CN106178117A - A kind of PLL DBM porous composite bone graft material, its preparation method and application - Google Patents
A kind of PLL DBM porous composite bone graft material, its preparation method and application Download PDFInfo
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- CN106178117A CN106178117A CN201610649256.4A CN201610649256A CN106178117A CN 106178117 A CN106178117 A CN 106178117A CN 201610649256 A CN201610649256 A CN 201610649256A CN 106178117 A CN106178117 A CN 106178117A
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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
- 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
- A61L27/3608—Bone, e.g. demineralised bone matrix [DBM], bone powder
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
-
- 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
- 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
- A61L27/365—Bones
-
- 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
- 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
-
- 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
- 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
- A61L27/56—Porous materials, e.g. foams or sponges
-
- 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
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
-
- 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 application relates to field of porous materials, particularly relates to a kind of PLL DBM porous composite bone graft material, its preparation method and application, and this material includes: have the mineral substrates of three-dimensional porous network structure；Being located at the biopolymer decorative layer on mineral substrates surface, described biopolymer is distributed at least some of interior of described three-dimensional porous network.By the way, the embodiment of the present invention is formed by the mineral substrates with three-dimensional porous network structure being carried out surface modification with biopolymer, high to the selectivity retention rate of medullary cell, with the bone grafting material of bone marrow stem cell rapid build, there is good biocompatibility and osteogenic activity, can meet as application demand to Bone Defect Repari height osteogenic activity bone grafting material of wound, osteopathia and emergency treatment Cranial defect.
Description
Technical field
The application relates to field of porous materials, particularly relates to a kind of PLL-DBM porous composite bone graft material, it prepares
Method and application.
Background technology
Free autogenous bone graft percutaneous injection treatment bone does not connect and Cranial defect are achieved with certain success, but the side that application is directly injected into
Method, Local Dry cell density is low, easily run off, and affects the treatment.Bone marrow stem cell beneficiation technologies is used for repairing bone defect and spinal column
Fusion, can obtain the effect suitable with autologous bone transplanting, it has also become the focus of Bone Defect Repari research field.Its core is being applicable to
The timbering material of Selective cell retention technology and the research of skeletonization effect thereof.
In consideration of it, the selectivity retention rate improving medullary cell becomes the technical problem that this area is urgently to be resolved hurrily.
Summary of the invention
The application is mainly solving the technical problems that provide a kind of PLL-DBM porous composite bone graft material, it is possible to increase
The selectivity retention rate of medullary cell, is suitable as bone marrow stem cell enrichment material, can meet wound, osteopathia and emergency treatment Cranial defect
To Bone Defect Repari treatment technology and the application demand of bone graft.
For solving above-mentioned technical problem, the technical scheme that the embodiment of the present invention uses is: provide a kind of PLL-DBM many
Hole composite bone graft material, compared with prior art, its difference is, this material includes:
There is the mineral substrates of three-dimensional porous network structure；With
Being located at the biopolymer decorative layer on mineral substrates surface, described biopolymer is distributed in described three-dimensional porous net
Network at least some of in；
Wherein, described mineral substrates is decalcified bone matrix, and described biopolymer is poly-l-lysine.
Preferably, this material is by carrying out surface with biopolymer to the mineral substrates with three-dimensional porous network structure
Modify and formed.
The biopolymer decorative layer being preferably located in mineral substrates hole has two grades of porous network structures.
Preferably, described decalcified bone matrix is 1000:(0.05～1 with the mass ratio of described poly-l-lysine).
For solving above-mentioned technical problem, another technical scheme that the embodiment of the present invention uses is: provide a kind of PLL-DBM
The preparation method of porous composite bone graft material, comprises the steps:
In aqueous medium, mineral substrates and biopolymer are mixed to form soak, make mineral substrates and biopolymerization
Thing is fully contacted；
Gained soak is placed enough time and to realize biopolymer, the surface of mineral substrates is modified；
Under temperature-42 DEG C～-47 DEG C, negative pressure 20～28Pa, gained soak is cooled down；
The soak of cooling is carried out lyophilization；
Wherein, described mineral substrates is decalcified bone matrix, and described biopolymer is poly-l-lysine.
Preferably, before cooling step, also include the step that soak is carried out negative pressure leaching.
Preferably, comprise the steps:
Decalcified bone matrix is carried out ultrasonic cleaning；
Poly-l-lysine distilled water is configured to the poly-l-lysine that concentration is 0.01～0.2mg/ml molten
Liquid, then decalcified bone matrix is placed in poly-l-lysine solution, form soak, wherein, decalcified bone matrix is left with poly
The ratio of rotation lysine solution is 1g:5ml；
Gained soak is placed enough time and to realize poly-l-lysine, the surface of decalcified bone matrix is modified；
The container being loaded with soak is positioned over and connects negative pressure leaching bottle and the aseptic drying basin of negative pressure leaching machine respectively
In, good seal joint and aseptic drying basin lid mouth, negative pressure leaching 4～8 hours at 18 DEG C～22 DEG C, until having no that bubble is from hole
Gap is overflowed；
Under temperature-42 DEG C～-47 DEG C, negative pressure 20～28Pa, gained soak is cooled down；
The soak of cooling is carried out lyophilization.
Present invention also offers above-mentioned PLL-DBM porous composite bone graft material as bone and/or regenerating bone or cartilage
The application of material.
The embodiment of the present application provides the benefit that: be different from the situation of prior art, and the porous of the embodiment of the present invention is combined
Bone grafting material is formed by the mineral substrates with three-dimensional porous network structure being carried out surface modification with biopolymer,
High to the selectivity retention rate of medullary cell, with the bone grafting material of bone marrow stem cell rapid build, there is good bio-compatible
Property and osteogenic activity, can meet as wound, osteopathia and emergency treatment Cranial defect to Bone Defect Repari height osteogenic activity bone grafting material
Application demand.
Accompanying drawing explanation
In order to be illustrated more clearly that the technical scheme of the embodiment of the present application, will make required in the embodiment of the present application below
Accompanying drawing be briefly described.It should be evident that drawings described below is only some embodiments of the application, for
From the point of view of those of ordinary skill in the art, on the premise of not paying creative work, it is also possible to obtain other according to these accompanying drawings
Accompanying drawing.
Fig. 1 is the outside drawing of the material of the embodiment of the present invention 1.
Fig. 2 is the 3 D video microscope observation figure of the material of the embodiment of the present invention 1.
Fig. 3 is the scanning electron microscope (SEM) photograph (× 600 times) of the material of the embodiment of the present invention 1.
Detailed description of the invention
In order to make the purpose of the application, technical scheme and advantage clearer, below in conjunction with drawings and Examples, right
The application is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the application, not
For limiting the application.
The PLL-DBM porous composite bone graft material of the embodiment of the present invention, including: there is the ore deposit of three-dimensional porous network structure
Thing base material；Be located at the biopolymer decorative layer on mineral substrates surface, described biopolymer is distributed in described three-dimensional porous
Network at least some of in, wherein, described mineral substrates is decalcified bone matrix, and described biopolymer is the left-handed bad ammonia of poly
Acid.This material is formed by the mineral substrates with three-dimensional porous network structure being carried out surface modification with biopolymer,
The biopolymer decorative layer being positioned in mineral substrates hole has two grades of porous network structures.
In the preferred version of the present embodiment, described decalcified bone matrix with the mass ratio of described poly-l-lysine is
1000:(0.05～1).
It should be noted that in this specification, poly-l-lysine (PLL), English is poly-L-lysine；Decalcification
Bone matrix (DBM), English is demineralized bone matrix, be one be by collagen protein, noncollagen protein and
(such as bone morphogenetic protein(BMP), the bone morphogenetic protein(BMP) in bone is held the somatomedin of low concentration by fine and close mineral composition, non-decalcification bone
Ability without induced osteogenesis, mechanical strength corresponding to calcium depletion difference is also different) etc. the complex nature bone graft materials of composition,
It is mainly derived from skull, femoral shaft and the tibial shaft of human or animal (pig, cattle, Canis familiaris L., rabbit etc.).
This material is by poly-l-lysine (poly-L-lysine, PLL) and decalcified bone matrix (demineralized
Bone matrix, DBM) in the immersion of preferred ratio, negative pressure leaching and lyophilization, decalcified bone matrix surface is modified
And optimize, promote sticking, breed and dividing of human marrow mesenchymal stem cell (bone marrow stromal cells, BMSCs)
Secrete the cell functions such as substrate, thus prepare the bone grafting material of high osteogenic activity.
The material of the present embodiment is prepared as follows method and obtains, first, in aqueous medium, by decalcified bone matrix and poly
Aminutrin is mixed to form soak, makes decalcified bone matrix and poly-l-lysine be fully contacted.Secondly, gained is soaked
Liquid is placed enough time and is modified the surface of decalcified bone matrix to realize poly-l-lysine.Again, in temperature-42 DEG C～-
47 DEG C, under negative pressure 20～28Pa, gained soak is cooled down.Finally, the soak of cooling is carried out lyophilization.
Preferably, before cooling step, also include the step that soak is carried out negative pressure leaching.
Specifically, the material of the present embodiment is prepared as follows method and obtains:
(1) decalcified bone matrix is carried out ultrasonic cleaning；
(2) poly-l-lysine distilled water is configured to the poly-l-lysine that concentration is 0.01～0.2mg/ml
Solution, then decalcified bone matrix is placed in poly-l-lysine solution, form soak, wherein, decalcified bone matrix and poly
The ratio of aminutrin solution is 1g:5ml；
(3) surface of decalcified bone matrix is repaiied by gained soak placement enough time to realize poly-l-lysine
Decorations；
(4) container being loaded with soak is positioned over connection negative pressure leaching bottle and the aseptic of negative pressure leaching machine respectively be dried
In ware, good seal joint and aseptic drying basin lid mouth, negative pressure leaching 4～8 hours at 18 DEG C～22 DEG C, until have no bubble from
Hole overflows；
(5) under temperature-42 DEG C～-47 DEG C, negative pressure 20～28Pa, gained soak is cooled down；
(6) soak of cooling is carried out lyophilization.
Embodiment 1
Present embodiments providing a kind of PLL-DBM porous composite bone graft material, this material is that bone marrow stem cell is enriched with material
Material, is to be constituted by DBM with PLL is compound, and the concentration of its PLL is 0.1% (0.1mg/ml), the volume=1g of the weight of DBM: PLL:
5ml.The preparation method step of described PLL-DBM is as follows:
(1) cleaning of DBM
Taking DBM to be placed in ultrasonic washing unit, again wash 3 times with aseptic double-distilled water, every 2 hours with changing distilled water one
Secondary, to PH7.3 ± 0.1 (acidometer mensuration).Draining 4h on aseptic operating platform, 38 DEG C of calorstats of juxtaposition dry 24h, envelope of weighing
Dress labelling are stand-by.
(2) PLL soaks DBM
Under sterile working, prepare the PLL of 0.1% and through 0.22 μm filter filtration sterilization with distilled water with PLL.Use respectively
State PLL immersion self-control DBM (DBM weight: PLL volume=1g:5ml) 15h, the every 3h concussion configured and shake up soak, make PLL
It is fully contacted with DBM.
(3) negative pressure leaching
Above-mentioned soaking container is placed in the aseptic drying basin connecting negative pressure leaching bottle, negative pressure leaching machine, good seal joint
With drying basin lid mouth, 20 DEG C of continuous negative pressure sucking filtration 5h, until having no that any bubble overflows from bone piece surface pore, taking out to soak and holding
It is overnight that-20 DEG C of refrigerator freezings put by device.
(4) lyophilization
Soaking container is placed in freezer dryer pallet, preset-42 DEG C～-47 DEG C of freezer dryer, negative pressure 20-
28Pa, starts evacuation, below vacuum 100MT after pre-cooling 30min.Lyophilization 48h, is shown in that material appears the most completely, white
Poly-D-lysine is during the overwhelming majority is adsorbed onto material, only remaining minimal amount poly-D-lysine white precipitate at the bottom of edge of materials and bottle
Thing, weighs material and encapsulates, labelled, and outside is sealed with layer of polyethylene bag again.
(5) sterilizing and preservation
Cobalt-60γray radiation sterilization, common dose is 15-25kGyCo, puts-20 DEG C of cryogenic refrigerators and preserves stand-by.Take few
Permitted DBM and sent aerobic, anaerobe, mycotic culture.
Embodiment 2
Present embodiments providing a kind of PLL-DBM porous composite bone graft material, compared with Example 1, difference is PLL
Concentration be 0.01% (0.01mg/ml).
Embodiment 3
Present embodiments providing a kind of PLL-DBM porous composite bone graft material, compared with Example 1, difference is PLL
Concentration be 0.05% (0.05mg/ml).
Embodiment 4
Present embodiments providing a kind of PLL-DBM porous composite bone graft material, compared with Example 1, difference is PLL
Concentration be 0.08% (0.08mg/ml).
Embodiment 5
Present embodiments providing a kind of PLL-DBM porous composite bone graft material, compared with Example 1, difference is PLL
Concentration be 0.15% (0.15mg/ml).
Embodiment 6
Present embodiments providing a kind of PLL-DBM porous composite bone graft material, compared with Example 1, difference is PLL
Concentration be 0.20% (0.20mg/ml).
Product Validation is carried out as a example by embodiment 1 products obtained therefrom:
Construction features and sign
PLL-DBM has three-dimensional mesh structure, and density is about 0.25～0.29g/ml, average out to (0.27 ± 0.02) g/
Ml, porosity is (73 ± 11) %.Material and pore surface have milky PLL of thickness about 2～7 μm to cover, and in hole
Form the mesh-structured of less, aperture about 20～65 μm, between major part Kong Yukong, have about 100 μm left and right apertures through, three
Dimension videomicroscopy observes visible material entirety and pore surface has uniform milky PLL to cover, and Raman spectrum analysis shows
The arbitrary site of material surfaces externally and internally is made up of DBM and poly-D-lysine, and histological observation PLL-DBM is table in collagen cavernous structure
There is light PLL coating in face with mesh-structured, refers to shown in Fig. 1 to Fig. 3, the visible mesh knot natural at decalcified bone matrix in figure
Structure defines more tiny poly-l-lysine mesh.
The cytotoxicity detection of PLL-DBM
With reference to the relevant standard of ISO10993 and GB/T10886, by PLL-DBM material and lixiviating solution thereof respectively with people's bone marrow
Stroma stem cell (BMSCs) co culture system in vitro, in cultivating 20h, 3d, 7d row morphological observation, MTT assay
Measure cultivation 1,3,5, the cell-proliferation activity of 7d, application spectrophotography carries out hemolytic test.Result is shown: with PLL-DBM and
The people BMSCs that its lixiviating solution co-cultures shows and preferably sticks and multiplication characteristic, and increases with lixiviating solution concentration and increase, carefully
Cellular toxicity is 0～1 grade.Lixiviating solution group hemolytic index is 2.17%, reaches standard-required.
PLL-DBM skeletonization effect detection
Implanting nude mice by subcutaneous after Adult Human Bone Marrow stem cell is combined PLL-DBM by bone marrow enriching apparatus, sheet taken the photograph by postoperative X line
All show that with histological examination its skeletonization is respond well, compare no significant difference with autologous bone group, and at each time point with obvious
It is better than DBM group.
PLL-DBM Selective cell retention effect detection
1. the observation to the selectivity retention effect of bone marrow nucleated cell, platelet and bone marrow stem cell: PLL-DBM is to having
The multiple that nucleus selectivity is detained is 3.18 ± 0.31 times, and adhesion rate reaches 53% ± 12%；Platelet selectivity is detained
Multiple is 3.88 ± 0.68 times, and adhesion rate reaches 34% ± 10%；The multiple being detained people's bone marrow stem cell selectivity is 5.25 ±
1.40 times, adhesion rate reaches 73% ± 13%, and selection rate reaches 1.41 ± 0.34.2. the selectivity facilitating skeletal growth factor is detained effect
The observation of fruit: PLL-DBM can significantly improve TGF-β 1 and PDGF content after processing with red bone marrow enrichment.
In sum, PLL-DBM is the decalcified bone matrix modified through poly-D-lysine surface, the selectivity to medullary cell
Retention rate is high, has good biocompatibility and osteogenic activity with the bone grafting material of bone marrow stem cell rapid build, can expire
Foot wound, osteopathia and the emergency treatment Cranial defect demand to Bone Defect Repari height osteogenic activity bone graft.
The foregoing is only embodiments herein, not thereby limit the scope of the claims of the present invention, every utilize this Shen
Please the equivalent structure made of description and accompanying drawing content or equivalence flow process conversion, or be directly or indirectly used in other relevant skills
Art field, is the most in like manner included in the scope of patent protection of the application.
Claims (8)
1. a PLL-DBM porous composite bone graft material, it is characterised in that this material includes:
There is the mineral substrates of three-dimensional porous network structure；With
Being located at the biopolymer decorative layer on mineral substrates surface, described biopolymer is distributed in described three-dimensional porous network
In at least some of；
Wherein, described mineral substrates is decalcified bone matrix, and described biopolymer is poly-l-lysine.
PLL-DBM porous composite bone graft material the most according to claim 1, it is characterised in that this material is by with life
Thing polymer carries out surface modification to the mineral substrates with three-dimensional porous network structure and is formed.
PLL-DBM porous composite bone graft material the most according to claim 1, it is characterised in that be positioned at mineral substrates hole
Biopolymer decorative layer in gap has two grades of porous network structures.
PLL-DBM porous composite bone graft material the most according to claim 1, it is characterised in that described decalcified bone matrix
It is 1000:(0.05～1 with the mass ratio of described poly-l-lysine).
5. the preparation method of a PLL-DBM porous composite bone graft material, it is characterised in that comprise the steps:
In aqueous medium, mineral substrates and biopolymer are mixed to form soak, make mineral substrates and biopolymer fill
Tap is touched；
Gained soak is placed enough time and to realize biopolymer, the surface of mineral substrates is modified；
Under temperature-42 DEG C～-47 DEG C, negative pressure 20～28Pa, gained soak is cooled down；
The soak of cooling is carried out lyophilization；
Wherein, described mineral substrates is decalcified bone matrix, and described biopolymer is poly-l-lysine.
Preparation method the most according to claim 5, it is characterised in that also included soak is carried out before cooling step
The step of negative pressure leaching.
Preparation method the most according to claim 5, it is characterised in that comprise the steps:
Decalcified bone matrix is carried out ultrasonic cleaning；
Poly-l-lysine distilled water is configured to the poly-l-lysine solution that concentration is 0.01～0.2mg/ml, then
Decalcified bone matrix is placed in poly-l-lysine solution, forms soak, wherein, decalcified bone matrix and the left-handed bad ammonia of poly
The ratio of acid solution is 1g:5ml；
Gained soak is placed enough time and to realize poly-l-lysine, the surface of decalcified bone matrix is modified；
The container being loaded with soak is positioned in the aseptic drying basin connecting negative pressure leaching bottle and negative pressure leaching machine respectively, close
Sealing joint and aseptic drying basin lid mouth, negative pressure leaching 4～8 hours at 18 DEG C～22 DEG C, until having no that bubble overflows from hole
Go out；
Under temperature-42 DEG C～-47 DEG C, negative pressure 20～28Pa, gained soak is cooled down；
The soak of cooling is carried out lyophilization.
8. the PLL-DBM porous composite bone graft material described in any one of Claims 1-4 is as bone and/or cartilage again
The application of raw material.
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| CN111035803A (en) * | 2019-11-07 | 2020-04-21 | 浙江大学 | Titanium implant material with anti-infection and osseointegration promoting functions and preparation method thereof |
| CN113633827A (en) * | 2021-08-16 | 2021-11-12 | 中国人民解放军总医院第四医学中心 | Silk woven meniscus implant and preparation method thereof |
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| CN109172863A (en) * | 2018-08-20 | 2019-01-11 | 中国人民解放军第二军医大学第二附属医院 | A kind of method that polycaprolactone-tricalcium phosphate bone tissue engineering scaffold carries out the modification of nanometer decalcifed bone matrix coating |
| CN109172863B (en) * | 2018-08-20 | 2021-04-27 | 中国人民解放军第二军医大学第二附属医院 | Method for modifying nano decalcification bone matrix particle coating of polycaprolactone-tricalcium phosphate bone tissue engineering scaffold |
| CN111035803A (en) * | 2019-11-07 | 2020-04-21 | 浙江大学 | Titanium implant material with anti-infection and osseointegration promoting functions and preparation method thereof |
| CN113633827A (en) * | 2021-08-16 | 2021-11-12 | 中国人民解放军总医院第四医学中心 | Silk woven meniscus implant and preparation method thereof |
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