CN104324418A - Nanofiber bone cartilage repairing stent for tissue engineering and preparation method thereof - Google Patents
Nanofiber bone cartilage repairing stent for tissue engineering and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a nanofiber bone cartilage repairing stent for tissue engineering and a preparation method thereof. The nanofiber bone cartilage repairing stent for tissue engineering is characterized by comprising a porous cartilage layer, a porous cartilage lower bone layer and a connecting layer between the porous cartilage layer and the porous cartilage lower bone layer, wherein the porous cartilage layer includes a PLLA-based composite degradable polymer material and is of a porous nanofiber structure which is 20-400mu m in pore diameter; the connecting layer includes a PLLA-based composite degradable polymer material and is of a porous nanofiber structure which is less than 5mu m in pore diameter; and the porous cartilage lower bone layer comprises a PLLA-based composite degradable polymer material containing 5-70% (w/w) of nano-hydroxyapatite and is of a porous nanofiber structure which is 50-500mu m in pore diameter. Microstructure of the bone cartilage repairing stent prepared by the invention is represented as bionic ECM nanofiber; the repairing stent can promote adhesion, propagation and differentiation of related bone cartilage cells on the stent, and can also promote transportation of nutrient substances in the stent and discharge of metabolic waste.
Description
Technical field
The invention belongs to osteochondral tissue engineering rack technical field, particularly a kind of organizational project nanofiber bone repair of cartilage support and preparation method thereof.
Background technology
The damage of articular cartilage and pathological changes are orthopaedic diseases common clinically, and are difficult to self-regeneration.And the damage of joint cartilage is usually along with the combined injuries of subchondral bone layer, also need while therefore repairing cartilage to repair subchondral bone.Therapeutic Method conventional clinically is at present autologous or allograph bone cartilage transplantation, achieve certain therapeutic effect, but also Shortcomings, autologous bone cartilage limited source, and secondary injury can be caused to patient, and there is the risk of immunologic rejection and disease propagation in allograph bone cartilage.The external structure bone repair of cartilage support that develops into of organizational project provides new approach.
Support is an important ingredient in organizational project, plays and supports the effect such as Growth of Cells, guide tissue regeneration.First desirable tissue engineering bracket should possess the nanofibrous structures of bionical natural extracellular matrix (ECM), to promote the adhesion of cell on support, propagation and differentiation, and promote the transport of nutrient substance in support and the discharge of metabolic waste.In addition, support also should have suitable pore structure, is applicable to cell to the growth in support, promotes the regeneration of tissue.Chinese patent (CN 102872480A) discloses a kind of two-phase osteochondral tissue engineering rack, and its cartilage layers and subchondral bone layer are loose structure, but microcosmic is not the nanofibrous structures of bionical ECM.And being the loose structure of connection due to double-layer structure, this makes the relevant cell of cartilage layers and subchondral bone layer be easy to migration mutually to grow into, affect the reparation of cartilage layers and subchondral bone layer function.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of organizational project nanofiber bone repair of cartilage support and preparation method thereof is provided.The method is based on bionic principle, and the nanofibrous structures of natural ECM fully simulated by the bone repair of cartilage support of preparation, promotes the adhesion of relevant cell on support, reproduction restraint; Nanofiber articulamentum fine and close is together defined between porous nano-fibre cartilage layers and porous nano-fibre subchondral bone layer, not only simulate the cartilaginous calcification layer of nature bone cartilage structure, and play the effect spatially isolating cartilage layers and subchondral bone layer, prevent the mutual migration of cartilage layers and subchondral bone layer relevant cell, ensure the regeneration of cartilage layers and subchondral bone layer function tissue.
In order to achieve the above object, the invention provides a kind of organizational project nanofiber bone repair of cartilage support, it is characterized in that, be made up of porous cartilage layers, porous subchondral bone layer and the articulamentum be located between porous cartilage layers and porous subchondral bone layer, described porous cartilage layers comprises PLLA base composite degradable polymeric material, for porous nano-fibre structure, aperture is at 20 ~ 400 μm; Articulamentum comprises PLLA base composite degradable polymeric material, is porous nano-fibre structure, aperture < 5 μm; Porous subchondral bone layer comprises the PLLA base composite degradable polymeric material containing 5 ~ 70% (w/w) nanometer hydroxyapatite, and be porous nano-fibre structure, aperture is 50 ~ 500 μm.
Present invention also offers the preparation method of above-mentioned organizational project nanofiber bone repair of cartilage support, it is characterized in that, concrete steps comprise:
The first step: under 40 ~ 80 DEG C of conditions, by PLLA and with it immiscible degradable polymeric material be dissolved in solvent I, obtain the uniform solution that concentration is 6 ~ 15g: 100mL, then the nanometer hydroxyapatite that mass fraction is 5 ~ 70% (w/w) (with the gross mass of nanometer hydroxyapatite and polymer for benchmark) is added wherein, obtain mixture solution, stir ultrasonic disperse, the mixed solution of gained is cast into and is preheating in 40 ~ 80 DEG C of moulds, be separated at-20 DEG C ~-80 DEG C and spend the night, to decorporate mould, obtain polymer gel, then immerse in solvent II and carry out solvent displacement, lyophilization, porous subchondral bone layer can be obtained,
Second step: under 40 ~ 80 DEG C of conditions, by PLLA and with it immiscible degradable polymeric material be dissolved in solvent I, obtain the homogeneous polymer solution that concentration is 6 ~ 15g: 100mL, be cast into and be preheating in 40 ~ 80 DEG C of moulds; Then porous subchondral bone layer is put into mould, pressing porous subchondral bone layer material is by itself and described polymer solution close contact; Then be separated at-20 DEG C ~-80 DEG C and spend the night, then immerse in solvent II and carry out solvent displacement, take out, lyophilization, forms articulamentum and porous cartilage layers successively under porous subchondral bone layer, can obtain organizational project nanofiber bone repair of cartilage support.
Preferably, in the described first step and second step is one or more compositions in poly-epsilon-caprolactone, polyglycolic acid, poly-β-hydroxybutyric acid, polyhydroxy-alkanoate, sebacic acid and propyl tri-alcohol ester, polyurethane, polyvinylpyrrolidone, polyethylene, polypropylene, polyethylene terephthalate, collagen, gelatin, fibroin albumen, Fibrinogen, cellulose and chitosan with the immiscible degradable polymeric material of PLLA.
Preferably, the solvent I in the described first step and second step is one or more the compositions in oxolane, hexafluoroisopropanol, trifluoroacetic acid, Isosorbide-5-Nitrae-dioxane, N, N-methyl nitrosourea, chloroform and dichloromethane.
Preferably, the solvent II in the described first step and second step is one or more the compositions in water, ethanol, methanol, hexane, cyclohexane extraction and acetone, and temperature is-20 DEG C ~ 0 DEG C, and solvent displacement total time is 2-3 days, changes 3-5 solvent II every day.
The present invention makes full use of the advantage that Thermal inactive technology can prepare three-dimensional manometer fibrous framework, adopt medically acceptable PLLA as main matrix, reach and promote the adhesion of cell on support, proliferation and growth etc., also promote the transport of nutrient substance in support and the discharge of metabolic waste simultaneously; The degradable polymeric material immiscible with PLLA that other is medically acceptable is introduced in system, prepare porous nano-fibre support, promote that bone cartilage relevant cell is to the growth in support, promote tissue regeneration, other polymer simultaneously introduced can also improve the physicochemical property of support, as degradation property, mechanical property, biological activity etc.; In bone repair of cartilage support prepared by the method, between porous cartilage layers and porous subchondral bone layer, yet forms both together fine and close articulamentum, cartilaginous calcification layer in bionical nature bone cartilage.
Compared with prior art, the invention has the beneficial effects as follows:
(1) the bone repair of cartilage support microcosmic that prepared by the present invention is the nanofibrous structures of bionical ECM, the adhesion of bone cartilage relevant cell on support, proliferation and growth can be promoted, also can promote the transport of nutrient substance in support and the discharge of metabolic waste, the growth for cell provides a desirable microenvironment;
(2) cartilage layers of bone repair of cartilage support prepared of the present invention and subchondral bone layer are loose structure, be applicable to cell to the growth in support, promote the regeneration of osteochondral tissue, the interpolation of porous subchondral bone layer hydroxyapatite simultaneously can induce the regeneration impelling subchondral bone layer;
(3) articulamentum of one deck densification is also had between the cartilage layers of bone repair of cartilage support prepared of the present invention and subchondral bone layer, cartilaginous calcification layer in bionical nature bone cartilage, spatially isolate porous cartilage layers and porous subchondral bone layer, prevent the mutual migration of cartilage layers and subchondral bone layer relevant cell, ensure the regeneration of cartilage layers and subchondral bone layer function tissue;
(4) the present invention can improve the overall physicochemical property of support by introducing other and the immiscible degradable polymeric material of PLLA in system;
(5) the present invention is simple to operate, and the height of cartilage layers and subchondral bone layer can with requiring to adjust with mould, and do not need complex device, can be mass-produced, preparation cost is cheap;
Accompanying drawing explanation
Fig. 1 is the SEM picture of the organizational project nanofiber bone repair of cartilage support entirety of preparation;
Fig. 2 is the SEM picture of organizational project nanofiber bone repair of cartilage support porous cartilage layers;
Fig. 3 is the SEM picture of the fine and close articulamentum of organizational project nanofiber bone repair of cartilage support;
Fig. 4 is the SEM picture of organizational project nanofiber bone repair of cartilage support porous subchondral bone layer.
Detailed description of the invention
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
As shown in Figure 1, organizational project nanofiber bone repair of cartilage support of the present invention, be made up of porous cartilage layers 1, porous subchondral bone layer 3 and the articulamentum 2 be located between porous cartilage layers 1 and porous subchondral bone layer 3, described porous cartilage layers 1 comprises PLLA base composite degradable polymeric material, for porous nano-fibre structure, aperture is at 20 ~ 60 μm, and thickness is 1mm; Articulamentum 2 comprises PLLA base composite degradable polymeric material, is porous nano-fibre structure, aperture < 5 μm, and thickness is 0.4mm; Porous subchondral bone layer 3 comprises the PLLA base composite degradable polymeric material containing 20% nanometer hydroxyapatite, and be porous nano-fibre structure, aperture is 60 ~ 400 μm, and thickness is 1.8mm.
The preparation method of the preparation method of described organizational project nanofiber bone repair of cartilage support is:
(1) by PLLA (viscosity: 1.92dl/g, purchased from Jinan Dai Gang biological engineering company limited) and polycaprolactone (PCL) (Mn:8 × 10
4purchased from Sigma) be dissolved in oxolane with mass ratio 6: 4 in 60 DEG C, stirring is mixed with the uniform solution of 10g: 100mL, then the nanometer hydroxyapatite of 20% (w/w) (with the gross mass of nanometer hydroxyapatite and polymer for benchmark) is added wherein, stir and ultrasonic disperse, then mixed solution is cast into the Teflon mould being preheating to 60 DEG C (barrel-shaped, band lid, internal diameter is about 18mm) in, make it be separated under being placed in rapidly-20 DEG C of conditions, spend the night;
(2) from low temperature, mould is taken out, and mould of decorporating, the gelatinous polymer of gained is immersed in deionization mixture of ice and water and carries out solvent and replace 2 days, change 3 deionization mixture of ice and water every day, take out polymer afterwards, lyophilization 2 days, namely obtains porous subchondral bone layer as shown in Figure 4;
(3) PLLA and PCL is dissolved in oxolane with mass ratio 7: 3 in 60 DEG C, stirring is mixed with the homogeneous polymer solution of 10g: 100mL, be cast in the Teflon mould being preheating to 60 DEG C, then obtained porous subchondral bone layer 3 is put into mould, flicking porous subchondral bone layer 3 material is by itself and described polymer solution close contact, then make it be separated under it being placed in rapidly-20 DEG C of conditions, spend the night;
(4) from low temperature, mould is taken out, and mould of decorporating, then carry out solvent in immersion deionization mixture of ice and water and replace 2 days, change 3 deionization mixture of ice and water every day, then support is taken out, lyophilization 2 days, forms articulamentum 2 as shown in Figure 3 and porous cartilage layers 1 as shown in Figure 2 successively, namely obtains nanofiber bone repair of cartilage support as shown in Figure 1 under porous subchondral bone layer 3.
Embodiment 2
Organizational project nanofiber bone repair of cartilage support of the present invention, be made up of porous cartilage layers, porous subchondral bone layer and the articulamentum be located between porous cartilage layers and porous subchondral bone layer, described porous cartilage layers comprises PLLA base composite degradable polymeric material, for porous nano-fibre structure, aperture is at 20 ~ 60 μm, and thickness is 1.5mm; Articulamentum comprises PLLA base composite degradable polymeric material, is porous nano-fibre structure, aperture < 5 μm, and thickness is 0.5mm; Porous subchondral bone layer comprises the PLLA base composite degradable polymeric material containing 30% nanometer hydroxyapatite, and be porous nano-fibre structure, aperture is 60 ~ 500 μm, and thickness is 2.5mm.
The preparation method of the preparation method of described organizational project nanofiber bone repair of cartilage support is:
(1) PLLA and PCL is dissolved in oxolane with mass ratio 5: 5 in 50 DEG C, stirring is mixed with the uniform solution of 12g: 100mL, then the nanometer hydroxyapatite of 30% (w/w) (with the gross mass of nanometer hydroxyapatite and polymer for benchmark) is added wherein, stir ultrasonic disperse, then mixed solution is cast into the Teflon mould being preheating to 50 DEG C (barrel-shaped, band lid, internal diameter is about 18mm) in, make it be separated under being placed in rapidly-80 DEG C of conditions, spend the night;
(2) from low temperature, take out mould, and mould of decorporating, the gelatinous polymer of gained is immersed in deionization mixture of ice and water and carries out solvent and replace 2 days, change 3 deionization mixture of ice and water every day, take out polymer afterwards, lyophilization 2 days, namely obtain porous subchondral bone layer 3;
(3) by PLLA and sebacic acid and propyl tri-alcohol ester (PGS) (Mn:1 × 10
4laboratory synthesizes) be dissolved in oxolane with mass ratio 7: 3 in 50 DEG C, stirring is mixed with the homogeneous polymer solution of 12g: 100mL, be cast in the Teflon mould being preheating to 50 DEG C, then obtained porous subchondral bone layer is put into mould, flicking porous subchondral bone layer 3 material, by itself and described polymer solution close contact, makes it be separated under then it being placed in rapidly-80 DEG C of conditions, spends the night;
(4) from low temperature, mould is taken out, and mould of decorporating, then carry out solvent in immersion deionization mixture of ice and water and replace 2 days, change 3 deionization frozen water every day, then support is taken out, lyophilization 2 days, forms articulamentum and porous cartilage layers successively under porous subchondral bone layer, namely obtains nanofiber bone repair of cartilage support.
Embodiment 3
Organizational project nanofiber bone repair of cartilage support of the present invention, be made up of porous cartilage layers, porous subchondral bone layer and the articulamentum be located between porous cartilage layers and porous subchondral bone layer, described porous cartilage layers comprises PLLA base composite degradable polymeric material, for porous nano-fibre structure, aperture is at 20 ~ 60 μm, and thickness is 2mm; Articulamentum comprises PLLA base composite degradable polymeric material, is porous nano-fibre structure, aperture < 5 μm, and thickness is 0.5mm; Porous subchondral bone layer comprises the PLLA base composite degradable polymeric material containing 40% nanometer hydroxyapatite, and be porous nano-fibre structure, aperture is 60 ~ 400 μm, and thickness is 2.5mm.
The preparation method of the preparation method of described organizational project nanofiber bone repair of cartilage support is:
(1) PLLA and polyurethane (PU) are dissolved in oxolane with mass ratio 6: 4 in 40 DEG C, stirring is mixed with the uniform solution of 8g: 100mL, then the nanometer hydroxyapatite of 40% (w/w) (with the gross mass of nanometer hydroxyapatite and polymer for benchmark) is added wherein, stir ultrasonic disperse, then mixed solution is cast into the Teflon mould being preheating to 40 DEG C (barrel-shaped, band lid, internal diameter is about 18mm) in, make it be separated under being placed in rapidly-80 DEG C of conditions, spend the night;
(2) from low temperature, take out mould, and mould of decorporating, gelatinous polymer is immersed in deionization mixture of ice and water and carries out solvent and replace 2 days, change 3 deionization frozen water every day, take out polymer afterwards, lyophilization 2 days, namely obtain porous subchondral bone layer 3;
(3) PLLA and PGS is dissolved in oxolane with mass ratio 7: 3 in 50 DEG C, stir the homogeneous polymer solution being mixed with 8g:100mL, be cast in the Teflon mould being preheating to 50 DEG C, then obtained porous subchondral bone layer is put into mould, flicking porous subchondral bone layer 3 material is by itself and described polymer solution close contact, then make it be separated under it being placed in rapidly-80 DEG C of conditions, spend the night;
(4) from low temperature, mould is taken out, and then mould of decorporating immerses in deionization mixture of ice and water and carries out solvent and replace 2 days, change 3 deionization frozen water every day, then support is taken out, lyophilization 2 days, under porous subchondral bone layer, form articulamentum and porous cartilage layers successively, namely obtain nanofiber bone repair of cartilage support.
Claims (5)
1. an organizational project nanofiber bone repair of cartilage support, it is characterized in that, be made up of porous cartilage layers (1), porous subchondral bone layer (3) and the articulamentum (2) be located between porous cartilage layers (1) and porous subchondral bone layer (3), described porous cartilage layers (1) comprises PLLA base composite degradable polymeric material, for porous nano-fibre structure, aperture is at 20 ~ 400 μm; Articulamentum (2) comprises PLLA base composite degradable polymeric material, is porous nano-fibre structure, aperture < 5 μm; Porous subchondral bone layer (3) comprises the PLLA base composite degradable polymeric material containing 5 ~ 70% (w/w) nanometer hydroxyapatite, and be porous nano-fibre structure, aperture is 50 ~ 500 μm.
2. the preparation method of organizational project nanofiber bone repair of cartilage support according to claim 1, it is characterized in that, concrete steps comprise:
The first step: under 40 ~ 80 DEG C of conditions, by PLLA and with it immiscible degradable polymeric material be dissolved in solvent I, obtain the uniform solution that concentration is 6 ~ 15g:100mL, then the nanometer hydroxyapatite that mass fraction is 5 ~ 70% (w/w) is added wherein, obtain mixture solution, stir ultrasonic disperse, the mixed solution of gained is cast into and is preheating in 40 ~ 80 DEG C of moulds, be separated at-20 DEG C ~-80 DEG C and spend the night, to decorporate mould, obtain polymer gel, then immerse in solvent II and carry out solvent displacement, lyophilization, porous subchondral bone layer (3) can be obtained,
Second step: under 40 ~ 80 DEG C of conditions, by PLLA and with it immiscible degradable polymeric material be dissolved in solvent I, obtain the homogeneous polymer solution that concentration is 6% ~ 15g:100mL, be cast into and be preheating in 40 ~ 80 DEG C of moulds; Then porous subchondral bone layer (3) is put into mould, pressing porous subchondral bone layer (3) material is by itself and described polymer solution close contact; Then be separated at-20 DEG C ~-80 DEG C and spend the night, then immerse in solvent II and carry out solvent displacement, take out, lyophilization, under porous subchondral bone layer (3), form articulamentum (2) and porous cartilage layers (1) successively, organizational project nanofiber bone repair of cartilage support can be obtained.
3. the preparation method of organizational project nanofiber bone repair of cartilage support as claimed in claim 2, it is characterized in that, in the described first step and second step is one or more compositions in poly-epsilon-caprolactone, polyglycolic acid, poly-β-hydroxybutyric acid, polyhydroxy-alkanoate, sebacic acid and propyl tri-alcohol ester, polyurethane, polyvinylpyrrolidone, polyethylene, polypropylene, polyethylene terephthalate, collagen, gelatin, fibroin albumen, Fibrinogen, cellulose and chitosan with the immiscible degradable polymeric material of PLLA.
4. the preparation method of organizational project nanofiber bone repair of cartilage support as claimed in claim 2, it is characterized in that, solvent I in the described first step and second step is oxolane, hexafluoroisopropanol, trifluoroacetic acid, 1, one or more compositions in 4-dioxane, N, N-dimethylformamide, chloroform and dichloromethane.
5. the preparation method of organizational project nanofiber bone repair of cartilage support as claimed in claim 2, it is characterized in that, solvent II in the described first step and second step is one or more the compositions in water, ethanol, methanol, hexane, cyclohexane extraction and acetone, temperature is-20 DEG C ~ 0 DEG C, solvent displacement total time is 2-3 days, changes 3-5 solvent II every day.
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| CN106178126A (en) * | 2016-07-16 | 2016-12-07 | 遵义市第人民医院 | A kind of reparation bone cartilage two-phase porous compound support frame and preparation method thereof |
| CN108042848A (en) * | 2018-01-23 | 2018-05-18 | 上海交通大学医学院附属第九人民医院 | Polyesters stent and its application in bone defect healing |
| CN108201632A (en) * | 2016-12-20 | 2018-06-26 | 重庆润泽医药有限公司 | A kind of articular cartilage repaiies scaffold |
| CN108404214A (en) * | 2018-06-01 | 2018-08-17 | 上海贝奥路生物材料有限公司 | A kind of bionical osteochondral composites and preparation method thereof |
| CN108451627A (en) * | 2018-04-04 | 2018-08-28 | 中国人民解放军总医院 | A kind of bone renovating material intermingling apparatus |
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| CN106178126A (en) * | 2016-07-16 | 2016-12-07 | 遵义市第人民医院 | A kind of reparation bone cartilage two-phase porous compound support frame and preparation method thereof |
| CN108201632A (en) * | 2016-12-20 | 2018-06-26 | 重庆润泽医药有限公司 | A kind of articular cartilage repaiies scaffold |
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| CN109876192A (en) * | 2019-03-13 | 2019-06-14 | 东华大学 | A kind of Bone Defect Repari film and preparation method thereof |
| CN110882416A (en) * | 2019-11-12 | 2020-03-17 | 东华大学 | Preparation method and application of bionic composite nanofiber scaffold material |
| CN113730658B (en) * | 2021-09-22 | 2022-08-16 | 太原理工大学 | Bionic bone cartilage integrated repair support and preparation method thereof |
| CN113730658A (en) * | 2021-09-22 | 2021-12-03 | 太原理工大学 | Bionic bone cartilage integrated repair support and preparation method thereof |
| CN114129775A (en) * | 2021-12-13 | 2022-03-04 | 上海大学 | Bionic cell-containing massive osteochondral biological scaffold and preparation method thereof |
| CN114129775B (en) * | 2021-12-13 | 2022-08-09 | 上海大学 | Bionic cell-containing massive osteochondral biological scaffold and preparation method thereof |
| CN115414532A (en) * | 2022-09-13 | 2022-12-02 | 华东交通大学 | Bacterial cellulose-based integrated osteochondral scaffold with interface barrier layer and preparation method |
| 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 |
| CN115671386A (en) * | 2022-10-27 | 2023-02-03 | 华中科技大学 | Polyether-ether-ketone support for integrally repairing cartilage and lower bone and manufacturing method thereof |
| CN115671386B (en) * | 2022-10-27 | 2024-03-19 | 华中科技大学 | Polyether-ether-ketone bracket for integrally repairing cartilage and lower bone and manufacturing method thereof |
| CN116392639A (en) * | 2023-02-17 | 2023-07-07 | 无锡市中医医院 | Full-layer repair double-layer bracket and preparation method and application thereof |
| CN116392639B (en) * | 2023-02-17 | 2024-02-13 | 无锡市中医医院 | Full-layer repair double-layer bracket and preparation method and application thereof |
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