CN111187443A - Collagen-sodium humate compound suitable for hemostatic material and preparation method thereof - Google Patents
Collagen-sodium humate compound suitable for hemostatic material and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a collagen-sodium humate compound suitable for a hemostatic material and a preparation method thereof. The invention takes natural collagen with good bioactivity and natural polymer sodium humate with the functions of stopping bleeding and promoting wound healing as main raw materials, prepares a composite solution with good compatibility by virtue of the charge shielding effect of salt under the condition of low temperature, prepares composite hydrogel through collagen self-assembly, and prepares the collagen-sodium humate composite through freeze drying. The prepared composite hydrogel and sponge improve the thermal stability, hemostasis, platelet coagulation promotion and other performances of single collagen hydrogel, and the hydrogel and sponge have the advantages of good porosity, hydrophilicity, antibiosis, anti-inflammatory and the like. The hydrogel and sponge can be used alone or in combination with other materials for hemostatic dressing.
Description
Technical Field
The invention belongs to the technical field of collagen-based medical materials, and particularly relates to a collagen-sodium humate compound suitable for a hemostatic material and a preparation method thereof.
Background
A dressing is a biomedical material used to treat skin lesions. Traditional gauze dressing only plays the effect of covering the wound, does not have the promotion effect and easily takes place the adhesion with the wound to the healing of wound, causes the damage to the wound when leading to changing the dressing. Accordingly, various novel modern dressings come into operation, most of the dressings can be well attached to the wound surface, play a part of the role of a skin barrier, provide a favorable microenvironment for wound healing, and are not bonded with the wound surface so as to avoid secondary injury caused by dressing replacement; meanwhile, has the important functions of diminishing inflammation, stopping bleeding, accelerating wound healing and the like. However, most of the dressings, particularly hydrogel dressing products, in the market at present are expensive, and thus many patients are prohibited.
Collagen is a natural hemostatic material, which has good biological activity, such as low antigenicity, biocompatibility, blood coagulation, and promotion of cell proliferation as a main component of extracellular matrix. However, in practice, it is necessary to ensure the integrity of the triple helical structure of collagen, and if the structure is damaged, the biological activity of collagen will be completely lost, and the unique biological properties of collagen will be lost. For example, gelatin or collagen hydrolysate has the advantages of no toxicity, no pollution, and biodegradability, although the amino acid composition is not significantly different from that of collagen, but the polypeptide chain thereof has a random coil structure rather than a triple helix conformation, and has physicochemical properties different from those of collagen and no biological activity. However, collagen alone has the disadvantages of poor thermal stability and poor mechanical properties in practical applications, and therefore needs to be modified by physical or chemical crosslinking.
Currently, there are three main methods for collagen modification: 1) physical modification: other exogenous substances are not introduced, more crosslinking is generated among collagen molecules, so that the stability of the collagen molecules is improved, the performance of the collagen is improved, and the good biocompatibility of the collagen can be ensured; 2) chemical crosslinking: the thermal stability and the mechanical property of the collagen can be obviously improved, but the residue of the cross-linking agent can cause inflammation or cytotoxicity, so the method is at the cost of reducing the biological property of the collagen; 3) blending with other polymers: the blending material takes advantages of both strong points and weak points, and can be fused with two materials. The mechanical properties are generally improved and blending is the simplest method of preparing new materials.
Materials blended with collagen can be generally classified into synthetic polymer materials and natural polymer materials. The natural polymer has excellent multifunctional property and good biocompatibility and biodegradability, and the biomaterial prepared by blending the collagen and the natural polymer has the advantages and performances that other materials cannot replace, such as collagen-chitosan and collagen-sodium alginate-carboxymethyl cellulose. However, chitosan has no biological activity and poor processability; alginic acid dressings are relatively expensive and most products are not self-adhesive.
Sodium humate is a natural multifunctional organic high molecular compound, is composed of aromatic and active functional groups such as hydroxyl, quinonyl, carboxyl, methoxyl and the like, has good physiological activity and functions of absorption, complexation, exchange and the like, particularly has good effects of stopping bleeding, diminishing inflammation, relieving pain and relieving itching on wounds, and is called as 'Wujin powder' for a long time.
The humic acid has rich resources, large reserve capacity, wide distribution and good grade, and is widely applied to the fields of light industry, medicine and health care and the like. The sodium humate contains a large amount of amino, carboxyl, quinonyl and other groups in molecules, and can generate chemical reaction with collagen to generate hydrogen bond and electrostatic action.
Disclosure of Invention
The invention aims to provide a collagen-sodium humate compound suitable for a hemostatic material and a preparation method thereof, which improve the thermal stability and mechanical property of collagen by adding sodium humate and endow the dressing with good hemostatic, antibacterial and anti-inflammatory effects by utilizing the slow release property of the sodium humate.
In order to achieve the above object, the present invention provides a method for preparing a collagen-sodium humate complex suitable for use as a hemostatic material, comprising the steps of:
respectively dissolving collagen sponge and sodium humate in a sodium salt solution to prepare a collagen solution and a sodium humate solution; adjusting the pH values of the collagen solution and the sodium humate solution to 4-9 by using sodium hydroxide or hydrochloric acid; the temperature of the solution is lower than 0-10 ℃; dropwise adding the sodium humate solution into the collagen solution, and stirring to obtain a composite solution; the composite solution is self-assembled at the temperature of 30-37 ℃ to prepare collagen-sodium humate composite hydrogel; or freeze-drying the composite solution to obtain the collagen-sodium humate composite sponge.
The salt solution is disodium hydrogen phosphate solution, sodium dihydrogen phosphate solution or sodium chloride solution, and the salt content is 80-200 mmol/L.
The preparation method comprises the following specific steps:
step one, preparing a collagen solution:
1-10 parts of collagen sponge are taken and dissolved in a salt solution to prepare a collagen solution with the mass fraction of 0.1-1.5%;
step two, preparing a sodium humate solution:
dissolving 0.25-40 parts of sodium humate in a salt solution at normal temperature to prepare a sodium humate solution with the concentration of 0.025-6%;
step three, preparing a collagen-sodium humate composite solution:
slowly dropwise adding the sodium humate solution prepared in the step two into the collagen solution prepared in the step one in stirring, wherein the mass ratio of the sodium humate to the collagen dry weight is 1: 0.05-20, the temperature is controlled to be below 10 ℃, the pH value is controlled to be 4-9, and stirring is carried out for 6-24 hours to prepare a collagen-sodium humate composite solution;
step four, preparing the collagen-sodium humate compound
And (3) placing the composite solution prepared in the third step at 30-37 ℃ for self-assembly to prepare composite hydrogel, or freeze-drying the composite solution by using a freeze dryer to obtain the composite sponge.
The collagen-sodium humate compound prepared by the preparation method is provided.
Compared with the prior art, the invention has the beneficial effects that:
1) the invention adopts the blending macromolecule which is natural macromolecule-sodium humate, and blends the natural macromolecule-sodium humate with collagen to prepare the composite hydrogel and the sponge, thereby not only improving the stability and the mechanical property of the collagen-based material, but also improving the hemostatic property of the collagen-based material and endowing the collagen-based material with the antibacterial and anti-inflammatory functions.
2) The invention adopts the low-temperature preparation condition to avoid the phenomenon of thermal denaturation of the collagen due to high temperature, and basically keeps the three-strand helical structure of the natural collagen to fully exert the biological performance of the natural collagen.
3) The natural collagen used in the invention can be extracted from the skin of livestock with edible value by an acid enzyme method, and the raw material source is wide; the used sodium humate has rich resources and low price, and the prepared collagen-sodium humate composite material is used for stopping bleeding and can generate high added value.
Drawings
FIG. 1 is an electropherogram of the collagen-sodium humate composite of example 1;
fig. 2 is a graph showing the thermal stability of the collagen-sodium humate composite of example 2.
In the figure, a-collagen and b-collagen-sodium humate.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention include, but are not limited to, the scope shown in the following examples.
The invention discloses a preparation method of a collagen-sodium humate compound suitable for a hemostatic material, which specifically comprises the following steps:
(1) dissolving 1-10 parts of collagen sponge in a salt solution to prepare a collagen solution with the concentration of 0.1-1.5%; controlling the salt content in the salt solution to be 80-200 mmol/L; the temperature is controlled below 10 ℃;
(2) dissolving 0.25-40 parts of sodium humate in a salt solution at normal temperature to prepare a sodium humate solution with the concentration of 0.025-6%; controlling the salt content in the salt solution to be 80-200 mmol/L;
(3) slowly dripping 0.025-6% sodium humate solution into the stirred collagen solution, wherein the dry weight ratio of the sodium humate to the collagen is 1: 0.05-20; controlling the temperature below 10 ℃, controlling the pH value at 4-9, and stirring for 6-24 hours to prepare a collagen-sodium humate composite solution;
(4) and (3) placing the composite solution at 30-37 ℃ for self-assembly to prepare composite hydrogel or freeze-drying by using a freeze dryer to obtain the composite sponge.
Preferably, the salt solution is at least one of disodium hydrogen phosphate, sodium dihydrogen phosphate or sodium chloride.
Preferably, the salt content in the salt solution is 110-130 mmol/L.
Preferably, the dry weight ratio of the sodium humate to the collagen is 1: 0.1-10; controlling the pH value to be 6-8, and stirring for 10-12 h.
Example 1:
1 part of collagen sponge is taken and dissolved in 100 parts of 120mmol/L sodium chloride solution at 4 ℃ to prepare collagen solution with the concentration of 1 percent, and the pH value is adjusted to 7.0 by adopting sodium hydroxide. Taking 1 part of sodium humate, dissolving in 100 parts of 120mmol/L sodium chloride solution at normal temperature to prepare 1% sodium humate solution, and adjusting the pH value to 7.0 by adopting sodium hydroxide; slowly dripping the sodium humate solution into the stirred collagen solution at 4 ℃, wherein the dry weight ratio of the sodium humate to the collagen is 1:1, and stirring for 12 hours to prepare a collagen-sodium humate composite solution; subsequently, the composite solution was placed at 37 ℃ for self-assembly to prepare a composite hydrogel.
FIG. 1 is an electropherogram of the collagen-sodium humate composite of example 1, wherein the collagen and the collagen-sodium humate composite have similar electrophoresis bands, and each of the three different bands includes two α chains (α 1 and α 2 chains, molecular weight of about 100 kDa) and 1 β chain (molecular weight of about 200 kDa), which are standard electrophoresis bands of collagen, and show that the three-strand helical structure of the collagen before and after the compounding has no significant change and does not affect the biological function of the collagen.
Example 2:
a collagen solution having a concentration of 1% was prepared by dissolving 1 part of collagen sponge in 125 parts of a solution containing 10mmol/L disodium hydrogenphosphate/sodium dihydrogenphosphate and 100mmol/L sodium chloride at 10 ℃ and the pH thereof was about 7.4. Dissolving 1.5 parts of sodium humate in a solution containing 10mmol/L disodium hydrogen phosphate/sodium dihydrogen phosphate and 100mmol/L sodium chloride at normal temperature to prepare a sodium humate solution with the concentration of 1.5 percent and the pH value of about 7.4; slowly dripping the sodium humate solution into the stirred collagen solution at 10 ℃, wherein the dry weight ratio of the sodium humate to the collagen is 6:4, and stirring for 20 hours to prepare a collagen-sodium humate composite solution; subsequently, the composite solution was placed at 35 ℃ for self-assembly to prepare a composite hydrogel.
Fig. 2 is a result of a thermal stability test of the collagen-sodium humate composite material in example 2, which shows that the thermal stability of the composite material is obviously higher than that of pure collagen, and the composite material is beneficial to later processing and use.
Example 3:
0.9 part of collagen sponge is taken and dissolved in 100 parts of 100mmol/L sodium chloride solution at the temperature of 6 ℃ to prepare collagen solution with the concentration of 0.9 percent, and the pH value is adjusted to 4.5 by adopting hydrochloric acid. Taking 0.1 part of sodium humate, dissolving in 100 parts of 100mmol/L sodium chloride solution at normal temperature to prepare 0.1% sodium humate solution, and adjusting the pH value to 4.5 by adopting sodium hydroxide; slowly dripping the sodium humate solution into the stirred collagen solution at 6 ℃, wherein the dry weight ratio of the sodium humate to the collagen is 1:9, and stirring for 12 hours to prepare a collagen-sodium humate composite solution; subsequently, the complex solution was lyophilized to obtain a complex sponge.
Example 4:
1.5 parts of collagen sponge was dissolved in 100 parts of 100mmol/L sodium dihydrogen phosphate solution at 6 ℃ to prepare a 1.5% collagen solution, and the pH was adjusted to 5.5 with sodium hydroxide. Taking 0.1 part of sodium humate, dissolving in 100 parts of 100mmol/L sodium dihydrogen phosphate solution at normal temperature to prepare 0.1% sodium humate solution, and adjusting pH to 5.5 by adopting sodium hydroxide; slowly dripping the sodium humate solution into the stirred collagen solution at 6 ℃, wherein the dry weight ratio of the sodium humate to the collagen is 1:15, and stirring for 8 hours to prepare a collagen-sodium humate composite solution; subsequently, the complex solution was lyophilized to obtain a complex sponge.
Example 5:
1 part of collagen sponge is taken and dissolved in 100 parts of 100mmol/L disodium hydrogen phosphate solution at 4 ℃ to prepare a collagen solution with the concentration of 1 percent, and the pH value is adjusted to 6.5 by hydrochloric acid. Taking 0.5 part of sodium humate, dissolving in 100 parts of 100mmol/L disodium hydrogen phosphate solution at normal temperature to prepare 1% sodium humate solution, and adjusting the pH value to 6.5 by adopting hydrochloric acid; slowly dripping the sodium humate solution into the stirred collagen solution at 4 ℃, wherein the dry weight ratio of the sodium humate to the collagen is 1:2, and stirring for 6 hours to obtain a collagen-sodium humate composite solution; subsequently, the composite solution was placed at 32 ℃ for self-assembly to prepare a composite hydrogel.
Claims (4)
1. A preparation method of a collagen-sodium humate compound suitable for a hemostatic material is characterized by comprising the following steps:
respectively dissolving collagen sponge and sodium humate in a sodium salt solution to prepare a collagen solution and a sodium humate solution; adjusting the pH values of the collagen solution and the sodium humate solution to 4-9 by using sodium hydroxide or hydrochloric acid; the temperature of the solution is lower than 0-10 ℃; dropwise adding the sodium humate solution into the collagen solution, and stirring to obtain a composite solution; the composite solution is self-assembled at the temperature of 30-37 ℃ to prepare the collagen-sodium humate composite hydrogel, or the composite solution is freeze-dried to prepare the collagen-sodium humate composite sponge.
2. The method for preparing a collagen-sodium humate complex for hemostatic materials as claimed in claim 1, wherein:
the salt solution is disodium hydrogen phosphate solution, sodium dihydrogen phosphate solution or sodium chloride solution, and the salt content is 80-200 mmol/L.
3. The method for preparing a collagen-sodium humate complex for hemostatic materials according to claim 1 or 2, wherein:
the method comprises the following specific steps:
step one, preparing a collagen solution:
1-10 parts of collagen sponge are taken and dissolved in a salt solution to prepare a collagen solution with the mass fraction of 0.1-1.5%;
step two, preparing a sodium humate solution:
dissolving 0.25-40 parts of sodium humate in a salt solution at normal temperature to prepare a sodium humate solution with the concentration of 0.025-6%;
step three, preparing a collagen-sodium humate composite solution:
slowly dropwise adding the sodium humate solution prepared in the step two into the collagen solution prepared in the step one in stirring, wherein the mass ratio of the sodium humate to the collagen dry weight is 1: 0.05-20, the temperature is controlled to be below 10 ℃, the pH value is controlled to be 4-9, and stirring is carried out for 6-24 hours to prepare a collagen-sodium humate composite solution;
step four, preparing the collagen-sodium humate compound
And (3) placing the composite solution prepared in the third step at 30-37 ℃ for self-assembly to prepare composite hydrogel, or freeze-drying the composite solution by using a freeze dryer to obtain the composite sponge.
4. A collagen-sodium humate complex suitable for use as a haemostatic material, prepared by the method of claims 1-3.
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| CN102908666A (en) * | 2012-10-11 | 2013-02-06 | 天津市赛瑞生物技术有限公司 | Collagen scaffold material for cornea |
| CN106349718A (en) * | 2016-10-25 | 2017-01-25 | 齐鲁工业大学 | Preparation method for antibacterial collagen |
| CN109337383A (en) * | 2018-09-30 | 2019-02-15 | 福建工程学院 | A kind of collagen-based selfreparing hydrogel and preparation method thereof |
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| CN111187443B (en) | 2022-05-10 |
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