CN116983461A - Hemostatic sponge system based on liposome modification - Google Patents
Hemostatic sponge system based on liposome modification Download PDFInfo
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- CN116983461A CN116983461A CN202310242013.9A CN202310242013A CN116983461A CN 116983461 A CN116983461 A CN 116983461A CN 202310242013 A CN202310242013 A CN 202310242013A CN 116983461 A CN116983461 A CN 116983461A
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- Prior art keywords
- alginate
- liposome
- polypeptide
- hemostatic
- hemostatic sponge
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- 230000002439 hemostatic effect Effects 0.000 title claims abstract description 40
- 239000002502 liposome Substances 0.000 title claims abstract description 31
- 230000004048 modification Effects 0.000 title description 3
- 238000012986 modification Methods 0.000 title description 3
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 claims abstract description 27
- 229940072056 alginate Drugs 0.000 claims abstract description 27
- 235000010443 alginic acid Nutrition 0.000 claims abstract description 27
- 229920000615 alginic acid Polymers 0.000 claims abstract description 27
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 13
- 108091005601 modified peptides Proteins 0.000 claims abstract description 8
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 7
- 238000004132 cross linking Methods 0.000 claims abstract description 6
- 229920001184 polypeptide Polymers 0.000 claims abstract description 6
- 238000004108 freeze drying Methods 0.000 claims abstract description 5
- 239000000017 hydrogel Substances 0.000 claims abstract description 5
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 5
- 238000013329 compounding Methods 0.000 claims abstract description 4
- 230000003993 interaction Effects 0.000 claims abstract description 4
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 239000000499 gel Substances 0.000 claims abstract description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 3
- 206010052428 Wound Diseases 0.000 claims description 12
- 208000027418 Wounds and injury Diseases 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 7
- 230000000740 bleeding effect Effects 0.000 claims description 4
- 238000007710 freezing Methods 0.000 claims description 4
- 230000008014 freezing Effects 0.000 claims description 4
- 230000001788 irregular Effects 0.000 claims description 4
- 238000011049 filling Methods 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims description 2
- 239000002131 composite material Substances 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 230000015271 coagulation Effects 0.000 abstract description 5
- 238000005345 coagulation Methods 0.000 abstract description 5
- 238000000034 method Methods 0.000 abstract description 3
- 230000035876 healing Effects 0.000 abstract 1
- 231100000956 nontoxicity Toxicity 0.000 abstract 1
- 230000002195 synergetic effect Effects 0.000 abstract 1
- 230000023555 blood coagulation Effects 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 4
- 206010053567 Coagulopathies Diseases 0.000 description 3
- 208000032843 Hemorrhage Diseases 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 230000035602 clotting Effects 0.000 description 3
- 230000008961 swelling Effects 0.000 description 3
- 206010060964 Arterial haemorrhage Diseases 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 229920001661 Chitosan Polymers 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 230000002522 swelling effect Effects 0.000 description 2
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- PHOQVHQSTUBQQK-SQOUGZDYSA-N D-glucono-1,5-lactone Chemical compound OC[C@H]1OC(=O)[C@H](O)[C@@H](O)[C@@H]1O PHOQVHQSTUBQQK-SQOUGZDYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 238000001266 bandaging Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000012137 double-staining Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 238000004442 gravimetric analysis Methods 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 239000012890 simulated body fluid Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
Classifications
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/08—Polysaccharides
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0036—Porous materials, e.g. foams or sponges
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/108—Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106
-
- 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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
Landscapes
- Health & Medical Sciences (AREA)
- Surgery (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Materials For Medical Uses (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a hemostatic sponge system constructed by compounding polypeptide modified liposome and alginate, which comprises the following steps: (1) grafting a fatty chain to the amino terminus of the platelet-binding peptide; (2) Inserting the modified peptide of step (1) into a liposome using hydrophobic interactions; (3) Uniformly mixing the liposome with an alginate solution, and forming hydrogel through double cross-linking; (4) And (3) freeze-drying the gel obtained in the step (3) to obtain the hemostatic sponge. The modified liposome and the alginate have synergistic hemostatic performance, so that the coagulation components at the wound can be effectively enriched to accelerate the coagulation process, and the biocompatibility and the non-toxicity of the alginate can promote the healing of the wound.
Description
Technical Field
The invention relates to a hemostatic sponge system constructed by compounding polypeptide modified liposome and alginate, which can be used for filling and stopping bleeding of grooves or irregular wounds.
Background
Effective hemostatic management is critical to global military and civilian wound care when bleeding loss due to trauma can be life threatening. Within minutes after uncontrolled bleeding, the survival rate of wounded can be improved by effective hemostatic intervention. Gauze and bandages are typical of traditional hemostatic materials and they do not exhibit an effective hemostatic effect in the face of extreme wound environments such as incompressible penetrating arterial bleeding wounds and irregular deep arterial bleeding. In recent years, development of novel hemostatic materials is faster and faster, and different forms of hemostatic materials are developed to cope with different types of wound environments, for example, patent CN202021887276 discloses a hemostatic bandaging device carrying polypeptide liposome, and a transmission assembly is used for improving medical staff to uniformly smear medicament on bandages; patent CN201510536381 discloses a hemostatic adhesive containing amino acid liposome and a preparation method thereof, which can actively provide nutrition or active substances for wound surfaces; patent CN202211268724 discloses a GelMA-DA/quaternized chitosan/glycerin composite hemostatic sponge material and a preparation method thereof, which have good adhesive capacity, proper mechanical property, good biocompatibility, obvious antibacterial property and excellent blood coagulation hemostatic property. Compared with the materials, firstly, different liposome modification means are selected, the hemostatic principle of the modified substances is completely different, in addition, compared with chitosan, the alginate used by the modified substances has undoubtedly excellent biocompatibility, and the high water absorption capacity of the sponge can effectively promote the aggregation of blood coagulation components in blood and accelerate the blood coagulation.
Disclosure of Invention
The invention aims to construct a hemostatic sponge system, which is prepared by embedding polypeptide modified liposome into double-crosslinked alginate. The invention provides a hemostasis method capable of filling irregular wounds such as grooves and the like.
In order to realize the technical scheme, the invention relates to a hemostatic sponge system constructed by compounding polypeptide modified liposome and alginate, which comprises the following steps:
(1) Grafting a fatty chain to the amino terminus of the platelet-binding peptide;
(2) Inserting the modified peptide of step (1) into a liposome using hydrophobic interactions;
(3) Uniformly mixing the liposome with an alginate solution, and forming hydrogel through double cross-linking;
(4) And (3) freeze-drying the gel obtained in the step (3) to obtain the hemostatic sponge.
The fatty chain to which the invention relates may be C 16 Or C 19 The platelet-binding peptide sequence was HHLGGAKQAGDV (seq_1).
The alginate concentration in the step (3) is 2% -10%, and the volume ratio of the liposome to the alginate is 1:5, 1:3 or 1:1.
The double crosslinking in the step (3) of the present invention is Ca 2+ Crosslinking and photocrosslinking, wherein the photocrosslinking is AlgMA-LP.
In the freeze-drying treatment in the step (4), the pre-freezing temperature of the sample is-80 ℃ and the pre-freezing time is 48 hours.
Compared with the prior art, the invention has the following advantages: (1) The material cost is low, the preparation method is simple, the use and operation are easy, and the large-scale production and the preservation are easy; (2) The prepared hemostatic dressing has good biocompatibility and no cytotoxicity; (3) The alginate hemostatic sponge has excellent swelling performance, can absorb excessive body fluid at a wound, and can maintain a proper physiological environment at the wound; (4) The liposome-platelet binding peptide can synergistically enrich coagulation components with alginate, and improve hemostatic efficiency.
Description of the drawings:
FIG. 1 is a modified peptide molecular structure of example 1 according to the present invention;
FIG. 2 is a modified peptide insertion liposome according to example 1 of the present invention;
FIG. 3 is the swelling characteristics of the hemostatic sponge of example 1 according to the present invention;
FIG. 4 is the clotting time of the hemostatic sponge of example 1 according to the present invention;
FIG. 5 is an experiment of hemostatic sponge cells according to example 1 of the present invention.
Detailed description of the invention
The invention is further illustrated by the following examples and the accompanying drawings.
Example 1:
(1) Grafting of fatty chains to platelet-binding peptides
Starting from the C-terminus, synthesis was performed by Fmoc solid phase synthesis, followed by grafting of fatty chain C on the N-terminal portion 16 Imparting hydrophobic character, and molecular structure is shown in figure 1.
(2) Insertion of modified peptides into liposomes
The liposome is dissolved in chloroform to form a film by utilizing hydrophobic interaction, then surfactant is added to dissolve, the liposome film is broken by ultrasonic, and finally the modified peptide is added to absorb the surfactant in liquid by utilizing macroporous resin, so that the self-assembly of the modified peptide and the liposome is completed, and the molecular structure is shown in figure 2.
(3) Preparation of liposome-peptide and alginate hydrogels
After mixing the liposome with 2% (w/v) alginate solution at a volume ratio of 1:5, 2mg of calcium carbonate powder, 10mg of AlgMA, 2mg of LP and 3mg of glucolactone were added, and stirred at room temperature for 1 hour, then the solution was poured into a cell culture dish having a diameter of 3cm, and after standing at room temperature for 3 hours, it was irradiated with 365nm ultraviolet lamp for 20 minutes.
(4) Preparation of hemostatic sponge
The hydrogel after ultraviolet irradiation was pre-frozen in a refrigerator at-80 ℃ for 48 hours, and then freeze-dried for 24 hours.
(5) Swelling Property of hemostatic sponge
Swelling behaviour of hemostatic sponges was quantitatively studied using gravimetric analysis. Sponge (weight m) 0 ) Immersed in a simulated body fluid at 37℃and the weight (m i ) And uses the formula r= (m i –m 0 )/m 0 (i=1, 2,3 …) the expansion ratio was calculated in hours, and the result is shown in fig. 3.
(6) Coagulation experiment
In vitro coagulation: 5mg of sponge and 800uL of antagonistic blood were placed in a 2mL centrifuge tube, incubated at 37℃and the blood clotting time was measured.
In vivo experiments: a cylindrical sponge with the diameter of 2cm is placed on the cut rat leg artery wound to be filled, the wound is not permeated with blood any more as a blood coagulation index, and the blood coagulation time is recorded. Wherein the living body adopts SD rat, and the blood coagulation experimental result is shown in figure 4.
(7) Cell experiment
NIH-3T3 cells were incubated with Dulbecco's modified Eagle's Medium (10% bovine serum added) at 37℃with 5% CO 2 Culturing in a humidity incubator. Cell culture media pre-incubated overnight with hemostatic sponges was used to culture NIH-3T3 cells for 24h. Cells were subjected to fluorescent photographing by the dead and live double staining method, and the cell state was recorded, and the result is shown in fig. 5.
Example 2:
this example is identical to example 1 except that the alginate concentration in step (3) is 5% and 10%.
Example 3:
this example and example 1 are identical except that the volume ratio of liposome to alginate in step (3) is 1:3 and 1:1.
Tests show that the hemostatic sponge prepared in example 2 has similar properties to example 1 except that the swelling property and clotting time are different from those of example 1; the hemostatic sponge prepared in example 3 has similar properties to example 1 except that the clotting time is somewhat different from that of example 1.
Claims (6)
1. A hemostatic sponge system constructed by compounding polypeptide modified liposome and alginate, which is characterized by comprising the following steps:
(1) Grafting a fatty chain to the amino terminus of the platelet-binding peptide;
(2) Inserting the modified peptide of step (1) into a liposome using hydrophobic interactions;
(3) Uniformly mixing the liposome with an alginate solution, and forming hydrogel through double cross-linking;
(4) And (3) freeze-drying the gel obtained in the step (3) to obtain the hemostatic sponge.
2. The hemostatic sponge system comprising polypeptide-modified liposomes complexed with alginate as claimed in claim 1, wherein the fatty chain is C 16 Or C 19 The platelet-binding peptide sequence was HHLGGAKQAGDV (seq_1).
3. The hemostatic sponge system constructed from polypeptide-modified liposomes complexed with alginate as claimed in claim 2, wherein the alginate concentration in step (3) is 2% -10% and the volume ratio of liposome to alginate is 1:5, 1:3 or 1:1.
4. A hemostatic sponge system constructed from polypeptide-modified liposomes complexed with alginate as claimed in claim 3, wherein the double cross-links in step (3) are Ca 2+ Crosslinking and photocrosslinking, wherein the photocrosslinking is AlgMA-LP.
5. A hemostatic sponge system constructed from polypeptide-modified liposomes complexed with alginate as claimed in claim 3, wherein the pre-freezing temperature of the sample is-80 ℃ and the pre-freezing time is 48 hours during the lyophilization treatment in step (4).
6. The hemostatic sponge system constructed from polypeptide-modified liposomes and alginate composite according to any one of claims 1-5, wherein the hemostatic material is effective in filling irregular wounds such as grooves and the like and stopping bleeding through the high water absorption capacity of polypeptide-targeted platelets and alginate sponge.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310242013.9A CN116983461A (en) | 2023-03-14 | 2023-03-14 | Hemostatic sponge system based on liposome modification |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310242013.9A CN116983461A (en) | 2023-03-14 | 2023-03-14 | Hemostatic sponge system based on liposome modification |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116983461A true CN116983461A (en) | 2023-11-03 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310242013.9A Pending CN116983461A (en) | 2023-03-14 | 2023-03-14 | Hemostatic sponge system based on liposome modification |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116983461A (en) |
-
2023
- 2023-03-14 CN CN202310242013.9A patent/CN116983461A/en active Pending
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