CN116585530B - Chloroplast composite hydrogel capable of efficiently producing oxygen and preparation method and application thereof - Google Patents
Chloroplast composite hydrogel capable of efficiently producing oxygen and preparation method and application thereof Download PDFInfo
- Publication number
- CN116585530B CN116585530B CN202310497716.6A CN202310497716A CN116585530B CN 116585530 B CN116585530 B CN 116585530B CN 202310497716 A CN202310497716 A CN 202310497716A CN 116585530 B CN116585530 B CN 116585530B
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- chloroplast
- oxygen
- composite hydrogel
- efficiency
- oxygen production
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
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- A—HUMAN NECESSITIES
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
Abstract
The invention belongs to the technical field of medical polymer materials, and discloses a chloroplast composite hydrogel for efficiently producing oxygen, a preparation method and application thereof, in particular to application in the aspects of chronic wound healing and tissue regeneration. The invention provides a chloroplast efficient oxygen generating system, which comprises chloroplast, a Hill reaction solution and an oxidant. The oxygen generating system is a mild and continuous oxygen generating system based on the Hill reaction. The invention also provides a chloroplast composite hydrogel for efficiently producing oxygen based on the oxygen producing system, which is obtained by compositing the chloroplast efficient oxygen producing system and a natural polysaccharide material. The chloroplast composite hydrogel has good biocompatibility, can self-generate oxygen through photosynthesis when being applied to the wound part of a patient, thereby promoting wound healing, and has the effects of green, mild and high-efficiency oxygen generation; can be applied to the biomedical field, such as preparing tissue engineering materials, in particular to preparing medical dressing for promoting chronic wound surfaces difficult to heal.
Description
Technical Field
The invention belongs to the technical field of medical polymer materials, and particularly relates to a chloroplast composite hydrogel for efficiently producing oxygen, a preparation method and application thereof, in particular to application in the aspects of chronic wound healing and tissue regeneration.
Background
The survival of most organisms on the earth depends on oxygen, and for human individuals, oxygen is inhaled into the body through a respiratory system and then transported to cells at various parts of the body to perform respiration to generate energy, so that oxygen is vital for the survival of the human; meanwhile, for cells, the cells consume a large amount of glucose to generate ATP under the anoxic condition, and once the sugar is supplied insufficiently or consumed completely, the cells die and necrose, which is particularly common in special wound sites accompanied by extrusion, vascular blockage and the like, such as burn, bedsore and diabetic wound sites. Chronic hypoxia in chronic wounds inhibits angiogenesis, re-epithelialization and synthesis of extracellular matrix, resulting in long-term non-healing of the wound, leading to tissue necrosis. Therefore, developing a dressing material capable of sustained oxygen release is critical for chronic wound healing.
Currently known oxygen delivery treatments are generally high pressure oxygen and local gaseous oxygen, which on the one hand may cause secondary damage to the wound due to the high pressure, and on the other hand, high efficiency oxygen generating materials are one of the best solutions for this situation because gaseous oxygen is difficult to penetrate the skin and cannot be absorbed by cells. The current oxygen generating systems applied to organisms mainly comprise platelets, peroxides and perfluorocarbons, but the oxygen generating functions of the oxygen generating systems are not controllable, substances which are harmful to cells can be generated, and the oxygen generating systems are not efficient. Microalgae gradually become new stars in the oxygen generating material due to controllable photosynthetic oxygen generation, but in actual use, essential nutrients are provided for survival, so that the possibility of bacteria infection of wounds is improved.
Chloroplast, a kind of plant cell used for photosynthesis, has the function of producing oxygen by utilizing solar energy to photolyze water. In 1937, it was found that in vitro chloroplasts were able to transfer electrons of oxidants to oxygen in water, thereby producing oxygen, a reaction also known as the hil reaction. The chloroplast is used as an intermediate of an oxygen production system, so that the method has the advantages of controllable oxygen production, high instantaneous oxygen production efficiency and the like. However, the double-layer membrane structure of chloroplast makes the reaction efficiency of the oxidant entering the membrane not high, and the generated active oxygen (mainly peroxide) can destroy the chloroplast itself, so that the oxygen production performance is greatly reduced, and the oxygen production efficiency is affected.
Therefore, the problem of high-efficiency oxygen production of chloroplasts is urgently needed to be solved, and a hydrogel dressing which utilizes chloroplasts to continuously and moderately produce oxygen is constructed, so that a new method is provided for solving the problem of difficult healing of anoxic wounds.
Disclosure of Invention
In order to overcome the defects and shortcomings of the prior art, the primary purpose of the invention is to provide a chloroplast efficient oxygen generating system.
The chloroplast high-efficiency oxygen generating system is a mild and continuous oxygen generating system based on Hill reaction, and all reagents have good biocompatibility and have green and high-efficiency oxygen generating effects.
The invention further aims to provide a chloroplast composite hydrogel for efficiently generating oxygen based on the chloroplast efficient oxygen generating system.
The chloroplast composite hydrogel has good biocompatibility, can self-generate oxygen through photosynthesis when being applied to the wound part of a patient, thereby promoting wound healing, and has the effects of green, mild and high-efficiency oxygen generation.
The invention also aims to provide the application of the chloroplast composite hydrogel in the biomedical field, such as the preparation of tissue engineering materials, and is particularly suitable for preparing medical dressings for promoting chronic difficult-to-heal wound surfaces.
The aim of the invention is achieved by the following scheme:
a high-efficiency oxygen generating system for chloroplast comprises chloroplast, hill reaction liquid and oxidant.
Wherein, the components of the Hill reaction liquid comprise: water is used as solvent, 40-60mmol/L tris-HCl,40-60mmol/L MgCl 2 ，80-120mmol/L NaCl。
Further, the pH of the Hill reaction liquid is preferably 7.0 to 8.0.
Further, the concentration of chloroplasts in the system may be 3-15. Mu.g/mL.
Further, the oxidizing agent may include potassium ferricyanide, ferric oxalate, and EDTA-Fe 3+ At least one of them.
Further, the concentration of the oxidizing agent in the system may be 1 to 100mmol/L, preferably 1 to 20mmol/L.
Further, the chloroplast is a chloroplast conventionally extracted from a green plant in the art.
Further, the green plant can be spinach, lettuce, rice, wheat, etc. The extraction method is conventional method, such as differential centrifugation.
The chloroplast high-efficiency oxygen generating system provided by the invention can generate oxygen through the Hill reaction of chloroplast mildly and efficiently under illumination. The chloroplast adopted can be obtained by green plant extraction with wide sources and low cost, the sources are wide and rich, the used reagents are simple, the biocompatibility is good, and the environment-friendly and efficient oxygen production effect is achieved.
Furthermore, the chloroplast high-efficiency oxygen generating system can also contain an antioxidant.
Further, the antioxidant may include at least one of anthocyanin, resveratrol, lipoic acid, catalase, and the like.
Further, the antioxidant content in the system may be 0.25 to 5wt%.
The antioxidant is added into the chloroplast high-efficiency oxygen production system, so that the generated active oxygen can be effectively removed, and the damage of the active oxygen to the chloroplast is reduced, thereby obviously improving the oxygen production efficiency.
Furthermore, the invention also provides a method for improving the oxygen production efficiency of the chloroplast high-efficiency oxygen production system, which comprises the steps of placing the chloroplast high-efficiency oxygen production system in an ultrasonic treatment state for illumination oxygen production; or after ultrasonic treatment, the light irradiation is carried out to produce oxygen.
Further, the time of the ultrasonic treatment may be 0 to 60 minutes, preferably 1 to 30 minutes.
Further, the ultrasonic treatment is a conventional ultrasonic treatment, for example, the ultrasonic power can be 10-50KHz.
Furthermore, the illumination is visible light.
The chloroplast high-efficiency oxygen generating system can effectively improve the permeability of chlorophyll cell membranes through ultrasonic treatment, thereby improving the oxygen generating efficiency of the system.
The invention also provides a chloroplast composite hydrogel for efficiently producing oxygen based on the chloroplast efficient oxygen production system.
The high-efficiency oxygen-generating chloroplast composite hydrogel disclosed by the invention comprises a chloroplast high-efficiency oxygen-generating system and a natural polysaccharide material; the specific components comprise chloroplast, hill reaction liquid, oxidant and natural polysaccharide material.
Further, in the hydrogel, the chloroplast content may be 3-15. Mu.g/mL.
Wherein, the chloroplast content can be calculated by characterization of chlorophyll concentration:
c is chloroplast concentration, A 652 Absorbance at 652nm, and dilution ratio at 30.
Further, the natural polysaccharide material may include at least one of alginic acid, hyaluronic acid, chitosan, collagen, gelatin, cellulose, and the like.
Further, the natural polysaccharide material may be used in the hydrogel in an amount of, for example, 1 to 30wt% based on the conventional hydrogel.
Further, the concentration of the oxidizing agent in the hydrogel may be 1 to 100mmol/L, preferably 1 to 20mmol/L.
Furthermore, the components of the chloroplast composite hydrogel for efficiently generating oxygen can also contain an antioxidant.
Further, the antioxidant may include at least one of anthocyanin, resveratrol, lipoic acid, catalase, and the like.
Further, the antioxidant content may be 1-10wt%.
According to the high-efficiency oxygen-producing chloroplast composite hydrogel, all the components are uniformly mixed, and the stable hydrogel can be formed based on crosslinking of natural polysaccharide materials. The crosslinking mechanism of the natural polysaccharide material may include anionic and cationic crosslinking, covalent crosslinking, hydrogen bonding crosslinking, or a mixture of the above mechanisms.
The invention also provides a method for improving the oxygen production efficiency of the chloroplast composite hydrogel, which comprises the steps of placing the hydrogel in an ultrasonic treatment state for illumination oxygen production; or after ultrasonic treatment, the light irradiation is carried out to produce oxygen.
Further, the time of the ultrasonic treatment may be 0 to 60 minutes, preferably 1 to 30 minutes.
Further, the ultrasonic treatment is a conventional ultrasonic treatment, for example, the ultrasonic power can be 10-50KHz.
Compared with the existing oxygen production system, the oxygen production mode is simple and easy to realize, no toxic byproducts are produced in the oxygen production process, and each component has good biocompatibility and does not increase cytotoxicity, and the high-efficiency oxygen production chloroplast composite hydrogel is applied to the biomedical field, such as preparation of tissue engineering materials, such as hydrogel dressing, in particular preparation of medical dressing for promoting chronic wound surfaces difficult to heal, and has remarkable advantages.
Compared with the prior art, the invention has the following advantages:
(1) Compared with the existing oxygen generating system, the chloroplast high-efficiency oxygen generating system has the advantages that the oxygen generating system is simple and easy to realize, and toxic byproducts are not generated in the oxygen generating process.
(2) According to the invention, chloroplasts are introduced into the field of tissue repair for the first time, and the chloroplast composite hydrogel provided by the invention is natural in components, simple and convenient to prepare and has excellent biocompatibility.
(3) The chloroplast composite hydrogel can realize mild, efficient and continuous oxygen production only by visible light illumination.
(4) The chloroplast composite hydrogel is used for preparing dressing, can effectively give consideration to oxygen production and antioxidation, can deeply improve cell activity from two aspects, and can promote cell proliferation, thereby achieving the purpose of rapid wound healing.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is an optical microscopic view of chlorophyll.
FIG. 2 is a graph showing the absorption spectrum of chlorophyll.
FIG. 3 is an oxygen production curve of the chloroplast high efficiency oxygen production system of the present invention.
FIG. 4 is a graph showing the effect of iron ion concentration on cell viability.
FIG. 5 is a graph showing the effect of ultrasound on the oxygen production efficiency of a chloroplast efficient oxygen production system.
FIG. 6 is a graph showing changes in the scavenging ability of four active oxygen species.
FIG. 7 is a graph showing the comparison of oxygen production before and after the chloroplast high-efficiency oxygen production system of the present invention is prepared into a hydrogel.
FIGS. 8 and 9 are graphs showing the effect of hydrogels of different chloroplast contents on cell proliferation capacity.
Fig. 10 is a graph showing wound healing effect in mice experiments.
Detailed Description
The present invention will be described in further detail with reference to examples, but embodiments of the present invention are not limited thereto. The materials referred to in the examples below are available commercially unless otherwise specified. The method is conventional unless otherwise specified.
Chloroplasts can be obtained by purchase or self-extraction. The isolated chloroplast can be obtained by a differential centrifugation method, and specifically can be obtained by extraction through the following steps:
(1) The component content of the extracting solution is as follows: 300-500mmol/L sucrose, 8-12mmol/L potassium chloride, 25-35mmol/L disodium hydrogen phosphate, 16-24mmol/L potassium dihydrogen phosphate.
(2) Cutting green plant such as herba Spinaciae leaves, grinding in extractive solution, filtering with gauze, centrifuging at 1500r/min for 3min for 2 times, and removing cells in the liquid; centrifuging at 4000r/min for 5min, and removing supernatant to obtain chloroplast; the extract can be added to obtain chloroplast suspension, and the chloroplast suspension is stored in a refrigerator at 4 ℃ in a dark place.
(3) The chloroplasts are seen to be elliptically shaped by light microscopy, as shown in FIG. 1. The absorption spectrum is shown in FIG. 2, and it can be seen that the wavelengths of light that can be absorbed are about 440nm and 650nm, both in the visible wavelength region.
Example 1: chloroplast high-efficiency oxygen production system
For comparison purposes, the following examples employ Hill reaction solutions having the same component contents, e.g., 50mmol/L tris-HCl and 50mmol/L MgCl, respectively 2 And 100mmol/L NaCl, the solvent being water.
And mixing chloroplast, a Hill reaction solution and an oxidant to obtain the chloroplast efficient oxygen production system. Wherein the chloroplast content is 3-15 mug/mL, and the oxidant content is 1-100mmol/L.
The high-efficiency oxygen generating system of chloroplast prepared by the method is placed under visible light, so that Hill reaction can be carried out to generate dissolved oxygen, and the oxygen generating rate is measured by an oxygen dissolving instrument, and the result is shown in figure 3. As can be seen from fig. 3, the oxygen production rate and the chlorophyll content are dose-dependent, and the oxygen production rate is correspondingly improved as the chloroplast concentration is increased (fig. 3 a); and the oxygen production rate and the content of the oxidant also have dose dependency, and the oxygen production rate is correspondingly improved along with the increase of the amount of the oxidant (figure 3 b); detecting the oxygen production amount of the chloroplast high-efficiency oxygen production system at different times (first day and second day after preparation), wherein the visible system can continuously produce oxygen under the illumination of visible light (figure 3 c); when the content of ferrous iron in the system is measured by using the phenanthroline hydrochloride method, it can be seen that ferric iron loses electrons to become ferrous iron in the system along with the change of illumination time, and the content of ferrous iron correspondingly increases along with the increase of the oxygen yield, so that the Hill reaction is proved to be continuously carried out (figure 3 d).
FIG. 4 is a graph showing the effect of different iron ion concentrations on cell viability. As can be seen from the figure, the iron ions used in the concentration range of the present invention have a promoting effect on cell activity and thus have no significant cytotoxicity.
Example 2: method for promoting chloroplast to produce oxygen in high-efficiency oxygen producing system
The chloroplast highly efficient oxygen producing system was prepared as in example 1.
And (5) placing the mixed chloroplast high-efficiency oxygen generating system in ultrasound for 5min, and then placing the chloroplast high-efficiency oxygen generating system in visible light for 15min, wherein the system generates a Hill reaction to generate dissolved oxygen, and measuring the oxygen generation rate by an oxygen dissolving instrument, so that the result is shown in figure 5. As can be seen from the graph, after ultrasonic treatment, the oxygen production of the system with different chloroplast concentrations is obviously improved, and the oxygen production is improved from 0.85mg/L to 1.55mg/L at the concentration of 3 mug/mL; at the concentration of 6 mug/mL, the oxygen production amount is increased from 1.44mg/L to 2.76mg/L; at the concentration of 12 mug/mL, the oxygen yield is increased from 2.03mg/L to 3.97mg/L, and the increasing trend is consistent, and almost reaches 200%. Therefore, the ultrasonic treatment can effectively improve the oxygen production efficiency of the chloroplast high-efficiency oxygen production system.
Example 3: high-efficiency chloroplast oxygen-generating system containing antioxidant
The preparation of the chloroplast high-efficiency oxygen production system is the same as that of the example 1, wherein the chloroplast content is 12 mug/mL, the potassium ferricyanide content is 10mmol/L, and the difference is that the system also contains 1mmol/L anthocyanin.
And (3) placing the prepared anthocyanin-containing chloroplast high-efficiency oxygen production system in visible light for illumination for 15min, so that Hill reaction can be performed to generate dissolved oxygen, and the oxygen production rate is measured by an oxygen dissolution instrument. The result shows that the high-efficiency oxygen production rate of the chloroplast containing anthocyanin is improved by 0.23mg/L compared with that of the chloroplast without anthocyanin.
Example 4: high-efficiency chloroplast oxygen-generating system containing antioxidant
The preparation of the chloroplast high-efficiency oxygen production system is the same as that of the example 1, wherein the chloroplast content is 12 mug/mL, the potassium ferricyanide content is 10mmol/L, and the difference is that the system also contains 1mmol/L resveratrol.
And (3) placing the prepared resveratrol-containing chloroplast high-efficiency oxygen generating system in visible light for illumination for 15min, so that Hill reaction can be performed to generate dissolved oxygen, and the oxygen generating rate is measured by an oxygen dissolving instrument. The result shows that the oxygen production rate of the high-efficiency oxygen production system of the chloroplast containing the resveratrol is improved by 0.45mg/L compared with that of the high-efficiency oxygen production system of the chloroplast without the resveratrol.
Example 5: high-efficiency chloroplast oxygen-generating system containing antioxidant
The preparation of the chloroplast high-efficiency oxygen production system is the same as that of example 1, wherein the chloroplast content is 12 mug/mL, the potassium ferricyanide content is 10mmol/L, and the difference is that lipoic acid of 1mmol/L is also contained in the system.
And (3) placing the prepared high-efficiency oxygen generating system of the chloroplast containing the lipoic acid under visible light for illumination for 15min, so that Hill reaction can be performed to generate dissolved oxygen, and the oxygen generating rate is measured by an oxygen dissolving instrument. The result shows that the oxygen production rate of the high-efficiency oxygen production system of the chloroplast without the lipoic acid is improved by 0.45mg/L.
Example 6: high-efficiency chloroplast oxygen-generating system containing antioxidant
The preparation of the chloroplast high-efficiency oxygen production system is the same as that of the example 1, wherein the chloroplast content is 12 mug/mL, the potassium ferricyanide content is 10mmol/L, and the difference is that catalase is also contained in the system.
The prepared high-efficiency oxygen-generating system of the chloroplast containing the catalase is placed under visible light for illumination for 15min, so that the Hill reaction can be carried out to generate dissolved oxygen, and the oxygen generation rate is measured by an oxygen dissolving instrument, so that the result shows that the oxygen generation rate of the high-efficiency oxygen-generating system of the chloroplast containing the catalase with the concentration of 0.5 weight percent is obviously improved compared with that of the high-efficiency oxygen-generating system of the chloroplast without the catalase, and the oxygen generation rate is improved by 1.03mg/L.
Meanwhile, the content of four active oxygen in the system is detected, and the result is shown in figure 6. The graph shows that the chloroplast high-efficiency oxygen generating system provided by the invention not only shows excellent oxygen generating capacity, but also has comprehensive active oxygen removing capacity, and has obvious removing effects on dpph, hydroxyl free radicals, superoxide anions and hydrogen peroxide, especially the hydroxyl free radicals and hydrogen peroxide, and the removing rate is more than 50%.
Example 7: chloroplast composite hydrogel
Preparation of chloroplast efficient oxygen production system the same as in example 6, natural polysaccharide material (gelatin) was added into the system, stirred well, after the solution was stable, placed into ice water bath to form gel. In the gel, the chloroplast content is 12 mug/mL, the potassium ferricyanide content is 1mmol/L, the catalase content is 0.5wt%, and the natural polysaccharide material (gelatin) content is 0.18g/mL.
After ultrasonic treatment of chloroplast composite hydrogel for 5min, the chloroplast composite hydrogel is placed under visible light for 15min, and the oxygen production rate is measured by an oxygen dissolving instrument. Wherein, the oxygen production rate of the chloroplast high-efficiency oxygen production system without adding gelatin is 2.28mg/L, and the oxygen release rate of the chloroplast composite hydrogel of the embodiment is 2.03mg/L, so that the hydrogel is considered to have no influence on the release of dissolved oxygen basically.
Example 8: chloroplast composite hydrogel
Preparation of efficient oxygen production System for chloroplasts As in example 6, adding natural polysaccharide material (alginic acid) into the System, stirring well, adding the solution into a mold with a syringe after the solution is stable, adding 4wt% CaCl 2 Spraying the aqueous solution to the surface of the covering solution to form glue. In the gel, the chloroplast content is 12 mug/mL, the potassium ferricyanide content is 1mmol/L, the catalase content is 0.5wt%, and the natural polysaccharide material (alginic acid) content is 0.02g/mL.
After ultrasonic treatment of chloroplast composite hydrogel for 5min, the chloroplast composite hydrogel is placed under visible light for 15min, and the oxygen production rate is measured by an oxygen dissolving instrument, and the result is shown in fig. 7. Wherein, the oxygen production rate of the chloroplast high-efficiency oxygen production system without adding alginic acid is 2.39mg/L, and the oxygen release rate of the chloroplast composite hydrogel of the embodiment is 2.22mg/L, so that the hydrogel is considered to have no influence on the release of dissolved oxygen basically.
The content of chloroplasts in the composite hydrogel is regulated to obtain three hydrogels with different chloroplast contents, namely 3 mug/mL, 6 mug/mL and 12 mug/mL, a group of blank groups without chloroplasts is arranged for cell culture experiments, a transwell cell is used for co-culture with cells, after the cells are planted in a pore plate, the transwell cell is covered, 200mL of the prepared hydrogel is placed in the cell, 800mL of culture solution is added, the four groups of cells are all under illumination, the hydrogel and the culture solution are replaced every day, and cck8 is tested in 1, 3 and 5 days of culture. Hydrogels were co-cultured with fibroblasts l929, the proliferation activity of the cells was tested with no hydrogel as a blank group, and live-dead staining fluorescence patterns were photographed, and the results are shown in fig. 8 and 9. The graph shows that the oxygen produced by the composite hydrogel promotes cell proliferation, simultaneously eliminates active oxygen produced by cells in the metabolic process, reduces the damage degree of the cells, obviously improves the proliferation activity of the cells, and has the most obvious effect that the proliferation activity percentage of a group of cells exceeds 120 percent.
Example 9: application of chloroplast composite hydrogel in diabetic wounds
Taking 12 male (8-10 weeks old) diabetic rats, anaesthetizing by intraperitoneal injection of 1% pentobarbital sodium (30 mg/kg), scraping back hair of the mice and washing back of the rats with 75% ethanol, and creating four circular full-thickness skin wounds on back of the rats with a mold at the same position of back of the miceRats which have been successfully wound-molded are randomly divided into A, B, C groups, wherein group A is pure calcium alginate hydrogel, group B is commercial gel which is successfully applied clinically, and group C is calcium alginate hydrogel with chloroplast content of 12 mug/mL prepared in example 8; the gel was changed daily and the wound healed as shown in fig. 10. From the graph, the chloroplast composite hydrogel can effectively give consideration to oxygen production and antioxidation, promote cell proliferation and achieveThe effect of the rapid wound healing is more pronounced than commercial gels.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (3)
1. A chloroplast composite hydrogel is characterized by comprising a chloroplast high-efficiency oxygen production system and a natural polysaccharide material, wherein the chloroplast high-efficiency oxygen production system and the natural polysaccharide material are compounded; also contains antioxidant;
the chloroplast high-efficiency oxygen-generating system comprises chloroplast, a Hill reaction solution and an oxidant; wherein, the components of the Hill reaction liquid comprise: water is used as solvent, 40-60mmol/L tris-HCl,40-60mmol/L MgCl 2 ，80-120mmol/L NaCl；
In the system, the concentration of chloroplast is 3-15 mug/mL; the concentration of the oxidant is 1-100mmol/L;
the oxidant comprises potassium ferricyanide, ferric oxalate and EDTA-Fe 3+ At least one of (a) and (b);
the antioxidant comprises at least one of anthocyanin, resveratrol, lipoic acid and catalase; the content of antioxidant in the system is 0.25-5wt%;
the natural polysaccharide material comprises at least one of alginic acid, hyaluronic acid, chitosan, collagen, gelatin and cellulose.
2. A method for improving the oxygen production efficiency of the chloroplast composite hydrogel of claim 1, which is characterized in that the chloroplast composite hydrogel of claim 1 is placed in an ultrasonic treatment state for light irradiation oxygen production; or after ultrasonic treatment, the light irradiation is carried out to produce oxygen.
3. The use of the chloroplast composite hydrogel of claim 1 in the preparation of tissue engineering materials.
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