CN106620815B - Based on G- tetrad/collagen gel dressing and its preparation and application - Google Patents
Based on G- tetrad/collagen gel dressing and its preparation and application Download PDFInfo
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- CN106620815B CN106620815B CN201611038109.XA CN201611038109A CN106620815B CN 106620815 B CN106620815 B CN 106620815B CN 201611038109 A CN201611038109 A CN 201611038109A CN 106620815 B CN106620815 B CN 106620815B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
- A61L15/325—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/20—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing organic materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/425—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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
- A61L2300/23—Carbohydrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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 discloses a kind of based on G- tetrad/collagen gel dressing and preparation method thereof, and guanosine, boric acid and potassium hydroxide are added in deionized water according to 1~3:1:1,90~98 DEG C is heated to, obtains G- tetrad liquid solution；35~40 DEG C are cooled to, collagen, which is added, makes its dispersion, makes the 2~10mg/mL of concentration of collagen in the solution, is cooled to room temperature, obtains described based on G- tetrad/collagen gel dressing.The preparation method reaction condition is simple, and synthesis step is few, and time-consuming short, production cost is low.The gel dressing shows good biocompatibility for wound healing, and good permeability, adhesion is strong, and water content is high, maintains wet environment, wound is made to be not easy to form a scab.Gel dressing of the present invention is free of any stimulating component, has no toxic side effect.The gel dressing is used for accelerating wound healing, has potential application.
Description
Technical field
The invention belongs to medical dressing field, it is related to a kind of gel dressing and preparation method thereof, it can more particularly to one kind
Promote wound healing based on G- tetrad/collagen gel dressing and preparation method thereof.
Background technique
Wound be normal skin the extraneous factor such as surgical operation, heat, external force, electric current, low temperature the effects of under caused by
Damage.If processing is not in time, wound may infect, or further deteriorate.The dressing being traditionally often used is gauze, cotton material
Equal materials, but these dressing hemostasis, moisturizing, prevent wound infection and promote in terms of effect difference strong man
Meaning.
Collagen is the main component of animal connective tissue, and the most functional egg of content in the mammalian body
It is white.Due to its low immunogenicity, degradability, biocompatibility and the characteristics such as hemagglutinin, collagen is by widely as giving birth to
Object material.Compared to other wound dressings, easy, the time-consuming short and bioactivity of gel dressing preparation containing collagen is good,
There is good result to wound healing.
Mainly there is also following problems for already present gel dressing: I. preparation aspect, there is also systems for the synthesis of gel
The deficiencies of standby complicated, time-consuming.II. aspect of performance, gel mechanical strength is poor and has certain bio-toxicity.III wound
Healing aspect, there are poor air permeability and wound healing time are long for part dressing.Therefore, easy, effective gel dressing is prepared
It, will be with important research significance and application value for the quick healing of wound.
Summary of the invention
For the above-mentioned deficiency of the prior art, it is an object of the invention to solve the problems, such as that preparing for gel dressing is many and diverse,
Overcome gel dressing poor air permeability, healing time length, the deficiency of poor biocompatibility；Select degradable, biocompatibility
As raw material, what is be prepared is had good G- tetrad and collagen based on G- tetrad/collagen gel dressing
Good mechanical performance keeps the moisturizing to wound, sticks and avoid infection, effectively and rapidly promotes the healing of wound.
Present invention preparation can promote reaction mechanism such as Fig. 1 of G- tetrad/collagen gel dressing of wound healing
It is shown:
Effect of the guanosine with neighbouring guanosine because of hydrogen bond is combined together, in this way four connected birds
Purine nucleosides and a K+ are coordinated, and form G- tetrad；Between G- tetrad together by borate bridging, space networks are formed
Shape structure.When collagen is added, collagen can be embedded, and be ultimately formed solidifying based on G- tetrad/collagen
Glue.
According to above-mentioned principle, the present invention is adopted the following technical scheme that:
A kind of preparation method based on G- tetrad/collagen gel dressing, comprising the following steps:
(1) guanosine, boric acid and potassium hydroxide are added to the water, heat, is completely dissolved, obtains to guanosine
G- tetrad liquid solution；
(2) after the G- tetrad liquid solution in step (1) is cooling, collagen is added, disperses collagen, obtains institute
It states based on G- tetrad/collagen gel dressing.
Wherein, in the step (1),
The guanosine can also be substituted with adenosine, cytidine.
The potassium hydroxide can also be substituted with sodium hydroxide, lithium hydroxide.
The molar ratio of the guanosine, boric acid and potassium hydroxide is 1~3:1:1；Preferably, 2:1:1.
The water is deionized water.
The temperature of the heating is 90~98 DEG C；Preferably 95 DEG C.
The time of the heating is 15-25；Preferably 15-20 minutes.
Wherein, in the step (2),
The temperature of the cooling is 35~40 DEG C.
The collagen can come from but be not limited to tilapia fishskin, gadus fish-skin.
Concentration after the collagen dispersion is 2~10mg/mL；Preferably 5mg/mL.
Specifically, the step includes:
(1) guanosine, boric acid and potassium hydroxide are add to deionized water according to the molar ratio of 1~3:1:1, are shaken
It swings 5~10 minutes, heating water bath is kept for constant temperature 15~25 minutes to 90~98 DEG C, is completely dissolved to guanosine, obtains G-
Tetrad.
(2) solution of above-mentioned steps (1) is progressively cooled between 35~40 DEG C, water-bath keeps constant temperature, and collagen egg is added
It is white, make the 2~10mg/mL of concentration of collagen in the solution, rocking disperses it rapidly, is cooled to room temperature, is made and is based on
G- tetrad/collagen gel dressing.
Wherein, in step (1), after guanosine is added, boric acid is first added, adds potassium hydroxide, and boron is added
After acid or potassium hydroxide, 20s is rocked rapidly, facilitates the dissolution of guanosine in this way.
Wherein, in step (1), the molar ratio of guanosine, boric acid and potassium hydroxide is 2:1:1, water bath heating temperature
It is best for 95 DEG C of reaction conditions；Dressing effect is best when the concentration of collagen in above-mentioned steps (2) is 5mg/mL.
It is applied the invention also provides prepared by the preparation method based on G- tetrad/collagen gel
Material.
Conveniently, permeability is good for the simple, preparation based on G- tetrad/collagen gel dressing composition of the invention,
Adhesion is strong, good mechanical property, good biocompatibility.
The invention also provides described based on G- tetrad/collagen gel dressing in the increasing for promoting wound cell
Application in growing, breaking up.
Wound healing is promoted in preparation based on G- tetrad/collagen gel dressing the invention also provides described
Application in product.
It is described that a kind of biomaterial can be used as based on G- tetrad/collagen gel dressing in the present invention.
In the present invention, the healing that wound can be accelerated based on G- tetrad/collagen gel dressing.
It is described that mouse wound healing is used for based on G- tetrad/collagen gel dressing in the present invention, have as follows
Feature: 1. hemostasis is fast, absorptive tissue juice, enhances the adhesive force in wound；2. the environment for maintaining wound relatively wet reduces
Infection probability；3. accelerating cell Proliferation, differentiation, promote the regeneration of skin.
It is described based on G- tetrad/collagen gel dressing in the present invention, including but not limited to applied as wound
Material.
It is described based on G- tetrad/collagen gel dressing in the present invention, including but not limited to it is used for mouse wound
Healing.
It is described based on G tetrad/collagen gel dressing in the present invention, including but not limited to for the negative of drug
It carries.
Preparation method provided by the present invention based on G- tetrad/collagen gel dressing, reaction condition is simple,
Synthesis step is few, and time-consuming short, production cost is low.It is provided by the present invention to be used for based on G- tetrad/collagen gel dressing
Wound healing has the advantages that good biocompatibility, good permeability, adhesion be strong, high-moisture, maintains the ring of wound moist
Border makes wound be not easy to form a scab.It is of the present invention based on G- tetrad/collagen gel dressing without any irritation at
Point, it has no toxic side effect.It is provided by the present invention that accelerating wound healing is used for based on G- tetrad/collagen gel dressing,
It has potential application.
Detailed description of the invention
Fig. 1 is reaction mechanism figure of the present invention preparation based on G- tetrad/collagen gel dressing.
Fig. 2 is the gel prepared in embodiment 2.
Fig. 3 is the healing state of control group, treatment group's (gel prepared in embodiment 2) wound at the 0th, 3,5,7 day.
Fig. 4 is the wound area difference of control group, treatment group's (gel prepared in embodiment 2) at the 0th, 3,5,7 day.
Fig. 5 is the histology of control group, treatment group's (gel prepared in embodiment 2) at the 3rd, 5,7 day.
Fig. 6 is the cytotoxicity experiment of control group, treatment group's (gel prepared in embodiment 2).
Specific embodiment
In conjunction with following specific embodiments and attached drawing, the present invention is described in further detail.Implement process of the invention,
Condition, experimental method etc. are among the general principles and common general knowledge in the art, this hair in addition to what is specifically mentioned below
It is bright that there are no special restrictions to content.
Embodiment 1
(1) guanosine, boric acid and potassium hydroxide are add to deionized water according to the molar ratio of 1:1:1, concussion 5
Minute, heating water bath constant temperature 20 minutes, is completely dissolved to 90 DEG C to guanosine.
(2) above-mentioned solution is progressively cooled to 37 DEG C, water-bath keeps constant temperature, and collagen is added, makes collagen molten
Concentration in liquid is 5mg/mL, and rocking disperses it rapidly, is cooled to room temperature, is made solidifying based on G- tetrad/collagen
Glue dressing.
Embodiment 2
(1) guanosine, boric acid and potassium hydroxide are add to deionized water according to the molar ratio of 2:1:1, are shaken
10 minutes, heating water bath was kept for constant temperature 15 minutes, is completely dissolved to guanosine to 95 DEG C.
(2) above-mentioned solution is progressively cooled between 40 DEG C, water-bath keeps constant temperature, and collagen is added, makes collagen egg
White concentration in the solution is 7mg/mL, and rocking disperses it rapidly, is cooled to room temperature, is made and is based on G- tetrad/collagen egg
White gel dressing.The gel of preparation is as shown in Figure 2.When the gel prepared handstand is placed, gel can be sticked in bottle wall, no
As aqueous solution can flow down；And after 15 days, gel, which stands upside down to place, can be still sticked in bottle wall.This is illustrated based on G- tetrad
Body/collagen gel dressing has good mechanical performance.
Embodiment 3:
The healing of mouse wound is used for the gel prepared in experimental example 2, as treatment group.
This experiment makes trauma scaring model in ICR back of mice using operation method.By being smeared in wound surface
Gel observes surface scars daily and calms down situation, the reparation effect of evaluation product to scar.
(1) modeling
Under 1% yellow Jackets 5mL/kg intraperitoneal injection of anesthesia, cropping is carried out to back of mice, it is de- with 4% vulcanized sodium
Hair, the circular incision that parallel vertebra two sides are about respectively 1.5cm as diameter.It is wiped and is stopped blooding with sterile cotton balls if slight bleeding
Afterwards, it sends back in mouse box.
(2) animal packet
The successful ICR mouse of modeling is randomly divided into a, b, c group, every group 3.
A control group: as a control group by the wound of right side of mice.
B treatment group: using the wound on the left of mouse as treatment group.
Control group is without any processing, it is allowed normally to heal.Gel dressing is applied on treatment group's surface of a wound, is smeared daily
Once, observation surface scars repair situation daily.When experiment was carried out to the 3rd day, a group mouse is handled, by wound system
It is made slice, is characterized for histology, the growing state of wound cells of superficial layer is observed.When experiment was carried out to the 5th day, to b group
Mouse is handled, and wound is fabricated to slice, is characterized for histology, and the growing state of wound cells of superficial layer is observed.Work as reality
When testing progress to the 7th day, c group mouse is handled, wound is fabricated to slice, for histology characterization, observation wound table
The growing state of confluent monolayer cells.
(3) result
As shown in figure 3, gel dressing is applied to mouse wound, dressing can be attached on wound, will not be fallen off as liquid,
This illustrates the stronger adhesiveness of dressing；After 4h, dressing can be air-dried, and illustrate good permeability.When performing the operation the 3rd day, mouse wound
Certain contraction and incrustation phenomenon occurs, but the healing that can obviously observe control group and treatment group mouse wound has significantly
Difference.Due to carrying out gel dressing to treatment group and smearing the scar Healing Rate for the treatment of significantly better than control group.By the 7th day, control
Treatment group mouse wound area is minimum, and completely, recovery effects are good for basic healing.
As shown in figure 4, by control group, treatment group's wound area in comparison in different time periods, hence it is evident that observe treatment
The wound healing rate of group outclass control group, illustrates based on G- tetrad/collagen gel dressing for wound healing
With good effect.
As shown in figure 5, due to having carried out the smearing of gel dressing to treatment group, collagen promotes wound cell
Proliferation, differentiation.Cell epithelia degree from histological observation to treatment group is better than control group, promotes treatment group's mouse skin
Regeneration is significantly faster than that control group.By the 7th day, treatment group's mouse skin substantially regenerated completely.
4 cytotoxicity experiment of embodiment
The gel prepared in experimental example 2 is filtered with 0.22 μm of sterilised membrane filter, removes the bacterium in gel.By degerming
Gel be used for fibroblast toxicity test, as treatment group；Blank is as a control group.
This experiment is using MTT (3- (4,5- dimethylthiazole -2) -2,5- diphenyltetrazolium bromide bromide) method measurement cell
Vigor.By measurement gel to the toxicity of cell, and then evaluate the biocompatibility of gel.
(1) edge hole of 96 orifice plates is filled with sterile PBS；Logarithmic phase cell is collected, adjustment cell suspended concentration is 50000
100 μ L cell suspensions are added in a/mL, every hole.
(2) 5%CO2, 37 DEG C of incubations, until cell monolayer is paved with bottom hole, 10 μ L 5mg/mL gels of addition.
(3) cell is put into incubator, in 5%CO2, 37 DEG C are incubated for for 24 hours, observe under inverted microscope.
(4) 20 μ L 5mg/mL MTT are added in every hole, continue to cultivate 4h.
(5) culture is terminated, culture solution in hole is sucked；150 μ L dimethyl sulfoxides are added in every hole, and shaking table low-speed oscillation 10 divides
Clock, later with the light absorption value in the microplate reader detection each hole OD490nm.
(6) simultaneously be arranged zeroing hole (culture medium, MTT, dimethyl sulfoxide), control wells (cell, with the gel of concentration, training
Support base, MTT, dimethyl sulfoxide).
(7) cell survival rate (cell viability) is calculated:
Cell survival rate=(gel group A value-zeroing hole A value)/(control wells A value-zeroing hole A value) × 100%
As shown in fig. 6, carrying out cytotoxicity experiment with fibroblast.Experimental result is shown: control group and treatment group
Cell survival rate is all close to 100%.This illustrates that gel dressing is very small to the toxicity of fibroblast, that is to say, that bright gel
The good biocompatibility of dressing.
It is only as described above several specific embodiments of the invention, is not intended to restrict the invention.It is all in essence of the invention
Within mind and principle, used using same or similar method is obtained based on G- tetrad/collagen gel dressing
In wound healing, all fall in the scope of protection of the present invention.
Claims (8)
1. a kind of preparation method based on G- tetrad/collagen gel dressing, which is characterized in that the specific step of this method
Suddenly are as follows:
(1) guanosine, boric acid and potassium hydroxide are added to the water, heat, after guanosine is completely dissolved, obtains G-
Tetrad liquid solution；Wherein, the molar ratio of the guanosine, boric acid and potassium hydroxide is 1~3:1:1；
(2) after the G- tetrad liquid solution of step (1) is cooling, collagen is added, disperses collagen, obtain described be based on
G- tetrad/collagen gel dressing；Concentration after the collagen dispersion is 2~10mg/mL.
2. preparation method as described in claim 1, which is characterized in that in the step (1), the guanosine gland is fast
Purine nucleosides or cytidine substitution；The potassium hydroxide sodium hydroxide or lithium hydroxide substitution.
3. preparation method as described in claim 1, which is characterized in that in the step (1), the temperature of the heating is 90~
98℃；The time of the heating is 15-25 minutes.
4. preparation method as described in claim 1, which is characterized in that in the step (2), the temperature of the cooling is 35~
40℃。
5. preparation method as described in claim 1, which is characterized in that in the step (2), the collagen comes from Rofe
Fish fish-skin, gadus fish-skin.
6. preparation method as described in claim 1 be prepared based on G- tetrad/collagen gel dressing.
7. promoting cell Proliferation, differentiation in preparation based on G- tetrad/collagen gel dressing as claimed in claim 6
Product in application.
8. promoting the production of wound healing in preparation based on G- tetrad/collagen gel dressing as claimed in claim 6
Application in product.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001079342A2 (en) * | 2000-04-18 | 2001-10-25 | Colbar R & D Ltd. | Cross-linked collagen matrices and methods for their preparation |
| CN101391115A (en) * | 2008-11-12 | 2009-03-25 | 乐普（北京）医疗器械股份有限公司 | Preparation method of biological activity protein or polypeptides coating biology bracket |
| CN103403158A (en) * | 2011-08-11 | 2013-11-20 | 松下电器产业株式会社 | Method for detecting G-quadruplex formation |
| CN104297429A (en) * | 2014-10-15 | 2015-01-21 | 广州甘蔗糖业研究所 | Monosaccharide sensitive type hydrogel and preparation method thereof |
| WO2015073360A2 (en) * | 2013-11-12 | 2015-05-21 | New England Biolabs Inc. | Dnmt inhibitors |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001079342A2 (en) * | 2000-04-18 | 2001-10-25 | Colbar R & D Ltd. | Cross-linked collagen matrices and methods for their preparation |
| CN101391115A (en) * | 2008-11-12 | 2009-03-25 | 乐普（北京）医疗器械股份有限公司 | Preparation method of biological activity protein or polypeptides coating biology bracket |
| CN103403158A (en) * | 2011-08-11 | 2013-11-20 | 松下电器产业株式会社 | Method for detecting G-quadruplex formation |
| WO2015073360A2 (en) * | 2013-11-12 | 2015-05-21 | New England Biolabs Inc. | Dnmt inhibitors |
| CN104297429A (en) * | 2014-10-15 | 2015-01-21 | 广州甘蔗糖业研究所 | Monosaccharide sensitive type hydrogel and preparation method thereof |
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