436 collagen Polymers 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
- DZYNKLUGCOSVKS-UHFFFAOYSA-N epigallocatechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3cc(O)c(O)c(O)c3 DZYNKLUGCOSVKS-UHFFFAOYSA-N 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 235000004515 gallic acid Nutrition 0.000 claims description 3
- 229940074391 gallic acid Drugs 0.000 claims description 3
- LVJJFMLUMNSUFN-UHFFFAOYSA-N gallocatechin gallate Natural products C1=C(O)C=C2OC(C=3C=C(O)C(O)=CC=3)C(O)CC2=C1OC(=O)C1=CC(O)=C(O)C(O)=C1 LVJJFMLUMNSUFN-UHFFFAOYSA-N 0.000 claims description 3
- 229920000159 gelatin Polymers 0.000 claims description 3
- 239000008273 gelatin Substances 0.000 claims description 3
- 235000019322 gelatine Nutrition 0.000 claims description 3
- 235000011852 gelatine desserts Nutrition 0.000 claims description 3
- 229920002674 hyaluronan Polymers 0.000 claims description 3
- 229960003160 hyaluronic acid Drugs 0.000 claims description 3
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims description 3
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims description 3
- 229920001223 polyethylene glycol Polymers 0.000 claims description 3
- 229940124350 antibacterial drug Drugs 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims 1
- 230000002209 hydrophobic effect Effects 0.000 abstract description 28
- 239000003106 tissue adhesive Substances 0.000 abstract description 10
- 230000008021 deposition Effects 0.000 abstract description 7
- 238000011065 in-situ storage Methods 0.000 abstract description 7
- 208000032843 Hemorrhage Diseases 0.000 description 23
- 208000034158 bleeding Diseases 0.000 description 20
- 206010052428 Wound Diseases 0.000 description 19
- 208000027418 Wounds and injury Diseases 0.000 description 19
- 230000023597 hemostasis Effects 0.000 description 16
- 210000004369 blood Anatomy 0.000 description 15
- 239000008280 blood Substances 0.000 description 15
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 150000001413 amino acids Chemical class 0.000 description 11
- 238000012360 testing method Methods 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 9
- 241000700157 Rattus norvegicus Species 0.000 description 8
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 230000036571 hydration Effects 0.000 description 7
- 238000006703 hydration reaction Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 210000004185 liver Anatomy 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- 241000255789 Bombyx mori Species 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 108010013296 Sericins Proteins 0.000 description 4
- 230000001846 repelling effect Effects 0.000 description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 208000031737 Tissue Adhesions Diseases 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 238000013168 hemostasis test Methods 0.000 description 3
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 2
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 125000003275 alpha amino acid group Chemical group 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003114 blood coagulation factor Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 230000002045 lasting effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 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
- 208000001953 Hypotension Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 229940019700 blood coagulation factors Drugs 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 230000036543 hypotension Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 230000004768 organ dysfunction Effects 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229920001864 tannin Polymers 0.000 description 1
- 235000018553 tannin Nutrition 0.000 description 1
- 239000001648 tannin Substances 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
Images
Landscapes
- Materials For Medical Uses (AREA)
Abstract
The invention discloses a silk fibroin adhesive hemostatic powder and a preparation method and application thereof. The silk fibroin adhesive hemostatic powder is dry powder, the size of the powder is 75-150 mu m, the hydrophobic property is strong, the tissue in-situ deposition performance is good under the wet physiological environment, the wet tissue adhesive strength is high, the hemostatic sealing speed is high, the silk fibroin adhesive hemostatic powder is suitable for the irregular-shaped and incompressible bleeding wound tissues under the wet physiological environment, the rapid hemostatic effect is realized, the effective liquid leakage sealing is realized, the wound tissue healing and repair is promoted, and the wound bacteriostasis is realized.
Description
Technical Field
The invention relates to the technical field of hemostatic powder, and particularly relates to silk fibroin adhesive hemostatic powder and a preparation method and application thereof.
Background
Excessive bleeding can lead to serious illnesses, including hypotension, organ dysfunction, and even death. The development of effective hemostatic materials is urgently needed clinically, especially for irregular-shaped, incompressible viscera, high-pressure arteriovenous bleeding wounds. An ideal hemostatic material needs to meet the following requirements: rapid and strong blood absorption to concentrate coagulation factors; the adhesion is strong; good mechanical properties, providing a physical barrier over the wound to maintain blood pressure; good blood compatibility and biocompatibility. The hemostatic powder can be directly deposited on an incompressible bleeding wound with an irregular shape, is gelatinized by absorbing blood, and further concentrates erythrocytes and blood coagulation factors to promote hemostasis, and has good adaptability. The hemostatic powder also has the advantages of convenient use and carrying, long-term storage and sterilization, and the like, and is widely applied in clinic. However, the conventional hemostatic powder has poor strength after being gelatinized, and is easily dispersed in a large-area bleeding process. In addition, upon contact with blood, the water in the blood can form a hydration layer at the interface between the hemostatic powder and the bleeding tissue, disrupting the adhesion between the hemostatic powder and the tissue, resulting in poor wet adhesion of the hemostatic powder to the bleeding tissue surface, leading to failure of hemostasis.
The silk fibroin is natural polymer fibrin extracted from silk, accounts for 70% -80% of the silk, and contains 18 amino acids. Structurally, silk fibroin is a naturally hydrophilic/hydrophobic block copolymer. The heavy chain (390 kDa), the light chain (26 kDa) and the glycoprotein (P25) together constitute silk fibroin, wherein the linkage between the light and heavy chains is mainly by means of disulfide bonds, while the glycoprotein is linked to them mainly by means of hydrophobic interactions. The heavy chain consists essentially of a repeating segment of hydrophobic amino acid repeats, such as (GAGAGS). The light chain comprises predominantly non-repetitive hydrophilic amino acid fragments. The silk fibroin has the characteristics of good biocompatibility, biodegradability, shape processability, functional plasticity and the like, and is widely applied to the field of biomedicine. However, at present, the hemostatic powder prepared from silk fibroin has poor strength and is easy to loosen after absorbing blood, and is easy to be dispersed in the process of large-area bleeding. Moreover, the hemostatic powder prepared from the existing silk fibroin still cannot overcome the obstruction of a blood interface hydration layer, has low adhesion performance with bleeding wound tissues, cannot meet the requirement of tissue adhesion strength in a wet physiological environment, and causes long hemostatic time. Therefore, the silk fibroin hemostatic powder with firm and durable adhesion performance in a wet physiological environment needs to be developed, and the defects that the existing hemostatic powder has poor strength after absorbing blood, is easy to loosen, has low adhesion performance with bleeding wound tissues, has low hemostatic speed and the like are overcome.
Disclosure of Invention
The invention aims to provide silk fibroin adhesive hemostatic powder and a preparation method and application thereof, and the prepared silk fibroin adhesive hemostatic powder has strong hydrophobic property, good tissue in-situ deposition property under a wet physiological environment, high wet tissue adhesive strength and high hemostatic sealing speed, is suitable for irregular and incompressible bleeding wound tissues under the wet physiological environment, realizes a quick hemostatic effect, effectively seals liquid leakage, promotes wound tissue healing and repair and inhibits bacteria of wounds.
Therefore, the invention provides a preparation method of silk fibroin adhesive hemostatic powder, which is prepared by uniformly mixing silk fibroin which is obtained after reduction reaction treatment by thiol reducing agents and polyphenol compounds; the powder size of the silk fibroin adhesive hemostatic powder is 75-150 μm.
Preferably, the method comprises the following steps:
(1) Dissolving degummed silk in lithium bromide or calcium chloride/absolute ethyl alcohol/water ternary solution, dialyzing with deionized water, and performing centrifugal filtration and vacuum freeze drying on the dialyzed solution to obtain silk fibroin powder;
(2) Dissolving the silk fibroin powder obtained in the step (1) in an aqueous solution containing thiol reducing agents, maintaining the pH value at 5.0, and placing the solution at room temperature for reduction reaction treatment to obtain a reduction reaction treated silk fibroin solution;
(3) Dissolving polyphenol compounds in deionized water, and stirring under the nitrogen protection environment to uniformly dissolve the polyphenol compounds to obtain polyphenol compound solution;
(4) Mixing the silk fibroin solution subjected to reduction reaction in the step (2) and the polyphenol compound solution in the step (3) with the same volume at room temperature, and maintaining the pH value to be 4.0-6.0 to obtain an insoluble aggregate;
(5) And (4) washing, freeze-drying, grinding, crushing and sieving the insoluble aggregate obtained in the step (4) to obtain the silk fibroin adhesive hemostatic powder.
Preferably, in the step (2), the concentration of the silk fibroin solution subjected to the reduction reaction is 1-10wt%, and the zeta potential value is more than +5mV.
Preferably, in the step (2), the thiol reducing agent includes, but is not limited to, tris (2-carboxyethyl) phosphine hydrochloride; the concentration of the thiol reducing agent is 10.0-50.0 mg/mL; the reduction reaction treatment time is 5-20 minutes at room temperature.
Preferably, in the step (3), the concentration of the polyphenol compound is 50.0-500.0mg/mL; the polyphenol compounds include, but are not limited to, epicatechin gallate, catechin, epicatechin, epigallocatechin, gallic acid, catechin gallate, epigallocatechin gallate, and tannic acid.
Preferably, in the step (4), bioactive molecules are further added during mixing at room temperature, and the bioactive molecules account for 0.1-10% of the weight of the silk fibroin adhesive hemostatic powder.
Preferably, the bioactive molecule comprises one or more of hydroxyapatite, bioglass, collagen, gelatin, antibacterial drugs, growth factors, chitosan, alginic acid, hyaluronic acid, polyethylene glycol and silicone oil.
Preferably, in the step (5), a screen with a mesh size of between 100 and 200 meshes is used for sieving.
The invention also provides the silk fibroin adhesive hemostatic powder prepared by the preparation method of the silk fibroin adhesive hemostatic powder.
The silk fibroin adhesive hemostatic powder disclosed by the invention is applied to the rapid hemostatic effect of irregular-shaped and incompressible bleeding wound tissues in a wet physiological environment, can effectively seal liquid leakage, can promote wound tissue healing and repair, and can inhibit bacteria of wounds.
Compared with the prior art, the invention has the advantages and positive effects that:
1) The invention utilizes thiol reducing agent to carry out reduction reaction treatment on silk fibroin, which can effectively decompose disulfide bonds in the silk fibroin and maintain free-SH groups of protein to obtain the silk fibroin treated by the reduction reaction, wherein the zeta potential value of the silk fibroin treated by the reduction reaction is more than +5mV. The exposure of hydrophobic amino acid can be induced by decomposing disulfide bonds in the silk fibroin, and the exposed hydrophobic amino acid can improve the hydrophobic effect of the silk fibroin adhesive hemostatic powder, so that the obstruction of a blood interface hydration layer in the process of repelling large-area hemorrhage can be effectively removed through high hydrophobic property, and the interface adhesion effect between the silk fibroin adhesive hemostatic powder and wound tissues can be improved.
2) The silk fibroin treated by the reduction reaction is assembled with the polyphenol compound together, so that the synergy of the high hydrophobicity effect and the strong tissue adhesion of the silk fibroin adhesive hemostatic powder under the wet physiological environment is realized, and the adhesion of an adhesive group and a bleeding tissue interface is facilitated. The silk fibroin adhesive hemostatic powder with the size of 75-150 mu m can quickly absorb blood to be gelatinized in a wet physiological environment, has good tissue in-situ deposition performance and high wet tissue adhesive strength, can not be dispersed in a large-area acute bleeding process, endows the silk fibroin adhesive hemostatic powder with strong, stable and lasting wet tissue adhesive performance in the wet physiological environment, realizes a quick and efficient tissue hemostatic sealing effect, and has the advantages of high hemostatic sealing speed and less tissue bleeding amount.
3) The silk fibroin adhesive hemostatic powder contains bioactive molecules, and under the synergistic effect of the bioactive molecules, the hemostatic powder has excellent antibacterial performance, and can reduce bacterial infection and inflammation, so that the healing and the regenerative repair of wound tissues are promoted.
4) The main components of the silk fibroin adhesive styptic powder are silk fibroin and polyphenol compounds which are both from natural compounds. Therefore, the invention has good biocompatibility and biodegradability, simple preparation process, low cost and great application potential.
5) The silk fibroin adhesive hemostatic powder prepared by the invention can overcome the obstruction caused by a blood interface hydration layer, has strong tissue in-situ deposition performance, high tissue adhesive strength in a wet physiological environment, the adhesive strength to wet pigskin tissue can reach more than 250kPa, and the wet tissue adhesive strength is obviously higher than that of the hemostatic powder prepared by the existing silk fibroin. The silk fibroin adhesive hemostatic powder can realize rapid hemostasis of liver bleeding tissues within 5 seconds, and the hemostasis speed is obviously higher than that of the hemostatic powder prepared from the existing silk fibroin.
Other features and advantages of the present invention will become more apparent upon reading of the following detailed description of the invention in conjunction with the accompanying drawings.
Drawings
FIG. 1 is a graph showing values of free-SH groups of silk fibroin according to example 1 of the present invention and comparative example 1;
FIG. 2 is a graph of zeta potential values of silk fibroin of example 1 of the present invention and comparative example 1;
FIG. 3 is a graph showing water contact angle values of the silk fibroin adhering hemostatic powders of example 1 of the present invention and comparative example 1.
Detailed Description
The following detailed description of specific embodiments of the present invention is provided to illustrate and explain the present invention and to be understood not to limit the present invention.
The invention provides a preparation method of silk fibroin adhering hemostatic powder, which is prepared from silk fibroin and polyphenol compound subjected to reduction reaction by thiol reducing agent; the zeta potential value of the silk fibroin is more than +5mV, and the size of the silk fibroin adhesion hemostatic powder is 75-150 μm.
Specifically, the preparation method of the silk fibroin adhesive hemostatic powder comprises the following steps:
(1) Dissolving degummed silk in lithium bromide or calcium chloride/absolute ethyl alcohol/water ternary solution, dialyzing with deionized water, and performing centrifugal filtration and vacuum freeze drying on the dialyzed solution to obtain silk fibroin powder.
(2) Dissolving the silk fibroin powder obtained in the step (1) in an aqueous solution containing thiol reducing agents, maintaining the pH value at 5.0, and placing the solution at room temperature for reduction reaction treatment to obtain a reduction reaction treated silk fibroin solution;
the concentration of the silk fibroin solution treated by the reduction reaction is 1-10wt%, and the zeta potential value is more than +5mV; the higher the zeta potential value is, the more hydrophobic amino acid in the silk fibroin is exposed, and the larger the capability of repelling a blood interface hydration layer in the process of repelling large-area hemorrhage through hydrophobic effect is, so that the interface adhesion effect between the silk fibroin adhesion hemostatic powder and wound tissues can be obviously improved.
The thiol reducing agent comprises, but is not limited to, tris (2-carboxyethyl) phosphine hydrochloride, and the concentration of the thiol reducing agent is 10.0-50.0 mg/mL; the tris (2-carboxyethyl) phosphine hydrochloride is a high-efficiency disulfide bond reducing agent, has good stability and solubility in an aqueous solution, and can fully and effectively reduce a silk fibroin solution.
The pH value is maintained to be 5.0, so that the reduction reaction treatment of the tris (2-carboxyethyl) phosphine hydrochloride on the silk fibroin is more sufficient, the exposure quantity of hydrophobic amino acid in the silk fibroin is increased, and the hydrophobic effect of the silk fibroin adhesive hemostatic powder is enhanced.
(3) Dissolving polyphenol compounds in deionized water, and stirring under the nitrogen protection environment to uniformly dissolve the polyphenol compounds to obtain polyphenol compound solution; the concentration of polyphenol compound is 50.0-500.0mg/mL; the polyphenol compounds include, but are not limited to, epicatechin gallate, catechin, epicatechin, epigallocatechin, gallic acid, catechin gallate, epigallocatechin gallate, and tannic acid.
(4) Mixing the silk fibroin solution subjected to the reduction reaction in the step (2) and the polyphenol compound solution in the step (3) at the same volume at room temperature, and maintaining the pH value at 4.0-6.0 to obtain insoluble aggregate.
The pH value is maintained to be 4.0-6.0 at room temperature, so that the silk fibroin solution and the polyphenol compound solution are more favorable for forming uniform insoluble aggregates when being mixed. In addition, the pH value is maintained to be 4.0-6.0 at room temperature, which is beneficial to the wet adhesion performance of the obtained silk fibroin adhesion hemostatic powder.
And (4) when the mixing is carried out at room temperature, one or more bioactive molecules are also contained, and the bioactive molecules account for 0.1-10% of the weight of the silk fibroin adhesive hemostatic powder. The bioactive molecule is one or more of hydroxyapatite, bioglass, collagen, gelatin, antibacterial agent, growth factor, chitosan, alginic acid, hyaluronic acid, polyethylene glycol, silicone oil, etc.
(5) And (4) washing, freeze-drying, grinding, crushing and sieving the insoluble aggregate obtained in the step (4) to obtain the silk fibroin adhesive hemostatic powder.
In the step (5), a sieve with 100 meshes to 200 meshes is adopted for sieving, and a sieve with 100 meshes to 200 meshes is adopted to ensure that the size of the hemostatic powder is between 75 and 150 mu m. The size of the silk fibroin hemostatic powder is 75-150 mu m, which is beneficial to ensuring that the contact area of the hemostatic powder and liquid is proper in the wet physiological environment, and stronger tissue in-situ deposition performance and adhesion performance are obtained.
1) The invention utilizes thiol reducing agent to carry out reduction reaction treatment on silk fibroin, which can effectively decompose disulfide bonds in the silk fibroin and maintain free-SH groups of protein to obtain the silk fibroin treated by the reduction reaction, wherein the zeta potential value of the silk fibroin treated by the reduction reaction is more than +5mV. The exposure of hydrophobic amino acid can be induced by decomposing disulfide bonds in the silk fibroin, and the exposed hydrophobic amino acid can improve the hydrophobic effect of the silk fibroin adhesive hemostatic powder, so that the obstruction of a blood interface hydration layer in the process of repelling large-area hemorrhage can be effectively removed through high hydrophobic property, and the interface adhesion effect between the silk fibroin adhesive hemostatic powder and wound tissues can be improved.
2) The silk fibroin treated by the reduction reaction is assembled with the polyphenol compound together, so that the synergy of the high hydrophobicity effect and the strong tissue adhesion of the silk fibroin adhesive hemostatic powder under the wet physiological environment is realized, and the adhesion of an adhesive group and a bleeding tissue interface is facilitated. The silk fibroin adhesive hemostatic powder with the size of 75-150 mu m can quickly absorb blood to be gelatinized in a wet physiological environment, has good tissue in-situ deposition performance and high wet tissue adhesive strength, can not be dispersed in a large-area acute bleeding process, endows the silk fibroin adhesive hemostatic powder with strong, stable and lasting wet tissue adhesive performance in the wet physiological environment, realizes a quick and efficient tissue hemostatic sealing effect, and has the advantages of high hemostatic sealing speed and less tissue bleeding amount.
3) The silk fibroin adhesive hemostatic powder contains bioactive molecules, and under the synergistic effect of the bioactive molecules, the hemostatic powder has excellent antibacterial performance, and can reduce bacterial infection and inflammation, so that the healing and the regenerative repair of wound tissues are promoted.
4) The main components of the silk fibroin adhesive styptic powder are silk fibroin and polyphenol compounds which are both from natural compounds. Therefore, the invention has good biocompatibility and biodegradability, simple preparation process, low cost and great application potential.
5) The silk fibroin adhesive hemostatic powder prepared by the invention can overcome the obstruction caused by a blood interface hydration layer, has strong tissue in-situ deposition performance, high tissue adhesive strength in a wet physiological environment, the adhesive strength to a wet pigskin tissue can reach more than 250kPa, and the wet tissue adhesive strength is obviously higher than that of the hemostatic powder prepared by the existing silk fibroin. The silk fibroin adhesive hemostatic powder can realize rapid hemostasis of liver bleeding tissues within 5 seconds, and the hemostasis speed is obviously higher than that of the hemostatic powder prepared from the existing silk fibroin.
The silk fibroin adhesive hemostatic powder disclosed by the invention is applied to the rapid hemostatic effect of irregular-shaped and incompressible bleeding wound tissues in a wet physiological environment, can effectively seal liquid leakage, can promote wound tissue healing and repair, and can inhibit bacteria of wounds.
Example 1
The preparation method of the silk fibroin adhesive hemostatic powder comprises the following steps:
(1) 50.0g of raw silkworm silk is put into 20.0L of 0.02mol/L sodium carbonate solution to be boiled for 20 minutes at 100 ℃, then is fully washed by deionized water to remove sericin, and is dried in a 50 ℃ oven to obtain the degummed silk. Dissolving 27g of degummed silk in 100.0mL of 9.3mol/L lithium bromide solution, treating for 240 minutes at 60 ℃, filling the dissolved solution into a dialysis bag after the degummed silk is completely dissolved, dialyzing with deionized water, centrifugally filtering the dialyzed solution, and placing the solution into a freeze dryer for vacuum freeze drying for 72 hours at-80 ℃ to obtain silk fibroin powder.
(2) Dissolving 300mg of tris (2-carboxyethyl) phosphine hydrochloride in 10mL of deionized water sufficiently, and maintaining the pH value at 5.0; then, 0.5g of silk fibroin powder was dissolved in 10mL of an aqueous solution containing tris (2-carboxyethyl) phosphine hydrochloride, and the solution was left at room temperature to be subjected to reduction treatment for 10 minutes, thereby obtaining a reduction-treated silk fibroin solution having a concentration of 5.0 wt%.
(3) Dissolving 5.0g of tannic acid in 10.0mL of deionized water, and stirring under the nitrogen protection environment to uniformly dissolve the tannic acid to obtain a tannic acid solution with the concentration of 500 mg/mL.
(4) Mixing the silk fibroin solution and the tannic acid solution at room temperature, maintaining pH at 4.5, and rapidly stirring to obtain insoluble aggregate.
(5) And (3) washing the insoluble aggregate obtained in the step (4) with deionized water for 3 times, placing the insoluble aggregate in a freeze dryer for vacuum freeze drying for 72 hours at the temperature of minus 80 ℃, grinding and crushing the freeze-dried solid, and sieving the crushed solid with a sieve of between 100 and 200 meshes to obtain hemostatic powder, namely the silk fibroin adhesive hemostatic powder. The hemostatic powder has a size of 75-150 μm.
And (3) testing the free-SH group content of the silk fibroin solution subjected to the reduction reaction treatment in the step (2), wherein the free-SH group content of the silk fibroin subjected to the reduction reaction treatment is 126.5 mu mol/g as shown in figure 1. And (3) testing the zeta potential value of the silk fibroin solution subjected to the reduction reaction treatment in the step (2), wherein the zeta potential value of the silk fibroin subjected to the reduction reaction treatment is +7.5mV as shown in figure 2. Therefore, the reduction reaction treatment of the silk fibroin by the tris (2-carboxyethyl) phosphine hydrochloride can effectively decompose disulfide bonds in the silk fibroin and maintain protein free-SH groups, so that the exposure of hydrophobic amino acid is induced, and the hydrophobic effect of the silk fibroin is improved.
And (3) testing the water contact angle of the silk fibroin adhesive hemostatic powder obtained in the step (5), wherein the water contact angle value of the silk fibroin adhesive hemostatic powder is 150 degrees as shown in fig. 3, which shows that the silk fibroin adhesive hemostatic powder has excellent high hydrophobic property.
The silk fibroin adhesive hemostatic powder of the embodiment has an adhesive strength of 280kPa to a wet pigskin tissue in a wet physiological environment, a Wistar rat is used as an animal hemostatic model, and the silk fibroin adhesive hemostatic powder is used for a Wistar rat liver hemostatic test, so that complete hemostasis can be realized within 5 seconds.
Comparative example 1
The preparation method of the silk fibroin adhesive styptic powder of the comparative example comprises the following steps:
(1) 50.0g of silkworm raw silk is put into 20.0L of 0.02mol/L sodium carbonate solution to be boiled for 20 minutes at 100 ℃, then the solution is fully washed by deionized water to remove sericin, and the solution is dried in a 50 ℃ oven to obtain the degummed silk. Dissolving 27g of degummed silk in 100.0mL of 9.3mol/L lithium bromide solution, treating at 60 ℃ for 240 minutes until the degummed silk is completely dissolved, filling the dissolved solution into a dialysis bag, dialyzing with deionized water, and centrifugally filtering the dialyzed solution to obtain a silk fibroin solution with the concentration of 5.0 wt%.
(2) Dissolving 5.0g of tannic acid in 10.0mL of deionized water, and stirring under the nitrogen protection environment to uniformly dissolve the tannic acid to obtain a tannic acid solution with the concentration of 500 mg/mL.
(3) Mixing the silk fibroin solution and the tannic acid solution with the same volume at room temperature, maintaining the pH value at 4.5, and rapidly stirring and uniformly mixing to obtain insoluble aggregate.
(4) And (3) washing the insoluble aggregate obtained in the step (3) with deionized water for 3 times, placing the insoluble aggregate in a freeze dryer for vacuum freeze drying for 72 hours at the temperature of minus 80 ℃, grinding and crushing the freeze-dried solid, and sieving the crushed solid with a sieve of between 100 and 200 meshes to obtain hemostatic powder, namely the silk fibroin adhesive hemostatic powder. The hemostatic powder has a size of 75-150 μm.
As shown in FIGS. 1 and 2, the silk fibroin of the comparative example had a free-SH group content of 3.8. Mu. Mol/g and a zeta potential value of-12 mV. As shown in FIG. 3, the silk fibroin adhesive hemostatic powder of the present comparative example had a water contact angle value of 80 °. Thus, it can be shown that, compared with example 1, in the comparative example, reduction reaction treatment of silk fibroin by tris (2-carboxyethyl) phosphine hydrochloride is not performed, exposure of hydrophobic amino acid of silk fibroin cannot be induced, and hydrophobic effect of the silk fibroin adhesive hemostatic powder cannot be improved.
The silk fibroin adhesive hemostatic powder of the comparative example has the adhesive strength of 15.2kPa to the wet pigskin tissue under the wet physiological environment. The Wistar rat is used as an animal hemostasis model, and the silk fibroin adhesion hemostasis powder is used for a Wistar rat liver hemostasis test, so that complete hemostasis can be realized within 65 seconds.
Example 2
The preparation method of the silk fibroin adhesive hemostatic powder comprises the following steps:
(1) 30.0g of raw silkworm silk is put into 12.0L of 0.02mol/L sodium carbonate solution to be boiled for 20 minutes at 100 ℃, then the raw silkworm silk is fully washed by deionized water to remove sericin, and the silk is dried in a 30 ℃ oven to obtain the degummed silk. Dissolving 13.5g of degummed silk in 50.0mL of 9.3mol/L lithium bromide solution, treating for 360 minutes at 50 ℃, filling the dissolved solution into a dialysis bag after the degummed silk is completely dissolved, dialyzing with deionized water, centrifugally filtering the dialyzed solution, and placing the solution into a freeze dryer for vacuum freeze drying at-80 ℃ for 24 hours to obtain silk fibroin powder.
(2) Dissolving 100mg of tris (2-carboxyethyl) phosphine hydrochloride in 10mL of deionized water, and fully dissolving the solution to maintain the pH value to be 5.0; then, 0.1g of silk fibroin powder was dissolved in 10mL of an aqueous solution containing tris (2-carboxyethyl) phosphine hydrochloride, and the solution was left at room temperature to be subjected to reduction treatment for 5 minutes, thereby obtaining a reduction-treated silk fibroin solution having a concentration of 1.0 wt%.
(3) Dissolving 0.5g of tannic acid in 10.0mL of deionized water, and stirring under the nitrogen protection environment to uniformly dissolve the tannic acid to obtain a tannic acid solution with the concentration of 50 mg/mL.
(4) Mixing the silk fibroin solution and the tannin solution which are subjected to the reduction reaction and have the same volume at room temperature, adding silicone oil which accounts for 10% of the weight of the hemostatic powder, maintaining the pH value at 5.5, and rapidly stirring and uniformly mixing to obtain an insoluble aggregate.
(5) And (3) washing the insoluble aggregate obtained in the step (4) with deionized water for 3 times, placing the insoluble aggregate in a freeze dryer for vacuum freeze drying for 24 hours at the temperature of minus 80 ℃, grinding and crushing the freeze-dried solid, and sieving the crushed solid with a sieve of between 100 and 200 meshes to obtain hemostatic powder, namely the silk fibroin adhesive hemostatic powder. The hemostatic powder has a size of 75-150 μm.
And (3) testing the free-SH group content of the silk fibroin solution subjected to the reduction reaction treatment in the step (2), wherein the free-SH group content of the silk fibroin subjected to the reduction reaction treatment is 116.2 mu mol/g. And (3) testing the zeta potential value of the silk fibroin solution subjected to the reduction reaction treatment in the step (2), wherein the zeta potential value of the silk fibroin subjected to the reduction reaction treatment is +6.5mV. Therefore, the reduction reaction treatment of the silk fibroin by the tris (2-carboxyethyl) phosphine hydrochloride can effectively decompose disulfide bonds in the silk fibroin and maintain protein free-SH groups, so that the exposure of hydrophobic amino acid is induced, and the hydrophobic effect of the silk fibroin is improved.
And (5) testing the water contact angle of the silk fibroin adhesive hemostatic powder in the step (5), wherein the water contact angle value of the silk fibroin adhesive hemostatic powder is 146 degrees, which shows that the silk fibroin adhesive hemostatic powder has excellent high hydrophobic property.
The silk fibroin adhesive hemostatic powder of the embodiment has the adhesive strength of 271kPa to the wet pigskin tissue under the wet physiological environment. The Wistar rat is used as an animal hemostasis model, and the silk fibroin adhesion hemostasis powder is used for a Wistar rat liver hemostasis test, so that complete hemostasis can be realized within 5 seconds.
Example 3
The preparation method of the silk fibroin adhesive hemostatic powder comprises the following steps:
(1) And (3) putting 25.0g of raw silkworm silks into 10.0L of 0.02mol/L sodium carbonate solution, boiling for 20 minutes at 100 ℃, fully washing with deionized water, removing sericin, and drying in a 60 ℃ oven to obtain the degummed silk. Dissolving 13.5g of degummed silk in 50.0mL of a ternary solution system with the molar ratio of calcium chloride/ethanol/water being 1.
(2) Dissolving 500mg of tris (2-carboxyethyl) phosphine hydrochloride in 10mL of deionized water, and fully dissolving to maintain the pH value to be 5.0; then, 1.0g of silk fibroin powder was dissolved in 10mL of an aqueous solution containing tris (2-carboxyethyl) phosphine hydrochloride, and the solution was left at room temperature to be subjected to reduction treatment for 20 minutes, thereby obtaining a reduction-treated silk fibroin solution having a concentration of 10.0 wt%.
(3) Dissolving 1.0g of epigallocatechin gallate in 10.0mL of deionized water, and stirring under nitrogen protection environment to dissolve uniformly to obtain epigallocatechin gallate solution with concentration of 100 mg/mL.
(4) Mixing the silk fibroin solution and the epigallocatechin gallate solution at room temperature, wherein the silk fibroin solution and the epigallocatechin gallate solution are subjected to the reduction reaction treatment in the same volume; adding growth factor which accounts for 0.1% of the weight of the hemostatic powder, maintaining the pH value at 5.0, and rapidly stirring and mixing to obtain insoluble aggregate.
(5) And (3) washing the insoluble aggregate obtained in the step (4) with deionized water for 3 times, placing the insoluble aggregate in a freeze dryer for vacuum freeze drying for 50 hours at the temperature of minus 80 ℃, grinding and crushing the freeze-dried solid, and sieving the crushed solid with a sieve of between 100 and 200 meshes to obtain hemostatic powder, namely the silk fibroin adhesive hemostatic powder. The hemostatic powder has a size of 75-150 μm.
And (3) testing the free-SH group content of the silk fibroin solution subjected to the reduction reaction treatment in the step (2), wherein the free-SH group content of the silk fibroin subjected to the reduction reaction treatment is 135.9 mu mol/g. And (3) testing the zeta potential value of the silk fibroin solution subjected to the reduction reaction treatment in the step (2), wherein the zeta potential value of the silk fibroin subjected to the reduction reaction treatment is +8.5mV. Therefore, the reduction reaction treatment of the silk fibroin by the tris (2-carboxyethyl) phosphine hydrochloride can effectively decompose disulfide bonds in the silk fibroin and maintain protein free-SH groups, so that the exposure of hydrophobic amino acid is induced, and the hydrophobic effect of the silk fibroin is improved.
And (5) testing the water contact angle of the silk fibroin adhering hemostatic powder in the step (5), wherein the water contact angle value of the silk fibroin adhering hemostatic powder is 155 degrees, which shows that the silk fibroin adhering hemostatic powder has excellent high hydrophobic property.
The silk fibroin adhesive hemostatic powder of the embodiment has the adhesive strength to the wet pigskin tissue under the wet physiological environment of 265kPa. The Wistar rat is used as an animal hemostasis model, and the silk fibroin adhesion hemostasis powder is used for a Wistar rat liver hemostasis test, so that complete hemostasis can be realized within 5 seconds.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (10)
1. A preparation method of silk fibroin adhesive hemostatic powder is characterized in that silk fibroin obtained after reduction reaction treatment by thiol reducing agents is used as a raw material and is uniformly mixed with polyphenol compounds to prepare the silk fibroin adhesive hemostatic powder; the powder size of the silk fibroin adhesive hemostatic powder is 75-150 μm.
2. The method for preparing the silk fibroin adhesive hemostatic powder of claim 1, comprising the steps of:
(1) Dissolving degummed silk in lithium bromide or calcium chloride/absolute ethyl alcohol/water ternary solution, dialyzing with deionized water, and performing centrifugal filtration and vacuum freeze drying on the dialyzed solution to obtain silk fibroin powder;
(2) Dissolving the silk fibroin powder obtained in the step (1) in an aqueous solution containing thiol reducing agents, maintaining the pH value at 5.0, and placing the solution at room temperature for reduction reaction treatment to obtain a reduction reaction treated silk fibroin solution;
(3) Dissolving polyphenol compounds in deionized water, and stirring in a nitrogen-protected environment to uniformly dissolve the polyphenol compounds to obtain polyphenol compound solution;
(4) Mixing the silk fibroin solution subjected to the reduction reaction in the step (2) and the polyphenol compound solution in the step (3) with the same volume at room temperature, and maintaining the pH value at 4.0-6.0 to obtain an insoluble aggregate;
(5) And (4) washing, freeze-drying, grinding, crushing and sieving the insoluble aggregate obtained in the step (4) to obtain the silk fibroin adhesive hemostatic powder.
3. The method for preparing the silk fibroin adhesive hemostatic powder of claim 2, wherein in the step (2), the concentration of the silk fibroin solution subjected to the reduction reaction is 1-10wt%, and the zeta potential value thereof is greater than +5mV.
4. The method for preparing the silk fibroin adhesive hemostatic powder of claim 2, wherein in the step (2), the thiol reducing agent includes, but is not limited to, tris (2-carboxyethyl) phosphine hydrochloride; the concentration of the thiol reducing agent is 10.0-50.0 mg/mL; the reduction reaction treatment time is 5-20 minutes at room temperature.
5. The method for preparing the silk fibroin adhesive hemostatic powder of claim 2, wherein in the step (3), the concentration of the polyphenol compound is 50.0-500.0mg/mL; the polyphenol compounds include, but are not limited to, epicatechin gallate, catechin, epicatechin, epigallocatechin, gallic acid, catechin gallate, epigallocatechin gallate, and tannic acid.
6. The method for preparing the silk fibroin adhesive hemostatic powder of claim 2, wherein in the step (4), bioactive molecules are further added during mixing at room temperature, and the bioactive molecules account for 0.1-10% of the weight of the silk fibroin adhesive hemostatic powder.
7. The method for preparing the silk fibroin adhesive hemostatic powder of claim 6, wherein the bioactive molecule comprises one or more of hydroxyapatite, bioglass, collagen, gelatin, antibacterial drugs, growth factors, chitosan, alginic acid, hyaluronic acid, polyethylene glycol, and silicone oil.
8. The method for preparing the silk fibroin adhesive hemostatic powder of claim 2, wherein in the step (5), a screen mesh of 100-200 meshes is adopted for sieving.
9. The silk fibroin adhesive hemostatic powder is characterized by being prepared by the preparation method of the silk fibroin adhesive hemostatic powder according to any one of claims 1-8.
10. The application of the silk fibroin adhesive hemostatic powder is characterized in that the silk fibroin adhesive hemostatic powder is applied to the rapid hemostatic effect of irregular-shaped and incompressible bleeding wound tissues in a wet physiological environment, the effective liquid leakage sealing is realized, the healing and repair of the wound tissues are promoted, and the wound bacteriostasis is realized.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211481795.3A CN115814144A (en) | 2022-11-24 | 2022-11-24 | Silk fibroin adhesive hemostatic powder and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211481795.3A CN115814144A (en) | 2022-11-24 | 2022-11-24 | Silk fibroin adhesive hemostatic powder and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115814144A true CN115814144A (en) | 2023-03-21 |
Family
ID=85531119
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211481795.3A Pending CN115814144A (en) | 2022-11-24 | 2022-11-24 | Silk fibroin adhesive hemostatic powder and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115814144A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110420348A (en) * | 2019-07-19 | 2019-11-08 | 南通纺织丝绸产业技术研究院 | A kind of fibroin albumen hemostatic material and preparation method thereof |
-
2022
- 2022-11-24 CN CN202211481795.3A patent/CN115814144A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110420348A (en) * | 2019-07-19 | 2019-11-08 | 南通纺织丝绸产业技术研究院 | A kind of fibroin albumen hemostatic material and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| HUAN LIU ET AL.: "Bio-Inspired Self-Hydrophobized Sericin Adhesive with Tough Underwater Adhesion Enables Wound Healing and Fluid Leakage Sealing", 《ADVANCED FUNCTIONAL MATERIALS》, vol. 32, no. 32, pages 2 - 3 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yuan et al. | A physicochemical double cross-linked multifunctional hydrogel for dynamic burn wound healing: shape adaptability, injectable self-healing property and enhanced adhesion | |
| Singh et al. | Chitin and chitosan: biopolymers for wound management | |
| EP1030698B1 (en) | Bi-composite collagen material, method for obtaining same and therapeutic applications | |
| EP2555804B1 (en) | Hemostatic sponge | |
| JP2021007754A (en) | In situ crosslinkable polymer composition and method for producing the same | |
| Kim et al. | Mussel-inspired poly (γ-glutamic acid)/nanosilicate composite hydrogels with enhanced mechanical properties, tissue adhesive properties, and skin tissue regeneration | |
| Zheng et al. | Recent progress in surgical adhesives for biomedical applications | |
| KR101865781B1 (en) | Hydrogel comprising oxidized polysaccharide and amine-modified hyaluronic acid for wound dressings and manufacturing method thereof | |
| KR102106487B1 (en) | Double crosslinked biocompatible hemostatic composition and preparation method thereof | |
| EP3308807B1 (en) | Porous structure and method for manufacturing same | |
| CN102580138A (en) | Polysaccharide composite film for arresting bleeding and preparation method thereof | |
| JP6501913B2 (en) | Composition for treating chronic wounds, method for producing the same, and dressing material for treating chronic wounds using the same | |
| CN104208741A (en) | Chitosan based adhesive bandage | |
| EP3558402A1 (en) | Injectable hydrogels and applications thereof | |
| EP2231134A1 (en) | Biocompatible and biodegradable biopolymer matrix | |
| KR102302405B1 (en) | Micro-beads and Method for producing thereof, Hemostatic dressings including the same | |
| CN106540310B (en) | Absorbable rapid hemostatic material and preparation method thereof | |
| CN115814144A (en) | Silk fibroin adhesive hemostatic powder and preparation method and application thereof | |
| CN104497345A (en) | Preparation method of hyaluronic acid-chitosan degradable dressing | |
| KR19990049107A (en) | Manufacturing method of polymer electrolyte composite sponge | |
| CN115737900A (en) | Silk fibroin medical tissue adhesive and preparation method and application thereof | |
| Xu et al. | Etamsylate loaded oxidized Konjac glucomannan-ε-polylysine injectable hydrogels for rapid hemostasis and wound healing | |
| KR100569227B1 (en) | Preparation methods of chitosan-gelatin-algin sponge using ultrasonication | |
| CN115779138B (en) | Silk fibroin hemostatic powder and preparation method and application thereof | |
| KR102494734B1 (en) | Bio absorbable hydrogel hemostatic |
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 |