CN104938476A - Osteochondral graft preservation liquid and preparation method thereof - Google Patents
Osteochondral graft preservation liquid and preparation method thereof Download PDFInfo
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- CN104938476A CN104938476A CN201510093497.0A CN201510093497A CN104938476A CN 104938476 A CN104938476 A CN 104938476A CN 201510093497 A CN201510093497 A CN 201510093497A CN 104938476 A CN104938476 A CN 104938476A
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Abstract
The invention discloses an osteochondral graft preservation liquid and a preparation method thereof. The osteochondral graft preservation liquid comprises the following raw materials: 80-125mmol of glucose, 0.1-2.0mmol of amino acid, 0.2-2.0mmol of an antioxidant, 5-60nmol of a basic fibroblast growth factor, 1-120mmol of inorganic salt, 1-9mmol of vitamin, 1-2mmol of sodium pyruvate, 50-70U of penicillin and the balance of deionized water. The method comprises the following steps: preparing a solution, adjusting the pH to be 7.30-7.50, filtering and degerming; cleaning fresh allogeneic osteochondral graft with a sterile saline solution for 3-5 times, and putting into the prepared osteochondral graft preservation liquid to soak; and storing at 4 DEG C. The osteochondral graft preservation liquid has the efficacies of resisting cartilage cells senescence in preservation, inhibiting cartilage cells apoptosis, keeping good biological activity of cartilage tissues, maintaining the survival rate of cartilage cells, prolonging the storage time and improving the storage effect of the osteochondral graft.
Description
Technical field
The invention belongs to biomedicine technical field, particularly relate to a kind of osteochondral graft conserving liquid and preparation method thereof.
Background technology
In recent years, along with China people growth in the living standard and the comprehensive of sports popularize, articular cartilage damage patient significantly increase.Current, the repairing and treating of articular cartilage damage does not also have very good method.Therefore, articular cartilage damage repairing and treating method becomes difficult medical problem urgently to be resolved hurrily in the world.
Because articular cartilage disorders can not be repaired naturally, if effectively do not treat, finally cause the generation of Osteoarthritis, function of joint is lost, severe patient can be disabled disease.Prosthetic replacement is applicable to the treatment of old Human Osteoarthritis, but does not but have desirable methods for the treatment of for young articular cartilage damage patient.
At present, the method for clinical treatment joint injury has: micro-fracture method, organizational project cure, Autologous Chondrocyte transplantation, joint replacement, articular cartilage grafting.In these methods of treatments, Bone and osterochordral allografts art can repair hyaline cartilage damage preferably, decreases second operation wound, becomes treatment and repairs the very promising method of articular defect.But the storage in vitro time of allograph bone cartilaginous tissue is shorter, limits clinical application.Therefore, continue to keep the activity of osteochondral graft and higher cartilage cell's survival rate, extension body outer holding time to become the current key technical problem being badly in need of solving.
In allograph bone cartilaginous tissue preservation process in vitro, along with the prolongation of holding time, articular chondrocyte apoptosis increases, cartilaginous tissue cell survival rate and tissue activity can obviously reduce, cartilage matrix degraded increases, have a strong impact on the survival of tissue after transplanting, the long-term in vitro limiting allograph bone cartilaginous tissue is preserved.Meanwhile, also have impact on the extensive use clinically of Bone and osterochordral allografts art.
The present invention is by the research and probe to different culture fluid preservation condition, through repeated multiple times experiment, finally develop the conserving liquid of the holding time effectively can improving osteochondral graft outside osteochondral graft preservation effect, extension body, thus for setting up cartilaginous tissue storehouse, expand Bone and osterochordral allografts to repair cartilage defect clinical technology and apply and establish good basis, also by for bringing glad tidings to the quick recovery of osteochondral defect patient.
Summary of the invention
The object of the present invention is to provide a kind of osteochondral graft conserving liquid and preparation method thereof, be intended to solve the activity of existing lasting maintenance osteochondral graft and cartilage cell's survival rate is low, the storage in vitro time is short problem.
The present invention realizes like this, a kind of osteochondral graft conserving liquid, this osteochondral graft conserving liquid every 1000ml osteochondral graft conserving liquid comprises: glucose 80 ~ 125mmol, amino acid 0.1 ~ 2.0mmol, antioxidant 0.2 ~ 2.0mmol, basic fibroblast growth factor 5 ~ 60nmol, mineral salt 1 ~ 120mmol, vitamin 1 ~ 9mmol, Sodium Pyruvate 1 ~ 2mmol, penicillin 50 ~ 70U, remaining be deionized water.
Further, the pH value of this bone cartilage preservation solution is 7.30 ~ 7.50; 4 DEG C of preservations.
Further, this bone cartilage preservation solution every 1000ml osteochondral graft conserving liquid is containing glucose 100mmol, amino acid 0.1mmol, antioxidant 0.2mmol, HBGH-2 5nmol, mineral salt 20mmol, vitamin 9mmol, Sodium Pyruvate 1mmol, penicillin 60U, remaining is deionized water.
Further, this bone cartilage preservation solution every 1000ml osteochondral graft conserving liquid containing glucose 110mmol, amino acid 0.5mmol, antioxidant 1.3mmol, basic fibroblast growth factor 5nmol, mineral salt 80mmol, vitamin 4mmol, Sodium Pyruvate 1.5mmol, penicillin 50U, remaining be deionized water.
Further, this bone cartilage preservation solution every 1000ml osteochondral graft conserving liquid containing glucose 125mmol, amino acid/11 .0mmol, antioxidant 2.0mmol, basic fibroblast growth factor 50nmol, mineral salt 120mmol, vitamin 1mmol, Sodium Pyruvate 1mmol, penicillin 55U, remaining be deionized water.
Another object of the present invention is to the preparation method providing a kind of osteochondral graft conserving liquid, the preparation method of this osteochondral graft conserving liquid comprises the steps:
Step one, gets glucose, amino acid, antioxidant, basic fibroblast growth factor, mineral salt, vitamin, Sodium Pyruvate, penicillin and deionized water;
Step 2, is mixed with solution by the raw material of step one, stirs, and regulates pH value to 7.30 ~ 7.50, filtration sterilization;
Step 3, fresh Bone and osterochordral allografts thing through stroke-physiological saline solution cleaning 3-5 time, puts into and prepares osteochondral graft conserving liquid and soak;
Step 4, puts into 4 DEG C of preservations by step 3 osteochondral graft conserving liquid.
Another object of the present invention is to provide the application of a kind of osteochondral graft conserving liquid in osteochondral tissue storehouse.
Another object of the present invention is to the application providing a kind of osteochondral graft conserving liquid at clinical Bone and osterochordral allografts art treatment articular cartilage damage.
Osteochondral graft conserving liquid provided by the invention and preparation method thereof, make cartilage cell's survival rate still remain on a higher level, there is effect that cartilage cell in opposing preservation process is old and feeble, suppress articular chondrocyte apoptosis, keep the good biological activity of cartilaginous tissue, reduce the degraded of cartilage matrix, extend the holding time, improve osteochondral graft preservation effect.The present invention can help the new technology spread of setting up osteochondral tissue storehouse and clinical Bone and osterochordral allografts art treatment articular cartilage damage.
Accompanying drawing explanation
Fig. 1 is preparation method's flow chart of the osteochondral graft conserving liquid that the embodiment of the present invention provides;
Fig. 2 is that the osteochondral graft conserving liquid that provides of the embodiment of the present invention preserves sheep knee joint osteochondral tissue double fluorescent staining look (EB/FDA) cartilage cell survival rate schematic diagram when preservation the 1st day, 21 days, 35 days.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Below in conjunction with drawings and the specific embodiments, application principle of the present invention is further described.
The osteochondral graft conserving liquid of the embodiment of the present invention comprises: every 1000ml osteochondral graft conserving liquid containing glucose 80 ~ 125mmol, amino acid 0.1 ~ 2.0mmol, antioxidant 0.2 ~ 2.0mmol, basic fibroblast growth factor (bFGF) 5 ~ 60nmol, mineral salt 1 ~ 120mmol, vitamin 1 ~ 9mmol, Sodium Pyruvate 1 ~ 2mmol, penicillin 50 ~ 70U, remaining be deionized water;
The pH value of this bone cartilage preservation solution is 7.30 ~ 7.50,4 DEG C of preservations.
As shown in Figure 1, the preparation method of the osteochondral graft conserving liquid of the embodiment of the present invention comprises the following steps:
S101: get glucose, amino acid, antioxidant, basic fibroblast growth factor, mineral salt, vitamin, Sodium Pyruvate, penicillin and deionized water;
S102: the raw material of S101 is mixed with solution, stirs, regulates pH value to 7.30 ~ 7.50, filtration sterilization;
S103: fresh Bone and osterochordral allografts thing through stroke-physiological saline solution cleaning 3-5 time, put into and prepare osteochondral graft conserving liquid and soak;
S104: S103 osteochondral graft conserving liquid is put into 4 DEG C of preservations.
Specific embodiments of the invention:
As shown in Figure 2:
Embodiment 1: every 1000ml osteochondral graft conserving liquid contains glucose 80mmol, amino acid 2.0mmol, antioxidant 0.5mmol, basic fibroblast growth factor (bFGF) 60nmol, mineral salt 1mmol, vitamin 6mmol, Sodium Pyruvate 2mmol, penicillin 70U.Remaining is deionized water, and the pH value of this bone cartilage preservation solution is 7.30 ~ 7.50.4 DEG C of preservations.When preservation the 21st day, 35 days, cartilage cell's survival rate of osteochondral tissue is detected.When preservation the 21st day, its cell survival rate was 85%.
Embodiment 2: every 1000ml osteochondral graft conserving liquid contains glucose 100mmol, amino acid 0.1mmol, antioxidant 0.2mmol, basic fibroblast growth factor (bFGF) 25nmol, mineral salt 20mmol, vitamin 9mmol, Sodium Pyruvate 1mmol, penicillin 60U.Remaining is deionized water, and the pH value of this bone cartilage preservation solution is 7.30 ~ 7.50.4 DEG C of preservations.When preservation the 21st day, 35 days, cartilage cell's survival rate of osteochondral tissue is detected.When preservation the 21st day, its cell survival rate was 86%.
Embodiment 3: every 1000ml osteochondral graft conserving liquid is containing glucose 110mmol, amino acid 0.5mmol, antioxidant 1.3mmol, basic fibroblast growth factor (bFGF) 5nmol, mineral salt 80mmol, vitamin 4mmol, Sodium Pyruvate 1.5mmol, penicillin 50U, remaining be deionized water, the pH value of this bone cartilage preservation solution is 7.30 ~ 7.50.4 DEG C of preservations.When preservation the 21st day, 35 days, cartilage cell's survival rate of osteochondral tissue is detected.When preservation the 21st day, its cell survival rate was 84%.
Embodiment 4: every 1000ml osteochondral graft conserving liquid is containing glucose 125mmol, amino acid/11 .0mmol, antioxidant 2.0mmol, basic fibroblast growth factor (bFGF) 50nmol, mineral salt 120mmol, vitamin 1mmol, Sodium Pyruvate 1mmol, penicillin 55U, remaining be deionized water, the pH value of this bone cartilage preservation solution is 7.30 ~ 7.50.4 DEG C of preservations.When preservation the 21st day, its cell survival rate was 85%.
In the present invention, mineral salt effectively can keep conserving liquid acid base equilibrium and Premeabilisation of cells pressure; Glucose, amino acid can effectively for cell provide energy and nutriment source; Antioxidant is highly resistant to cell ageing; Basic fibroblast growth factor effectively can promote chondrocytes for regeneration.Adjust pH is 7.30 ~ 7.50 be suitable for histiocytic growth in vitro, preserves to be beneficial to for 4 DEG C and reduces histocyte metabolic rate, alleviate to organize and damage, extend the holding time.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. an osteochondral graft conserving liquid, it is characterized in that, the osteochondral graft conserving liquid of every 1000ml comprises: glucose 80 ~ 125mmol, amino acid 0.1 ~ 2.0mmol, antioxidant 0.2 ~ 2.0mmol, basic fibroblast growth factor 5 ~ 60nmol, mineral salt 1 ~ 120mmol, vitamin 1 ~ 9mmol, Sodium Pyruvate 1 ~ 2mmol, penicillin 50 ~ 70U, remaining for deionized water.
2. osteochondral graft conserving liquid as claimed in claim 1, it is characterized in that, the pH value of this bone cartilage preservation solution is 7.30 ~ 7.50; 4 DEG C of preservations.
3. osteochondral graft conserving liquid as claimed in claim 1, it is characterized in that, every 1000ml osteochondral graft conserving liquid is containing glucose 100mmol, amino acid 0.1mmol, antioxidant 0.2mmol, HBGH-2 5nmol, mineral salt 20mmol, vitamin 9mmol, Sodium Pyruvate 1mmol, penicillin 60U, remaining is deionized water.
4. osteochondral graft conserving liquid as claimed in claim 1, it is characterized in that, every 1000ml osteochondral graft conserving liquid containing glucose 110mmol, amino acid 0.5mmol, antioxidant 1.3mmol, basic fibroblast growth factor 5nmol, mineral salt 80mmol, vitamin 4mmol, Sodium Pyruvate 1.5mmol, penicillin 50U, remaining be deionized water.
5. osteochondral graft conserving liquid as claimed in claim 1, it is characterized in that, every 1000ml osteochondral graft conserving liquid containing glucose 125mmol, amino acid/11 .0mmol, antioxidant 2.0mmol, basic fibroblast growth factor 50nmol, mineral salt 120mmol, vitamin 1mmol, Sodium Pyruvate 1mmol, penicillin 55U, remaining be deionized water.
6. an osteochondral graft conserving liquid as claimed in claim 1, is characterized in that, the preparation method of this osteochondral graft conserving liquid comprises the steps:
Step one, gets glucose, amino acid, antioxidant, basic fibroblast growth factor, mineral salt, vitamin, Sodium Pyruvate, penicillin and deionized water;
Step 2, is mixed with solution by the raw material of step one, stirs, and regulates pH value to 7.30 ~ 7.50, filtration sterilization;
Step 3, fresh Bone and osterochordral allografts thing through stroke-physiological saline solution cleaning 3-5 time, puts into and prepares osteochondral graft conserving liquid and soak;
Step 4, puts into 4 DEG C of preservations by step 3 osteochondral graft conserving liquid.
7. the application of an osteochondral graft conserving liquid as claimed in claim 1 in osteochondral tissue storehouse.
8. an osteochondral graft conserving liquid as claimed in claim 1 is in the application of clinical Bone and osterochordral allografts art treatment articular cartilage damage.
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Cited By (8)
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| CN106614526A (en) * | 2016-12-30 | 2017-05-10 | 潍坊医学院 | Vitrified cryopreservation method for cartilage |
| CN108041023A (en) * | 2017-12-14 | 2018-05-18 | 泰山医学院 | A kind of mechanical stimulation method for improving articular cartilage storage in vitro effect |
| CN109392892A (en) * | 2018-12-14 | 2019-03-01 | 广州悦清再生医学科技有限公司 | A kind of biological collagen sill saves liquid and application |
| 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 |
| CN110558312A (en) * | 2019-10-09 | 2019-12-13 | 科瑞百奥泰州生物技术有限公司 | Cartilage tissue low-temperature refrigeration preservation solution and application thereof |
| CN114831108A (en) * | 2022-05-20 | 2022-08-02 | 杭州联川生物技术股份有限公司 | Fresh tissue preservation solution and preparation method and application thereof |
| CN116439231A (en) * | 2023-06-09 | 2023-07-18 | 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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Cited By (11)
| 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 |
| CN108041023A (en) * | 2017-12-14 | 2018-05-18 | 泰山医学院 | A kind of mechanical stimulation method for improving articular cartilage storage in vitro effect |
| 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 |
| CN109392892A (en) * | 2018-12-14 | 2019-03-01 | 广州悦清再生医学科技有限公司 | A kind of biological collagen sill saves liquid and application |
| 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 |
| CN110558312A (en) * | 2019-10-09 | 2019-12-13 | 科瑞百奥泰州生物技术有限公司 | Cartilage tissue low-temperature refrigeration preservation solution and application thereof |
| CN114831108A (en) * | 2022-05-20 | 2022-08-02 | 杭州联川生物技术股份有限公司 | Fresh tissue preservation solution and preparation method and application thereof |
| CN116439231A (en) * | 2023-06-09 | 2023-07-18 | 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 |
| 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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