CN110476954A - Collagen tissue material preservation liquid and its application - Google Patents
Collagen tissue material preservation liquid and its application Download PDFInfo
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- CN110476954A CN110476954A CN201910858282.1A CN201910858282A CN110476954A CN 110476954 A CN110476954 A CN 110476954A CN 201910858282 A CN201910858282 A CN 201910858282A CN 110476954 A CN110476954 A CN 110476954A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
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- General Health & Medical Sciences (AREA)
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Abstract
The present invention provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: thickness maintains 25 ~ 95 parts of agent, and transparency maintains 3 ~ 6 parts of agent, 25 ~ 55 parts of solvent.Collagen tissue material preservation liquid provided by the invention is suitable for the storage and transport that artificial collagen tissue material is especially artificial de- cell corneal material, in irradiation process, to maintain the original collagen fiber structure of corneal material and transparency, avoid the collagenous fibres of corneal material because irradiation damages or changes, and in long-term preservation, it is able to maintain that corneal thickness and transparency.
Description
Technical field
The invention belongs to medical material Techniques of preserving fields, and in particular to a kind of collagen tissue material preservation liquid and its answer
With.
Background technique
Corneal blindness is second-biggest-in-the-world blinding disease, and according to the statistics of the World Health Organization, whole world disease of cornea patient is about
About 5,000,000 people of 60000000 people, China, and increase 100,000 newly every year.Traditional therapeutic scheme mostly be using donor's cornea transplant into
Row treatment, but clinically donor wretched insufficiency, the donor's cornea transfer operation that China can carry out every year is less than 3000, cornea
The scarcity of donor makes vast majority of people lose the chance for restoring bright.The rise of organizational project canthus membrane technology be treat it is various can
The canthus membrane disease of blindness can be caused to bring hope.However, the preservation problem of tissue engineering comea becomes limitation tissue at present
The critical issue of engineering cornea industrialization, early-stage study person is saved frequently with glycerol, while adding some antioxidant, with
Protect cornea in irradiation process from destruction, such as glutathione, mercaptoethanol, but after irradiation, these are anti-oxidant
Often recurring structure recombinates substance, or is changed into other substances, and penetrating odor is had after being irradiated such as mercaptoethanol and is generated,
The operation of serious shadow clinician, the influence of the diagonal film properties of penetrating odor at present not yet studies have reported that, in addition, by
The preservation liquid of antioxidant is added, after being irradiated, is only capable of maintaining corneal thickness and transparency in short term, when long-term preservation can still lead
Cornea appearance, the transparency even variation of performance are caused, cornea appearance or structure in irradiation process can not be fundamentally solved and change
The problem of.
Summary of the invention
In view of the above problems, the present invention provides a kind of collagen tissue material preservation liquid, it is suitable for artificial collagen tissue material
The storage and transport for especially manually taking off cell corneal material, in irradiation process, to maintain the original collagen of corneal material fine
Structure and transparency are tieed up, avoids the collagenous fibres of corneal material because irradiation damages or changes, and in long-term preservation, it can
Maintain corneal thickness and transparency.
The purpose of the present invention is achieved through the following technical solutions:
The present invention provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: thickness maintenance agent 25 ~
95 parts, transparency maintains 3 ~ 6 parts of agent, 25 ~ 55 parts of solvent.
Preferably, the thickness maintains agent to be selected from albumin and glycerol.
Preferably, the albumin is selected from human serum albumin, and the human serum albumin is freeze dried powder.
Preferably, according to parts by weight, the human serum albumin content is 10 ~ 50 parts, and the glycerol content is 15 ~ 45
Part.
Preferably, the human serum albumin content is 22 ~ 45 parts, it is highly preferred that the human serum albumin content is 25 ~ 40
Part.
Preferably, the glycerol content is 20 ~ 35 parts.
Preferably, the transparency maintains agent to be selected from Sodium Hyaluronate.
Preferably, the Sodium Hyaluronate is low-numerator sodium hyaluronate, the molecular weight of the low-numerator sodium hyaluronate
For 20 ~ 800,000 Da.
Preferably, the solvent is selected from purified water.
Another aspect of the present invention provides above-mentioned collagen tissue material preservation liquid in saving artificial collagen tissue material
Using.
Preferably, the collagen tissue material is selected from artificial collagen tissue material, and the artificial collagen tissue material is selected from
Cell-eliminating coanea matrix, acellular conjunctiva matrix, de- cell stroma of sclera or de- cell amnion.
Compared with the existing technology, the beneficial effects of the present invention are:
The present invention uses human serum albumin to maintain agent as thickness, and human serum albumin is usually used in shock caused by blood loss, burn etc.,
Hypoproteinemia caused by oedema caused by cirrhosis or nephrosis and many reasons etc., pharmacological action are to increase blood volume
With maintenance plasma colloid osmotic pressure, albumin accounts for the 80% of plasma colloid osmotic pressure, main to adjust moisture between tissue and blood vessel
Dynamic equilibrium.It is slower through speed in film compared with salt and moisture since albumin molecule amount is higher, make the glue of albumin
The static pressure of body osmotic pressure and capillary is contended with, and maintains normal and constant blood volume with this.Use human serum albumin as thickness
Agent is maintained, cornea can be maintained to save the colloid osmotic pressure in liquid, to maintain thickness of the cornea in preservation and transportational process
Stablize.
Human serum albumin used in the present invention also can be used as corneal material irradiation protective agent can in irradiation process
It protects cornea by the damage of irradiated rays, not maintain original natural collagen fibre stable structure, and maintains the transparency of cornea.
Glycerol is employed herein as thickness and maintains agent, glycerol no cytotoxicity, molecular weight is small, and it is soluble easily in water, and
Frequently as cryoprotector, corneal material collagenous fibres is avoided to generate damage from freezing and temperature-rise period.In addition, numerous studies
Report, certain density glycerol have a degree of dehydration to cornea collagenous fibres, so that corneal thickness is maintained to stablize,
Oedema will not occur during storage and transport, avoid the swelling of corneal collagen fiber and cause inside collagenous fibres and collagen is fine
Structure between dimension is destroyed.Meanwhile glycerol has antibacterial and virally inactivated effect, can inhibit to save to a certain extent thin in liquid
The growth of bacterium.
Human serum albumin and glycerol are used as thickness to maintain agent by the present invention simultaneously, and specific mechanism is, first with glycerol
Dehydration a degree of dehydration, then the colloid by albumin solution maintenance corneal material are carried out to cornea collagenous fibres
Osmotic pressure avoids corneal collagen caused by long-term preservation to guarantee that corneal material thickness during storage and transport is stablized
Fiber oedema and the cornea cosmetic variation occurred.Meanwhile the presence of human serum albumin, can protect cornea in irradiation process from
Damage of the ray to collagenous fibres maintains the stable structure of cornea, maintains corneal transparence.
The present invention uses low-numerator sodium hyaluronate to maintain agent as transparency.The present invention saves in liquid and adds low molecular weight
Sodium Hyaluronate, Sodium Hyaluronate are widely present in animal and human body, are the main constituents of people's vitreous body of eye, in spy
There is dehydrating effect when fixed concentration, polymer hyaluronic acid sodium molecule amount is higher, is easy film forming, and it is more sticky to form solution, not side
Just the preservation of cornea, and the Sodium Hyaluronate of low molecular weight is guaranteeing the front lower of solution viscosity, additionally it is possible to the thickness of cornea is maintained,
It is one of comparatively ideal component.In irradiation process, the irradiation of ray is likely to carry out the structure of collagenous fibres or albumin
It destroys, so as to cause collagenous degeneration, cornea is caused to turn yellow, transparency decline, and low-numerator sodium hyaluronate can be with collagenous fibres
Portion fractures point recombined, to maintain collagen fiber structure integrality, maintain corneal material in irradiation process
In transparency.
Preservation liquid constituent used herein can maintain corneal thickness to stablize, while can protect corneal material
Structural intergrity and the transparency in irradiation process, are beneficial to the storage and transport of artificial cell-eliminating coanea matrix.
Preservation liquid of the present invention, constituent is simple, and preparing process is easy to operate, is not only applicable to manually take off cell angle
The storage and transport of membrane matrix are equally applicable to the storage and transport of various artificial collagen tissue materials.
Detailed description of the invention
Fig. 1 is the de- cell cornea base that 6 months are stored at room temperature using collagen tissue material preservation liquid in the embodiment of the present invention
Matter state diagram；
Fig. 2 is the cell-eliminating coanea matrix state diagram that 2 months are stored at room temperature using 95% glycerol preservation solution；
Fig. 3 is the cell-eliminating coanea matrix state diagram that 6 months are stored at room temperature using 95% glycerol preservation solution.
Specific embodiment
The present invention is specifically described below by embodiment, embodiment is only used for carrying out furtherly the present invention
It is bright, it should not be understood as limiting the scope of the present invention, person skilled in art can be according to the content of foregoing invention to this
Some modifications and adaptations for being not required to carry out by creative work are made in invention.
The present invention provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: thickness maintains agent
25 ~ 95 parts, transparency maintains 3 ~ 6 parts of agent, 25 ~ 55 parts of solvent.
In some embodiments, it is 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 that the thickness, which maintains the parts by weight of agent,
Part, 32 parts, 35 parts, 37 parts, 38 parts, 40 parts, 41 parts, 42 parts, 43 parts, 45 parts, 47 parts, 52 parts, 54 parts, 56 parts, 57 parts, 58
Part, 59 parts, 60 parts, 62 parts, 63 parts, 64 parts, 65 parts, 67 parts, 70 parts, 71 parts, 72 parts, 73 parts, 72 parts, 75 parts, 78 parts, 80
Part, 82 parts, 85 parts, 87 parts, 88 parts, 89 parts, 90 parts, 91 parts, 92 parts, 93 parts, 94 parts or 95 parts.
In some embodiments, it is 3 parts, 4 parts, 5 parts or 6 parts that the transparency, which maintains the parts by weight of agent,.
In some embodiments, the thickness maintains agent to be selected from albumin and glycerol.
In some embodiments, the albumin is selected from human serum albumin, and the human serum albumin is freeze dried powder.
Human serum albumin content of the present invention is 10 ~ 50 parts, and the glycerol content is 15 ~ 45 parts.
In some embodiments, according to parts by weight, the human serum albumin content be 10 parts, 11 parts, 12 parts, 13 parts,
14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29
Part, 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts, 37 parts, 38 parts, 39 parts, 40 parts, 41 parts, 42 parts, 43 parts, 44
Part, 45 parts, 46 parts, 47 parts, 48 parts, 49 or 50 parts, the glycerol content be 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts,
21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36
Part, 37 parts, 38 parts, 39 parts, 40 parts, 41 parts, 42 parts, 43 parts, 44 parts or 45 parts.
Transparency described in the embodiment of the present invention maintains agent to be selected from Sodium Hyaluronate.
Sodium Hyaluronate described in the embodiment of the present invention is low-numerator sodium hyaluronate, the low-numerator sodium hyaluronate
Molecular weight is 20 ~ 800,000 Da.
Solvent described in the embodiment of the present invention is selected from purified water.
The embodiment of the present invention also provides above-mentioned collagen tissue material preservation liquid in saving artificial collagen tissue material
Using.
Artificial collagen tissue material described in the embodiment of the present invention is selected from artificial collagen tissue material, the artificial collagen tissue
Material is selected from cell-eliminating coanea matrix, acellular conjunctiva matrix, de- cell stroma of sclera or de- cell amnion.
It is illustrated below with reference to specific embodiment:
Embodiment 1
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 40
Part, 30 parts of glycerol, 5 parts of low-numerator sodium hyaluronate and 25 parts of purified water.
Embodiment 2
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 30
Part, 30 parts of glycerol, 5 parts of low-numerator sodium hyaluronate and 35 parts of purified water.
Embodiment 3
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 22
Part, 20 parts of glycerol, 3 parts of low-numerator sodium hyaluronate and 55 parts of purified water.
Embodiment 4
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 25
Part, 35 parts of glycerol, 5 parts of low-numerator sodium hyaluronate and 35 parts of purified water.
Embodiment 5
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 10
Part, 45 parts of glycerol, 3 parts of low-numerator sodium hyaluronate and 42 parts of purified water.
Embodiment 6
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 50
Part, 15 parts of glycerol, 6 parts of low-numerator sodium hyaluronate and 29 parts of purified water.
Embodiment 7
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 35
Part, 20 parts of glycerol, 5 parts of low-numerator sodium hyaluronate and 40 parts of purified water.
Embodiment 8
The present embodiment provides a kind of collagen tissue material preservation liquid, according to parts by weight, including following components: human serum albumin 45
Part, 20 parts of glycerol, 3 parts of low-numerator sodium hyaluronate and 32 parts of purified water.
Compliance test result:
By de- cell cornea product dehydrate to original corneal thickness, the preceding elastic force of specific thicknesses is cut using 400um cutter head later
Layer and fraction matrix layer, and corneal thickness is measured with feeler, record thickness is numbered.The cornea sample cut is divided into
Two groups, at the same be put into collagen tissue material preservation liquid described in any embodiment of the present invention and the glycerol preservation solution of 95% concentration in,
Carry out electron beam irradiation sterilization.The performance parameters such as thickness, appearance and the transparency of cornea are measured after a period of time, obtain being put into guarantor
The thickness change ratio (table 1) of different time after liquid storage.As can be seen from the table, traditional glycerol preservation solution, although can tie up
Hold the thickness of cornea, but cornea has flavescence to be hardened trend.Collagen tissue material preservation liquid described in the embodiment of the present invention can have
The thickness and transparency and elasticity of the maintenance cornea of effect, and operation consent only need to be rinsed slightly, it is simple to operate.
1 corneal graft of table is put into the relevant parameter variation after different preservation liquid
Through testing, collagen tissue material preservation liquid prepared by the embodiment of the present invention becomes after irradiation or electron beam sterilization without color
Change, and has no irritating odor.With reference to Fig. 1-Fig. 3, Fig. 1 is to be protected using collagen tissue material preservation liquid room temperature in the embodiment of the present invention
Deposit cell-eliminating coanea matrix state diagram in 6 months；Fig. 2 is the de- cell cornea that 2 months are stored at room temperature using 95% glycerol preservation solution
Substrate status figure；Fig. 3 is the cell-eliminating coanea matrix state diagram that 6 months are stored at room temperature using 95% glycerol preservation solution.This preservation liquid
In 4 DEG C cornea preservation 6 months, corneal transparence is good, no yellowing phenomenon, and is able to maintain good thickness, toughness and elasticity, multiple
It is not in Swelling after water.
In conclusion the collagen tissue material preservation liquid of preparation of the embodiment of the present invention, can maintain de- cell angle for a long time
The thickness and transparency of membrane matrix, can be widely used in medical field.Collagen tissue material preservation liquid of the present invention is to cutting
The de- cell cornea product cut, there is certain dehydrating effect, is able to maintain that the transparency and thickness of cornea.
Although embodiment of the present invention is described in conjunction with attached drawing above, the invention is not limited to above-mentioned
Specific embodiments and applications field, above-mentioned specific embodiment is only schematical, directiveness, rather than is limited
Property.Those skilled in the art are under the enlightenment of specification, in the range for not departing from the claims in the present invention and being protected
In the case where, a variety of forms can also be made, these belong to the column of protection of the invention.
Claims (9)
1. collagen tissue material preservation liquid, which is characterized in that according to parts by weight, including following components: thickness maintains agent 25 ~ 95
Part, transparency maintains 3 ~ 6 parts of agent, 25 ~ 55 parts of solvent.
2. collagen tissue material preservation liquid according to claim 1, which is characterized in that the thickness maintains agent to be selected from white egg
White and glycerol.
3. collagen tissue material preservation liquid according to claim 1, which is characterized in that the albumin is selected from the white egg of people's blood
White, the human serum albumin is freeze dried powder.
4. collagen tissue material preservation liquid according to claim 1, which is characterized in that according to parts by weight, people's blood
Albumin content is 10 ~ 50 parts, and the glycerol content is 15 ~ 45 parts.
5. collagen tissue material preservation liquid according to claim 1, which is characterized in that transparency maintains agent to be selected from hyalomitome
Sour sodium.
6. collagen tissue material preservation liquid according to claim 1, which is characterized in that the Sodium Hyaluronate is low molecule
Sodium Hyaluronate, the molecular weight of the low-numerator sodium hyaluronate are 20 ~ 800,000 Da.
7. collagen tissue material preservation liquid according to claim 1, which is characterized in that the solvent is selected from purified water.
8. collagen tissue material preservation liquid described in claim 1 is saving the application in collagen tissue material.
9. application according to claim 8, the collagen tissue material is selected from artificial collagen tissue material, the artificial rubber
Basic stitch material is selected from cell-eliminating coanea matrix, acellular conjunctiva matrix, de- cell stroma of sclera or de- cell amnion.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111802378A (en) * | 2020-07-24 | 2020-10-23 | 镇江雷音再生医学科技有限公司 | SMILE (Small Scale Integrated Circuit) -derived protective solution for human corneal lens and preparation method thereof |
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| CN111802378A (en) * | 2020-07-24 | 2020-10-23 | 镇江雷音再生医学科技有限公司 | SMILE (Small Scale Integrated Circuit) -derived protective solution for human corneal lens and preparation method thereof |
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Application publication date: 20191122 |