CN111214694A - Dressing with functions of stopping bleeding and accelerating wound healing and preparation method thereof - Google Patents

Dressing with functions of stopping bleeding and accelerating wound healing and preparation method thereof Download PDF

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CN111214694A
CN111214694A CN202010243275.3A CN202010243275A CN111214694A CN 111214694 A CN111214694 A CN 111214694A CN 202010243275 A CN202010243275 A CN 202010243275A CN 111214694 A CN111214694 A CN 111214694A
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polydeoxyribonucleotide
smpdrn
hemostatic
wound healing
dressing
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CN111214694B (en
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王超云
高原
董书萍
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Hengyu Biopharmaceutical Shandong Co ltd
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Shandong Big Fish Biotechnology Co Ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L15/00Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/20Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing organic materials
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L15/00Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42Use of materials characterised by their function or physical properties
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L15/00Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42Use of materials characterised by their function or physical properties
    • A61L15/44Medicaments
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/258Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/402Anaestetics, analgesics, e.g. lidocaine
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404Biocides, antimicrobial agents, antiseptic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2400/00Materials characterised by their function or physical properties
    • A61L2400/04Materials for stopping bleeding

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Abstract

The invention discloses a dressing with the functions of stopping bleeding and accelerating wound healing, which comprises small molecular poly-deoxyribonucleotide (SMPDRN) extracted from salmon. The SMPDRN is extracted from salmon tissues, and the salmon tissues comprise spermary, semen and eggs of salmon. The experiment of the invention proves that the invention can achieve the effects of stopping bleeding, easing pain, diminishing inflammation, promoting wound healing and the like. The small-molecule polydeoxyribonucleotide (SMPDRN) used in the invention is obtained by the preparation method provided by the invention, has the characteristics of molecular weight concentration in a high-efficiency area, relatively simple components, high purity, simple process, stable components and the like, meets the preparation requirement of the hemostatic dressing, ensures that the hemostatic dressing has stable curative effect and ideal safety, overcomes the difficulty of applying animal preparations to the preparation of the hemostatic dressing, and develops new application of the small-molecule polydeoxyribonucleotide in the field of medicine.

Description

Dressing with functions of stopping bleeding and accelerating wound healing and preparation method thereof
Technical Field
The invention relates to a dressing with the functions of stopping bleeding and accelerating wound healing and a preparation method thereof.
Background
Trauma blood loss can cause pain, coma and shock of patients, and the trauma can cause local inflammation, cause healing delay and form abnormal scar and keloid; the serious infection causes toxemia and septicemia, and seriously endangers human health and life safety. It is statistically calculated that massive blood loss is the main cause of war injury to members who are reduced by 50% of the war. In addition, excessive blood loss is also a key factor causing the rise of casualties in peace time. The rapid hemostasis is realized, the synthesis of collagen is accelerated, the collagen is re-epithelialized, and the epidermis is sealed, so that the blood loss can be reduced, and the treatment time is saved; on the other hand, the stable internal environment is maintained, the infection is reduced, the normal proceeding of healing is ensured, and a good prognosis effect is obtained.
At present, the variety of the hemostatic drugs is various, more than 70 hemostatic drugs are reported only in Chinese patent documents, and the number of the hemostatic drugs is estimated to be hundreds in addition to the products sold in the market, and the hemostatic drugs are designed according to four stages of wound healing. Although they have some hemostatic effect, the limitations are also prominent: only has the efficacy of one or two periods in the wound healing process, and can not take multiple periods into consideration and play a role together; the medicine has complex formula and more components; some pharmaceutical compositions contain toxic ingredients; the hemostasis speed is slow, and the wound healing is slow; the use and carrying are inconvenient; the components are special and expensive.
With the intensive research on wound healing, analgesia, infection resistance and healing promotion become necessary for the development of hemostatic materials. The search for a novel rapid hemostatic material is a problem which needs to be solved urgently and is highly valued by various national and military medicines in the world. Italian fishermen find that the semen in the spermary of the salmon is extruded and periodically smeared on the wound, so that the wound fester can be effectively inhibited, and the wound healing is promoted. The functional substances in the medical composition have great potential in the medical treatment of wound blood loss.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a medical auxiliary material taking salmon micromolecule polydeoxyribonucleotide as an active ingredient.
The invention takes the spermary, the semen and the ovum of the salmon as materials, and obtains Small Molecular Polydeoxyribonucleotide (SMPDRN) by utilizing the unique extraction process, wherein the SMPDRN is n which is more than or equal to 50bp and less than or equal to 500bp in length. The inventor finds that the SMPDRN has obvious effects of stopping bleeding, promoting collagen synthesis, accelerating wound healing, relieving pain and resisting inflammation through cell, animal and human body experiments. The substance has the characteristics of rapid hemostasis, anti-infection and pain relief, and medical materials such as hemostatic patches, wet tissues, gels and the like produced by taking the substance as dressing are favorable for solving the comprehensive problem of clinical wound hemostasis and improving the life quality of patients.
The specific technical scheme of the invention is as follows:
one of the objects of the present invention is to provide a dressing having hemostatic and wound healing accelerating functions, comprising Small Molecule Polydeoxyribonucleotides (SMPDRNs) extracted from salmon.
The salmon comprises Atlantic salmon, Pacific salmon or rainbow trout.
The SMPDRN is extracted from salmon tissues, and the salmon tissues comprise spermary, semen and eggs of salmon. The experiment of the invention proves that the invention can achieve the effects of stopping bleeding, easing pain, diminishing inflammation, promoting wound healing and the like.
Preferably, the hemostatic dressing provided by the invention can further comprise other multiple medicines, and the other medicines can be at least one of antibacterial medicines and analgesics.
More preferably, the antibacterial agent is at least one of borneol, maculownin and erythromycin; the analgesic is at least one of morphine and fentanyl.
The pain drug is at least one of morphine and fentanyl.
Preferably, the dressing comprises a drug carrier and a liquid medicine, and the drug carrier can be medical gauze, non-woven fabric or gel. If the dressing is a band-aid, the dressing carrier is a gauze or non-woven fabric drug core, and the quantity of SMPDRN is 0.3-20.5 mg per square centimeter of drug core; if the dressing is a wound healing wet tissue, the drug carrier is non-woven fabric, and the amount of SMPDRN is 14-260 mg per gram of wet tissue carrier; if the dressing is gel, the content of SMPDRN in the gel system is 0.2-30 wt%.
The molecular weight of SMPDRN in the dressing is 50 bp-500 bp. Preferably, the mass of the fragment with the molecular weight of 100 bp-500 bp accounts for more than 90 wt% of the total mass of the SMPDRN.
The dressing can contain antibacterial drugs or/and analgesic drugs. Administration can be by transdermal, intramuscular or subcutaneous, or mucocutaneous routes. Can be in the form of patch, colloid preparation, emulsion, solution, powder, etc. Can be prepared by a conventional method of pharmacy.
The SMPDRN single body or mixture has the effects of hemostasis, immunoregulation, antibiosis, anti-inflammation, peripheral analgesia and the like; the SMPDRN single body or mixture has the effects of stopping bleeding, resisting bacteria and diminishing inflammation on a wound part; the SMPDRN single body or mixture can induce the growth of fibroblasts and promote the synthesis of collagen at a wound part; SMPDRN promotes the proliferation and migration of endothelial cells and keratinocytes.
Preferably, the preparation method of the small molecule polydeoxyribonucleotide comprises the steps of tissue lysis, neutralization, centrifugation, alcohol precipitation and centrifugal purification. Drying of the final product may also be included, preferably low temperature vacuum drying.
In order to obtain the desired molecular fragment, the applicant further optimizes the original SMPDRN extraction process based on the invention patent "a small molecule polydeoxyribonucleotide and its preparation and application" of application No. 201911191251.1 (published) to obtain SMPDRN with hemostatic and wound healing accelerating effects, wherein the preparation method comprises the following steps:
(1) cracking salmon tissues by using an alkali cracking solution, wherein the alkali cracking solution contains 1-3 mol/L NaOH, 50-200 mM NaCl and 5-25 mg/ml proteinase K;
(2) adding Tris-HCl into the reaction system obtained in the step (1);
(3) adding HCL into the reaction system obtained in the step (2);
(4) centrifuging the reaction system obtained in the step (3), and collecting a supernatant;
(5) adding isopropanol into the supernatant obtained in the step (4), and standing for more than 30 min;
(6) and (5) centrifuging the reaction system obtained in the step (5), and reserving the precipitate to obtain the micromolecule polydeoxyribonucleotide.
Further preferred preparation methods are disclosed by comparison with the preparation method of SMPDRN as it is originally in 201911191251.1, as shown in the following table:
Figure BDA0002433260730000041
Figure BDA0002433260730000051
the preparation method of SMPDRN can further use a vacuum freeze dryer to dry to obtain precipitates.
The invention also aims to provide a preparation method of a plurality of medical dressings which take salmon small molecular polydeoxyribonucleotide as an effective component, and the medical dressings comprise band-aid, hemostatic gel and wound healing wet tissue.
A preparation method of a band-aid uses salmon small molecular poly-deoxyribonucleotide as an effective component, and comprises the following steps:
(1) preparing an antibacterial liquid;
(2) dissolving small molecular poly-deoxyribose nucleotide in normal saline to fully dissolve the small molecular poly-deoxyribose nucleotide, slowly passing the drug core through the solution, extruding, rotating and drying at the temperature of below 70 ℃, spraying the dried drug core on the antibacterial liquid obtained in the step (1), cutting, rewinding and packaging to obtain a finished product.
Preferably, the specific working conditions of step (1) of the preparation method of the adhesive bandage are as follows: weighing 0.5-1.0 g of borneol, adding 80-120 mL of 75-95 wt% ethanol, and fully dissolving to obtain borneol liquid.
Preferably, the specific working conditions of step (1) of the preparation method of the adhesive bandage are as follows: 0.01-0.5 g of motipocin is weighed, and 70-130 mL of sterile water is added to fully dissolve the motipocin to prepare the antibiotic solution.
Preferably, the specific working conditions of step (2) of the preparation method of the adhesive bandage are as follows:
weighing 0.4-28 g of small molecular poly-deoxyribonucleotide (SMPDRN), adding 70-600 mL of normal saline to fully dissolve the small molecular poly-deoxyribonucleotide, slowly passing the drug core through the solution to ensure that the drug core per square centimeter contains 0.3-20.5 mg of SMPDRN, extruding, rotating and drying below 70 ℃, installing the dried drug core on an integrated machine for preparing the band-aid, spraying the antibacterial liquid obtained in the step (1), cutting, rewinding and packaging to obtain a finished product.
Preferably, the specification of the band-aid can be 1.5cm × 2.3cm or 4.5cm × 6.0cm or 2.5cm × 4.7cm or 7.2cm × 2.5 cm.
A preparation method of hemostatic gel, which uses salmon small molecular polydeoxyribonucleotide as an effective component, comprises the following steps:
(1) dissolving hyaluronic acid in deionized water to prepare 1-10 wt% solution, and filtering to remove insoluble substances for later use;
(2) preparing 0.6-90 wt% aqueous solution of polydeoxyribonucleotide;
(3) preparing 0.8-2 wt% of gel aqueous solution;
(4) mixing the solutions obtained in the steps (1), (2) and (3), stirring, and homogenizing to obtain a gel system;
(5) and (4) subpackaging, sealing and sterilizing the mixture obtained in the step (4).
Preferably, the specific working conditions of the step (4) in the preparation method of the hemostatic gel are that the solutions obtained in the step (1), the step (2) and the step (3) are mixed in equal mass proportion.
Preferably, the gel in step (3) in the preparation method of the hemostatic gel is one or more of carbomer, sodium carboxymethylcellulose, polyethylene glycol, polyvinyl alcohol, acacia, gelatin, guar gum, polyvinyl pyrrolidone, sodium alginate and polylactic acid. Further preferred is carbomer or a mixture of carbomer and polyethylene glycol.
Preferably, antibacterial agent accounting for 0-2 wt% of the gel system and analgesic agent accounting for 0-2 wt% of the gel system can be added according to requirements.
Preferably, antioxidants and preservatives can be added to the hemostatic gel as desired.
Further preferably, the mass ratio of the antioxidant to the preservative to the gel system is 0.05:0.02: 100.
Preferably, the molecular weight of the hyaluronic acid is 1 × 106~2.2×106Da。
A preparation method of a wound healing wet tissue, which uses salmon micromolecule polydeoxyribonucleotide as an effective component, comprises the following steps:
the non-woven fabric is used as a wet tissue carrier, and wet tissue immersion liquid with the weight 1-5 times that of the wet tissue carrier is sprayed and dripped on the wet tissue carrier;
the wet tissue immersion liquid comprises the following components in parts by weight: 3-7 parts of glycerol, 2-5 parts of alkyl glycoside, 2-4 parts of squalane, 1-10 parts of micromolecular polydeoxyribonucleotide, 0.05-0.5 part of antibacterial agent, 0.1-0.5 part of surfactant, 0.1-0.6 part of mildew preventive and 60-90 parts of deionized water.
Preferably, the antibacterial agent is one or a mixture of two of the mopidacin and the erythromycin.
Preferably, the surfactant is one or a mixture of fatty alcohol-polyoxyethylene ether sodium sulfate and lauryl alcohol sodium sulfate.
Preferably, the mildew preventive is PB 205.
The invention has the following beneficial effects:
the invention applies the salmon micromolecule polydeoxyribonucleotide to the preparation of the hemostatic dressing, so that the hemostatic dressing has the effects of hemostasis of wound parts, immunoregulation, antibiosis, anti-inflammation, peripheral analgesia and the like. The small-molecule polydeoxyribonucleotide (SMPDRN) used in the invention is obtained by the preparation method provided by the invention, has the characteristics of molecular weight concentration in a high-efficiency area, relatively simple components, high purity, simple process, stable components and the like, meets the preparation requirement of the hemostatic dressing, ensures that the hemostatic dressing has stable curative effect and ideal safety, overcomes the difficulty of applying animal preparations to the preparation of the hemostatic dressing, and develops new application of the small-molecule polydeoxyribonucleotide in the field of medicine.
Drawings
FIG. 1 is an agarose gel electrophoresis chart of SMPDRN obtained in the original preparation method and the SMPDRNs obtained in the examples 1 and 2 of the invention;
fig. 2 is a wound healing experiment of the SMPDRN hemostatic gel of example 7;
fig. 3 is a wound healing experiment of the SMPDRN wound dressing of example 2;
FIG. 4 is a comparison of wound healing effects of SMPDRN from example 2 and SMPDRN from original preparation method;
FIG. 5 is a graph of the effect of SMPDRN from example 2 on the level of inflammatory factor IL-1 β in mouse wounds;
in FIG. 1, lanes 1 and 2 show SMPDRN obtained from the original preparation process under application No. 201911191251.1; lane 3 is the SMPDRN obtained in example 1 of the present invention, and lane 4 is the SMPDRN obtained in example 2 of the present invention.
Detailed Description
The principles and features of this invention are described below in conjunction with examples, which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
The reagents and materials used in the examples are commercially available common reagents and materials. Salmon used in the examples was pacific salmon (salmon from the pacific county of black dragon river).
Example 1
A band-aid comprises a drug carrier and a liquid medicine, wherein the liquid medicine contains micromolecular poly-deoxyribonucleotide and borneol.
The preparation method of the adhesive bandage comprises the following steps:
1. the preparation method of the salmon Small Molecular Polydeoxyribonucleotide (SMPDRN) used in the hemostatic dressing comprises the following steps:
(1) accurately weighing 100g of salmon testis, adding 600ml of alkali lysis solution (200mmol of NaCl,2mol/L of NaOH and 20mg/ml of protein K), quickly crushing tissues, reversing and uniformly mixing, and carrying out constant-temperature water bath at 95 ℃ for 20 min.
(2) Cooling in ice to below 25 deg.C, adding 2mol/L Tris-HCl (pH8.0), adding 1/2 accounting for the volume of the reaction system obtained in step (1), and reversing and mixing.
(3) Adding 1mol/L HCL, adding 1/3 accounting for the volume of the reaction system obtained in the step (2), and reversing and mixing.
(4) Centrifuging at 6000rpm for 15min at 0-4 ℃, and collecting supernatant.
(5) And (4) adding isopropanol with the same volume as the supernatant obtained in the step (4), reversing, uniformly mixing, and standing for 30 min.
(6) Centrifuging at 8000rpm for 15min at 0-4 ℃, decanting the supernatant, adding 75 wt% ethanol into the precipitate, blowing gently to cause the precipitate to rise, slightly reversing for several times, centrifuging at 8000rpm for 15min, removing the upper liquid relatively, sucking the residual ethanol with a gun head, and keeping the precipitate.
(7) Taking a small amount of precipitate obtained in step (6), dissolving with TE solution, measuring its concentration, and performing agarose gel electrophoresis.
(8) And (4) drying the precipitate obtained in the step (6) in a vacuum freeze-drying instrument to obtain the SMPDRN freeze-dried powder.
As shown in FIG. 1, lane 3 in the figure is an electropherogram of the SMPDRN obtained in example 1, and the molecular weight length of the electropherogram is 50 bp-500 bp; wherein the mass of the 100 bp-500 bp fragment accounts for more than 90 percent of the total mass.
2. Preparing the adhesive bandage with the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
(1) preparing an antibacterial liquid: weighing 0.5-1.0 g of borneol, adding 80-120 mL of 75-95 wt% ethanol, and fully dissolving to obtain borneol liquid.
(2) Weighing 0.4g of micromolecule polydeoxyribonucleotide, adding 100mL of normal saline to fully dissolve the micromolecule polydeoxyribonucleotide, slowly passing the medicine core through the solution, extruding, rotating and drying at the temperature of below 70 ℃ to enable the medicine core to contain 0.3mg of SMPDRN per square centimeter, extruding, rotating and drying at the temperature of below 70 ℃, installing the dried medicine core on an integrated machine for preparing the band-aid, spraying the antibacterial liquid obtained in the step (1), slitting, rewinding and packaging to obtain a finished product, wherein the specification of the obtained band-aid is 1.5cm multiplied by 2.3cm, and calculating to obtain the SMPDRN contained in each band-aid.
Example 2
A band-aid comprises a drug carrier and a liquid medicine, wherein the liquid medicine contains micromolecular polydeoxyribonucleotide and mupidoticin.
The preparation method of the adhesive bandage comprises the following steps:
1. the preparation method of the salmon Small Molecular Polydeoxyribonucleotide (SMPDRN) used in the hemostatic dressing comprises the following steps:
(1) weighing 100g salmon sperm accurately, adding 600ml alkali lysis solution (200mmol NaCl,2mol/L NaOH,20mg/ml protein K), quickly crushing tissue, reversing and mixing uniformly, and performing constant temperature water bath at 95 ℃ for 40 min.
(2) Cooling in ice to below 25 deg.C, adding 2mol/L Tris-HCl (pH8.0), adding 1/2 accounting for the volume of the reaction system obtained in step (1), and reversing and mixing.
(3) Adding 1mol/L HCL, adding 1/3 accounting for the volume of the reaction system obtained in the step (2), and reversing and mixing.
(4) Centrifuging at 6000rpm for 15min at 0-4 ℃, and collecting supernatant.
(5) And (4) adding isopropanol with the same volume as the supernatant obtained in the step (4), reversing, uniformly mixing, and standing for 30 min.
(6) Centrifuging at 8000rpm for 15min at 0-4 ℃, decanting the supernatant, adding 75 wt% ethanol into the precipitate, blowing gently to cause the precipitate to rise, slightly reversing for several times, centrifuging at 8000rpm for 15min, removing the upper liquid relatively, sucking the residual ethanol with a gun head, and keeping the precipitate.
(7) Taking a small amount of precipitate obtained in step (6), dissolving with TE solution, measuring its concentration, and performing agarose gel electrophoresis.
(8) And (5) drying the precipitate obtained in the step (6) in a vacuum freeze-drying instrument to obtain the SMPDRN freeze-dried powder.
As shown in FIG. 1, lane 4 in the figure is an electropherogram of the SMPDRN obtained in example 2, the molecular weight length is 50bp to 500bp, and the mass of the fragment of 100bp to 500bp accounts for more than 90% of the total mass of the SMPDRN. Example 2 differs from example 1 in the preparation of the SMPDRN only by the time of heating in step (1) thereof.
2. Preparing the adhesive bandage with the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
(1) preparing an antibacterial liquid: 0.1g of motpidoxin is weighed and added with 100mL of sterile water to be fully dissolved, thus obtaining the antibiotic solution.
(2) Weighing 8g of micromolecule polydeoxyribonucleotide, adding 200mL of normal saline to fully dissolve the micromolecule polydeoxyribonucleotide, slowly passing the medicine core through the solution, extruding, rotationally drying at the temperature of below 70 ℃ to ensure that each square centimeter of the medicine core contains 6mg of SMPDRN, extruding, rotationally drying at the temperature of below 70 ℃, installing the dried medicine core on an integrated machine for preparing the band-aid, spraying the antibacterial liquid obtained in the step (1), cutting, rewinding and packaging to obtain a finished product, wherein the specification of the band-aid is 1.5cm multiplied by 2.3cm, and calculating to obtain the band-aid containing 20.7mg of SMPDRN of each band-aid.
Example 3
A band-aid comprises a drug carrier and a liquid medicine, wherein the liquid medicine contains micromolecular polydeoxyribonucleotide and mupidoticin.
The preparation method of the adhesive bandage comprises the following steps:
1. the preparation method of the salmon Small Molecular Polydeoxyribonucleotide (SMPDRN) used in the hemostatic dressing comprises the following steps:
(1) accurately weighing 100g of salmon ovary tissue, adding 800ml of alkaline lysis solution (50mmol of NaCl,1mol/LNaOH and 5mg/ml of protein K), quickly crushing the tissue, reversing and uniformly mixing, and carrying out constant-temperature water bath at 90 ℃ for 40 min.
(2) Cooling in ice to below 25 deg.C, adding 1mol/L Tris-HCl (pH8.0), adding 1/2 accounting for the volume of the reaction system obtained in step (1), and reversing and mixing.
(3) Adding 4mol/L of HCL, adding 1/5 accounting for the volume of the reaction system obtained in the step (2), and reversing and mixing.
(4) Centrifuging at 6000rpm for 20min at 18-25 ℃, and collecting supernatant.
(5) And (4) adding isopropanol with the same volume as the supernatant obtained in the step (4), reversing, uniformly mixing, and standing for 2 h. .
(6) Centrifuging at 8000rpm for 20min at 18-25 ℃, pouring out supernatant, adding 75 wt% ethanol into the precipitate, slightly blowing to enable the precipitate to rise, slightly reversing for several times, centrifuging at 6000rpm for 20min, removing upper liquid relatively, sucking residual ethanol by using a gun head, and keeping the precipitate.
(7) And (5) drying the precipitate obtained in the step (6) in a vacuum freeze-drying instrument to obtain the SMPDRN freeze-dried powder.
2. Preparing the adhesive bandage with the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
(1) preparing an antibacterial liquid: 0.01g of motpidoxin is weighed and added with 70mL of sterile water to be fully dissolved, thus obtaining the antibiotic solution.
(2) Weighing 28g of micromolecule polydeoxyribonucleotide, adding 600mL of normal saline to fully dissolve the micromolecule polydeoxyribonucleotide, slowly passing the medicine core through the solution, extruding, rotationally drying at the temperature of below 70 ℃ to enable the medicine core to contain 20.5mg of SMPDRN per square centimeter, extruding, rotationally drying at the temperature of below 70 ℃, installing the dried medicine core on an integrated machine for preparing the band-aid, spraying the antibacterial liquid obtained in the step (1), cutting, rewinding and packaging to obtain a finished product, wherein the specification of the obtained band-aid is 2.5cm multiplied by 4.7cm, and calculating to obtain the band-aid containing 241mg of SMPDRN per band-aid.
Example 4
A band-aid comprises a drug carrier and a liquid medicine, wherein the liquid medicine contains micromolecular polydeoxyribonucleotide and mupidoticin.
The preparation method of the adhesive bandage comprises the following steps:
1. the preparation method of the salmon Small Molecular Polydeoxyribonucleotide (SMPDRN) used in the hemostatic dressing comprises the following steps:
(1) accurately weighing 100g of salmon testis tissue, adding 1000ml of alkaline lysis buffer (100mmol of NaCl,3mol/LNaOH and 15mg/ml of protein K), quickly crushing the tissue, reversing and uniformly mixing, and carrying out constant temperature water bath at 90 ℃ for 40 min.
(2) Cooling in ice to below 25 deg.C, adding 3mol/L Tris-HCl (pH8.0), adding 1/3 accounting for the volume of the reaction system obtained in step (1), and reversing and mixing.
(3) Adding 1mol/L HCL, adding the HCL in a volume ratio of 1:1 to the volume of the reaction system obtained in the step (2), and reversing and mixing the mixture.
(4) Centrifuging at 10000rpm for 10min at 18-25 ℃, and collecting supernatant.
(5) Adding isopropanol with the same volume as the supernatant obtained in the step (4), reversing and mixing evenly, and standing for 3 hours at the temperature of below 0 ℃.
(6) Centrifuging at 10000rpm for 10min at 18-25 ℃, pouring off supernatant, adding 75 wt% ethanol into the precipitate, slightly blowing to enable the precipitate to be upwelled, slightly reversing for several times, centrifuging at 10000rpm for 10min, removing upper liquid relatively, sucking off residual ethanol by using a gun head, and keeping the precipitate.
(7) And (5) precipitating the precipitate obtained in the step (6), and drying in a vacuum freeze-drying instrument to obtain the SMPDRN freeze-dried powder.
2. Preparing the adhesive bandage with the SMPDRN obtained in the step 1 as an effective component.
(1) Preparing an antibacterial liquid: 0.5g of motpidoxin is weighed and added with 100mL of sterile water to be fully dissolved, thus obtaining the antibiotic solution.
(2) Weighing 28g of micromolecule polydeoxyribonucleotide, adding 500mL of normal saline to fully dissolve the micromolecule polydeoxyribonucleotide, slowly passing the medicine core through the solution, extruding, rotating and drying at the temperature of below 70 ℃ to enable the medicine core to contain 20.5mg of SMPDRN per square centimeter, extruding, rotating and drying at the temperature of below 70 ℃, installing the dried medicine core on an integrated machine for preparing the band-aid, spraying the antibacterial liquid obtained in the step (1), slitting, rewinding and packaging to obtain a finished product, wherein the specification of the band-aid is 4.5cm multiplied by 6.0cm, and calculating to obtain the band-aid containing 553mg of SMPDRN per band-aid.
Example 5
A hemostatic gel contains small molecule polydeoxyribonucleotide as effective component.
The preparation method of the adhesive bandage comprises the following steps:
1. salmon Small Molecule Polydeoxyribonucleotide (SMPDRN) used in hemostatic dressings was prepared in the same manner as in example 2.
2. Preparing the hemostatic gel taking the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
(1) dissolving hyaluronic acid in deionized water to prepare 1 wt% solution, and filtering to remove insoluble substances for later use;
(2) preparing an aqueous solution with the concentration of small molecule polydeoxyribonucleotide of 0.6 wt%, the concentration of mupidoxin of 2 wt% and the concentration of fentanyl of 1.5 wt%;
(3) preparing a gel aqueous solution with the concentration of 2 wt%, wherein the concentration of carbomer is 1.5 wt%, and the concentration of polyethylene glycol is 0.5 wt%;
(4) mixing the solutions obtained in the steps (1), (2) and (3) in equal mass proportion, stirring, and homogenizing to obtain a gel system;
(5) and (4) subpackaging, sealing and sterilizing the mixture obtained in the step (4).
The content of the small molecule polydeoxyribonucleotide in the hemostatic gel of example 5 was 0.2 wt%.
Example 6
A hemostatic gel contains small molecule polydeoxyribonucleotide as effective component.
The preparation method of the hemostatic gel comprises the following steps:
1. salmon Small Molecule Polydeoxyribonucleotide (SMPDRN) used in hemostatic dressings was prepared in the same manner as in example 2.
2. Preparing the hemostatic gel taking the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
(1) dissolving hyaluronic acid in deionized water to prepare a 10 wt% solution, and filtering to remove insoluble substances for later use;
(2) preparing 90 wt% aqueous solution of polydeoxyribonucleotide;
(3) preparing 0.8 wt% carbomer glue solution;
(4) mixing the solutions obtained in the steps (1), (2) and (3) in equal mass proportion, stirring, and homogenizing to obtain a gel system;
(5) and (4) subpackaging, sealing and sterilizing the mixture obtained in the step (4).
The content of the small molecule polydeoxyribonucleotide in the hemostatic gel of example 6 was 30 wt%.
Example 7
A hemostatic gel contains small molecule polydeoxyribonucleotide as effective component.
The preparation method of the hemostatic gel comprises the following steps:
1. salmon Small Molecule Polydeoxyribonucleotide (SMPDRN) used in hemostatic dressings was prepared in the same manner as in example 2.
2. Preparing the hemostatic gel taking the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
(1) dissolving hyaluronic acid in deionized water to prepare a 3 wt% solution, and filtering to remove insoluble substances for later use;
(2) preparing 30 wt% aqueous solution of polydeoxyribonucleotide;
(3) preparing a carbomer glue solution with the concentration of 1.2 wt%;
(4) mixing the solutions obtained in the steps (1), (2) and (3) in equal mass proportion, stirring, and homogenizing to obtain a gel system;
(5) and (4) subpackaging, sealing and sterilizing the mixture obtained in the step (4).
The content of the small molecule polydeoxyribonucleotide in the hemostatic gel of example 7 was 10 wt%.
Example 8
A wet towel for wound healing comprises micromolecular polydeoxyribonucleotide as effective component.
The preparation method of the wound healing wet tissue comprises the following steps:
1. salmon Small Molecule Polydeoxyribonucleotide (SMPDRN) used in hemostatic dressings was prepared in the same manner as in example 2.
2. Preparing the wound healing wet tissue taking the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
non-woven fabrics are used as a wet tissue carrier, and wet tissue immersion liquid with the weight 1 time of that of the wet tissue carrier is sprayed and dripped on the wet tissue carrier;
the wet tissue immersion liquid comprises the following components in parts by weight: 3 parts of glycerol, 2 parts of alkyl glycoside, 2 parts of squalane, 1 part of micromolecular polydeoxyribonucleotide, 0.05 part of mupitoxin, 0.1 part of fatty alcohol-polyoxyethylene ether sodium sulfate, a mildew preventive PB 2050.1 part and 60 parts of deionized water.
The quantity of SMPDRN was calculated to be 14.7mg per gram of wet wipe carrier.
Example 9
A wet towel for wound healing comprises micromolecular polydeoxyribonucleotide as effective component.
The preparation method of the wound healing wet tissue comprises the following steps:
1. salmon Small Molecule Polydeoxyribonucleotide (SMPDRN) used in hemostatic dressings was prepared in the same manner as in example 2.
2. Preparing the wound healing wet tissue taking the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
non-woven fabrics are used as a wet tissue carrier, and wet tissue soaking liquid with the weight 3 times that of the wet tissue carrier is sprayed and dripped on the wet tissue carrier;
the wet tissue immersion liquid comprises the following components in parts by weight: 7 parts of glycerol, 5 parts of alkyl glycoside, 4 parts of squalane, 10 parts of micromolecular polydeoxyribonucleotide, 0.5 part of erythromycin, 0.5 part of sodium lauryl sulfate, a mildew preventive PB 2050.5 part and 90 parts of deionized water.
The quantity of SMPDRN was calculated to be 255.3mg per gram of wet wipe carrier.
Example 10
A wet towel for wound healing comprises micromolecular polydeoxyribonucleotide as effective component.
The preparation method of the wound healing wet tissue comprises the following steps:
1. salmon Small Molecule Polydeoxyribonucleotide (SMPDRN) used in hemostatic dressings was prepared in the same manner as in example 2.
2. Preparing the wound healing wet tissue taking the SMPDRN obtained in the step 1 as an effective component, and comprising the following steps:
non-woven fabrics are used as a wet tissue carrier, and wet tissue soaking liquid with the weight 5 times that of the wet tissue carrier is sprayed and dripped on the wet tissue carrier;
the wet tissue immersion liquid comprises the following components in parts by weight: 6 parts of glycerol, 3 parts of alkyl glycoside, 3 parts of squalane, 3 parts of micromolecular polydeoxyribonucleotide, 0.05 part of mupidoxin, 0.1 part of erythromycin, 0.2 part of sodium lauryl sulfate, a mildew preventive PB 2050.2 part and 70 parts of deionized water.
The quantity of SMPDRN was calculated to be 175.3mg per gram of wet wipe carrier.
Experiment of
The inventor proves that the SMPDRN and the medical dressing taking the SMPDRN as the functional component have the effects of quickly stopping bleeding, promoting wound healing, improving the immunity of the organism, resisting inflammation and relieving pain through the following experiments, and have high safety.
Experiment 1: SMPDRN toxicology test
A 30 wt% aqueous solution of SMPDRN (SMPDRN prepared according to example 2) was prepared and SMPDRN wound dressings and wet wipes were produced according to the methods of examples 1 and 4 and examples 8 and 9, respectively, to obtain a large dose of the dressing (example 4), a low dose of the dressing (example 1) and a large dose of the wet wipes (example 9), and a low dose of the dressing (example 8). And observing the toxic reaction of the solution, the band-aid and the wet tissue to the rabbit cornea, the intact skin and the damaged skin.
The experimental results show that the SMPDRN solution and the SMPDRN dressing with low dose and high dose do not cause anaphylactic reaction and toxic adverse reaction.
Experiment 2: hemostasis and wound healing experiments
The medical dressing of the invention is subjected to human body wound experiment
Wangzhi, male 49 years old, voluntarily participated in the wound healing test of this product, using the hemostatic gel prepared in example 7. The results of the experiment are shown in FIG. 2. In fig. 2, the left side wound was coated with saline and the right side wound was coated with SMPDRN hemostatic gel, and the wound was observed for changes over 0-7 days.
For a 40 year old female, the SMPDRN band-aid prepared by the method of example 2 (SMPDRN content 6 mg/cm) was used due to inadvertent finger laceration by a razor blade, and the wound healing was as shown in fig. 3. Fig. 3 shows the wound after 0h, 2min, 30min, 1 day, 4 days in sequence.
As can be seen from the figures 2 and 3 and the feedback of use, the adhesive bandage containing the SMPDRN gel has the effects of quickly stopping bleeding and promoting the healing of surface wounds of a human body.
The SMPDRN obtained in example 2 of the invention and the SMPDRN obtained in the invention with the application number of 201911191251.1 are used for carrying out a mouse wound healing experiment, as shown in figure 4, the SMPDRN prepared by the invention can obviously promote wound healing, and the effect is slightly better than that of the SMPDRN obtained in the invention with the application number of 201911191251.1.
Experiment 3: effect of SMPDRN on clotting time in mice
The SMPDRN prepared in the method of example 2 was used as the subject. Healthy Kunming mice with half male and female bodies and the weight of 20 +/-3 g are selected and divided into a normal control group, a 0.02 wt% SMPDRN low dose group, a 0.1 wt% SMPDRN medium dose group, a 0.5 wt% SMPDRN high dose group, a 0.5 wt% SMPDRN original process group (201911191251.1) and Yunnan white drug powder (200mg/kg), and 10 mice are selected. Except SMPDRN administration by intraperitoneal injection, normal group and Yunnan white drug group are administered by intragastric administration for 3 days continuously, 1h after the last administration, a glass capillary with the inner diameter of 1mm is inserted into orbital venous plexus of a mouse for blood sampling, the capillary is broken every 15s after being filled with the blood sampling, the capillary is pulled leftwards and rightwards slowly, and the time of appearing the blood coagulation streak is the blood coagulation time. 1h after the last administration, the mice were cut 0.3cm from the tail tip, and blood was aspirated from the side of the tail every 10s with a weighed filter paper at the time of recording until the filter paper could not aspirate, and bleeding time was recorded. The bleeding time is determined by the time when the blood flow naturally stops as follows. Data were t-tested using the sps 10.0 statistical software. The results are shown in Table 1.
TABLE 1 influence of SMPDRN on clotting time, bleeding time
Figure BDA0002433260730000181
Figure BDA0002433260730000191
Note that P <0.05 was significantly different and P <0.01 was significantly different compared to the normal group; # P <0.05 was significantly different compared to the 0.1% SMPDRN group.
As can be seen from table 1, SMPDRN can effectively shorten the clotting time and bleeding time of mice, and exhibits significant effect at lower dose compared to the normal group. Compared with the same dosage, the SMPDRN obtained by the new process has better blood coagulation and hemostasis effects than the SMPDRN obtained by the original process.
Experiment 4: anti-inflammatory and analgesic effects of SMPDRN
The SMPDRN prepared in the method of example 2 was used as the subject.
1. Anti-inflammatory action
(1) Selecting healthy and clean Kunming mice of each male and female half, dividing the mice into a control group, a 0.02 wt% SMPDRN low dose group, a 0.1 wt% SMPDRN middle dose group, a 0.5 wt% SMPDRN high dose group, a 0.5 wt% SMPDRN group (201911191251.1) of the prior art and an indomethacin group (15mg/kg), wherein each group comprises 10 mice, except for the SMPDRN groups, the indomethacin group is administrated by intraperitoneal injection, the indomethacin group is administrated by gastric lavage for 5 days continuously, 0.05ml of dimethylbenzene is accurately dripped into the right ear of each mouse after the last administration, the mice are killed after 1h, the left and right ear pieces are punched by a puncher and are respectively weighed, the swelling degree is expressed by the weight difference between the two ears, and the inhibition rate is calculated.
The inhibition rate (%). ratio (1-difference between the weight of both ears in the administration group/difference between the weight of both ears in the control group) × 100%
TABLE 2 anti-inflammatory action of SMPDRN
Figure BDA0002433260730000192
Figure BDA0002433260730000201
Note that P <0.05 was significantly different and P <0.01 was very poorly significant compared to the control group; compared with the 0.02% SMPDRN group, # P <0.05 shows significant difference, and # P <0.01 shows very poor significance.
(2) 36 healthy and clean Sprague Dawley rats are taken and divided into a control group and an SMPDRN (0.1 wt%) group, wherein 18 rats are taken in each group, the same part (back) is selected, hair is shaved by an electric hair clipper, the naked skin is wiped by an alcohol cotton ball, the skin is cut by a scalpel, the wound size is the same, the control group coats the wound by physiological saline, the SMPDRN group coats the wound by 0.1 wt% SMPDRN, the skin of the rat wound of 2mm is respectively cut into pieces after 3 days and 7 days of medicine taking, homogenization is carried out, and the content of an inflammatory factor IL-1 β is detected by a kit of Nanjing institute of bioengineering, and the result is shown in figure 5.
2. Analgesic effect
(1) Animal experiments: selecting healthy and clean Kunming mice of which the male and the female are half respectively, continuously administering for 3 days in groups, injecting 0.6% acetic acid solution 0.1ml/10g into the abdominal cavity of each mouse 30min after the last administration, observing and recording the number of mice with writhing in each group within 20min after the acetic acid injection and the number of writhing, and calculating the drug writhing inhibition rate.
The drug writhing inhibition ratio (%) is (number of writhing of control group-number of writhing of administration group)/number of writhing of control group × 100%
Data were t-tested using the sps 10.0 statistical software.
TABLE 3 effects of SMPDRN on torsos
Figure BDA0002433260730000202
Note that P <0.05 was significantly different and P <0.01 was very poorly significant compared to the control group; compared with the 0.02% SMPDRN group, # P <0.05 shows significant difference, and # P <0.01 shows very poor significance.
(2) Human body experiment: wangzhi, male 49 years old, when cooking in the morning, the back of the hand is scratched by a sharp instrument carelessly, and the wound appears obvious inflammation symptom at night: the wound surface is red and swollen and painful, pus flows out when being squeezed, the pain is relieved immediately when the SMPDRN solution with the concentration of 0.5% is smeared, the wound is found to be convergent on the next morning, the pus disappears, and the wound surface is slightly red and free of pain.
As can be seen from tables 2, 3 and 5, the SMPDRN can effectively inhibit the production of inflammatory factors, reduce the auricle swelling of mice caused by xylene, and significantly reduce the twisting action induced by acetic acid, which indicates that the SMPDRN has better anti-inflammatory and analgesic effects. In addition, human body experiments show that the SMPDRN has the effects of resisting inflammation, reducing swelling and easing pain. Compared with the SMPDRN prepared by the original process, the SMPDRN prepared by the process has more obvious effects on analgesia and anti-inflammation under the condition of the same dosage.
Experiment 5: hemostatic healing test effect of hemostatic gel with SMPDRN as active ingredient
The hemostatic gel prepared in the manner of example 7 was used as the subject.
The experimental population is 100 subjects, including 50 female subjects and 50 male subjects, and the age is between 40 and 60 years. 100 subjects with hemostatic wound healing were randomly divided into 50 subjects (using the product of example 7 of the present invention), 50 controls (using a commercially available hemostatic gel product), and male and female halves. After each subject continuously observes for 7 days, the effect of the tested group is obviously good and the effect of the control group, and the specific test result is shown in table 4.
TABLE 4 clinical Effect of SMPDRN wipes
Figure BDA0002433260730000211
As can be seen from table 4, the gel prepared by the present invention is superior to the control group in both hemostatic and accelerated wound healing effects.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (10)

1. A dressing having hemostatic and wound healing promoting effects, comprising a small molecule polydeoxyribonucleotide extracted from salmon.
2. The dressing having hemostatic and wound healing accelerating functions according to claim 1, further comprising at least one of an antibacterial drug and an analgesic drug.
3. The dressing having hemostatic and wound healing accelerating functions according to claim 2, wherein the antibacterial agent is at least one of borneol, macadoxin and erythromycin; the analgesic is at least one of morphine and fentanyl.
4. The dressing having hemostatic and wound healing accelerating functions according to claim 1, wherein the dressing is a wound patch, hemostatic gel or wound healing wet tissue; the content of the micromolecule polydeoxyribonucleotide in the band-aid is 0.3-20.5 mg per square centimeter of the medicine core; the content of micromolecular polydeoxyribonucleotide in the hemostatic gel is 0.2-30 wt%; the content of the micromolecule polydeoxyribonucleotide in the wound healing wet tissue is 14-260 mg per gram of the wet tissue carrier.
5. The dressing having hemostatic and wound healing accelerating functions as claimed in claim 1, wherein the preparation method of the small molecule polydeoxyribonucleotide comprises the steps of tissue lysis, neutralization, centrifugation, alcohol precipitation and centrifugal purification.
6. The dressing having hemostatic and wound healing accelerating functions according to claim 5, wherein the preparation method of the small molecule polydeoxyribonucleotide comprises the following steps:
(1) cracking salmon tissues by using an alkali cracking solution, wherein the alkali cracking solution contains 1-3 mol/L NaOH, 50-200 mM NaCl and 5-25 mg/ml proteinase K;
(2) adding Tris-HCl into the reaction system obtained in the step (1);
(3) adding HCL into the reaction system obtained in the step (2);
(4) centrifuging the reaction system obtained in the step (3), and collecting a supernatant;
(5) adding isopropanol into the supernatant obtained in the step (4), and standing for more than 30 min;
(6) and (5) centrifuging the reaction system obtained in the step (5), and reserving the precipitate to obtain the micromolecule polydeoxyribonucleotide.
7. The dressing having hemostatic and wound healing accelerating functions according to claim 6,
the working conditions of the step (1) are as follows: adding the alkaline lysis solution into salmon tissue, wherein the dosage ratio of the salmon tissue to the alkaline lysis solution is 1g (6-10) mL; crushing the tissue, reversing and uniformly mixing, and carrying out constant-temperature water bath at 90-100 ℃ for 20-40 min;
the working conditions of the step (2) are as follows: carrying out ice-bath on the reaction system obtained in the step (1) to below 25 ℃, adding 1-3 mol/L Tris-HCl according to the volume ratio of 1/3-1 of the total volume, and reversing and uniformly mixing;
the working conditions of the step (3) are as follows: adding 1-4 mol/L HCL according to the volume ratio of 1/3-1 of the total volume of the reaction system obtained in the step (2), and reversing and uniformly mixing;
the working conditions of the step (4) are as follows: centrifuging the reaction system obtained in the step (3) at 6000-10000 rpm above 0 ℃ for 10-20 min, and collecting the supernatant;
the working conditions of the step (5) are as follows: adding isopropanol with the same volume as the supernatant obtained in the step (4), reversing and uniformly mixing, and standing for more than 30 min;
the working conditions of the step (6) are as follows: centrifuging the reaction solution obtained in the step (5) at 6000-10000 rpm above 0 ℃ for 10-20 min, pouring off the supernatant to leave precipitates, adding 70-80 wt% of ethanol to suspend the precipitates, centrifuging at 6000-10000 rpm for 10-20 min, and keeping the precipitates.
8. A method for preparing a band-aid using the small-molecule polydeoxyribonucleotide according to any one of claims 1 to 7, comprising the steps of:
(1) preparing an antibacterial liquid;
(2) dissolving small molecular poly-deoxyribose nucleotide in normal saline to fully dissolve the small molecular poly-deoxyribose nucleotide, slowly passing the drug core through the solution, extruding, rotating and drying at the temperature of below 70 ℃, spraying the dried drug core on the antibacterial liquid obtained in the step (1), cutting, rewinding and packaging to obtain a finished product.
9. A method for preparing hemostatic gel using the small molecule polydeoxyribonucleotide according to any one of claims 1 to 7, comprising the steps of:
(1) dissolving hyaluronic acid in deionized water to prepare 1-10 wt% solution, and filtering to remove insoluble substances for later use;
(2) preparing 0.6-90 wt% aqueous solution of polydeoxyribonucleotide;
(3) preparing 0.8-2 wt% of gel aqueous solution;
(4) mixing the solutions obtained in the steps (1), (2) and (3) in equal mass proportion, stirring and homogenizing;
(5) and (4) subpackaging, sealing and sterilizing the mixture obtained in the step (4).
10. A method for preparing a wet tissue for wound healing by using the small molecule polydeoxyribonucleotide as claimed in any one of claims 1 to 7, comprising the steps of:
the non-woven fabric is used as a wet tissue carrier, and wet tissue immersion liquid with the weight 1-5 times that of the wet tissue carrier is sprayed and dripped on the wet tissue carrier;
the wet tissue immersion liquid comprises the following components in parts by weight: 3-7 parts of glycerol, 2-5 parts of alkyl glycoside, 2-4 parts of squalane, 1-10 parts of micromolecular polydeoxyribonucleotide, 0.05-0.5 part of antibacterial agent, 0.1-0.5 part of surfactant, 0.1-0.6 part of mildew preventive and 60-90 parts of deionized water.
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