CN103845758A - Novel nano bioceramic coated artificial ligament and preparation method of ligament - Google Patents
Novel nano bioceramic coated artificial ligament and preparation method of ligament Download PDFInfo
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Abstract
The invention relates to a novel nano bioceramic coated artificial ligament and a preparation method of the ligament. The novel nano bioceramic coated artificial ligament comprises an artificial ligament and a Ca-Mg-Si bioceramic coating which is uniformly coated on the surface of the artificial ligament, wherein the grain size of ceramic particles of the Ca-Mg-Si bioceramic coating is 20-80nm. By coating the Ca-Mg-Si bioceramic coating on the surface of the artificial ligament, the artificial ligament coated by the coating has multifunctional characteristics of good biological activity, bone formability, bone graft union and the like by means of excellent bone induction capacity of the Ca-Mg-Si bioceramic, so that the artificial ligament becomes a novel graft material with the biological activity.
Description
Technical field
The present invention relates to artificial ligament of a kind of novel nano-biological cerimic coating and its production and use, belong to technical field of biological material.
Background technology
After anterior cruciate ligament (ACL) damage, be difficult to spontaneous recovery, and expectant treatment is invalid, functional rehabilitation is incomplete, after athlete plays again, very easily repeatedly hinders again.The treatment of ACL damage is focus and the difficult point of domestic and international sports medical science research.At present, ACL reconstruction operations is the main method for the treatment of ACL damage, the postoperative kneed stability [1] of recovering.ACL rebuilds graft and selects to comprise autologous tendon, allosome tendon and artificial ligament.Autologous tendon graft application is more extensive, but has tendon for district's infringement, delay reconditioning and rebuild the problems such as overhaul technology graft source shortage.Tendon allograft thing, although avoid sacrificing autologous tendon tissue, originates more and more difficult, has pathophoresis and immune rejection equivalent risk [2].Because autologous tendon and allogeneic tendon exist above-mentioned shortcoming, surgeon and researcher attempt to work out a kind of Utopian artificial ligament that possesses high strength and biocompatibility, are used for rebuilding ACL operation as graft.In order to address this problem, polyethylene terephthalate (PET) artificial ligament is used as ACL and rebuilds graft.But it is postoperative that LARS ligament is rebuild ACL, distinct issues are " graft knitting are bad ".Graft knitting needs scar tissue on graft bone interface to reduce and new osteogenesis, reaches graft bone and fits tightly, thereby guarantee the stability in graft Gu road.Also do not have at present Histological research to confirm the postoperative artificial ligament [3] that can effectively induce osseous tissue to grow in bone road of LARS ligament reconstruction ACL.Artificial ligament not yet solves the firmly problem of healing of artificial ligament graft Yu Gu road.Therefore, accelerate and promote graft knitting significant to the success or failure of PET artificial ligament reconstruction ACL operation.The bad topmost reason of PET artificial ligament graft knitting is because material surface hydrophobicity and lack corresponding biological activity, is unfavorable for that osteoblast is in the sticking, breed and break up of material surface, and skeletonization on graft bone interface.In order to address this problem, in recent years, researcher adopts different biocompatibility or bio-active material coating modification PET artificial ligament to promote graft knitting, mainly comprises [4,5] such as hydroxyapatite, bioactivity glass, organic polymers.But the main problem existing of current PET artificial ligament coating research: (1) coating process is single, coating is inhomogeneous, less stable, experimental result repeatability are poor; (2) coating result is slower, and graft knitting is not ideal; (3) lack relevant different coating thickness, degree of crystallinity and grain size for skeletonization impact in cell in vitro and body.Therefore, if can use a kind of new method uniformly and stably bioactive materials to be coated on to PET artificial ligament surface, the biological activity that improves graft surface promotes skeletonization, and improve the biology performance of coating by the parameter of regulation and control coating, will play very important effect for selecting best coating schemes to solve the bad problem of artificial ligament graft knitting so.Previous research shows, in recent years, pulsed laser technique (PLD) is widely used in material preparation [6].Ca-Mg-Si system pottery, such as akermanite (Akermanite, AKT), be the bioceramic of a kind of calcic, magnesium, silicon, has excellent bone inducibility, is widely used [7] in bone tissue engineer.
List of references
1.Levy?BA.Is?early?reconstruction?necessary?for?all?anterior?cruciate?ligament?tears?N?Engl?J?Med2010,363,386-388.
2.Hu?J,Qu?J,Xu?D,Zhou?J,Lu?H.Allograft?versus?autograft?for?anterior?cruciate?ligament?reconstruction:an?up-to-date?meta-analysis?of?prospective?studies.Int?Orthop2013,37,311-320.
Korea Spro increase refined, Guo Xiucheng, Xu Dong. the experimentation that artificial ligament is combined with bone interface. the Orthopedic Journal of China 2009,17,1407-1410.
4.Li?H,Ge?Y,Wu?Y,Jiang?J,Gao?K,Zhang?P,Wu?L,Chen?S.Hydroxyapatite?coating?enhances?polyethylene?terephthalate?artificial?ligament?graft?osseointegration?in?the?bone?tunnel.Int?Orthop2011,35,1561-1567.
5.Li?H,Chen?S,Wu?Y,Jiang?J,Ge?Y,Gao?K,Zhang?P,Wu?L.Enhancement?of?the?osseointegration?of?a?polyethylene?terephthalate?artificial?ligament?graft?in?a?bone?tunnel?using58S?bioglass.Int?Orthop2012,36,191-197.
6.Eisenhawer?B,Sivakov?V,Christiansen?S,Falk?F.A?time-resolved?numerical?study?of?the?vapor-liquid-solid?growth?kinetics?describing?the?initial?nucleation?phase?as?well?as?pulsed?deposition?processes.Nano?Lett2013,13,873-883.
7.Gu?H,Guo?F,Zhou?X,Gong?L,Zhang?Y,Zhai?W,Chen?L,Cen?L,Yin?S,Chang?J,Cui?L.The?stimulation?of?osteogenic?differentiation?of?human?adipose-derived?stem?cells?by?ionic?products?from?akermanite?dissolution?via?activation?of?the?ERK?pathway.Biomaterials2011,32,7023-7033.。
Summary of the invention
In the face of the problem that prior art exists, the object of the present invention is to provide artificial ligament of a kind of novel nano-biological cerimic coating and preparation method thereof.
At this, on the one hand, the invention provides the artificial ligament of a kind of nano-biological cerimic coating, comprise artificial ligament and be evenly coated in the Ca-Mg-Si bio-ceramic coating on described artificial ligament surface, the particle diameter of the ceramic particle of described Ca-Mg-Si bio-ceramic coating is 20～80nm.
In the present invention, by applying Ca-Mg-Si bio-ceramic coating on artificial ligament surface, can be by means of the excellent bone inducibility of Ca-Mg-Si bioceramic, make the artificial ligament that is coated with this coating there is the multi-functional characteristics such as good biological activity, osteogenic and graft knitting, become a kind of novel bioactive graft material that has.Again, the particle diameter of the ceramic particle by making Ca-Mg-Si bio-ceramic coating is Nano grade, be more conducive to osteoblast material surface stick, breed and break up and graft bone interface on skeletonization.
Preferably, the thickness of described Ca-Mg-Si bio-ceramic coating is 20～100nm.
Preferably, described Ca-Mg-Si bioceramic is akermanite bioceramic, Bredigites biological ceramic, mussite and manganolite bioceramic.
Preferably, described Ca-Mg-Si bio-ceramic coating makes by pulsed laser deposition.Such Ca-Mg-Si bio-ceramic coating coating is even, Stability Analysis of Structures, is conducive to improve its biological activity.
Preferably, described artificial ligament is polyethylene terephthalate artificial ligament.
On the other hand, the present invention also provides the preparation method of the artificial ligament of above-mentioned nano-biological cerimic coating, comprising: utilize pulsed laser deposition, take Ca-Mg-Si pottery as target, at artificial ligament surface deposition Ca-Mg-Si bio-ceramic coating.
In the present invention, by adopting pulsed laser deposition, can make coating evenly, the Ca-Mg-Si bio-ceramic coating of good stability, thereby make the artificial ligament of the nano-biological cerimic coating making there is better biological activity.And the method technique is simple, parameter is easy to control, favorable repeatability.In addition, can control coating layer thickness, degree of crystallinity and grain size etc. by controlling technological parameter.
Preferably, the preparation of described Ca-Mg-Si pottery comprises the steps: that taking ethyl orthosilicate, soluble calcium salt and solubility magnesium salt by the stoichiometric proportion of Ca-Mg-Si pottery is raw material, makes xerogel through sol-gal process; And gained xerogel is calcined and obtained Ca-Mg-Si pottery at 1000～1400 ℃.By adopting sol-gal process, can be easy to obtain superior in quality Ca-Mg-Si pottery using as target.
Preferably, the technological parameter of described pulsed laser deposition comprises: laser frequency 5～20Hz, and atmosphere pressure is 5～30Pa, depositing temperature is 25～100 ℃, distance between matrix and target is 5～10cm, and laser beam becomes 30～60 degree angles, sedimentation time 5～40 minutes with target material surface.
In the present invention, by regulation and control depositing temperature, atmosphere pressure, target-substrate distance and sedimentation time, can control thickness, degree of crystallinity and the grain size of coating.
Preferably, stating technological parameter also comprises: in deposition process, Ca-Mg-Si pottery is with the speed rotation of 1000～3000r/hr.By means of this, can improve the uniformity of deposit film and prevent that target is breakdown.
The present invention has prepared the artificial ligament of nano-biological cerimic coating by pulse laser sediment method, have technique fairly simple, and condition is easy to the advantages such as control.And there is the multi-functional characteristics such as good biological activity, osteogenic and graft knitting containing the artificial ligament of nano-biological cerimic as human body implantation of ligament thing material.The artificial ligament of the nano-biological cerimic coating that therefore, prepared by the present invention has very strong Practical significance.
Accompanying drawing explanation
Fig. 1 be bio-ceramic coating artificial ligament (AKT-PET) (b, d) and without the SEM figure of coating PET artificial ligament (a, c), wherein (a) and (b) are respectively the partial enlarged drawing of (c), (d), show that bioceramic granular size is Nano grade;
Fig. 2 is bio-ceramic coating artificial ligament (AKT-PET) and the XRD figure without coating PET artificial ligament;
Fig. 3 is that bio-ceramic coating artificial ligament (AKT-PET) (f) and without the EDS of coating PET artificial ligament (e) is schemed;
Fig. 4 is that the bone marrow stroma stem cell of rabbit is at bio-ceramic coating artificial ligament (AKT-PET) and without the growing multiplication situation of coating PET artificial ligament;
Fig. 5 is that the bone marrow stroma stem cell of rabbit sticks to bio-ceramic coating artificial ligament (AKT-PET) and without on coating PET artificial ligament.PET (a, c, e), AKT-PET (b, d, f); 1 day (a, b), 3 days (c, d); (e) be and (f) high-amplification-factor scanning electron microscope (SEM) photograph;
Fig. 6 is bone marrow mesenchymal stem cells at blank PET ligament group and bio-ceramic coating group (AKT-PET) surface medium because of expression (Real time-PCR): (a) alkali phosphatase (ALP); (b) osteopontin (OPN); (c) Runx2; (d) VEGF (VEGF);
Fig. 7 is bio-ceramic coating artificial ligament (AKT-PET) and without tunnel situation behind the implantation tibial bone tunnel of coating PET artificial ligament;
Fig. 8 is bio-ceramic coating artificial ligament (AKT-PET) and the biomechanical property without coating PET artificial ligament;
Fig. 9 is histology's Image Display graft knitting situation, without coating PET artificial ligament (a, b, c) and bio-ceramic coating artificial ligament (AKT-PET) (d, e, f).
The specific embodiment
Further illustrate the present invention below in conjunction with accompanying drawing and following embodiment, should be understood that accompanying drawing and following embodiment are only for the present invention is described, and unrestricted the present invention.
One aspect of the present invention provides a kind of artificial ligament of nano-biological cerimic coating, and it comprises artificial ligament and is evenly coated in the Ca-Mg-Si bio-ceramic coating on described artificial ligament surface.Fig. 1 illustrate an example of the present invention bio-ceramic coating artificial ligament (AKT-PET) (b, d) and without the SEM figure of coating PET artificial ligament (a, c).As shown in Figure 1, the bio-ceramic coating granular size in AKT-PET is Nano grade, for example, be 20～80nm.The thickness of bio-ceramic coating is 20～100nm.
In the present invention, described Ca-Mg-Si bioceramic is the pottery of the ternary component that contains Ca, Mg and Si, includes but not limited to akermanite bioceramic, Bredigites biological ceramic, mussite bioceramic, manganolite.
In a preferred example, described Ca-Mg-Si bioceramic makes by pulsed laser deposition.Can make thus Ca-Mg-Si bio-ceramic coating apply even, Stability Analysis of Structures, be conducive to improve its biological activity.
Artificial ligament in the present invention includes but not limited to polyethylene terephthalate artificial ligament.In addition, should be understood that the matrix as coated bio-ceramic coating, be not limited to artificial ligament, can be also other graft material need to osteogenic and/or knitting, the such as tissue renovation material of bone, cartilage, tooth etc.
Fig. 2 illustrates bio-ceramic coating artificial ligament (AKT-PET) and the XRD figure without coating PET artificial ligament, and as shown in Figure 2, the PET of coating and pure PET do not have significant difference, and the ceramic coating that means generation is unformed phase.Fig. 3 illustrates bio-ceramic coating artificial ligament (AKT-PET) (f) and schemes without the EDS of coating PET artificial ligament (e), as shown in Figure 3, in AKT-PET, contain C, O, Ca, Mg, Si element, illustrate that it contains PET artificial ligament and Ca-Mg-Si bio-ceramic coating.
By applying Ca-Mg-Si bio-ceramic coating on artificial ligament surface, can be by means of the excellent bone inducibility of Ca-Mg-Si bioceramic, make the artificial ligament that is coated with this coating there is the multi-functional characteristics (referring to biological activity test described later) such as good biological activity, osteogenic and graft knitting, can become a kind of novel bioactive graft material that has.Again, the particle diameter of the ceramic particle by making Ca-Mg-Si bio-ceramic coating is Nano grade, be more conducive to osteoblast material surface stick, breed and break up and graft bone interface on skeletonization.
The present invention provides the preparation method of the artificial ligament of above-mentioned nano-biological cerimic coating on the other hand, utilizes pulsed laser deposition, take Ca-Mg-Si pottery as target, at artificial ligament surface deposition Ca-Mg-Si bio-ceramic coating.
Ca-Mg-Si pottery as target can be prepared by sol-gal process.In one example, taking ethyl orthosilicate, soluble calcium salt and solubility magnesium salt by the stoichiometric proportion of Ca-Mg-Si pottery is raw material, makes xerogel through sol-gal process; And gained xerogel is calcined and obtained Ca-Mg-Si pottery at 1000～1400 ℃.Wherein soluble calcium salt includes but not limited to lime nitrate, calcium chloride etc.Solubility magnesium salt includes but not limited to magnesium nitrate, magnesium chloride etc.
In pulsed laser deposition, for example, can adopt KrF laser instrument (wavelength 248nm, pulse width 20ns).Laser frequency can be 5～20Hz.Distance between matrix and target can be 5～10cm.Laser beam can become 30～60 degree angles with target material surface.Atmosphere pressure can be 5～30Pa.Depositing temperature can be 25～100 ℃.Sedimentation time can be 5～40 minutes.Can control by regulation and control depositing temperature, atmosphere pressure, target-substrate distance and sedimentation time thickness, degree of crystallinity and the grain size of coating.For improving the uniformity of deposit film and preventing that target is breakdown, the speed rotation that in deposition process, Ca-Mg-Si pottery can 1000～3000r/hr.
Below, as example, an embodiment of the invention are described.
1. pulse laser deposition is prepared bioceramic AKT coating and sign
Test is PET artificial ligament material with the matrix material of pld (pulsed laser deposition), and its source is the postoperative residue stump of LARS (Surgical Implants and Devices, Arc-sur-Tille, France) artificial ligament.Adopt 75% ethanol ultrasonic cleaning 10 minutes, then spend the night as for 60 degree hyperthermia dryings.Ligament is cut as 3cm × 3cm to square under superclean bench environment, packed.
Target for pld (pulsed laser deposition) is Ca-Mg-Si system pottery.Adopt ethyl orthosilicate (TEOS), calcium nitrate tetrahydrate [Ca (NO
3)
24H
2o] and magnesium nitrate hexahydrate [Mg (NO
3)
26H
2o] be raw material, through colloidal sol, gel, ageing, the process such as dry, and calcining obtains pure Ca-Mg-Si ceramic powder at 1300 ℃.
Experiment adopts KrF laser instrument (wavelength 248nm, pulse width 20ns).Laser frequency 5-10Hz, the distance between artificial ligament and ceramic target is 5-10cm, laser beam becomes 30-60 degree angle with target material surface.For improving the uniformity of deposit film and preventing that target is breakdown, in deposition process, Ca-Mg-Si pottery is with the speed rotation of 2400r/hr.Adopt molecular pump to be evacuated down to 20Pa to settling chamber, use its pressure of mechanical pump control.Thickness, degree of crystallinity and the grain size of coating mainly controlled in this test by regulation and control depositing temperature (room temperature), atmosphere pressure (20Pa), target-substrate distance (5-10cm) and sedimentation time (5-40 minute).
The coating obtaining is analyzed and is characterized thing phase composition, the surface microstructure of coating material by wide-angle x-ray diffraction (XRD), scanning electron microscope (SEM), energy spectrum analysis (EDS) and atomic force microscope (AFM) etc.Referring to Fig. 1～3, known bioceramic AKT coating is in PET artificial ligament material surface, and bioceramic granular size is Nano grade.In the material making, contain C, O, Ca, Mg, Si element.
2. the cultivation of rabbit bone marrow mesenchymal stem and impact and Related Mechanism thereof at adhesion, propagation, differentiation and the bone related gene expression of graft surface
The separation of 2.1 rabbit BMSC and cultivation
One monthly age new zealand rabbit (Shanghai Slac Experimental Animal Co., Ltd.), anaesthetizes with pentobarbital; Use marrow puncture needle to carry out femur puncture, extract about 0.5mL bone marrow; 0.5mL bone marrow is transferred to 10mL centrifuge tube; Use the DMEM culture medium of adding 10%FBS that Cell sap in centrifuge tube is mended to 10ml, centrifugal 5 minutes, collecting precipitation; Use 10ml complete medium gravity treatment cell, inoculating cell is to 96 porocyte culture plates, normal condition (37 ℃ and 5%CO
2) cultivate, name cell is P1 generation.The visible a large amount of round cell of microscopic examination; After normal cultivation 3d, use the complete culture solution of 10ml DMEM+10%FBS to change liquid to cell, Microscopic observation, visible spindle cell, circle have protrusion cell, round cell, continue to cultivate; After changing liquid, cultivate 5d, Microscopic observation, has Fusoid cells to become to assemble shape growth, has multiple clone's speckles to generate at the bottom of ware; Use the method for trypsinization, cell is carried out to 1/2 processings of going down to posterity, in cell called after P2 generation after going down to posterity, after going down to posterity, continue cultivation; After going down to posterity, P2 is for cell culture 48 hours, Microscopic observation cell shuttle type, and cell central authorities are full and have overshooting shape, and growth is evenly.
2.2CCK-8 detects cytoactive
After inoculation, normal condition is cultivated, and get each 5 of each group of material in 1d, 3d, 7d, CCK-8 method detects cytoactive: before experiment, clip material becomes " 1cm × 1cm " size, put into slowly rinsing of 24 porocyte culture plates (500 μ l serum-free DMEM culture medium are added in every hole), remove the residual cell that does not attach growth of material surface; Washed material is reentered into 96 new orifice plates (every hole adds the MEM culture medium of 100 μ l serum-frees in advance); Every hole adds the CCK-8 solution (note not generating bubble in hole, they can affect the reading of OD value) of 10 μ l; By culture plate in incubator (37 ℃ and 5%CO
2concentration) hatch 4 hours; Then liquid rotating in 96 well culture plates is moved to another 96 orifice plate; Upper machine, uses microplate reader to be determined at the absorbance at 450nm place; Data analysis.Result is referring to Fig. 4,5, and the bone marrow stroma stem cell of known rabbit is better than the artificial ligament without coating PET in bio-ceramic coating artificial ligament (AKT-PET) tactophily propagation situation.Show by being conducive to osteoblast sticking, breeding and breaking up on its surface at artificial ligament surface coating bio-ceramic coating.
2.3Real Time-PCR detects cell sample
Main relevant osteogenesis gene (RUNX2, OPN) and the one-tenth vascularization gene (VEGF, HIF-1a) of detecting.Concise and to the point step is as follows: sample process; The extracting of cell sample Total RNA; Carry out reverse transcription with reverse transcriptase Reverse Transcriptase M-MLV (D2640A, Takara) and synthesize cDNA; Real time pcr amplification system and reaction condition; Data analysis.Result is referring to Fig. 6, known compared to without coating PET ligament, and AKT-PET more can promote alkali phosphatase (ALP), relevant osteogenesis gene (as RUNX2, OPN) and become the expression of vascularization gene (VEGF, HIF-1a).
3. goat Healing in Anterior Cruciate Ligament Reconstruction experiment
All animal feedings, at university's Animal Lab., are carried out in strict accordance with national experimental animal feeding and use regulations.Experiment adopts 48 bull goats, every right anterior cruciate ligament of knee joint reconstruction model operation of goat row.Be divided into two groups (PET artificial ligament graft group, AKT-PET artificial ligament graft groups), 24 every group.Adopt ketalar (5mg/kg) intramuscular injection anesthesia, then adopt pentobarbital (25mg/kg) intravenous anesthesia.After anaesthetizing successfully, supine position is on operating-table.In knee joint region, after preserved skin and sterilization, along the long stringer otch of knee joint center line outer row 10cm, successively separate, expose anterior cruciate ligament.Then complete resection ACL, drills through respectively diameter 5mm femur and tibial tunnel in ACL at femur and tibia stop.Graft free end is drawn in to osseous tunnel from inside to outside.In tunnel face, ligament stump being carried out to deadman respectively fixes.Rinse, layer-by-layer suture, closes wound.Postoperative, put into cage, permission activity.In the time of 6 weeks and March, dead 12 of every component other places, cut specimen and detect.
3.1Micro-CT scanning
Carrying out before biomechanics test, specimen can first be carried out micro-CT scanning (Siemens Inveon Micro PET/CT scanner (Siemens Medical Solution, Germany), sweep parameter is as follows: voltage=80kV, current=500uA, exposure time=800ms, binning=2, FOV=61.05mm × 40.70mm.On preprocessor software, adopt the cylindrical area-of-interest (ROI) of the high 5mm of diameter 3mm to measure the area of new bone amount in osseous tunnel.Result is referring to Fig. 7, and known compared to without coating PET ligament, AKT-PET more can promote new osteogenesis.
3.2 biomechanics inspections
After sacrifice of animal, cut graft specimen, retain bone road end length 1cm.Put into liquid nitrogen container ,-30 degree are preserved.Mechanics thaws under room temperature 24 hours before detecting.Biomechanics test adopts Instron(8874) testing of materials device (Instron Co.USA).Graft is fixed to specific fixation clamp.Adopt the pre-tractive of 20N pulling force 5 minutes.Then with the draw speed tractive of 2mm/min until ligament pull-out or break.Record stretching-inflection curves and final state (pull out or break), calculate maximum pull intensity and elastic modelling quantity.Result is referring to Fig. 8, and known compared to without coating PET ligament, the maximum pull intensity of AKT-PET is larger, has better biomechanical property.
3.3 histologic analysis
After sacrifice of animal, cut and move down plant specimen, put into 10% formalin solution 48 hours.Then adopt EDTA decalcification processing, paraffin embedding, adopts RM2155 microtome (Leica, Germany) to be parallel to osseous tunnel direction and cuts the thick section of 5 μ m.Adopt HE dyeing, histological examination is done in Masson dyeing and Picro-Sirius red dyeing.Adopt Olympus inverted microscope (IX71SBF-2, Olympus Co., Japan) to observe graft bone interface situation.DP Manager(Olympus Optical Co., Japan) take pictures.Result is referring to Fig. 8, and known compared to without coating PET ligament, the knitting ability of AKT-PET is stronger.
From testing above, the artificial ligament of nano-biological cerimic coating of the present invention is conducive to osteoblast sticking, breeding and breaking up on its surface, can promote alkali phosphatase (ALP), relevant osteogenesis gene (as RUNX2, OPN) and become vascularization gene (VEGF, HIF-1a) expression, promote new osteogenesis, and there is excellent knitting ability, and there is in addition good biomechanical property, therefore can be used as novel bioactive implanting material.
Further exemplify embodiment below to describe the present invention in detail.Should understand equally; following examples are only used to further illustrate the present invention; can not be interpreted as limiting the scope of the invention, some nonessential improvement that those skilled in the art's foregoing according to the present invention is made and adjustment all belong to protection scope of the present invention.The technological parameter that following example is concrete etc. is only also an example in OK range, and those skilled in the art can be done in suitable scope and be selected by explanation herein, and do not really want to be defined in the below concrete numerical value of example.
Example 1:
Experiment adopts KrF laser instrument (wavelength 248nm, pulse width 20ns).Laser frequency 5Hz, the distance between artificial ligament and ceramic target is 5.5cm, laser beam becomes miter angle with target material surface.For improving the uniformity of deposit film and preventing that target is breakdown, in deposition process, Ca-Mg-Si pottery adopts molecular pump to be evacuated down to 20Pa to settling chamber with the speed rotation of 2400r/hr, uses its pressure of mechanical pump control.Thickness, degree of crystallinity and the grain size of coating mainly controlled in this test by regulation and control depositing temperature (room temperature), atmosphere pressure (20Pa), target-substrate distance (5.5cm) and sedimentation time (40 minutes).Then carry out material sign, become vascularization, osteogenic and graft knitting performance study.
Example 2:
Experiment adopts KrF laser instrument (wavelength 248nm, pulse width 20ns).Laser frequency 6Hz, the distance between artificial ligament and ceramic target is 6cm, laser beam becomes 30 degree angles with target material surface.For improving the uniformity of deposit film and preventing that target is breakdown, in deposition process, Ca-Mg-Si pottery adopts molecular pump to be evacuated down to 20Pa to settling chamber with the speed rotation of 2400r/hr, uses its pressure of mechanical pump control.Thickness, degree of crystallinity and the grain size of coating mainly controlled in this test by regulation and control depositing temperature (room temperature), atmosphere pressure (20Pa), target-substrate distance (6cm) and sedimentation time (10 minutes).
Claims (9)
1. an artificial ligament for nano-biological cerimic coating, is characterized in that, comprises artificial ligament and is evenly coated in the Ca-Mg-Si bio-ceramic coating on described artificial ligament surface, and the particle diameter of the ceramic particle of described Ca-Mg-Si bio-ceramic coating is 20～80nm.
2. the artificial ligament of nano-biological cerimic coating according to claim 1, is characterized in that, the thickness of described Ca-Mg-Si bio-ceramic coating is 20～100nm.
3. the artificial ligament of nano-biological cerimic coating according to claim 1 and 2, is characterized in that, described Ca-Mg-Si bioceramic is akermanite bioceramic, Bredigites biological ceramic, mussite bioceramic and manganolite.
4. according to the artificial ligament of the nano-biological cerimic coating described in any one in claims 1 to 3, it is characterized in that, described Ca-Mg-Si bio-ceramic coating makes by pulsed laser deposition.
5. according to the artificial ligament of the nano-biological cerimic coating described in any one in claim 1 to 4, it is characterized in that, described artificial ligament is polyethylene terephthalate artificial ligament.
6. the preparation method of the artificial ligament of the coating of the nano-biological cerimic described in any one in a claim 1 to 5, it is characterized in that, comprise: utilize pulsed laser deposition, take Ca-Mg-Si pottery as target, at artificial ligament surface deposition Ca-Mg-Si bio-ceramic coating.
7. preparation method according to claim 6, it is characterized in that, the preparation of described Ca-Mg-Si pottery comprises the steps: that taking ethyl orthosilicate, soluble calcium salt and solubility magnesium salt by the stoichiometric proportion of Ca-Mg-Si pottery is raw material, makes xerogel through sol-gal process; And gained xerogel is calcined and obtained Ca-Mg-Si pottery at 1000～1400 ℃.
8. according to the preparation method described in claim 6 or 7, it is characterized in that, the technological parameter of described pulsed laser deposition comprises: laser frequency 5～20Hz, atmosphere pressure is 5～30Pa, depositing temperature is 25～100 ℃, distance between matrix and target is 5～10 cm, and laser beam becomes 30～60 degree angles, sedimentation time 5～40 minutes with target material surface.
9. preparation method according to claim 8, is characterized in that, described technological parameter also comprises: in deposition process, Ca-Mg-Si pottery is with the speed rotation of 1000～3000r/hr.
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| CN104117095A (en) * | 2014-07-29 | 2014-10-29 | 复旦大学附属华山医院 | Method for preparing strontium/fibroin bionic coating modified artificial ligament |
| CN106606805A (en) * | 2015-10-22 | 2017-05-03 | 中国科学院上海硅酸盐研究所 | Novel nanometer bioglass-coated patterned electrospun fiber membrane and its preparation method and use |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104117094A (en) * | 2014-07-29 | 2014-10-29 | 复旦大学附属华山医院 | Preparation method of silver/fibroin/hydroxyapatite biomimetic coating-modified artificial ligament |
| CN104117095A (en) * | 2014-07-29 | 2014-10-29 | 复旦大学附属华山医院 | Method for preparing strontium/fibroin bionic coating modified artificial ligament |
| CN104117095B (en) * | 2014-07-29 | 2015-12-30 | 复旦大学附属华山医院 | Strontium/fibroin bionic coating modifies the preparation method of artificial ligament |
| CN104117094B (en) * | 2014-07-29 | 2016-05-04 | 复旦大学附属华山医院 | Silver/silk-fibroin/hydroxyapatite bionic coating is modified artificial ligament preparation method |
| CN106606805A (en) * | 2015-10-22 | 2017-05-03 | 中国科学院上海硅酸盐研究所 | Novel nanometer bioglass-coated patterned electrospun fiber membrane and its preparation method and use |
| CN106606805B (en) * | 2015-10-22 | 2020-05-19 | 中国科学院上海硅酸盐研究所 | Nano-bioglass coated patterned electrospun fibrous membrane and preparation method and application thereof |
| CN107149696A (en) * | 2016-03-03 | 2017-09-12 | 中国科学院上海硅酸盐研究所 | A kind of new eggshell membrane of cupric bio-vitric nano coating coating and its preparation method and application |
| CN107149696B (en) * | 2016-03-03 | 2020-01-21 | 中国科学院上海硅酸盐研究所 | Novel egg membrane coated with copper-containing bioglass nano coating and preparation method and application thereof |
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