CN104225686A - Mechano growth factor and magnesium alloy-based compound bone repair material and preparation method thereof - Google Patents
Mechano growth factor and magnesium alloy-based compound bone repair material and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a mechano growth factor and magnesium alloy-based compound bone repair material and a preparation method thereof. The preparation method comprises the following steps: loading a mechano growth factor on the surface of a magnesium alloy, wherein the mechano growth factor is a mechano growth factor 24 peptide, and by removing the oxide layer on the surface, the magnesium alloy matrix has a layer of stable positive charge; selecting electrolyte materials with different charges as carriers and orderly placing the magnesium alloy matrix in a carrier A, the mechano growth factor, a carrier A and a carrier B, wherein the carriers A are electronegative polyelectrolytes containing carboxyl functional groups, the carrier B is an electropositive polyelectrolyte containing an amino functional group, and the carriers A and B have biocompatibility and are completely degraded in vivo and alternately deposited on the surface of the matrix through electrostatic interaction. According to the mechano growth factor and magnesium alloy-based compound bone repair material and the preparation method thereof, the mechano growth factor is loaded on the surface of the magnesium alloy and then implanted in vivo to be released in time so as to stimulate the growth of the cells, and then the problem caused by under stressing is overcome. Both the he polyelectrolyte solution for modification and the mechano growth factor can be completely degraded in vivo and the biocompatibility is good.
Description
Technical field
The invention belongs to technical field of biological material, particularly adopt Mechano growth factor to carry out surface modification to magnesium alloy.
Background technology
Magnesium alloy is as a kind of embedded material having great potential, lightweight, density is little, has high strength/weight ratio; Elastic modelling quantity (45GPa), closer to normal bone tissues, can avoid stress-shielding effect on the impact of area of new bone and plasticity; And can degradation in vivo, second operation can be avoided to bring misery to patient, and its catabolite magnesium ion is the cation that body burden enriches, and major storage, in skeleton, is the cofactor of multiple enzyme, is one of ribosomal key structure; Magnesium ion can also stimulate generation, the induced osteogenesis of fracture end os osseum crust, promotes union of fracture, and stimulates Chondrogenesis.The bone implant material that these characteristics make magnesium and alloy thereof likely become desirable.But magnesium alloy degradation rate is too fast etc., and problem causes that magnesium alloy will be degradable before impaired bones organization healing after implanting, thus good supporting role can not be played.And Mechano growth factor to be one can promote osteoblastic proliferation, repair the active factors of damage, so carry out to magnesium alloy the quickly-healing that surface modification can promote injured bone with Mechano growth factor.Meanwhile, Mechano growth factor has biological activity, carrier must be selected to carry load at Mg alloy surface, and carrier can form protecting film at Mg alloy surface, thus can suppress the fast degradation of magnesium alloy.
LBL self-assembly (layer-by-layer assembly, LBL) technology says the interaction of band positive and negative charge based on electrolyte zwitterion, by alternatively stacked for the polyelectrolyte of oppositely charged ultrathin membrane preparation method.This invention adopts the Process of Surface Modification of this kind of technology realization to magnesium alloy.
Summary of the invention
The object of the invention is for the too fast problem of magnesium alloy degradation rate, there is provided one to utilize layer-by-layer to prepare the method for load at Mg alloy surface Mechano growth factor 24 peptide (aminoacid sequence is as shown in SEQ ID No.1), thus magnesium alloy is played a role better as embedded material.
Technical scheme of the present invention is as follows:
Mechano growth factor surface modification magnesium alloy of the present invention is at Mg alloy surface carrying somatomedin.
Described Mechano growth factor is Mechano growth factor 24 peptide.
Mechano growth factor surface modification magnesium alloy preparation method of the present invention, comprises the following steps:
(1) before modification, magnesium alloy needs to use sand papering, removes the oxide layer on surface.
(2) pretreatment is carried out to magnesium alloy, on the one hand make negative charge on matrix surface band so that uploading of polyelectrolyte and Mechano growth factor 24 peptide, the erosion-resisting protecting film of one deck can be covered at matrix surface on the other hand.Preprocess method can select basification and fluorination treatment.
(3) for making matrix surface with the stable positive charge of one deck, matrix is immersed in polyethylenimine solution, because magnesium alloy is very active in liquid environment, so soak time should be less than 10 minutes.
(4) electrolyte of the different electric charge of zone of preference is as carrier, magnesium alloy substrate is placed in successively carrier A-Mechano growth factor-carrier A-carrier B.
Wherein carrier A is that electronegative polyelectrolyte containing carboxyl functional group (comprises sodium alginate, kayexalate etc.), carrier B is that the positively charged polyelectrolyte containing amidine functional group (comprises chitosan, PLL bromide etc.), and carrier A, B have biocompatibility, in vivo can be degradable, by electrostatic interaction, alternating deposit is at matrix surface.This upload process repeats m time, often carries out 1 time and can be regarded as one deck, altogether load m layer Mechano growth factor, the upper carrying capacity of Mechano growth factor and upload the number of plies and be directly proportional.M is preferably 20-40.
Preprocess method described in step (2) is more preferably: magnesium alloy substrate is immersed 60 DEG C, in the sodium hydroxide solution of 4-6mol/L 2 hours, then uses washed with de-ionized water, dry at 60 DEG C more afterwards.
Step (3) is preferably the polyethylenimine solution 10 minutes pretreated magnesium alloy substrate being placed in 3-6g/L, with deionized water rinsing 1 minute after taking out.
Step (4) concrete steps are preferred: the polyelectrolyte solution used in the present invention can select chitosan (CS) and sodium alginate (ALG), wherein chitosan is the polyelectrolyte solution of positively charged, and sodium alginate is electronegative electrolyte solution.These two kinds of materials in vivo can be degradable, and a part is by body absorption, and another part can excrete smoothly.Often complete uploading of a polyelectrolyte solution, need with deionized water rinsing 1 minute, then carry out uploading of polyelectrolyte next time.That so under the prerequisite rinsed well, washing time is more short better because magnesium alloy is very active in water equally.
Because magnesium alloy is very active in polyelectrolyte solution, electrochemical reaction can be there is, so uploading of above-mentioned polyelectrolyte solution needs to be undertaken by sol evenning machine (model can be selected according to specimen size).
A the magnesium alloy substrate of completing steps (3) is positioned in sol evenning machine by (), by the chitosan solution of 2.5g/L
Dripping is coated on matrix, washes 1 minute.
B the Mechano growth factor of 0.36g/L drips and is coated on matrix by (), wash 1 minute.
C () repeats step (a).
D the chitosan of 2g/L drips and is coated on matrix by (), wash 1 minute.
E more than (), four steps repeat m time.
In the inventive method, according to conventional LBL self-assembly method, select chitosan (CS) and sodium alginate (ALG) to do electrolyte, both structures, as Fig. 1, their strand contain positive charged group (-NH3 respectively
+) and negative electricity group (-COO
-).Because after soaking in polymine (PEI) solution, matrix surface is positively charged, so replace by sodium alginate (ALG), Mechano growth factor 24 peptide, sodium alginate (ALG), chitosan (CS) by electrostatic interaction, load is at magnesium alloy matrix surface.
The present invention has following beneficial effect:
Compare with unmodified magnesium alloy, can the growth of stimulating osteoblast, and magnesium alloy degradation rate in vivo can be reduced.Modification magnesium alloy of the present invention is expected to overcome the relevant issues that in bone tissue restoration or regenerative therapy process, understrressing causes.Magnesium alloy is as orthopedic transplant material, elastic modelling quantity is still a little more than normal bone tissues, certain stress-shielding effect can be produced, cause osteoporosis, fracture delayed union, does not even heal, by Mechano growth factor is loaded to Mg alloy surface, implant and discharge in time thus the growth of stimulating osteoblast, overcome the problem that understrressing causes.And modified magnesium alloy biocompatibility is good, and the polyelectrolyte solution of modification and Mechano growth factor in vivo all can be degradable, the material making health produce rejection can not be produced.
Accompanying drawing explanation
The structural formula of Fig. 1 chitosan (CS) and sodium alginate (ALG)
A is the structural representation of chitosan, can obtain the polyelectrolyte that proton becomes positively charged in water.B is the structural representation of sodium alginate, can be ionized into electronegative polyelectrolyte in aqueous.
The infrared spectrogram of Fig. 2 Mechano growth factor 24 peptide modification magnesium alloy coating (A) and three kinds of pure materials (chitosan, sodium alginate, Mechano growth factor 24 peptide) (B).
The scanning electron microscope diagram of Fig. 3 Mechano growth factor 24 peptide modification Mg alloy surface
A is the scanning electron microscope (SEM) photograph of load 20 layers of Mechano growth factor 24 peptide modification magnesium alloy.B is the scanning electron microscope (SEM) photograph of load 40 layers of Mechano growth factor 24 peptide modification magnesium alloy.
The atomic force microscope figure of Fig. 4 Mechano growth factor 24 peptide modification Mg alloy surface, wherein:
A is the plane graph of the atomic force microscope of load 20 layers of Mechano growth factor 24 peptide modification magnesium alloy;
B is the graphics of the atomic force microscope of load 20 layers of Mechano growth factor 24 peptide modification magnesium alloy;
C is the plane graph of the atomic force microscope of load 40 layers of Mechano growth factor 24 peptide modification magnesium alloy;
D is the graphics of the atomic force microscope of load 40 layers of Mechano growth factor 24 peptide modification magnesium alloy.
Detailed description of the invention
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.Mechano growth factor 24 peptide used in the present invention is purchased from Mai Motuopu bio tech ltd, Wuxi.
Embodiment 1 is based on the preparation (m=20) of Mechano growth factor (chitosan, sodium alginate do carrier) modification magnesium alloy materials.
1,5mol/L sodium hydroxide solution is prepared in advance, the polyethylenimine solution of 5g/L, the chitosan polyelectrolyte solution of 2g/L, the sodium alginate electrolyte of 2.5g/L, Mechano growth factor 24 peptide solution of 0.36g/L.Because the chitosan bought is insoluble in water, be soluble in acetum, so first being diluted by the acetum of 3g/L is 1%, then add Chitosan powder, be finally configured to the chitosan polyelectrolyte solution of 2g/L.
2, with 400#, 600#, 1200#, 1500#, 2000# silicon carbide paper, magnesium alloy matrix surface is progressively polished successively, by the matrix deionized water rinsing after polishing smooth, put into dehydrated alcohol ultrasonic cleaning subsequently 10 minutes, natural air drying.
3, the matrix cleaned up is put into 60 DEG C, the sodium hydroxide solution of 5mol/L soaks 2 hours, takes out rear washed with de-ionized water, then be placed in 60 DEG C of drying baker dryings.
4, magnesium alloy substrate is placed in the polyethylenimine solution that configures 10 minutes, takes out rear deionized water rinsing, natural air drying.
5, be positioned on sol evenning machine by magnesium alloy substrate, regulate sol evenning machine rotating speed to be 1000r/min, the cycle is 1min.Open sol evenning machine, be coated on matrix by a sodium alginate (ALG) dripping, sol evenning machine rotates after 1 minute and stops; Again open sol evenning machine, be coated on matrix, play the effect of flushing by a deionized water drop, same sol evenning machine rotates after 1 minute and stops.Repeat said process afterwards, need sodium alginate (ALG) to change Mechano growth factor 24 peptide, sodium alginate (ALG), chitosan (CS) into.Like this, the deposition of one deck Mechano growth factor 24 peptide is just completed.
6, repeat the method 20 times of step 5, realize the preparation at magnesium alloy matrix surface load 20 layers of Mechano growth factor 24 peptide.
Embodiment 2 is based on the preparation (m=40) of Mechano growth factor (chitosan, sodium alginate do carrier) modification magnesium alloy materials.
1,5mol/L sodium hydroxide solution is prepared in advance, the polyethylenimine solution of 5g/L, the chitosan polyelectrolyte solution of 2g/L, the sodium alginate electrolyte of 2.5g/L, Mechano growth factor 24 peptide solution of 0.36g/L.Because the chitosan bought is insoluble in water, be soluble in acetum, so first being diluted by the acetum of 3g/L is 1%, then add Chitosan powder, be finally configured to the chitosan polyelectrolyte solution of 2g/L.
2, with 400#, 600#, 1200#, 1500#, 2000# silicon carbide paper, magnesium alloy matrix surface is progressively polished successively, by the matrix deionized water rinsing after polishing smooth, put into dehydrated alcohol ultrasonic cleaning subsequently 10 minutes, natural air drying.
3, the matrix cleaned up is put into 60 DEG C, the sodium hydroxide solution of 5mol/L soaks 2 hours, takes out rear washed with de-ionized water, then be placed in 60 DEG C of drying baker dryings.
4, magnesium alloy substrate is placed in the polyethylenimine solution that configures 10 minutes, takes out rear deionized water rinsing, natural air drying.
5, be positioned on sol evenning machine by magnesium alloy substrate, regulate sol evenning machine rotating speed to be 1000r/min, the cycle is 1min.Open sol evenning machine, be coated on matrix by a sodium alginate (ALG) dripping, sol evenning machine rotates after 1 minute and stops; Again open sol evenning machine, be coated on matrix, play the effect of flushing by a deionized water drop, same sol evenning machine rotates after 1 minute and stops.Repeat said process afterwards, need sodium alginate (ALG) to change Mechano growth factor 24 peptide, sodium alginate (ALG), chitosan (CS) into.Like this, the deposition of one deck Mechano growth factor 24 peptide is just completed.
6, repeat the method 40 times of step 5, realize the preparation at magnesium alloy matrix surface load 40 layers of Mechano growth factor 24 peptide.
Embodiment 3 is based on the preparation (m=20) of Mechano growth factor (PLL bromide, kayexalate do carrier) modification magnesium alloy materials.
1,5mol/L sodium hydroxide solution is prepared in advance, the polyethylenimine solution of 5g/L, the PLL bromide polyelectrolyte solution of 2.5g/L, the kayexalate electrolyte of 5g/L, Mechano growth factor 24 peptide solution of 0.36g/L.
2, with 400#, 600#, 1200#, 1500#, 2000# silicon carbide paper, magnesium alloy matrix surface is progressively polished successively, by the matrix deionized water rinsing after polishing smooth, put into dehydrated alcohol ultrasonic cleaning subsequently 10 minutes, natural air drying.
3, the matrix cleaned up is put into 60 DEG C, the sodium hydroxide solution of 5mol/L soaks 2 hours, takes out rear washed with de-ionized water, then be placed in 60 DEG C of drying baker dryings.
4, magnesium alloy substrate is placed in the polyethylenimine solution that configures 10 minutes, takes out rear deionized water rinsing, natural air drying.
5, be positioned on sol evenning machine by magnesium alloy substrate, regulate sol evenning machine rotating speed to be 1000r/min, the cycle is 1min.Open sol evenning machine, be coated on matrix by a kayexalate (PSS) dripping, sol evenning machine rotates after 1 minute and stops; Again open sol evenning machine, be coated on matrix, play the effect of flushing by a deionized water drop, same sol evenning machine rotates after 1 minute and stops.Repeat said process afterwards, need kayexalate (PSS) to change Mechano growth factor 24 peptide, kayexalate (PSS), PLL bromide (PLL) into.Like this, the deposition of one deck Mechano growth factor 24 peptide is just completed.
6, repeat the method 20 times of step 5, realize the preparation at magnesium alloy matrix surface load 20 layers of Mechano growth factor 24 peptide.
Embodiment 4 is based on the Qualitative Identification of Mechano growth factor modification magnesium alloy materials.
The rete of the Mg alloy surface having prepared the load of 20 layers of Mechano growth factor 24 peptide is hung and carries out infrared spectrum analysis (a), compare can find out with the chitosan (b) of pure material, sodium alginate (c), Mechano growth factor (d) infrared spectrogram, as shown in Figure 2, owing to also having TYR in Mechano growth factor 24 peptide of pure material, thus containing phenyl ring, crest can be produced at 1453 places; Sodium alginate is a kind of electronegative polyelectrolyte, containing carboxyl, produces asymmetric and symmetrical stretching vibration absworption peak at 1609,1418 places; Chitosan is a kind of polyelectrolyte with stable positive charge, and containing primary amine, 3443,1588 places are the stretching vibration of primary amine groups respectively and become angle of throw absworption peak.In the infrared spectrogram of scraped coating, can see that producing crests at 1449 places proves containing phenyl ring in coated substance, and to only have in Mechano growth factor 24 peptide be containing phenyl ring, thus can the load of proof growth factor-2 4 peptide at Mg alloy surface; And at 1654 and 1590 crests going out to produce because carboxyl and amino condensation are that amide groups causes, thus prove sodium alginate and chitosan also load at Mg alloy surface.
Observe under the magnesium alloy having prepared the load of 20 and 40 layers of Mechano growth factor 24 peptide is placed in scanning electron microscope, result as shown in Figure 3, can find out that the modification Mg alloy surface of load 20 layers of Mechano growth factor 24 peptide defines more uniform polyelectrolyte layer, and more smooth at the modification Mg alloy surface polyelectrolyte layer of load 40 layers of Mechano growth factor 24 peptide.
Observe under the magnesium alloy having prepared 20 layers and the load of 40 layers of Mechano growth factor 24 peptide is placed in atomic force microscope, result as shown in Figure 4.Modification Mg alloy surface roughness (RMS) value of load 20 layers of Mechano growth factor 24 peptide is 251, the modification Mg alloy surface RMS value of load 40 layers of Mechano growth factor 24 peptide is 165, the modification Mg alloy surface island of load 40 layers of Mechano growth factor 24 peptide obviously reduces, surface is more smooth, thus biocompatibility is better.
Claims (9)
1. a Mechano growth factor surface modification magnesium alloy, is characterized in that at Mg alloy surface carrying somatomedin.
2. alloy as claimed in claim 1, is characterized in that described Mechano growth factor is Mechano growth factor 24 peptide.
3. the Mechano growth factor surface modification magnesium alloy preparation method of claim 1 or 2, is characterized in that comprising the following steps:
(1) magnesium alloy substrate removes the oxide layer on surface;
(2) by step (1) magnesium alloy matrix surface with the stable positive charge of one deck;
(3) electrolyte of the different electric charge of zone of preference is as carrier, magnesium alloy substrate is placed in successively carrier A-Mechano growth factor-carrier A-carrier B; Wherein carrier A is the electronegative polyelectrolyte containing carboxyl functional group, carrier B is the positively charged polyelectrolyte containing amidine functional group, and carrier A, carrier B all have biocompatibility, degradable in vivo, by electrostatic interaction, alternating deposit is at matrix surface.
4. method as claimed in claim 3, it is characterized in that carrier A-Mechano growth factor-carrier A-carrier B is one deck assembling, upload process repeats m time, load m layer Mechano growth factor, the upper carrying capacity of Mechano growth factor with upload the number of plies and be directly proportional.
5. method as claimed in claim 3, is characterized in that described m is 20-40 time.
6. method as claimed in claim 3, is characterized in that the preprocess method described in described step (1) is: magnesium alloy substrate is immersed 60 DEG C, in the sodium hydroxide solution of 4-6mol/L 2 hours, then uses washed with de-ionized water, drier at 60 DEG C.
7. method as claimed in claim 3, is characterized in that pretreated magnesium alloy substrate is placed in the polyethylenimine solution 10 minutes of 3-6g/L by described step (2), uses deionized water rinsing after taking-up.
8. method as claimed in claim 3, it is characterized in that described step (3) electrolyte is chitosan (CS) and sodium alginate (ALG), wherein chitosan is the polyelectrolyte solution of positively charged, and sodium alginate is electronegative electrolyte solution.
9. Mechano growth factor surface modification magnesium alloy is as the application of orthopedic implanting material.
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Cited By (1)
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| CN113304316A (en) * | 2021-05-27 | 2021-08-27 | 南京医科大学附属口腔医院 | Treatment method for promoting bone formation activation on surface of zirconia implant |
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