CN104938476B - A kind of osteochondral graft saves liquid and preparation method thereof - Google Patents
A kind of osteochondral graft saves liquid and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of osteochondral grafts to save liquid and preparation method thereof, comprising: 80~125mmol of glucose, 0.1~2.0mmol of amino acid, 0.2~2.0mmol of antioxidant, 5~60nmol of basic fibroblast growth factor, 1~120mmol of inorganic salts, 1~9mmol of vitamin, 1~2mmol of Sodium Pyruvate, 50~70U of penicillin, it is remaining be deionized water；It is configured to solution, adjusting pH value to 7.30~7.50, filtration sterilization；Fresh Bone and osterochordral allografts object is cleaned 3-5 times through sterile saline, is put into have prepared osteochondral graft and saved and be impregnated in liquid；4 DEG C of preservations.The present invention resists cartilage cell's aging in preservation, the good biological activity for inhibiting articular chondrocyte apoptosis, keeping cartilaginous tissue, maintains cartilage cell's survival rate, the extension holding time, improves the effect of osteochondral graft preservation effect.
Description
Technical field
The invention belongs to biomedicine technical fields more particularly to a kind of osteochondral graft to save liquid and its preparation side
Method.
Background technique
In recent years, with comprehensive universal, the articular cartilage damage of China's improvement of people's living standards and sports
Patient significantly increases.Currently, there are no very ideal methods for the repairing and treating of articular cartilage damage.Therefore, articular cartilage is damaged
Hurt repairing and treating method as difficult medical problem urgently to be resolved in the world.
Since articular cartilage disorders cannot be repaired naturally, if eventually leading to the hair of Osteoarthritis without effectively treating
It is raw, lose function of joint, serious person can disable disease.Prosthetic replacement is suitble to the treatment of old Human Osteoarthritis, however
For young articular cartilage damage patient but without ideal treatment method.
Currently, the method for clinical treatment joint injury has: micro fractures method, organizational project cure, Autologous Chondrocyte
Transplantation, joint replacement, articular cartilage grafting.In these treatment methods, Bone and osterochordral allografts art can be compared with
Good reparation hyaline cartilage damage, reduces second operation wound, becomes treatment and repairs the very promising method of articular defect.
However, the storage in vitro time of allosome osteochondral tissue is shorter, clinical application is limited.Therefore, bone cartilage is persistently kept
The activity of graft and higher cartilage cell's survival rate, extension body outer holding time become current key technology urgently to be solved
Problem.
During allosome osteochondral tissue saves in vitro, with the extension of holding time, articular chondrocyte apoptosis increases, soft
Bone and its cells survival rate and tissue activity can be substantially reduced, cartilage matrix degradation increase, seriously affect transplanting after organize deposit
Living, the long-term in vitro for limiting allosome osteochondral tissue saves.Meanwhile Bone and osterochordral allografts art is also affected in clinic
On extensive use.
The present invention is finally developed by the research and probe to different culture solution preservation conditions by repeated multiple times experiment
The preservation liquid that can effectively improve the holding time of the outer osteochondral graft of osteochondral graft preservation effect, extension body, from
And be to establish cartilaginous tissue library, expand Bone and osterochordral allografts reparation cartilage defect clinical technology popularization and application and establishes well
Basis, also will be to bring glad tidings to the quick recovery of osteochondral defect patient.
Summary of the invention
The purpose of the present invention is to provide a kind of osteochondral grafts to save liquid and preparation method thereof, it is intended to solve existing
Persistently keep the problem that the activity of osteochondral graft is low with cartilage cell's survival rate, the storage in vitro time is short.
The invention is realized in this way a kind of osteochondral graft saves liquid, it is every which saves liquid
It includes: 80~125mmol of glucose, 0.1~2.0mmol of amino acid, antioxidant 0.2 that 1000ml osteochondral graft, which saves liquid,
~2.0mmol, 5~60nmol of basic fibroblast growth factor, 1~120mmol of inorganic salts, 1~9mmol of vitamin, third
1~2mmol of ketone acid sodium, 50~70U of penicillin, it is remaining be deionized water.
Further, the pH value of the bone cartilage preservation solution is 7.30~7.50；4 DEG C of preservations.
Further, the every 1000ml osteochondral graft of the bone cartilage preservation solution saves liquid 100mmol containing glucose, amino acid
0.1mmol, antioxidant 0.2mmol, basic fibroblast growth factor 2 5nmol, inorganic salts 20mmol, vitamin
9mmol, Sodium Pyruvate 1mmol, penicillin 60U, remaining is deionized water.
Further, the every 1000ml osteochondral graft of the bone cartilage preservation solution saves liquid 110mmol containing glucose, amino acid
0.5mmol, antioxidant 1.3mmol, basic fibroblast growth factor 5nmol, inorganic salts 80mmol, vitamin 4mmol,
Sodium Pyruvate 1.5mmol, penicillin 50U, it is remaining be deionized water.
Further, the every 1000ml osteochondral graft of the bone cartilage preservation solution saves liquid 125mmol containing glucose, amino acid
1.0mmol, antioxidant 2.0mmol, basic fibroblast growth factor 50nmol, inorganic salts 120mmol, vitamin
1mmol, Sodium Pyruvate 1mmol, penicillin 55U, it is remaining be deionized water.
Another object of the present invention is to provide the preparation method that a kind of osteochondral graft saves liquid, the bone cartilage transplantations
The preparation method that object saves liquid includes the following steps:
Step 1, take glucose, amino acid, antioxidant, basic fibroblast growth factor, inorganic salts, vitamin,
Sodium Pyruvate, penicillin and deionized water；
The raw material of step 1 is configured to solution, stirred evenly by step 2, is adjusted pH value to 7.30~7.50, is crossed and filter out
Bacterium；
Step 3 cleans fresh Bone and osterochordral allografts object 3-5 times through sterile saline, is put into and has prepared
Osteochondral graft saves to be impregnated in liquid；
Step 3 osteochondral graft preservation liquid is put into 4 DEG C of preservations by step 4.
Another object of the present invention is to provide a kind of osteochondral grafts to save liquid in the application in osteochondral tissue library.
Liquid is saved another object of the present invention is to provide a kind of osteochondral graft to move in clinical homogeneous allogenic bone cartilage
Plant the application of art treatment articular cartilage damage.
Osteochondral graft provided by the invention saves liquid and preparation method thereof, and cartilage cell's survival rate is made to remain at one
A higher level has cartilage cell's aging during resisting preservation, inhibits articular chondrocyte apoptosis, keeps the good of cartilaginous tissue
Biological activity, the degradation for reducing cartilage matrix extend the holding time, improve the effect of osteochondral graft preservation effect.This
Invention can help to establish the new skill in osteochondral tissue library and clinical Bone and osterochordral allografts art treatment articular cartilage damage
Art is promoted.
Detailed description of the invention
Fig. 1 is the preparation method flow chart that osteochondral graft provided in an embodiment of the present invention saves liquid；
Fig. 2 is that osteochondral graft preservation liquid preservation sheep knee joint osteochondral tissue provided in an embodiment of the present invention is saving
Double fluorescent staining color (EB/FDA) cartilage cell's survival rate schematic diagram when the 1st day, 21 days, 35 days.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
With reference to the accompanying drawing and specific embodiment is further described application principle of the invention.
It includes: that every 1000ml osteochondral graft saves liquid containing grape that the osteochondral graft of the embodiment of the present invention, which saves liquid,
80~125mmol of sugar, 0.1~2.0mmol of amino acid, 0.2~2.0mmol of antioxidant, basic fibroblast growth factor
(bFGF) 5~60nmol, 1~120mmol of inorganic salts, 1~9mmol of vitamin, 1~2mmol of Sodium Pyruvate, penicillin 50~
70U, it is remaining be deionized water；
The pH value of the bone cartilage preservation solution is 7.30~7.50,4 DEG C of preservations.
As shown in Figure 1, the embodiment of the present invention osteochondral graft save liquid preparation method the following steps are included:
S101: glucose, amino acid, antioxidant, basic fibroblast growth factor, inorganic salts, vitamin, third are taken
Ketone acid sodium, penicillin and deionized water；
S102: being configured to solution for the raw material of S101, stir evenly, adjusting pH value to 7.30~7.50, filtration sterilization；
S103: fresh Bone and osterochordral allografts object is cleaned 3-5 times through sterile saline, is put into and has prepared bone
Cartilage graft saves to be impregnated in liquid；
S104: S103 osteochondral graft preservation liquid is put into 4 DEG C of preservations.
Specific embodiments of the present invention:
It is as shown in Figure 2:
Embodiment 1: every 1000ml osteochondral graft saves liquid 80mmol containing glucose, amino acid 2.0mmol, anti-oxidant
Agent 0.5mmol, basic fibroblast growth factor (bFGF) 60nmol, inorganic salts 1mmol, vitamin 6mmol, Sodium Pyruvate
2mmol, penicillin 70U.Remaining is deionized water, and the pH value of the bone cartilage preservation solution is 7.30~7.50.4 DEG C of preservations.It is saving
When the 21st day, 35 days, cartilage cell's survival rate of osteochondral tissue is detected.When saving the 21st day, cell survival
Rate is 85%.
Embodiment 2: every 1000ml osteochondral graft saves liquid 100mmol containing glucose, amino acid 0.1mmol, antioxygen
Agent 0.2mmol, basic fibroblast growth factor (bFGF) 25nmol, inorganic salts 20mmol, vitamin 9mmol, acetone
Sour sodium 1mmol, penicillin 60U.Remaining is deionized water, and the pH value of the bone cartilage preservation solution is 7.30~7.50.4 DEG C of preservations.?
When saving the 21st day, 35 days, cartilage cell's survival rate of osteochondral tissue is detected.When saving the 21st day, cell
Survival rate is 86%.
Embodiment 3: every 1000ml osteochondral graft saves liquid 110mmol containing glucose, amino acid 0.5mmol, antioxygen
Agent 1.3mmol, basic fibroblast growth factor (bFGF) 5nmol, inorganic salts 80mmol, vitamin 4mmol, pyruvic acid
Sodium 1.5mmol, penicillin 50U, it is remaining be deionized water, the pH value of the bone cartilage preservation solution is 7.30~7.50.4 DEG C of preservations.?
When saving the 21st day, 35 days, cartilage cell's survival rate of osteochondral tissue is detected.When saving the 21st day, cell
Survival rate is 84%.
Embodiment 4: every 1000ml osteochondral graft saves liquid 125mmol containing glucose, amino acid 1 .0mmol, antioxygen
Agent 2.0mmol, basic fibroblast growth factor (bFGF) 50nmol, inorganic salts 120mmol, vitamin 1mmol, acetone
Sour sodium 1mmol, penicillin 55U, it is remaining be deionized water, the pH value of the bone cartilage preservation solution is 7.30~7.50.4 DEG C of preservations.?
When saving the 21st day, cell survival rate 85%.
Inorganic salts can effectively keep saving liquid acid-base balance and Premeabilisation of cells pressure in the present invention；Glucose, amino acid can have
Effect provides energy and nutriment source for cell；Antioxidant is highly resistant to cell ageing；Basic fibroblast growth
The factor can effectively facilitate chondrocytes for regeneration.Adjusting pH value is 7.30~7.50 suitable for histiocytic growth in vitro, and 4 DEG C are preserved
Conducive to histocyte metabolic rate is reduced, mitigates tissue and damage again, extend the holding time.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (6)
1. a kind of osteochondral graft saves liquid, which is characterized in that it includes: grape that the osteochondral graft of every 1000ml, which saves liquid,
80~125mmol of sugar, 0.1~2.0mmol of amino acid, 0.2~2.0mmol of antioxidant, basic fibroblast growth factor 5
~60nmol, 1~120mmol of inorganic salts, 1~9mmol of vitamin, 1~2mmol of Sodium Pyruvate, 50~70U of penicillin, Yu Wei
Deionized water；
The pH value of the bone cartilage preservation solution is 7.30~7.50；4 DEG C of preservations.
2. osteochondral graft as described in claim 1 saves liquid, which is characterized in that every 1000ml osteochondral graft saves
Liquid 100mmol containing glucose, amino acid 0.1mmol, antioxidant 0.2mmol, basic fibroblast growth factor 2 5nmol,
Inorganic salts 20mmol, vitamin 9mmol, Sodium Pyruvate 1mmol, penicillin 60U, remaining is deionized water.
3. osteochondral graft as described in claim 1 saves liquid, which is characterized in that every 1000ml osteochondral graft saves
Liquid 110mmol containing glucose, amino acid 0.5mmol, antioxidant 1.3mmol, basic fibroblast growth factor 5nmol,
Inorganic salts 80mmol, vitamin 4mmol, Sodium Pyruvate 1.5mmol, penicillin 50U, it is remaining be deionized water.
4. osteochondral graft as described in claim 1 saves liquid, which is characterized in that every 1000ml osteochondral graft saves
Liquid 125mmol containing glucose, amino acid 1 .0mmol, antioxidant 2.0mmol, basic fibroblast growth factor 50nmol,
Inorganic salts 120mmol, vitamin 1mmol, Sodium Pyruvate 1mmol, penicillin 55U, it is remaining be deionized water.
5. a kind of osteochondral graft as described in claim 1 saves liquid, which is characterized in that the osteochondral graft saves liquid
Preparation method include the following steps:
Step 1 takes glucose, amino acid, antioxidant, basic fibroblast growth factor, inorganic salts, vitamin, acetone
Sour sodium, penicillin and deionized water；
The raw material of step 1 is configured to solution, stirred evenly by step 2, adjusting pH value to 7.30~7.50, filtration sterilization；
Step 3 cleans fresh Bone and osterochordral allografts object 3-5 times through sterile saline, is put into that have prepared bone soft
Bone graft saves to be impregnated in liquid；
Step 3 osteochondral graft preservation liquid is put into 4 DEG C of preservations by step 4.
6. a kind of osteochondral graft as described in claim 1 saves liquid in the application in osteochondral tissue library.
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106614526A (en) * | 2016-12-30 | 2017-05-10 | 潍坊医学院 | Vitrified cryopreservation method for cartilage |
| CN108041023B (en) * | 2017-12-14 | 2021-04-16 | 泰山医学院 | Mechanical stimulation method for improving in-vitro preservation effect of articular cartilage |
| CN109392892B (en) * | 2018-12-14 | 2021-07-23 | 广州悦清再生医学科技有限公司 | Biological collagen-based material preserving fluid and application thereof |
| CN109566601A (en) * | 2018-12-29 | 2019-04-05 | 中国人民解放军第四军医大学 | A kind of cartilage preservation solution and its application method |
| CN109644989A (en) * | 2019-01-28 | 2019-04-19 | 徐艳红 | A kind of articular cartilage frozen solution and cartilage freezing and storing method |
| CN110558312B (en) * | 2019-10-09 | 2020-07-28 | 科瑞百奥泰州生物技术有限公司 | Cartilage tissue low-temperature refrigeration preservation solution and application thereof |
| CN114831108B (en) * | 2022-05-20 | 2023-05-12 | 杭州联川生物技术股份有限公司 | Fresh tissue preservation solution and preparation method and application thereof |
| CN116439231B (en) * | 2023-06-09 | 2023-09-19 | 山东第一医科大学(山东省医学科学院) | Ultrasonic stimulation method for improving preservation effect of in-vitro bone cartilage tissue |
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| US20140120068A1 (en) * | 2012-11-01 | 2014-05-01 | Animal Cell Therapies, Inc. | Solutions, systems and methods for cell, tissue and organ preservation |
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