CN111803704B

CN111803704B - Nursing hemostatic gel and preparation method thereof

Info

Publication number: CN111803704B
Application number: CN202010729742.3A
Authority: CN
Inventors: 王久英; 李欣; 王珺; 周杰; 安烜玉; 韩鹏飞
Original assignee: Mudanjiang Medical University
Current assignee: Mudanjiang Medical University
Priority date: 2020-07-27
Filing date: 2020-07-27
Publication date: 2022-03-18
Anticipated expiration: 2040-07-27
Also published as: CN111803704A

Abstract

The invention belongs to the field of biomedical materials, and particularly relates to a hemostatic gel for nursing and a preparation method thereof. The nursing hemostatic gel is prepared by taking carboxymethyl chitosan with a hemostatic effect, paeoniflorin, antibacterial peptide with a bacteriostatic effect and zinc oxide as active ingredients and matching with gel matrix materials such as locust bean gum, sodium carboxymethylcellulose and gelatin. The nursing hemostatic gel has a good sealing effect on wounds, can quickly activate platelets, enables blood to finish coagulation in a short time, and has a good hemostatic effect and a wound-preventing mouth-feel dyeing effect. The nursing hemostatic gel is suitable for various patients, can meet the requirements of different clinical scenes, and has positive clinical medical significance.

Description

Nursing hemostatic gel and preparation method thereof

Technical Field

The invention belongs to the field of biomedical materials, and particularly relates to a hemostatic gel for nursing and a preparation method thereof.

Background

When an accidental wound occurs, a human body has a spontaneous blood coagulation mechanism so as to reduce bleeding and alleviate body injury. The normal blood coagulation of a human body depends on the contraction of a blood vessel wall, a series of complex biochemical and cell biological reactions are started, and finally blood clots are formed to finish the blood coagulation. Blood is circulated and exchanged throughout the operation of the body. When the blood volume of the wounded is lost to 20%, about 800ml, shock symptoms may appear, and even the wounded may be life threatening.

With frequent occurrence of unstable factors such as natural disasters and the like, accidents of accidental injuries are continuously increased, and the demand of people for rapid hemostatic materials is also sharply increased. Hemostatic materials are always hot spots of domestic and foreign research, the application of currently developed hemostatic products in the treatment of accidents and clinical operations is wide, the hemostatic products have good effects on hemostasis of partial bleeding and small amount of bleeding of wounds, and the hemostatic, wound healing and repair functions are emphasized. The absorbable hemostatic material is a medical material which acts on a wound bleeding part, achieves the aim of hemostasis by accelerating the blood coagulation process of a human body, and can be degraded and absorbed by the human body within a certain time. There are many kinds of existing hemostatic materials, and there are many classification methods according to different classification bases: according to the structural characteristics, the material can be divided into hydrocolloids, powder, foams, liquids and films; they can be classified according to the hemostatic mechanism into coagulation dependent and coagulation independent mechanisms; the absorbable hemostatic material can be classified into collagen, gelatin, fibrin glue, oxidized cellulose, polysaccharide, etc. The ideal hemostatic material needs to meet the following requirements: (1) the hemostatic effect is obvious, and especially when the aorta is in acute hemorrhage, the hemostatic effect can be ensured to be effective within 2 minutes; (2) the cleaning and safety are realized, the cleaning and safety agent is non-toxic, harmless and non-irritant to human bodies, has no cytotoxicity and immunogenicity, is sterile and pathogen-free, and does not increase the infection probability; (3) the wound healing promoting agent has no negative effect on the wound, does not affect the healing of tissues, and can promote the healing of the wound and repair the wound; (4) the price is cheap, portable, can be widely used in each occasion, and the size is suitable, is convenient for storage and transportation.

Traumatic bleeding, resulting from accidental injury, presents a significant challenge to hemostatic technology. With the aging of the population, more and more age-related anticoagulation patients appear, such as patients who suffer from myocardial infarction due to aging and take anticoagulation medicines, and the like, and the challenge is brought to the hemostasis technology. With the improvement of medical care and nursing level, the cost of medical expenses is greatly increased, and no ideal hemostatic agent or hemostatic gel exists at present. In the face of the current challenges of traumatic bleeding and hemostasis in surgical clinic, a novel nursing hemostatic gel is urgently needed to meet the clinical requirement of trauma hemostasis.

Disclosure of Invention

Aiming at the defects of the prior art, the technical problem to be solved by the invention is to provide a nursing hemostatic gel and a preparation method thereof. The hemostatic gel is a biological material with functions of hemostasis and promotion of wound repair, has good hemostatic and coagulation promoting effects, is good in cell compatibility, and provides a new solution for hemostasis and promotion of wound repair on the surface of a wound.

In order to solve the technical problems, the invention adopts the technical scheme that:

a hemostatic gel for nursing comprises the following components in percentage by weight: 2-6% of carboxymethyl chitosan, 1-5% of paeoniflorin, 0.5-2% of antibacterial peptide, 1-2% of nano zinc oxide, 2-4% of glycerol, 5-10% of locust bean gum, 5-10% of sodium carboxymethyl cellulose, 10-30% of gelatin and the balance of purified water.

Preferably, the nursing hemostatic gel comprises the following components in percentage by weight: 3-5% of carboxymethyl chitosan, 2-4% of paeoniflorin, 1-2% of antibacterial peptide, 1-1.5% of nano zinc oxide, 2-3% of glycerol, 6-8% of locust bean gum, 6-8% of sodium carboxymethyl cellulose, 15-25% of gelatin and the balance of purified water.

Preferably, the nursing hemostatic gel comprises the following components in percentage by weight: 4% of carboxymethyl chitosan, 3% of paeoniflorin, 1.5% of antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethyl cellulose, 20% of gelatin and the balance of purified water.

Preferably, the molecular weight of the carboxymethyl chitosan is 4000-6000, and the substitution degree is 80-90%.

Preferably, the antibacterial peptide is extracted from fly maggot protein, and the molecular weight is 8000-12000.

Preferably, the particle size of the nano zinc oxide is between 20 and 80 nm.

Preferably, the galactomannan content in the locust bean gum is more than 85%.

Further, the invention also provides a preparation method of the hemostatic gel for nursing, which comprises the following steps:

(1) weighing the components according to the weight ratio;

(2) mixing carboxymethyl chitosan, glycerol, locust bean gum, sodium carboxymethylcellulose, gelatin and purified water, heating to 80-90 deg.C in water bath, stirring, and cooling to room temperature to obtain gel base material;

(3) adding paeoniflorin, antibacterial peptide and nano zinc oxide into the gel base material, uniformly mixing, performing vacuum defoaming, performing ultrasonic oscillation for 1-2 hours, and performing irradiation sterilization to obtain the nursing hemostatic gel.

Preferably, the radiation sterilization uses Co-60 gamma radiation, and the radiation dose is 10-20 kGy.

Furthermore, the invention also provides application of the nursing hemostatic gel in preparing hemostatic medical materials.

The amino group of the carboxymethyl chitosan carries positive charges, and when the high-density positive charges on the molecular chain contact the negative charges on the surfaces of the erythrocytes, charge interaction occurs, so that a large number of erythrocytes are gathered on the macromolecular skeleton of the carboxymethyl chitosan. Meanwhile, a large amount of electronegative phosphatidylserine on the surface of the platelet can also be electrostatically attracted with the carboxymethyl chitosan with positive charge, and when the chitosan is contacted with blood, a large amount of red blood cells and platelets can be rapidly attracted to be adsorbed on the surface of the material, so that a network structure complex of the hemostatic material and the red blood cells, the platelets and plasma proteins is formed. The carboxymethyl chitosan forms a network structure of high-viscoelasticity gel after absorbing blood, is blocked at a vascular fracture, does not depend on blood coagulation waterfall reaction, and plays a role in stopping bleeding. Carboxymethyl chitosan is used for stopping bleeding by virtue of physical action, and is also suitable for animals and people with blood coagulation disorder.

Radix Paeoniae Rubra is dried root of Paeonia lactiflora pall of Ranunculaceae. Most of the identified compounds in red peony root are monoterpene components, mainly including paeoniflorin, paeonol, and the like. Paeoniflorin has the following pharmacological actions: anti-inflammatory and immunoregulatory effects, inducing apoptosis, protecting nervous system, and improving sleep. In addition, researches show that the paeoniflorin accords with the five principles of the Lipinski medicines and has certain hemostatic activity by predicting hemostatic active ingredients by adopting a molecular docking method.

The antibacterial peptide is a basic polypeptide substance with antibacterial activity generated after organisms are induced, and the molecular weight of the basic polypeptide substance is usually less than 10 kDa. The antibacterial peptide has strong antibacterial ability and wide antibacterial range, and can resist various microorganisms, such as gram-positive bacteria, gram-negative bacteria and certain fungi. The fly and larva mainly grow in the severe environment containing a large amount of bacteria, such as saprophytic substances, excrement, garbage and the like, the body surface of the fly and larva is provided with a plurality of pathogenic bacteria, a plurality of diseases can be transmitted, but the fly and larva can not die due to the infection of the pathogenic bacteria, and the fly has strong immunity to pathogenic microorganisms and can generate strong antibacterial substances. Researches find that the housefly antibacterial peptide has strong adaptability to temperature change, can resist high osmotic pressure environment such as high-concentration salt solution and extreme acid-base solution. The housefly antibacterial peptide has high solubility in water, alkaline aqueous solution and broad-spectrum antibacterial capability.

The zinc oxide is a multifunctional semiconductor oxide with good chemical stability, thermal stability and antimicrobial activity, is white and regular hexagonal wurtzite structure, is not decomposed at high temperature, and is low in price and rich in resources. It has been reported in the prior art that zinc oxide has a certain antibacterial activity against gram-negative bacteria such as pseudomonas aeruginosa, escherichia coli and gram-positive bacteria such as bacillus subtilis and staphylococcus aureus, and has the effect of resisting the external microbial infection at the wound.

Compared with the prior art, the invention has the beneficial effects that: the nursing hemostatic gel is prepared by taking carboxymethyl chitosan with a hemostatic effect, paeoniflorin, antibacterial peptide with a bacteriostatic effect and zinc oxide as active ingredients and matching with gel matrix materials such as locust bean gum, sodium carboxymethylcellulose and gelatin. The nursing hemostatic gel has a good sealing effect on wounds, can quickly activate platelets, enables blood to finish coagulation in a short time, and has a good hemostatic effect and a wound-preventing mouth-feel dyeing effect. The use process is simple and comfortable, and the product has good flexibility and biocompatibility. The raw materials are safe and non-toxic, and the gel after use can be biodegraded without causing any harm to human bodies and environment. The nursing hemostatic gel is suitable for various patients, can meet the requirements of different clinical scenes, and has positive clinical medical significance.

Detailed Description

The present invention is further illustrated by the following examples, which are provided for the understanding of the present invention, but are not intended to limit the scope of the present invention, and should not be construed as limiting the scope of the present invention in any way.

Example 1

A hemostatic gel for nursing comprises the following components in percentage by weight: 4% of carboxymethyl chitosan, 3% of paeoniflorin, 1.5% of fly maggot antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethyl cellulose, 20% of gelatin and the balance of purified water.

The preparation method of the hemostatic gel for nursing comprises the following steps:

(1) weighing the components according to the weight ratio;

(3) adding paeoniflorin, fly maggot antibacterial peptide and nano zinc oxide into the gel base material, uniformly mixing, performing vacuum defoaming, performing ultrasonic oscillation for 1-2 hours, and performing irradiation sterilization to obtain the hemostatic gel for nursing.

Example 2

A hemostatic gel for nursing comprises the following components in percentage by weight: 2% of carboxymethyl chitosan, 5% of paeoniflorin, 0.5% of antibacterial peptide, 2% of nano zinc oxide, 4% of glycerol, 5% of locust bean gum, 5% of sodium carboxymethyl cellulose, 30% of gelatin and the balance of purified water.

The preparation method of the hemostatic gel for nursing is the same as that of the example 1.

Example 3

The nursing hemostatic gel is characterized by comprising the following components in percentage by weight: 6% of carboxymethyl chitosan, 1% of paeoniflorin, 2% of antibacterial peptide, 1% of nano zinc oxide, 2% of glycerol, 10% of locust bean gum, 10% of sodium carboxymethyl cellulose, 10% of gelatin and the balance of purified water.

Comparative example 1

A hemostatic gel for nursing comprises the following components in percentage by weight: 7% of carboxymethyl chitosan, 1.5% of housefly maggot antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethyl cellulose, 20% of gelatin and the balance of purified water.

Comparative example 2

A hemostatic gel for nursing comprises the following components in percentage by weight: 7% of paeoniflorin, 1.5% of housefly maggot antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethylcellulose, 20% of gelatin and the balance of purified water.

Comparative example 3

A hemostatic gel for nursing comprises the following components in percentage by weight: 4% of hydroxyethyl chitosan, 3% of paeoniflorin, 1.5% of maggot antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethyl cellulose, 20% of gelatin and the balance of purified water.

Comparative example 4

A hemostatic gel for nursing comprises the following components in percentage by weight: 4% of hydroxypropyl chitosan, 3% of paeoniflorin, 1.5% of maggot antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethylcellulose, 20% of gelatin and the balance of purified water.

Mouse wound hemostasis test

35 mice were randomly divided into 7 groups, corresponding to the hemostatic gels for nursing use of examples 1-3 and comparative examples 1-4, respectively, and 5 mice were each group. The mouse was lacerated with a sterile blade 5mm from the tail end, and immediately after bleeding, 0.2ml of the corresponding hemostatic gel was applied to the wound and timing was started. The observation was continued and the bleeding stopped when the wound no longer bleeds. The blood time(s) from the start of bleeding to the time when bleeding stops. Another 5 mice were used as natural control groups, no drug was administered to the natural control group, the remaining treatment methods were the same as the experimental groups, and the hemostasis time(s) was recorded after the wound stopped bleeding. The results of the experiment are shown in table 1.

TABLE 1 Effect of nursing hemostatic gels on hemostatic time of mouse wounds

As can be seen from the experimental results in Table 1, compared with the natural control group, the nursing hemostatic gel of the present invention can significantly shorten the hemostatic time of the wound of the mouse, and has a good hemostatic effect. Under the premise of keeping the total amount of the hemostatic components unchanged, the hemostatic effect of the hemostatic gel (comparative examples 1-2) using a single hemostatic component is obviously inferior to that of the hemostatic gel using the single hemostatic component and the hemostatic gel using the single hemostatic gel in combination, which shows that the combined use of carboxymethyl chitosan and paeoniflorin can achieve a synergistic effect in hemostasis. In addition, the type of chitosan derivative is important, and when hydroxyethyl chitosan, hydroxypropyl chitosan and paeoniflorin are used (comparative examples 3-4), the hemostatic effect is not the same as that when carboxymethyl chitosan and paeoniflorin are used.

Blood coagulation index measurement experiment

Mice were decapped, whole blood was collected and stored in tubes (pre-coated with 3.8% heparin sodium), centrifuged at 12000rpm for 5min at room temperature, and the upper plasma was aspirated and the experiment was performed within 1 hour. Mu.l of the above blood plasma was taken, 50. mu.l of the hemostatic gel of examples 1 to 3 and comparative examples 1 to 4 and physiological saline (same volume, negative control group) were added thereto, mixed, and then incubated at 37 ℃. Prothrombin Time (PT), Thrombin Time (TT), Activated Partial Thrombin Time (APTT), fibrinogen amount (FIB) were measured by a semi-automatic plasma coagulation analyzer. 3 replicates of each gel were run. The results of the experiment are shown in table 2.

TABLE 2 Effect of nursing hemostatic gels on blood coagulation indices in mice

As can be seen from the experimental results in table 2, compared with the negative control group, the Prothrombin Time (PT) of the hemostatic gels for nursing use in examples 1 to 3 of the present invention did not change significantly, the Thrombin Time (TT) and the Activated Partial Thrombin Time (APTT) were significantly decreased, and the fibrinogen amount (FIB) was significantly increased. The hemostatic gels using a single hemostatic ingredient (comparative examples 1-2) and different types of chitosan derivatives (comparative examples 3-4) had a weak effect of promoting plasma coagulation, indicating that the type selection and reasonable compatibility of the hemostatic ingredients are crucial to the hemostatic effect.

The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications or equivalents may be made to the technical solution without departing from the principle of the present invention, and these modifications or equivalents should also be regarded as the protection scope of the present invention.

Claims

1. The nursing hemostatic gel is characterized by comprising the following components in percentage by weight: 2-6% of carboxymethyl chitosan, 1-5% of paeoniflorin, 0.5-2% of antibacterial peptide, 1-2% of nano zinc oxide, 2-4% of glycerol, 5-10% of locust bean gum, 5-10% of sodium carboxymethyl cellulose, 10-30% of gelatin and the balance of purified water; the molecular weight of the carboxymethyl chitosan is 4000-6000, and the substitution degree is 80-90%.

2. The nursing hemostatic gel according to claim 1, which is composed of the following components in percentage by weight: 3-5% of carboxymethyl chitosan, 2-4% of paeoniflorin, 1-2% of antibacterial peptide, 1-1.5% of nano zinc oxide, 2-3% of glycerol, 6-8% of locust bean gum, 6-8% of sodium carboxymethyl cellulose, 15-25% of gelatin and the balance of purified water.

3. The nursing hemostatic gel according to claim 2, which is composed of the following components in percentage by weight: 4% of carboxymethyl chitosan, 3% of paeoniflorin, 1.5% of antibacterial peptide, 1% of nano zinc oxide, 3% of glycerol, 7% of locust bean gum, 7% of sodium carboxymethyl cellulose, 20% of gelatin and the balance of purified water.

4. The hemostatic gel for nursing use according to any one of claims 1-3, wherein the antibacterial peptide is extracted from fly maggot protein and has a molecular weight of 8000-12000.

5. Hemostatic gel for care according to any of claims 1-3, wherein the nano zinc oxide has a particle size of between 20-80 nm.

6. A nursing haemostatic gel according to any of claims 1-3, wherein the galactomannan content of the locust bean gum is above 85%.

7. A process for the preparation of a hemostatic gel for care according to any one of claims 1 to 6, comprising the steps of: (1) weighing the components according to the weight ratio; (2) mixing carboxymethyl chitosan, glycerol, locust bean gum, sodium carboxymethylcellulose, gelatin and purified water, heating to 80-90 deg.C in water bath, stirring, and cooling to room temperature to obtain gel base material; (3) adding paeoniflorin, antibacterial peptide and nano zinc oxide into the gel base material, uniformly mixing, performing vacuum defoaming, performing ultrasonic oscillation for 1-2 hours, and performing irradiation sterilization to obtain the nursing hemostatic gel.

8. The method for preparing hemostatic gel for nursing use according to claim 7, wherein the radiation sterilization uses Co-60 gamma radiation at a dose of 10-20 kGy.

9. Use of a haemostatic gel for care according to any of claims 1-6 in the preparation of a haemostatic medical material.