CN104117097B - Integrated bone cartilage frame with bionical interfacial structure and preparation method thereof - Google Patents
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Abstract
The invention discloses a kind of integrated bone cartilage frame with bionical interfacial structure and preparation method thereof, belong to prosthetic material field.The present invention is made up of cartilage layers, calcification layer and subchondral bone layer from top to bottom successively, and cartilage layers is the vertical orientated property support of silk fibroin protein solution through crystallographic orientation, lyophilizing; Calcification layer is the compact area that fibroin albumen and hydroxyapatite are formed, calcification layer be that organic that a lyophilizing is shaped is connected between cartilage layers, subchondral bone layer; Subchondral bone layer is made up of fibroin albumen and hydroxyapatite, through the three-dimensional porous structure support of the drilling of paraffin microsphere, lyophilizing, dewaxing.The present invention has good biocompatibility and degradability, calcification layer is connected with organic between cartilage layers and subchondral bone layer, play isolation bone, the effect of cartilage microenvironment, be beneficial to cell better differentiation and proliferation under two kinds of microenvironments of plantation, and technology of preparing controllability is strong, without the need to other cross-linking agent or toxic reagent, application potential is huge.
Description
Technical field
The invention belongs to prosthetic material field, be specifically related to a kind of integrated bone cartilage frame with bionical interfacial structure and preparation method thereof.
Background technology
The joint part osteocartilaginous and subchondralo bone injury caused by reasons such as wound, disease, regressions is Orthopedic Clinical common disease, particularly common in China, and in ascendant trend year by year.At present this cartilage is merged to the injury in treating difficulty of subchondral bone, effect still can not be satisfactory, needs the permanent reconstruction measure that research is desirable badly, and expectant treatment can only respite pain, can not stop advancing of disease; Bone and cartilage autotransplantation art limited source, is difficult to the defect of repairing larger area; There is immunological rejection and pathophorous possibility in allograph bone cartilage grafting; Artificial joint replacement somewhat expensive, revision rate is higher.Developing rapidly of tissue engineering technique, the Regeneration and Repair for ossa articularia cartilage injury provides new solution.
Once had employing two kinds of different stent materials to build tissue engineered bone, cartilaginous tissue respectively, the method be then assembled to together builds tissue engineered bone-cartilage complex.The method of assembling comprises fibrin gel bonding, hurtless measure suture method or polylactic acid and is adhesively fixed, but the bone-cartilage complex constructed exists bone, cartilage portion in conjunction with not good enough problem.
Tissue engineering relates to a cross discipline of cytobiology, material science, engineering and bioreactor, utilize ultimate principle and the method for life sciences and engineering, build needed by human body will organize, for repairing, substituting because of wound, disease and non-functional tissue or organ.The main contents of Tissue Engineering Study comprise: seed cell, timbering material, somatomedin.Seed cell comprises chondrocyte, bone marrow stroma stem cell, fat-derived stem cells and through genetic engineering modified cell etc.The domestic and international research about organizational project bone-cartilage at present mainly concentrates on choosing of timbering material and seed cell and skeletonization with in connected mode becoming cartilage portion etc.,
Chinese patent application 201010140115.2 discloses " a kind of method for preparing double-layer bionic cartilage tissue engineering scaffold ".First substrate is prepared, double-layer bionic cartilage frame is obtained again at substrate surface, the double-layer bionic cartilage frame of substrate surface is infiltrated finally by NaOH solution, peel off from substrate and namely obtain double-layer bionic cartilage frame, double-layer bionic cartilage frame thickness prepared by the present invention is thin, good mechanical properties, one-shot forming, be connected between upright opening with circular port closely, there is not the problem mutually departed from; Supporting structure is close to natural cartilage structure, and cartilage frame directly can be combined with the substrate containing polylactic acid PLA/bone meal and form bone cartilage frame, and for the reparation of defect while of bone cartilage, cartilage layers part still has double-decker.
Chinese patent application 201210088049.8 discloses one " a kind of tissue engineering bone/cartilage timbering material and preparation method thereof and device ".Cartilage support material of the present invention is followed successively by top layer, middle level and calcification layer from top to bottom; Top layer is made up of lyophilizing cartilage matrix and II Collagen Type VI, and matrix fiber is parallel distribution in the horizontal direction; Middle level is made up of lyophilizing cartilage matrix, GAG, X-type collagen and II Collagen Type VI and TGF-β, and matrix fiber is longitudinal arrangement; Calcification layer is made up of lyophilizing cartilage matrix, GAG and X-type collagen and calcined bone powder, and its fiber is three-dimensional staggered distribution.Be conducive to timbering material and implant fusion that is rear and subchondral bone, promote the regeneration of subchondral bone; After compound chondrocyte, can be used in full-thickness cartilage defects reparation, the cartilaginous tissue of regeneration has the space structure consistent with natural cartilage and protein component; Be suitable for the reparation of the heavy burden position articular cartilage such as knee joint, the neocartilage that regeneration comprcssive strength is larger, improve wearability and the anti-pressure ability on cartilage frame surface.
Summary of the invention
The present invention, in order to overcome the bone that exists in prior art and cartilage portion proposes in conjunction with not good enough shortcoming, its objective is and provides a kind of integrated bone cartilage frame with bionical interfacial structure and preparation method thereof.
Technical scheme of the present invention is:
A kind of integrated bone cartilage frame with bionical interfacial structure, described support is made up of cartilage layers 1, calcification layer 2 and subchondral bone layer 3 from top to bottom successively, the vertical orientated property support that described cartilage layers 1 is formed for fibroin albumen, described calcification layer 2 is the compact area that fibroin albumen and hydroxyapatite are formed, and described subchondral bone layer 3 is the three-dimensional porous structure support that fibroin albumen and hydroxyapatite are formed; Calcification layer 2 is connected with organic between cartilage layers 1, subchondral bone layer 3.
Fibroin albumen in described calcification layer 2 and subchondral bone layer 3 and the mass ratio of hydroxyapatite are 1:1.
The pore diameter range of the three-dimensional porous structure of described subchondral bone layer 3 is 250 μm ~ 425 μm.
The thickness of described calcification layer 2 is less than 500 μm.
Described cartilage layers 1 is 1:1 with the thickness proportion of subchondral bone layer 3.
There is a preparation method for the integrated bone cartilage frame of bionical interfacial structure, comprise the following steps:
A. paraffin microsphere template preparation
Paraffin microsphere is put into polyethylene mould, dry for standby;
B. the preparation of calcification layer 2 and subchondral bone layer 3
Be the mixing of 10% silk fibroin protein solution and 10% hydroxyapatite solution 1:1 in mass ratio by mass fraction, add in paraffin microsphere template, aspiration vacuum, mixed liquor is fully filled, and exceeds paraffin microsphere template certain distance, and after cooled with liquid nitrogen, room temperature leaves standstill;
C. the preparation of cartilage layers 1
Until calcification layer 2 surface micro-melt after, be that the silk fibroin protein solution 2mL of 6% slowly adds on the calcification layer 2 in mould by mass fraction ,-80 DEG C of lyophilization molding;
D. fibroin allosteric and dewaxing
Take out from polyethylene mould after molding, soaked in absolute ethyl alcohol, filter wax, dries, obtains bone cartilage frame.
The preparation of described silk fibroin protein solution comprises the following steps:
1. natural silk degumming: natural silk is added the Na of 0.02mol/L by 1:200 in mass ratio
2cO
3in solution, boil, repeatedly wash, change Na
2cO
3repeat after solution to boil, wash, then ventilate and dry, obtain fibroin silk;
2. fibroin silk lysate: fibroin silk adds in the lithium-bromide solution of 0.8g/mL by 1:4 in mass ratio, in 60 DEG C of water-baths, magnetic agitation obtains;
3. dialysis desalting: pouring fibroin silk lysate into molecular cut off is in the bag filter of 12000 ~ 14000, dialyse 3 days, every day changes water, centrifugal, get supernatant, proceeding to molecular cut off is in the bag filter of 3500, dialyse in the polyglycol solution of mass fraction 15%, centrifuging and taking supernatant and get final product;
The preparation of described paraffin microsphere comprises the following steps:
1. preparing mass fraction is the poly-vinyl alcohol solution of 5%, heating and melting;
2. take 60g paraffin mass, chopping, adds in 1080ml distilled water, then to add 120ml mass fraction be 5% polyvinyl alcohol, heated and stirred, cooling, and 20 ~ 80 eye mesh screens stack successively, sieve;
3. sieve paraffin microsphere water rinses repeatedly, ethanol dewaters, dry, get 40 ~ 60 order paraffin microspheres for subsequent use.
Described filter wax technology application apparatus,Soxhlet's, solvent is normal hexane or cyclohexane extraction.
The invention has the beneficial effects as follows:
A kind of integrated bone cartilage frame with bionical interfacial structure is constructed in the present invention, make its structure closer to physiological environment, it is the new model in the regeneration of a kind of osteochondro tissue and the field of reconstruction, providing new approaches for building tissue engineering bone cartilage compound, providing experimental basis to further clinical practice.The material adopted belongs to natural material, have good biocompatibility and degradability, and material source is extensive.Calcification layer and cartilage layers and subchondral bone layer is organic is connected, namely the silk fibroin solution itself utilizing non-occurred conformation to change dissolves each other, bone-cartilage two parts support well can be coupled together, preparation simultaneously connects calcification layer, play isolation bone, the effect of cartilage microenvironment, be beneficial to cell better differentiation and proliferation under two kinds of microenvironments of plantation.This support technology of preparing controllability is strong simultaneously, and without the need to other cross-linking agent or toxic reagent, application potential is huge.The present invention on composition, form, mechanical characteristic close to " hyaline cartilage-interfacial structure-subchondral bone " extra-cellular matrix structure of human synovial, and combine closely avoid layering.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention;
Fig. 2 is the structure chart of the present invention under scanning electron microscopic observation;
Fig. 3 is the cartilage layers longitudinal section figure under scanning electron microscopic observation;
Fig. 4 is the cartilage layers cross-sectional view under scanning electron microscopic observation;
Fig. 5 is the subchondral bone layer figure under scanning electron microscopic observation;
Fig. 6 is cartilage portion HE colored graph in the present invention;
Fig. 7 is subchondral bone part HE colored graph in the present invention;
Fig. 8 is cartilage portion Toluidine blue staining figure in the present invention;
Fig. 9 is subchondral bone part calcium tuberosity colored graph in the present invention;
Figure 10 be under scanning electron microscope cell adhesion at cartilage frame orientation microcellular structure exterior view;
Figure 11 is that under scanning electron microscope, cell fills the structure chart in subchondral bone brace aperture;
Figure 12 is that LIVE/DEAD dyeing showed cell fills the structure chart in cartilage frame orientation texture;
Figure 13 is that LIVE/DEAD dyeing showed cell adheres to cartilage frame microcellular structure figure;
Figure 14 is that LIVE/DEAD dyeing showed cell fills structure chart in subchondral bone brace aperture.
Wherein:
1. cartilage layers 2. calcification layer 3. subchondral bone layer.
Detailed description of the invention
Below in conjunction with Figure of description and embodiment, the present invention is described in detail:
As shown in Figure 1, a kind of integrated bone cartilage frame with bionical interfacial structure, described support is made up of cartilage layers 1, calcification layer 2 and subchondral bone layer 3 from top to bottom successively, the vertical orientated property support that described cartilage layers 1 is formed for fibroin albumen; Described calcification layer 2 is the compact area that fibroin albumen and hydroxyapatite are formed, and calcification layer 2 is connected with organic between cartilage layers 1, subchondral bone layer 3; Described subchondral bone layer 3 is the three-dimensional porous structure support that fibroin albumen and hydroxyapatite are formed.
Described cartilage layers 1 be 6% silk fibroin protein solution form through crystallographic orientation, lyophilizing.
Fibroin albumen in described calcification layer 2 and subchondral bone layer 3 and the mass ratio of hydroxyapatite are 1:1, and calcification layer 2 and cartilage layers 1,3 lyophilizing of subchondral bone layer are shaped.The thickness of described calcification layer 2 is less than 500 μm.
The pore diameter range of the three-dimensional porous structure of described subchondral bone layer 3 is 250 μm ~ 425 μm, and subchondral bone layer 3 adopts the method for paraffin microsphere drilling, through lyophilizing, dewaxes and get final product.
Described cartilage layers 1 is 1:1 with the thickness proportion of subchondral bone layer 3.
There is a preparation method for the integrated bone cartilage frame of bionical interfacial structure, comprise the following steps:
A. paraffin microsphere template preparation
0.5g40 ~ 60 object paraffin microsphere is placed in polyethylene mould, flattens gently, be placed in 55 DEG C of baking box 70min and melt solid, room temperature cools;
B. the preparation of calcification layer 2 and subchondral bone layer 3
Get mass fraction respectively and be 10% silk fibroin protein solution and each 1g mixing of hydroxyapatite solution, be added drop-wise to and fill in the polyethylene mould of paraffin microsphere, vacuum draw, mixed liquor is made to fill between paraffin microsphere, suck surplus liquid, only stay paraffin microsphere surface thin layer, namely mixed liquor exceeds paraffin microsphere template 300 μm, after cooled with liquid nitrogen, room temperature leaves standstill 20min;
C. the preparation of cartilage layers 1
Until calcification layer 2 surface is micro-melt after, be that the silk fibroin protein solution of 6% drips to above silk fibroin protein solution and hydroxyapatite mixed liquor gently by mass fraction, height is advisable to drip a full mould, about 2mL; The metal derby of-80 DEG C of coolings is placed on mould, is placed horizontally at-80 DEG C of refrigerator overnight, lyophilizing;
D. fibroin allosteric and dewaxing
The support of lyophilizing is taken out from polyethylene mould, soaked in absolute ethyl alcohol 2h, be put into after drying in the apparatus,Soxhlet's filling cyclohexane extraction and filter wax 48h, slough paraffin, dry and get final product.
The preparation of wherein said paraffin microsphere comprises the following steps:
1. preparing mass fraction is the poly-vinyl alcohol solution of 5%, heating and melting 2h;
2. take 60g paraffin mass, chopping, adds in 1080ml distilled water, then to add 120ml mass fraction be 5% poly-vinyl alcohol solution, heated and stirred, and cooling is sieved, and screen sizes stacks successively from 20 order ~ 80 orders;
3. the paraffin microsphere tap water in screen cloth rinsed repeatedly, ethanol dewaters, dry, class wrapping by size, get 40 ~ 60 order paraffin microspheres for subsequent use.
The preparation that wherein said mass fraction is the silk fibroin protein solution of 10% comprises the following steps:
1. natural silk degumming: Na 40g natural silk being added 8kg0.02mol/L
2cO
3in solution, boil 30min, distillation washing 10 times, changes Na
2cO
3solution, repeats aforesaid operations, and cleaning is placed on ventilation and dries, and obtains fibroin silk;
2. fibroin silk dissolves: take 80g lithium bromide standardize solution distilled water 100ml and dissolve, take the fibroin silk 20g dried, slowly fill in lithium-bromide solution, 60 DEG C of water-bath magnetic agitation 4h;
3. dialysis desalting: pouring fibroin silk lysate into molecular cut off is in the bag filter of 13000, dialyse 3 days, change distilled water every day 3 times, the centrifugal 15min of 11000rpm, get supernatant, pouring molecular cut off into is in the bag filter of 3500, and dialyse 10h in the polyglycol solution of mass fraction 15%, the centrifugal 15min of 11000rpm, gets supernatant and is silk fibroin protein solution;
4. silk fibroin protein solution measurement of concetration: accurately measure 5 groups of 1ml silk fibroin protein solutions, be placed in culture dish, 55 DEG C of dried overnight, with the empty culture dish of analytical balance Measurement accuracy, dry before and the weight of culture dish after drying, calculate concentration and the density of silk fibroin protein solution.
5. according to fibroin albumen concentration measurement, add appropriate distilled water, be mixed with the silk fibroin protein solution of 10%, stir;
Wherein the preparation method of 10% silk fibroin protein solution and 10% hydroxyapatite solution mixed liquor is as follows:
Adopt nano-grade hydroxy apatite, take the hydroxyapatite powder of 1kg, making mass fraction according to mass ratio 1:10 interpolation distilled water 10kg is 10% hydroxyapatite solution, is placed in shaking table 1h and mixes; According to the ratio of mass ratio 1:1, to get mass fraction be 10% silk fibroin protein solution and mass fraction is each 1kg mixing of 10% hydroxyapatite solution, utilize ultrasonator the two to be combined, obtain 10% silk fibroin protein solution and 10% hydroxyapatite solution mixed liquor.
There is in the present invention the Stereo microscope of the integrated bone cartilage frame of bionical interfacial structure, scanning electron microscope detect:
Fig. 2 is the structure chart of the present invention of (× 100) under scanning electron microscopic observation, as shown in Figure 2, observe 3 part levels under integrated bracket Stereo microscope of the present invention obvious, integrated bracket calcification layer 2 is obvious, cartilage layers 1 and calcification layer 2, is connected between calcification layer 2 with subchondral bone layer 3 tight.
Fig. 3 is the cartilage layers longitudinal section figure of (× 500) under scanning electron microscopic observation, Fig. 4 is the cartilage layers cross-sectional view of (× 500) under scanning electron microscopic observation.
As shown in Figure 3,4, cartilage layers has obvious bionical orientation microcellular structure, is evenly distributed;
Fig. 5 is the subchondral bone layer figure of (× 250) under scanning electron microscopic observation, and as shown in Figure 5, subchondral bone layer 3 has good three-dimensional porous structure, porous nickel, connective good.
The present invention has the biocompatibility of the integrated bone cartilage frame of bionical interfacial structure:
By fat stem cell respectively to after cartilage and osteogenic induction, plantation is to cartilage, the subchondral bone part of support respectively, to dye observation through histopathology dyeing, scanning electron microscope, LIVE/DEAD.
Fig. 6 is cartilage portion HE colored graph (× 200) in the present invention, Fig. 7 is subchondral bone part HE colored graph (× 100) in the present invention, Fig. 8 is that in the present invention, cartilage portion Toluidine blue staining figure, Fig. 9 are subchondral bone part calcium tuberosity colored graph (× 100) in the present invention.
As shown in Fig. 6 ~ 9, the fat stem cell uniform adhesion of histological stain display through inducing is in the hole of support, and chondrocyte is spherical in shape, secretes the substrate such as a large amount of proteoglycans, and osteoblast is fusiformis, has a large amount of calcium tuberosity to deposit.
Figure 10 is that under scanning electron microscope, (× 1000) cell adhesion is at cartilage frame orientation microcellular structure exterior view, and Figure 11 is that under scanning electron microscope, (× 500) cell fills the structure chart in subchondral bone brace aperture.
As shown in Figure 10,11, scanning electron microscopic observation, cell uniform adhesion is on support;
Figure 12 is that LIVE/DEAD dyeing showed cell fills the structure chart in cartilage frame orientation texture, Figure 13 is that LIVE/DEAD dyeing showed cell adheres to cartilage frame microcellular structure figure, and Figure 14 is that LIVE/DEAD dyeing showed cell fills structure chart in subchondral bone brace aperture.
As shown in Figure 12 ~ 14, (× 200) are observed in LIVE/DEAD dyeing, and the fat stem cell through induction is green fluorescence respectively on cartilage frame and subchondral bone support, represents cell survival good.
Major advantage of the present invention is as follows:
1. " hyaline cartilage-interfacial structure-subchondral bone " natural structure according to bionics principle simulation joint is proposed, the design concept of development " bone support-interfacial structure-cartilage frame "; And the SF solution proposing to utilize non-occurred conformation to change itself dissolves each other, and by three portion support compact siro spinning technology, has both isolated bone, cartilage microenvironment, and has strengthened again the mechanical strength of cartilage, bone parts.
2. cartilage layers 1 is bionical orientation microcellular structure, subchondral bone layer 3 possesses the Macro-micro structure of high connectivity and porosity, interfacial structure composition is similar to natural cartilage calcification layer 2, making its structure function closer to physiology, is the new model in the regeneration of a kind of bone-cartilage complex tissue and the field of reconstruction.
3. cartilage layers 1, subchondral bone layer 3 have good pore structure and connectedness, are conducive to the transfer that cell migration enters internal stent and nutrient substance and metabolite.
Claims (7)
1. there is a preparation method for the integrated bone cartilage frame of bionical interfacial structure, it is characterized in that: comprise the following steps:
A. paraffin microsphere template preparation
Paraffin microsphere is put into polyethylene mould, dry for standby;
B. the preparation of calcification layer (2) and subchondral bone layer (3)
Be the mixing of 10% silk fibroin protein solution and 10% hydroxyapatite solution 1:1 in mass ratio by mass fraction, add in paraffin microsphere template, aspiration vacuum, mixed liquor is fully filled, and exceeds paraffin microsphere template certain distance, and after cooled with liquid nitrogen, room temperature leaves standstill;
C. the preparation of cartilage layers (1)
Until calcification layer (2) surface micro-melt after, be that the silk fibroin protein solution 2mL of 6% slowly adds on the calcification layer (2) in mould by mass fraction, by-80 DEG C cooling metal derbies be placed on mould, be placed horizontally at-80 DEG C of lyophilization molding;
D. fibroin allosteric and dewaxing
Take out from polyethylene mould after molding, soaked in absolute ethyl alcohol, filter wax, dries, obtains bone cartilage frame; Described bone cartilage frame is made up of cartilage layers (1), calcification layer (2) and subchondral bone layer (3) from top to bottom successively, the vertical orientated property support that described cartilage layers (1) is formed for fibroin albumen, the compact area that described calcification layer (2) is formed for fibroin albumen and hydroxyapatite, the three-dimensional porous structure support that described subchondral bone layer (3) is formed for fibroin albumen and hydroxyapatite; Calcification layer (2) is connected with organic between cartilage layers (1), subchondral bone layer (3).
2. a kind of preparation method with the integrated bone cartilage frame of bionical interfacial structure according to claim 1, is characterized in that: the pore diameter range of the three-dimensional porous structure of described subchondral bone layer (3) is 250 μm-425 μm.
3. a kind of preparation method with the integrated bone cartilage frame of bionical interfacial structure according to claim 1, is characterized in that: the thickness of described calcification layer (2) is less than 500 μm.
4. a kind of preparation method with the integrated bone cartilage frame of bionical interfacial structure according to claim 1, is characterized in that: described cartilage layers (1) is 1:1 with the thickness proportion of subchondral bone layer (3).
5. a kind of preparation method with the integrated bone cartilage frame of bionical interfacial structure according to claim 1, is characterized in that: the preparation of described silk fibroin protein solution comprises the following steps:
1. natural silk degumming: natural silk is added the Na of 0.02mol/L by 1:200 in mass ratio
2cO
3in solution, boil, repeatedly wash, change Na
2cO
3repeat after solution to boil, wash, then ventilate and dry, obtain fibroin silk;
2. fibroin silk lysate: fibroin silk adds in the lithium-bromide solution of 0.8g/mL by 1:4 in mass ratio, in 60 DEG C of water-baths, magnetic agitation obtains;
3. dialysis desalting: pouring fibroin silk lysate into molecular cut off is in the bag filter of 12000-14000, dialyse 3 days, every day changes water, centrifugal, get supernatant, proceeding to molecular cut off is in the bag filter of 3500, dialyse in the polyglycol solution of mass fraction 15%, centrifuging and taking supernatant and get final product.
6. a kind of preparation method with the integrated bone cartilage frame of bionical interfacial structure according to claim 1, is characterized in that: the preparation of described paraffin microsphere comprises the following steps:
1. preparing mass fraction is the poly-vinyl alcohol solution of 5%, heating and melting;
2. take 60g paraffin mass, chopping, adds in 1080ml distilled water, then to add 120ml mass fraction be 5% polyvinyl alcohol, heated and stirred, cooling, and 20-80 eye mesh screen stacks successively, sieves;
3. sieve paraffin microsphere water rinses repeatedly, ethanol dewaters, dry, get 40-60 order paraffin microsphere for subsequent use.
7. a kind of preparation method with the integrated bone cartilage frame of bionical interfacial structure according to claim 1, is characterized in that: described filter wax technology application apparatus,Soxhlet's, solvent is normal hexane or cyclohexane extraction.
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| CN106039400B (en) * | 2016-05-27 | 2019-10-11 | 浙江大学 | Ice crystal template prepares the methods and applications of regular lamellar structure three dimensional biological bracket |
| US10022231B2 (en) * | 2016-07-22 | 2018-07-17 | Cytex Therapeutics, Inc. | Articular cartilage repair |
| CN108478871B (en) * | 2018-04-17 | 2020-10-13 | 四川大学 | Integrated bone-cartilage repair scaffold and preparation method thereof |
| CN108578780B (en) * | 2018-05-06 | 2020-12-08 | 西北工业大学 | Preparation method of artificial bone scaffold carrying silver ions and having mechanical gradient |
| CN108404214B (en) * | 2018-06-01 | 2021-05-14 | 上海贝奥路生物材料有限公司 | Bionic bone cartilage complex and preparation method thereof |
| CN108914368A (en) * | 2018-07-24 | 2018-11-30 | 高昕文 | A kind of preparation method of facial mask fabric nonwoven cloth |
| CN109172870B (en) * | 2018-07-26 | 2021-05-18 | 南开大学 | Oriented full-layer cartilage scaffold containing calcified layer and preparation method thereof |
| CN110101917B (en) * | 2019-05-16 | 2022-05-17 | 南开大学 | Calcified layer-containing cartilage scaffold with slow-release double growth factors and preparation method thereof |
| CN115414532B (en) * | 2022-09-13 | 2023-12-19 | 华东交通大学 | Bacterial cellulose-based integrated osteochondral scaffold with interface barrier layer and preparation method thereof |
| CN115382021B (en) * | 2022-09-22 | 2024-01-05 | 诺一迈尔（苏州）医学科技有限公司 | Composite artificial cartilage bracket and preparation method thereof |
| CN115554475B (en) * | 2022-09-29 | 2023-09-05 | 中国人民解放军总医院第一医学中心 | Osteochondral scaffold and preparation method thereof |
| CN115414529B (en) * | 2022-09-30 | 2024-04-02 | 重庆生物智能制造研究院 | Preparation method of three-layer 3D printing bone cartilage scaffold |
| CN116271209B (en) * | 2023-03-03 | 2024-07-02 | 北京爱康宜诚医疗器材有限公司 | Cartilage repair product, preparation method and application thereof |
| CN117230103B (en) * | 2023-09-25 | 2024-04-19 | 爱光生物医药(南昌)有限公司 | 17-Type humanized collagen and protein scaffold, filling material and application thereof |
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| CN100522265C (en) * | 2007-03-08 | 2009-08-05 | 中国人民解放军第三军医大学第一附属医院 | Integral engineering rack of interface osteochondro tissue with bionic function |
| CN101810885B (en) * | 2010-04-06 | 2013-03-06 | 清华大学 | Method for preparing double-layer bionic cartilage tissue engineering scaffold |
| GB201021438D0 (en) * | 2010-12-16 | 2011-02-02 | Imp Innovations Ltd | Layered fibrous construct |
| CN102580156B (en) * | 2012-03-29 | 2014-10-08 | 陕西博鸿生物科技有限公司 | Tissue engineering cartilage framework material, as well as preparation method and device thereof |
| CN103028145B (en) * | 2012-10-24 | 2014-06-18 | 浙江大学 | Silk fibroin base integrated osteochondral two-layer bracket, preparation and application thereof |
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