CN111903663A - Method for preserving scleral tissue - Google Patents
Method for preserving scleral tissue Download PDFInfo
- Publication number
- CN111903663A CN111903663A CN202010823467.1A CN202010823467A CN111903663A CN 111903663 A CN111903663 A CN 111903663A CN 202010823467 A CN202010823467 A CN 202010823467A CN 111903663 A CN111903663 A CN 111903663A
- Authority
- CN
- China
- Prior art keywords
- preserving
- scleral tissue
- tissue
- conjunctiva
- scleral
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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Classifications
-
- 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
-
- 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
- A01N1/0231—Chemically defined matrices, e.g. alginate gels, for immobilising, holding or storing cells, tissue or organs for preservation purposes; Chemically altering or fixing cells, tissue or organs, e.g. by cross-linking, for preservation purposes
Abstract
The invention discloses a method for preserving scleral tissues, which is characterized in that virus-free rinsed tissues carrying fresh sclera and conjunctiva are sterilized and then subjected to air exposure culture; the air exposure culture is to cover the conjunctiva on the scleral tissue and then place the scleral tissue on a water-permeable membrane, and a preservation solution or a cell culture medium which can provide tissue nutrients is arranged below the membrane. The method for preserving the scleral tissue can ensure that the conjunctiva and the scleral tissue are preserved in the environment of the artificial wet house, is similar to the surface survival environment of normal physiological eyeballs in human bodies, and has the advantages of little damage to scleral cell structures and complete preservation of the structures. Compared with the existing scleral tissue preservation method, the method has the advantages of simple and reliable manufacturing process, easy implementation, relatively low cost, easy popularization and convenient transportation, and can provide important technical support for industrialization of the conjunctiva and sclera of tissue engineering.
Description
Technical Field
The invention relates to the technical field of ophthalmic medical treatment, in particular to a scleral tissue preservation method.
Background
During the preservation period of the traditional sclera material, the traditional sclera material is mainly preserved according to the preservation method of the cornea material, and the specific method is as follows: after eye tissues such as conjunctiva, extraocular muscles, iris, ciliary body, choroid, optic nerve and the like are removed, the storage method is complex in flow and difficult to operate, and the clinical application of active tissues is limited to a certain extent.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a scleral tissue preservation method which is simple in manufacturing process, reliable and easy to implement.
In order to achieve the above purpose, the solution of the invention is:
a method for preserving sclera tissue comprises sterilizing fresh sclera and conjunctiva tissue without virus by repeated rinsing, and air-exposing and culturing; the air exposure culture is to cover the conjunctiva on the scleral tissue and then place the scleral tissue on a water-permeable membrane, and a preservation solution or a cell culture medium which can provide tissue nutrients is arranged below the membrane.
After the scheme is adopted, the method for preserving the scleral tissue can ensure that the conjunctiva and the scleral tissue are preserved in the environment of the artificial wet house, is similar to the surface survival environment of normal physiological eyeballs in human bodies, and has the advantages of little damage to scleral cell structures and complete preservation of the structures. Compared with the existing scleral tissue preservation method, the method has the advantages of simple and reliable manufacturing process, easy implementation, relatively low cost, easy popularization and convenient transportation, and can provide important technical support for industrialization of the conjunctiva and sclera of tissue engineering.
Further, the disinfection is to soak and wash fresh sclera and conjunctiva with phosphate buffer solution containing antibiotics, wherein the soaking time is 5-10min, and the PBS is washed for 3 times and 5 min.
Further, the antibiotic comprises any one or any combination of 5 million U/L penicillin, 8 million U/L tobramycin and 100mg/L streptomycin.
Further, the film laying comprises any one of a nitrocellulose film, a cellulose acetate film and a microporous filter film.
Further, the pore diameter of the film laying is as follows: 0.1 μm, 0.22 μm, 0.45 μm, 0.65 μm, 1.0 μm, 3.0 μm, 5.0 μm, and the thickness of the coating film is: 100-140 μm. Therefore, the conjunctiva and the sclera tissues can be fully flattened, a certain stretching degree is kept, and meanwhile, the conjunctiva and the sclera tissues can be fully contacted with nutrient solution.
Further, the preservation solution comprises Optisol metaphase preservation solution, DMEM medium, SHEM medium, serum or one or more combination of various cytokines.
Further, the air exposure culture condition is 0-37 ℃, and the oxygen concentration is 0-20%.
Detailed Description
In order to further explain the technical solution of the present invention, the present invention is explained in detail by the following specific examples.
A method for preserving sclera tissue comprises the steps of sterilizing fresh sclera and conjunctiva tissue carried by virus-free rinsed repeatedly, and preserving the conjunctiva tissue in an air exposure way, wherein the air exposure culture is to cover the conjunctiva on the sclera tissue and then place the sclera tissue on a water-permeable film laying, and preserving solution or cell culture medium capable of providing tissue nutrients is arranged below the film laying. The sclera tissue preservation method can ensure that the preserved conjunctiva and the sclera tissue are in an artificial wet room environment, is similar to the surface survival environment of normal physiological eyeballs in a human body, and has little damage to the sclera cell structure.
The disinfection is carried out by soaking fresh sclera and conjunctiva in phosphate buffer solution containing antibiotics for 5-10min, and washing with PBS for 3 times and 5 min. The antibiotic comprises one or the combination of more than two of 5 million U/L penicillin, 8 million U/L tobramycin and 100mg/L streptomycin.
The film laying comprises one of a nitrocellulose film, a cellulose acetate film, a microporous filter film or other water permeable materials, and the optional pore diameter is as follows: 0.1 μm, 0.22 μm, 0.45 μm, 0.65 μm, 1.0 μm, 3.0 μm, 5.0 μm. Appearance: white. Thickness: 100-140 μm. The technology can ensure that conjunctiva and sclera tissues are fully flattened, and can be fully contacted with nutrient solution while keeping a certain stretching degree.
The preservation solution comprises Optisol metaphase preservation solution, DMEM medium, SHEM medium, serum or one or more combinations of various cytokines. The formula is shown later.
The air exposure culture conditions are 0-37 ℃ and the oxygen concentration is 0-20%.
The above embodiments are not intended to limit the form and style of the present invention, and any suitable changes or modifications made by those skilled in the art should be considered as not departing from the scope of the present invention.
Claims (7)
1. A method for preserving scleral tissue, comprising: sterilizing the repeatedly rinsed virus-free sclera and conjunctiva tissues carrying fresh sclera and conjunctiva tissues and then carrying out air exposure culture; the air exposure culture is to cover the conjunctiva on the scleral tissue and then place the scleral tissue on a water-permeable membrane, and a preservation solution or a cell culture medium which can provide tissue nutrients is arranged below the membrane.
2. A method of preserving scleral tissue as in claim 1, wherein: the disinfection is carried out by soaking fresh sclera and conjunctiva in phosphate buffer solution containing antibiotics for 5-10min and washing with phosphate buffer solution for 3 times and 5 min.
3. A method of preserving scleral tissue as in claim 2, wherein: the antibiotic comprises any one or any combination of 5 million U/L penicillin, 8 million U/L tobramycin and 100mg/L streptomycin.
4. A method of preserving scleral tissue as in claim 1, wherein: the film laying comprises any one of a nitrocellulose film, a cellulose acetate film and a microporous filter film.
5. A method of preserving scleral tissue as claimed in claim 1 or 4, wherein: the pore diameter of the film laying is as follows: 0.1 μm, 0.22 μm, 0.45 μm, 0.65 μm, 1.0 μm, 3.0 μm, 5.0 μm, and the thickness of the coating film is: 100-140 μm.
6. A method of preserving scleral tissue as in claim 1, wherein: the preservation solution comprises Optisol metaphase preservation solution, DMEM medium, SHEM medium, serum or one or more combinations of various cytokines.
7. A method of preserving scleral tissue as in claim 1, wherein: the air exposure culture conditions are 0-37 ℃ and the oxygen concentration is 0-20%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010823467.1A CN111903663A (en) | 2020-08-17 | 2020-08-17 | Method for preserving scleral tissue |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010823467.1A CN111903663A (en) | 2020-08-17 | 2020-08-17 | Method for preserving scleral tissue |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111903663A true CN111903663A (en) | 2020-11-10 |
Family
ID=73278112
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010823467.1A Pending CN111903663A (en) | 2020-08-17 | 2020-08-17 | Method for preserving scleral tissue |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111903663A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726370A (en) * | 2012-06-29 | 2012-10-17 | 厦门大学附属厦门眼科中心 | Preservation method for corneal limbus tissue |
| US20200121829A1 (en) * | 2014-05-12 | 2020-04-23 | Gholam A. Peyman | Method Of Corneal And Scleral Inlay Crosslinking And Preservation |
-
2020
- 2020-08-17 CN CN202010823467.1A patent/CN111903663A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726370A (en) * | 2012-06-29 | 2012-10-17 | 厦门大学附属厦门眼科中心 | Preservation method for corneal limbus tissue |
| US20200121829A1 (en) * | 2014-05-12 | 2020-04-23 | Gholam A. Peyman | Method Of Corneal And Scleral Inlay Crosslinking And Preservation |
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| Date | Code | Title | Description |
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| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: No. 336, Xiahe Road, Siming District, Xiamen City, Fujian Province, 361000 Applicant after: Xiamen Eye Center Co.,Ltd. Applicant after: Zhenjiang rehabilitation Eye Hospital Co.,Ltd. Address before: No. 336, Xiahe Road, Siming District, Xiamen City, Fujian Province, 361000 Applicant before: XIAMEN EYE CENTER OF XIAMEN UNIVERSITY Co.,Ltd. Applicant before: Zhenjiang rehabilitation Eye Hospital Co.,Ltd. |
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| CB02 | Change of applicant information | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201110 |
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| RJ01 | Rejection of invention patent application after publication |