CN110237302A - A kind of preparation method of articular cartilage repair materials-autologous platelet rich plasma combination hyaluronic acid gel - Google Patents
A kind of preparation method of articular cartilage repair materials-autologous platelet rich plasma combination hyaluronic acid gel Download PDFInfo
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
- A61L27/3645—Connective tissue
- A61L27/3654—Cartilage, e.g. meniscus
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
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- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- 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
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
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- A—HUMAN NECESSITIES
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- 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
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- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus
Abstract
The present invention relates to a kind of preparation method of articular cartilage repair materials-autologous platelet rich plasma combination hyaluronic acid gel, the preparation including (1) hyaluronic acid gel；(2) extraction of autologous platelet rich plasma.The present invention is repaired by situ autologous stem cells of raising, the shortcomings that so as to avoid the low survival rate of cell of in vitro culture and low differentiation potential.Hyaluronic acid gel and platelet rich plasma are injected into defect point by the present invention, secondary operation are not necessarily to, to realize cartilage defect repair.It is thus greatly improved in clinical replicability.
Description
Technical field
The present invention relates to medical material fields, and in particular to a kind of preparation method-of articular cartilage repair materials is self rich
Thrombocyte plasma combination hyaluronic acid gel.
Background technique
Osteoarthritis is a kind of disease of joint degeneration, with the continuous progress of domestic aging of population, therefore bone
Arthritis becomes the main reason for influencing life of elderly person quality.Due to clinically there is no effective alleviation Osteoarthritis at present
Method, can only finally alleviate arthralgia caused by it by joint replacement.But a large amount of joint replacement to family and
Society brings serious financial burden, thus Osteoarthritis is the significant problem of urgent need to resolve.
Cartilage defect is the main performance of Osteoarthritis, and clinically commonly the method for the treatment of cartilage defect is main at present
Including micro fractures, self or allograph bone cartilage transplantation, chondrocyte cell transplantation, and these methods can not regenerate it is natural transparent
Cartilage, the group repaired is woven in ingredient and performance significantly lower than natural cartilage, thus can only alleviate cartilage to a certain extent
Regression.In recent years, it is had been greatly developed using tissue engineering technique repairing articular cartilage defect, such as utilizes cartilage cell
In vitro culture forms cartilaginous tissue, is subsequently implanted at cartilage defect, or cartilage cell is implanted into cartilage in conjunction with three-dimensional rack and is lacked
It is repaired at damage.No matter which kind of organizational project recovery technique all be unable to do without three elements, i.e. bracket, growth factor, seed cell.
Its repair process can provide stem cell for bracket and be adhered to, and the space three-dimensional structure for growing and breaking up, growth factor can promote
Stem cell secretes cartilage matrix at cartilage differentiation, is deposited in three-dimensional rack, gradually degrades with after-poppet, to regenerate
Natural cartilage.Below advantage of the method used in the present invention compared with other existing methods will be illustrated in terms of three from this.
Firstly, currently used tissue engineering bracket is PLA, PGA or collagen scaffold, the cartilaginous element that this bracket generates
A mainly Collagen Type VI, and the main component of native Zona cartilage is Type Ⅱ collagen, because natural cartilage can not be regenerated.This hair
Bright use is injectable hyaluronic acid gel bracket, and the outstanding advantage which has mainly includes, 1, locally injecting enters
It is i.e. curable after then being irradiated by ultraviolet point light source at cartilage defect, therefore do not limited by cartilage defect shape；2. transparent
Matter acid is the proper constituent in natural cartilage, thus hyaluronic acid gel has good biocompatibility, and has good
Good degradability.3, hyaluronic acid gel is the higher bracket of water content, can imitate the three-dimensional space knot of normal cartilage
Structure, so that the Growth and Differentiation for cell provides good microenvironment.
Growth factor can promote stem cell to break up to cartilage direction, to generate cartilage correlation matrix.Due to normal at present
Growth factor is external synthesis or allosome growth factor, and these growth factors have certain immunogenicity, and valence
Lattice are expensive, to limit its application in vivo.The present invention is used be centrifuged by autoblood after the rich platelet that obtains
Blood plasma, rich in there are many growth factors in platelet rich plasma, including TGF-beta, VEGF, PDGF, IGF etc., these growths because
Son can promote stem cell to be broken up to cartilage direction.And these growth factors are sustainable to be discharged from blood platelet, thus in cartilage
Defect persistently stimulates Chondrogenesis.
The reparation of cartilage be unable to do without the participation of cell, and the cell for participating in repairing after cartilage defect mainly has between derived from bone marrow
Mesenchymal stem cells, the stem cell of synovial origin or adipose-derived stem cell.Tissue engineering technique is thin by vitro culture at present
Born of the same parents, are subsequently implanted defect, but the cell survival rate after being implanted into is lower, cause its differentiation potential to drop after especially in vitro
It is low, it is limited so as to cause repair ability.And the present invention is soft by injecting hyaluronic acid gel and autologous platelet rich plasma
Bone defect position, wherein the growth factor of platelet rich plasma release can raise autologous stem cells and migrate to defect, then
It is grown in hydrogel scaffold, to Chondrocyte Differentiation under the growth factor effect of platelet rich plasma release, to repair
Defect cartilage.
Summary of the invention
The purpose of the present invention is aiming at the problems existing in the prior art, provide a kind of preparation of articular cartilage repair materials
Method-autologous platelet rich plasma combination hyaluronic acid gel is repaired by autologous stem cells in situ of raising, to keep away
The low survival rate of cell of in vitro culture is exempted from and the shortcomings that low differentiation potential.
To achieve the above object, present invention provide the technical scheme that
A kind of preparation method of articular cartilage repair materials-autologous platelet rich plasma combination hyaluronic acid gel, packet
Include following steps:
One, the preparation of hyaluronic acid gel:
1) it takes appropriate Sodium Hyaluronate to be dissolved in distilled water, under cryogenic conditions 4-20 degree, stirs 12h；
2) appropriate methacrylic acid anhydride is added into acquired solution, is 8-9 in pH, temperature is reacted under conditions of being 4 degree
24h；
3) products therefrom is precipitated in acetone, is cleaned with ethyl alcohol, dissolved in deionized water, deionized water dialysis 48h
Afterwards, the Sodium Hyaluronate after being modified；
4) Sodium Hyaluronate after modification is dissolved in distilled water by 2% concentration, is added according to 0.1% concentration light-initiated
Liquid hydrogel is injected cartilage defect by agent, and ultraviolet point light source irradiates lower gel in-situ, cured hyalomitome can be obtained
Acid hydrogel；
Two, the extraction of autologous platelet rich plasma:
1) appropriate sodium citrate anticoagulant is added and is extracted in centrifuge tube；
2) it is added in whole blood and is mixed in the centrifuge tube of sodium citrate anticoagulant in advance, mix well；
3) first time centrifugation, centrifugal speed 2400r/min, centrifugation time 10min are carried out；
4) after being centrifuged for the first time, blood is divided into three layers, the tunica albuginea layer of haemocyte, middle layer including bottom, surface layer serum,
The haemocyte of bottom is extracted out；
5) it puts into a centrifuge progress second to be centrifuged, after second is centrifuged, is equally divided into three layers, using syringe by table
The serum layer extraction of layer, can be obtained the platelet rich plasma of concentration；
6) will in the concentration platelet rich plasma injection articular cavity of extraction, can be obtained autologous platelet rich plasma combine it is saturating
Bright matter acid hydrogel cartilage repair systems.
The photoinitiator uses Irgacure 2959.
In step 1,2g Sodium Hyaluronate is dissolved in every 100ml distilled water, and the methacrylic acid anhydride of 1.6ml is then added.
Step 1: 4) ultraviolet ray intensity is 15 μ W/cm2, irradiation time 30s in.
In step 2, the volume ratio of sodium citrate anticoagulant and whole blood is 1:9.
Compared with prior art, the beneficial effects of the present invention are:
(1) safely and effectively, the hyaluronic acid gel water content that the present invention uses is higher, can sufficiently simulate natural cartilage
Three-dimensional structure, so that the microenvironment of good cell-ECM and cell-ECM matrix interaction is provided for cell, and
And hyaluronic acid has the stretching, extension for promoting stem cell at cartilage differentiation potential, and passing through inhibition cell, to be conducive to cell point
Secrete more Type Ⅱ collagen albumen.Hyaluronic acid is natural cartilage constituent, and catabolite is nontoxic, thus has good life
Object compatibility.In addition, it is autologous platelet rich plasma that the present invention, which uses, it is immunized caused by exogenous growth factors to eliminate
Pathogenic problems, and autologous platelet rich plasma growth factor rich in there are many, to can promote stem cell to cartilage direction point
Change, has greatly facilitated the reparation of cartilage defect.
(2) cheap, the present invention can significantly reduce repair of cartilage cost, firstly, hyaluronic acid is a kind of cheap
Material, it is from a wealth of sources, and hyaluronic acid gel preparation method is simple to operation；Secondly, autologous platelet rich plasma is
The ingredient obtained after being centrifuged by autoblood, due to avoiding and using exogenous growth factor from patient itself, from
And significantly reduce the procurement cost of growth factor.
(3) simple to operation, currently used method is to open articular cavity by biggish notch, and repair materials are inserted
Defect, this method wound are larger and expensive.The present invention uses injectable hyaluronic acid gel, passes through note
Hydrogel is injected defect by emitter, then under the irradiation of ultraviolet point light source can in-situ solidifying, realize that wound is small, easy behaviour
The demand of work.And autologous platelet rich plasma can be extracted by platelet rich plasma in the market and is set with, and be carried out referring to step
Extraction, operation is simple.
(4) one-step method repair cartilage defect, currently used tissue engineering technique, either Autologous Chondrocyte transplanting or
The transplanting of matrix Induction of chondrocytes is required to one-stage operation and obtains cartilaginous tissue, Porcine Chondrocytes Cultured in vitro, subsequent secondary operation
It implants, and this operation undoubtedly brings bigger wound to patient.And the present invention passes through one-step method for hyaluronic acid water-setting
Glue and platelet rich plasma are injected into defect point, secondary operation are not necessarily to, to realize cartilage defect repair.Thus greatly improve
It is in clinical replicability.
Detailed description of the invention
Fig. 1: it is in a liquid state before hyaluronic acid gel solidification.
Fig. 2: being in solid-state after hyaluronic acid gel solidification.
Fig. 3: the extraction schematic diagram of autologous platelet rich plasma.
Fig. 4: sample is repaired and substantially sees assessment, and display experimental group cartilage defect repair effect is substantially better than control group.
Fig. 5: sample repairs HE dyeing assessment, and display experimental group cartilage defect repair effect is substantially better than control group.
Fig. 6: sample repairs HE dyeing assessment, and display experimental group cartilage defect repair effect is substantially better than control group.
Fig. 7: sample repairs the assessment of Type Ⅱ collagen immunohistochemical staining, and display experimental group cartilage defect repair effect is obviously excellent
In control group, the regenerated cartilage of experimental group, which is rich in, Type Ⅱ collagen.
Specific embodiment
The present invention is further explained in the light of specific embodiments.
A kind of preparation method of the cartilage repair material in autologous platelet rich plasma combination hyaluronic acid gel joint, packet
Include the preparation of (1) hyaluronic acid gel and the extraction of (2) autologous platelet rich plasma.
(1) preparation of hyaluronic acid gel, includes the following steps:
1. 2g Sodium Hyaluronate is dissolved in 100ml distilled water, (4-20 degree) stirs 12h under cryogenic conditions；2. to gained
The methacrylic acid anhydride of 1.6ml is added in solution, is 8-9 in PH, temperature is reacted for 24 hours under conditions of being 4 degree；3. by products therefrom
It precipitates, is cleaned with ethyl alcohol in acetone, dissolved in deionized water, the hyalomitome after deionized water dialysis 48h, after being modified
Sour sodium.4. the Sodium Hyaluronate after modification is dissolved in distilled water by 2% concentration, photoinitiator is added according to 0.1% concentration
Liquid hydrogel is injected cartilage defect (Fig. 1) by Irgacure 2959, and ultraviolet ray intensity is 15 μ W/cm2, irradiation time
For 30s, cured hyaluronic acid gel (Fig. 2) can be obtained.
(2) extraction of autologous platelet rich plasma, includes the following steps:
Suit, which is extracted, using the platelet rich plasma of Shandong Wei Gao Co., Ltd in this patent extracts rich platelet blood
Slurry.It is extracted in centrifuge tube specific steps are as follows: 1. 5ml sodium citrate anticoagulant is added；2. using blood taking needle from vein
45ml whole blood is collected, is then added and is mixed in the centrifuge tube of sodium citrate anticoagulant in advance, mix well to obtain containing 50ml liquid
The centrifuge tube of body；3. 50ml water is added in another identical centrifuge tube, first time centrifugation, centrifugal speed 2400r/ are then carried out
Min, centrifugation time 10min；4. blood is divided into three layers, the tunica albuginea layer of haemocyte, middle layer including bottom after being centrifuged for the first time
The serum on (i.e. platelet rich plasma layer), surface layer, the hollow tube that can connect bottom of the tube that syringe insertion centrifuge tube is carried,
Then the haemocyte of bottom is extracted out, about 20ml；5. the water of another centrifuge tube removal same volume is subsequently placed into centrifuge
It carries out second to be centrifuged, after second is centrifuged, is equally divided into three layers, the serum layer on surface layer is extracted out using syringe, can be obtained
To the platelet rich plasma of concentration, about 5ml.
See that Fig. 3, A, platelet rich plasma obtain schematic diagram, left hand view represents whole blood and mixes with anti-coagulants；Middle graph represents
Blood is divided into three layers after being centrifuged for the first time；Right part of flg represents the concentration platelet rich plasma obtained after second of centrifugation；B is transparent
Matter acid hydrogel scanning electron microscope (SEM) photograph, scale: 5 microns；C, hyaluronic acid gel (m-HA) and autologous platelet rich plasma (PRP)
Using schematic diagram, UV irradiation: ultraviolet light, PRP injection: injecting platelet-rich plasma, Syringe:
Syringe, HA scaffold with PRP: hyaluronic acid gel bracket combination platelet rich plasma；D, articular cartilage in art
Defect makes picture, scale: 5 millimeters.
Embodiment 1:
(1) preparation of hyaluronic acid gel, it is saturating after selecting in this patent optimal proportion embodiment to be modified
Bright matter acid sodium is used for cartilage defect repair；
(2) extraction of autologous platelet rich plasma uses the platelet rich plasma of Shandong Wei Gao Co., Ltd in this patent
It extracts suit and extracts platelet rich plasma, platelet rich plasma is concentrated in the final about 5ml that obtains；
(3) Sodium Hyaluronate after modification will be dissolved in distilled water by 2% concentration, light is added according to 0.1% concentration
Liquid hydrogel is injected cartilage defect by initiator Irgacure 2959, and ultraviolet ray intensity is 15 μ W/cm2, when irradiation
Between be 30s, obtain cured hyaluronic acid gel；
(4) by the concentration platelet rich plasma injection articular cavity of extraction, autologous platelet rich plasma combination can be obtained
Hyaluronic acid gel cartilage repair systems.
Test example 1:
The autologous platelet rich plasma combination hyaluronic acid gel cartilage repair systems are verified to close in Ba-Ma mini pig knee
Save the repairing effect of condylus medialis femoris weight bearing area's full-thickness cartilage defects and osteochondral defect.
This research uses Ba-Ma mini pig as animal model, makes holostrome respectively in knee joint femoral entocondyle weight bearing area
Cartilage defect and osteochondral defect, production defect size are divided into four kinds, including diameter 6.5mm, 8.5mm and depth be 2mm,
Hyaluronic acid gel is injected at cartilage defect experimental group by the defect of 5mm, then solid under the irradiation of ultraviolet point light source
It is melted into glue, suit is extracted using platelet rich plasma and extracts autologous platelet rich plasma, be subsequently injected into knee joint cavity, suture is cut
Mouthful；Control animals, the only reparation of local use hyaluronic acid gel or space management.Postoperative miniature pig freely bears a heavy burden June, with
After carry out substantially see assessment.
Fig. 4 is that sample reparation is substantially seen, and 8.5FT: diameter 8.5mm full-thickness cartilage defects, 6.5FT: diameter 6.5mm holostrome is soft
Bone defect, 8.5X5: diameter 8.5mm, depth 5mm osteochondral defect, 6.5X5: diameter 6.5mm, depth 5mm osteochondral defect, HA
+ PRP: hyaluronic acid gel combination platelet rich plasma processing group, HA: hyaluronic acid gel processing group, Blank: blank
Control group.
Sample carries out HE dyeing (Fig. 5,6) and Type Ⅱ collagen immunohistochemical staining (Fig. 7) assesses defect repair.As the result is shown
Experimental group cartilage defect repair effect is substantially better than control group:
Fig. 5: sample repairs HE dyeing.A, diameter 8.5mm full-thickness cartilage defects each group repairing effect；B, diameter 6.5mm are complete
Layer cartilage defect each group repairing effect；Cartilage interface: cartilage interface；Repaired area: neocartilage is repaired
Multiple region；Osteochondral interface: bone cartilage interface；HA+PRP: hyaluronic acid gel combination rich platelet blood
Starch processing group；HA: hyaluronic acid gel processing group；Blank: blank control group；RA: restoring area, IF: interface, NA: normal
Region, SB: subchondral bone.
Fig. 6: sample repairs HE dyeing.A, diameter 8.5mm, depth 5mm osteochondral defect each group repairing effect；B, diameter
6.5mm, depth 5mm osteochondral defect each group repairing effect；Cartilage interface: cartilage interface；Repaired
Area: neocartilage restoring area；Osteochondral interface: bone cartilage interface；HA+PRP: hyaluronic acid water-setting
Cementing conjunction platelet rich plasma processing group；HA: hyaluronic acid gel processing group；Blank: blank control group；RA: area is repaired
Domain, IF: interface, NA: normal region, SB: subchondral bone.
Fig. 7: sample repairs Type Ⅱ collagen immunohistochemical staining.A, diameter 8.5mm full-thickness cartilage defects each group repairing effect；
B, diameter 6.5mm full-thickness cartilage defects each group repairing effect；C, diameter 8.5mm, depth 5mm osteochondral defect each group reparation effect
Fruit；D, diameter 6.5mm, depth 5mm osteochondral defect each group repairing effect；Cartilage interface: cartilage interface；
Repaired area: neocartilage restoring area；HA+PRP: hyaluronic acid gel combination platelet rich plasma processing group；
HA: hyaluronic acid gel processing group；Blank: blank control group；RA: restoring area, IF: interface, NA: normal region.
The above is only presently preferred embodiments of the present invention, is not intended to limit the present invention in any form, any ripe
Professional and technical personnel is known, without departing from the scope of the present invention, according to the technical essence of the invention, to the above reality
Any simple modifications, equivalent substitutions and improvements etc. made by example are applied, it is fallen within the scope of protection of the technical scheme of the present invention
It is interior.
Claims (5)
1. a kind of preparation method of articular cartilage repair materials-autologous platelet rich plasma combination hyaluronic acid gel, special
Sign is: the following steps are included:
One, the preparation of hyaluronic acid gel:
1) it takes appropriate Sodium Hyaluronate to be dissolved in distilled water, under cryogenic conditions 4-20 degree, stirs 12h；
2) appropriate methacrylic acid anhydride is added into acquired solution, is 8-9 in pH, temperature is reacted for 24 hours under conditions of being 4 degree；
3) products therefrom is precipitated in acetone, is cleaned with ethyl alcohol, dissolution in deionized water, after deionized water dialysis 48h, obtains
Sodium Hyaluronate after to modification；
4) Sodium Hyaluronate after modification is dissolved in distilled water by 2% concentration, photoinitiator is added according to 0.1% concentration,
Liquid hydrogel is injected into cartilage defect, ultraviolet point light source irradiates lower gel in-situ, cured hyaluronic acid can be obtained
Hydrogel；
Two, the extraction of autologous platelet rich plasma:
1) appropriate sodium citrate anticoagulant is added and is extracted in centrifuge tube；
2) it is added in whole blood and is mixed in the centrifuge tube of sodium citrate anticoagulant in advance, mix well；
3) first time centrifugation, centrifugal speed 2400r/min, centrifugation time 10min are carried out；
4) after being centrifuged for the first time, blood is divided into three layers, the tunica albuginea layer of haemocyte, middle layer including bottom, surface layer serum, the bottom of by
The haemocyte extraction of layer；
5) it puts into a centrifuge progress second to be centrifuged, after second is centrifuged, is equally divided into three layers, using syringe by surface layer
The extraction of serum layer, can be obtained the platelet rich plasma of concentration；
6) by the concentration platelet rich plasma injection articular cavity of extraction, autologous platelet rich plasma combination hyalomitome can be obtained
Acid hydrogel cartilage repair systems.
2. preparation method-autologous platelet rich plasma of articular cartilage repair materials according to claim 1 combines transparent
Matter acid hydrogel, it is characterised in that: the photoinitiator uses Irgacure 2959.
3. preparation method-autologous platelet rich plasma of articular cartilage repair materials according to claim 1 combines transparent
Matter acid hydrogel, it is characterised in that: in step 1,2g Sodium Hyaluronate is dissolved in every 100ml distilled water, 1.6ml is then added
Methacrylic acid anhydride.
4. preparation method-autologous platelet rich plasma of articular cartilage repair materials according to claim 1 combines transparent
Matter acid hydrogel, it is characterised in that: Step 1: ultraviolet ray intensity is 15 μ W/cm2, irradiation time 30s in 4).
5. preparation method-autologous platelet rich plasma of articular cartilage repair materials according to claim 1 combines transparent
Matter acid hydrogel, it is characterised in that: in step 2, the volume ratio of sodium citrate anticoagulant and whole blood is 1:9.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112915264A (en) * | 2021-01-21 | 2021-06-08 | 中国人民解放军空军军医大学 | Preparation method for gelatin-sodium alginate-PRP composite material |
| CN114099543A (en) * | 2021-10-29 | 2022-03-01 | 南京国青血液净化科技有限公司 | Preparation method of prp serum for repairing sports injury |
| CN114522272A (en) * | 2022-03-04 | 2022-05-24 | 南京鼓楼医院 | Bionic platelet hydrogel for wound repair and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104587531A (en) * | 2014-12-25 | 2015-05-06 | 南京臻泉医药科技有限公司 | Preparation method for gel scaffold for repairing articular cartilage injuries |
| CN109701073A (en) * | 2019-01-24 | 2019-05-03 | 广州贝奥吉因生物科技有限公司 | A kind of injectable cartilage repair hydrogel and preparation method thereof |
-
2019
- 2019-07-22 CN CN201910660987.2A patent/CN110237302A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104587531A (en) * | 2014-12-25 | 2015-05-06 | 南京臻泉医药科技有限公司 | Preparation method for gel scaffold for repairing articular cartilage injuries |
| CN109701073A (en) * | 2019-01-24 | 2019-05-03 | 广州贝奥吉因生物科技有限公司 | A kind of injectable cartilage repair hydrogel and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| JASON A. BURDICK ET AL: "Controlled Degradation and Mechanical Behavior of Photopolymerized Hyaluronic Acid Networks", 《BIOMACTOMOLECULES》 * |
| 曹永平: "《膝关节骨关节炎曹永平2018观点》", 30 November 2017 * |
| 范猛: "《骨性关节炎的防治与康复》", 31 August 2017 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112915264A (en) * | 2021-01-21 | 2021-06-08 | 中国人民解放军空军军医大学 | Preparation method for gelatin-sodium alginate-PRP composite material |
| CN114099543A (en) * | 2021-10-29 | 2022-03-01 | 南京国青血液净化科技有限公司 | Preparation method of prp serum for repairing sports injury |
| CN114522272A (en) * | 2022-03-04 | 2022-05-24 | 南京鼓楼医院 | Bionic platelet hydrogel for wound repair and preparation method thereof |
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