CN104906637B - Porous polymethyl methacrylate base compound rest bone grafting material for carrying medicine of a kind of injectable and preparation method thereof - Google Patents
Porous polymethyl methacrylate base compound rest bone grafting material for carrying medicine of a kind of injectable and preparation method thereof Download PDFInfo
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Abstract
The invention discloses porous polymethyl methacrylate base compound rest bone grafting material for carrying medicine of a kind of injectable and preparation method thereof, belong to organic functional material preparation field.The polymethyl methacrylate base compound rest bone grafting material of the present invention uses polymethyl methacrylate（PMMA）As the support for providing mechanical support, for chitosan-based thermo-sensitive gel as pore former and the carrier of bone guided material and medicine, both are mutually mixed the bone cement compound for the porous three-dimensional structure that may make up new injectable.The preparation method of this support bone grafting material is simple, reaction temperature is suitable, good biocompatibility, mechanical property matching, with good biomineralization function and corresponding antibacterial, anti-inflammatory or the ability such as antitumor, there is wide prospect in terms of the clinical practice that the bone tissue in future is rebuild.
Description
Technical field
The invention belongs to organic functional material preparation field, and in particular to a kind of poly- methyl-prop of injectable-porous-load medicine
E pioic acid methyl ester base compound rest bone grafting material and preparation method thereof.
Background technology
Bone grafting is directed to repairing a wide range of Cranial defect caused by wound, tumour, infection etc. always, to recover
It is body function, attractive in appearance, it is one of most common transfer operation.Bone collection is divided into autologous bone transplanting, allograph bone by graft source
Transplanting and artificial bone substitute materials transplanting.Autologous bone transplanting is the goldstandard of Bone Defect Repari and bone remoulding, yet with limited source,
Secondary insult can be caused, it is impossible to which the shortcomings of providing incipient stability, its clinical practice are very restricted.Allogenic bone transplantation expense
Height, limited amount.Meanwhile allograph bone has antigenicity, particularly when planting body is larger, often because of immunological rejection or outer disease
Poison, which is propagated, causes graft failure.Therefore, the research of artificial bone alternate material is focus in recent years.Wherein, injectable bone substitutes
Material（injectable bone substitutes, IBSs）Due to its minimally invasive surgical pathway and to complicated irregular scarce
The advantages that damaging effective filling in area, make its in clinical practice can effective reduction of patient discomfort, shortening cure time, subtract
Small treatment cost, therefore IBSs obtains increasing concern in bone tissue engineer field.
Polymethyl methacrylate（Polymethylmethacrylate, PMMA）As a kind of IBS, its is cheap,
With certain mechanical strength, at room temperature can rapid shaping, it is and small with the rejection of body, since the 1960s just
It is widely used in bone defect healing.However, in long-term clinical practice, PMMA some drawbacks gradually expose, such as machine
Tool performance and nature bone mismatch, Integrated implant is poor, polymeric reaction temperature is higher, infect etc. so that PMMA implants are in acceptor
Occur little by little loosening during the continuous reconstruction of bone, come off and surrounding tissue is downright bad, and graft failure may be ultimately caused.
Therefore, carry out that itself is modified or to add new biomaterial imperative to PMMA.
Porous three-dimensional stereochemical structure is preferable stent model in recent years bone tissue engineer, it is possible to provide roomy surface area
And space, enter beneficial to cell adherence growth, extrtacellular matrix deposition, nutrition and oxygen and metabolite is discharged, be also beneficial to
Blood vessel and nerve are grown into and medicament slow release, so that acceptor and more healthy, the firm combination of timbering material.Based on this, by PMMA
Be made porous carrier, interconnected loose structure will make PMMA obtain with mechanical property as bone photo, be easy to Bone Ingrowth.Separately
Outside, using the material with bone guided function as filler, porous PMMA bone guided function will be improved.In addition, it is local
The intervention of drug sustained release system can effectively play the effect of loaded medicine, reach antibacterial, anti-inflammatory or the effect such as antitumor,
Improve bone collection success rate.
Chitosan（CS）It is the part deacetylation product of chitin, is the unique a large amount of existing alkalescence of current nature
Polysaccharide, no biotoxicity, there is good histocompatbility, biodegradability and antibiotic property, obtained in medical science, field of biology
In-depth study is arrived and has been widely applied.Recent study is found, by adding basic salt（Such as sodium glycero-phosphate：GP）, contain
Hydroxy polymer（Such as chitosan quaternary ammonium salt：HTCC）, chitosan is performed the derivatization（Such as hydroxyl butyl chitosan：HBC）And grafting
Reaction（Such as polyethylene glycol grafted chitosan：PEG-g-CS）Or and high-molecular compound（Such as polyvinyl alcohol：PVA；Methyl cellulose
Element：MC）Blending etc., can obtain the chitosan-based physical gel with temperature sensitivity, due to its syringeability, bio-compatible
Property, degradability and artificial cell support arm function etc., have obtained increasing concern.This complex solution can be in room
Temperature keeps liquid during less than room temperature for a long time, and when temperature is increased to physiological body temperature（37℃）When gelation immediately.In addition, shell
Glycan base thermo-sensitive gel can realize local medicament slow release as bone guided material and the carrier of medicine.
The content of the invention
The defects of it is an object of the invention to overcome prior art and deficiency, there is provided a kind of injectable-porous-load medicine gathers
Methyl methacrylate base compound rest bone grafting material and preparation method thereof.
The purpose of the present invention is achieved through the following technical solutions：
The polymethyl methacrylate base compound rest bone grafting material of a kind of injectable-porous-load medicine, for poly- methyl-prop
The new injectable that e pioic acid methyl ester and the chitosan-based thermo-sensitive gel of load bone guiding material and medicine are mixed to get it is porous
The bone cement compound of three-dimensional structure.
Described polymethyl methacrylate is self-curing resin, preferably presses the pulvis of polymethyl methacrylate and liquor
Mass ratio 1:1 ratio is stirred to obtain, and it is homogeneous dough plastic.
Described chitosan-based thermo-sensitive gel includes：Basic salt containing 1-5wt%（Such as sodium glycero-phosphate：GP；Sodium acid carbonate；Phosphorus
Sour sodium）Chitosan gel rubber, hydroxy polymer containing 1-10wt%（Such as chitosan quaternary ammonium salt：HTCC）Chitosan gel rubber, shell is gathered
Sugar performs the derivatization（Such as hydroxyl butyl chitosan：HBC）, graft reaction（Such as polyethylene glycol grafted chitosan：PEG-g-CS；It is poly- different
Propylacrylamide；Poloxamer grafted chitosan）Or the high-molecular compound with concentration for 1-10wt%（Such as polyvinyl alcohol：
PVA；Methylcellulose：MC）One kind in the chitosan gel rubber that blending etc. obtains.
Described bone guided material includes calcium phosphate ceramic（Such as hydroxyapatite）, calcium phosphate bone cement（Such as β-tricresyl phosphate
Calcium）, calcium silicon materials（Such as bio-vitric）Etc. one of which or several compositions.Described chitosan-based thermo-sensitive gel load bone
The amount of guiding material is preferably 5-100mg/mL.
Described medicine includes the chemistry such as all kinds of antibacterials, anti-inflammatory drug, antineoplastic, Peptides and proteins
One or more of compositions in synthetic drug.Described antibacterials include various antibiotic, sulfamido, imidazoles, nitre
One or more of compositions in the chemical synthetic drugs such as base imidazoles, quinolones；Described anti-inflammatory drug includes all kinds of
Steroidal anti-inflammatory medicine and NSAIDs（Such as aspirin, phenylbutazone, celecoxib）Deng one kind in chemical synthetic drug or
Several compositions, drugloading rate are；Described antineoplastic includes alkylating agent, antimetabolite, antitumor antibiotics class, autonomic drug
One or more of compositions in the chemical synthetic drugs such as class, hormone and miscellany, drugloading rate are.Described peptide and protein
Class medicine includes bovine serum albumin(BSA), insulin, growth factor, hematopoietin, interferon, therapeutic monoclonal antibodies
Deng one or more of compositions in chemical synthetic drug.The amount of described chitosan-based thermo-sensitive gel carrying medicament is preferably
1-150mg/mL.It is furthermore preferred that when described medicine is antibacterials, the drugloading rate of chitosan-based thermo-sensitive gel is 1-50mg/
mL；When medicine is anti-inflammatory drug, the drugloading rate of chitosan-based thermo-sensitive gel is 1-100mg/mL；When medicine is antineoplastic, shell
The drugloading rate of glycan base thermo-sensitive gel is 10-150mg/mL；It is chitosan-based temperature sensitive solidifying when medicine is polypeptide or protein drug
The drugloading rate of glue is 1-85mg/mL.
In described compound rest bone grafting material, polymethyl methacrylate and load bone guiding material and the shell of medicine
The volume ratio of glycan base thermo-sensitive gel is preferably 3-12:10.
The preparation method of the polymethyl methacrylate base compound rest bone grafting material of above-mentioned injectable-porous-load medicine,
Comprise the following steps：
（1）Prepare one kind in following chitosan-based composite solution：Basic salt containing 1-5wt%（Such as sodium glycero-phosphate：GP；Carbon
Sour hydrogen sodium；Sodium phosphate）Chitosan-based composite solution, hydroxyl polymer-containing containing 1-10wt%（Such as chitosan quaternary ammonium salt：HTCC）'s
Chitosan-based composite solution, is performed the derivatization to chitosan（Such as hydroxyl butyl chitosan：HBC）And graft reaction（Such as polyethylene glycol
Grafted chitosan：PEG-g-CS；PNIPAM；Poloxamer grafted chitosan）Or the height with concentration for 1-10wt%
Molecular compound（Such as polyvinyl alcohol：PVA；Methylcellulose：MC）Blending etc. obtains chitosan-based composite solution.
（2）Bone guided material and medicine are added into well mixed chitosan-based composite solution, and supersound process makes it
It is well mixed.
（3）Composite solution is placed in incubator and forms it into gel.
（4）The pulvis of polymethyl methacrylate and liquor are stirred to form homogeneous dough plastic.
（5）Will（4）Gained dough plastic with（3）Gained gel is well mixed, and is injected into mould, cured,
It is stripped, is dried to obtain the polymethyl methacrylate base compound rest bone grafting material of-porous-the load medicine of injectable.
Step（2）It is excellent to add bone guided material in the mixed solution of bone guided material and medicine, the final concentration difference of medicine
Elect 5-100mg/mL, 1-150mg/mL as.
Step（3）The middle temperature for forming gel is preferably 30-60 DEG C.
Step（4）The pulvis of middle polymethyl methacrylate and the mass ratio of liquor are preferably 1:1.
Step（5）The volume ratio of middle dough plastic and gel is preferably 3-12:10.
The polymethyl methacrylate base compound rest bone grafting material of injectable provided by the invention-porous-load medicine with
PMMA is as the support for providing mechanical support, and chitosan-based thermo-sensitive gel is as pore former and the load of bone guided material and medicine
Body, both are mutually mixed the porous bone cement compound for forming new injectable.The compound rest bone grafting material have with
Under several features：（1）As IBS, effective filling due to its minimally invasive surgical pathway and to complicated irregular defective region etc.
Advantage, make its in clinical practice can effective reduction of patient discomfort, shorten cure time, reduce treatment cost.（2）It is right
Existing conventional dense, solid PMMA are modified, and are formed the three-dimensional porous rack with multiple advantage, are easy to cell adherence to give birth to
Length, extrtacellular matrix deposition, nutrition and oxygen enter and metabolite discharge, is also beneficial to blood vessel and nerve grows into and delayed with medicine
Release.（3）The equalization point of porous compound support frame material mechanical property is found, close to human body cancellous bone characteristic.（4）Bone guided material
Insert, be easy to further guide mineral deposition, the adhesion of reinforcement material-bone interface.（5）Jie of local drug sustained release system
The effect of entering effectively to play loaded medicine, reach antibacterial, anti-inflammatory or the effect such as antitumor, improve bone collection success
Rate.The material preparation method is simple, good biocompatibility, mechanical property matching, also with good biomineralization function and resists
Bacterium, anti-inflammatory or anti-tumor capacity, there is wide prospect in terms of the clinical practice that the bone tissue in future is rebuild.
The present invention has following remarkable result relative to prior art：
1st, this compound rest bone grafting material preparation method is simple, and the time is short, and cost is low.
2nd, due to the syringeability of this material, using minimally invasive surgical pathway, can effectively alleviate in clinical practice
The discomfort of patient, shorten cure time, reduce treatment cost.
3rd, compared with traditional PMMA, during the Clinical practice in future, because the addition of pore former CS-GP gels is dropped
Low PMMA polymerisation, heat release is reduced, reduce the risk of surrounding tissue necrosis.It is in addition, hard to material in vivo from being injected into
Working time between change is suitable, is easy to material to be sufficient filling with Cranial defect area, and operator has the time behaviour of abundance in art
Make.
4th, compared with traditional PMMA, this porous three-dimensional three-dimensional bracket material has and mechanical property as bone photo, good
Bone guided and biocompatibility, so as to benefit vast Cranial defect patient and produce far-reaching social influence.
Brief description of the drawings
Fig. 1 is the antibacterial ring size experiment that embodiment 1 obtains（ZOI）Result figure.
Fig. 2 is the antibacterial ring size experiment that embodiment 2 obtains（ZOI）Result figure.
Fig. 3 is the Micro-CT scanning for the PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials that embodiment 3 obtains（µ-
CT）Result figure, data are by the minitype CTs of μ-CT 50 collection by Scanco Medical companies of Switzerland.
Fig. 4 is the SEM in the PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials cross section that embodiment 3 obtains
Figure（×250）, the photo of ESEM shoots to obtain by the sigma Flied emission transmission electron microscopes of German Carl Zeiss Inc..
Fig. 5 is the obtained PMMA/CS-GP/Nano-HA/GM compound rests bone grafting material of embodiment 3 in being soaked in SBF
The SEM figures in its cross section of 28d（×20000）, the photo of ESEM transmits by the sigma Flied emissions of German Carl Zeiss Inc.
Electronic Speculum shoots to obtain.
Fig. 6 is the obtained PMMA/CS-GP/Nano-HA/GM compound rests bone grafting material of embodiment 3 in being soaked in SBF
28d mechanical property result figure, data by MTS Systm Corp. of the U.S. MTS810 ceramic test system acquisitions.
Fig. 7 is the obtained PMMA/CS-GP/Nano-HA/GM compound rests bone grafting material of embodiment 4 in being soaked in SBF
28d mechanical property result figure, data by MTS Systm Corp. of the U.S. MTS810 ceramic test system acquisitions.
Fig. 8 is the obtained PMMA/CS-GP/Nano-HA/GM compound rests bone grafting material of embodiment 5 in being soaked in SBF
28d mechanical property result figure, data by MTS Systm Corp. of the U.S. MTS810 ceramic test system acquisitions.
Fig. 9 is PMMA/CS-GP/Nano-HA/GM compound rests bone grafting material that embodiment 6 obtains to MC3T3 cells
Toxicity data figure.
Figure 10 is PMMA/CS-GP/Nano-HA/GM compound rests bone grafting material that embodiment 7 obtains to BMSC cells
Toxicity data figure.
Embodiment
Further detailed description is done to the present invention with reference to embodiment, and illustrate the present invention outstanding feature and significantly
It is progressive.Following embodiments are only that the explanation present invention without limiting the scope of the invention, other any without departing from this hair
Bright Spirit Essence with made under principle change, modification, replacement, combine, simplification, should be equivalent substitute mode, all wrap
It is contained within protection scope of the present invention.Unless otherwise specified, technological means used in embodiment is those skilled in the art
Known conventional meanses.
Embodiment 1
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg gentamicin sulphates（GM）With 300mg hydroxyapatites（HA）In well mixed CS-GP
In solution, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/GM mixed solutions.
（5）CS-GP/Nano-HA/GM mixed solutions are placed in 37 DEG C of insulating box, CS-GP/Nano- is formed after 5min
HA/GM gels.
（6）Tabletting after CS-GP/Nano-HA/GM gel refrigeration dryings, a diameter of 13mm, it is 1mg/mL to be placed in and scribble concentration
Escherichia coli suspension LB agar mediums center, observed after cultivating 24h in 37 DEG C of incubators, and it is straight to measure antibacterial ring size
Footpath.
As a result see Fig. 1, have the transparent antibacterial ring size that a loop diameter is 4.39cm around gel film, it was confirmed that the present embodiment obtains
The good biocidal property of the CS-GP/Nano-HA/GM gels arrived.
Embodiment 2
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg vancomycins（VM）With 300mg hydroxyapatites（HA）In well mixed CS-GP solution
In, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/VM mixed solutions.
（5）CS-GP/Nano-HA/VM mixed solutions are placed in 37 DEG C of insulating box, gel is formed after 5min.
（6）Tabletting after CS-GP/Nano-HA/VM gel refrigeration dryings, a diameter of 13mm, it is 1mg/mL to be placed in and scribble concentration
Staphylococcus aureus suspension MH agar mediums center, observed after cultivating 24h in 37 DEG C of incubators, and measure suppression
Collarium diameter.
As a result see Fig. 2, have the transparent antibacterial ring size that a loop diameter is 4.53cm around gel film, it was confirmed that the present embodiment obtains
The good biocidal property of the CS-GP/Nano-HA/VM gels arrived.
Embodiment 3
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg gentamicin sulphates（GM）With 300mg hydroxyapatites（HA）In well mixed CS-GP
In solution, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/GM mixed solutions.
（5）CS-GP/Nano-HA/GM mixed solutions are placed in 37 DEG C of insulating box, CS-GP/Nano- is formed after 5min
HA/GM gels.
（6）By PMMA pulvis and liquor according to mass ratio 1:1 ratio is stirred, and it is plastic to form homogeneous dough
Thing.
（7）By this dough plastic according to 3:4 volume ratio is sufficiently mixed with above-mentioned CS-GP/Nano-HA/GM gels
Uniformly, it is subsequently injected into the cylinder mold of polytetrafluoroethylene (PTFE)（Internal diameter 6mm, high 12mm）It is interior, so as to moulding.
（8）Overnight after solidification, the demoulding, PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are dried to obtain.
With the minitype CTs of μ-CT 50（Scanco Medical companies of Switzerland）Collection gained compound rest bone grafting material shows
Micro- CT results, are as a result shown in Fig. 3, from three-dimensional reconstruction result it can be seen that material surface has many irregular holes, and are inlaid with
The nano HA particle of white；Cross section scanning result then shows that irregular aperture is interconnected, and nano HA distribution of particles is in material
Expect in support.
With sigma Flied emission transmission electron microscopes（German Carl Zeiss Inc.）Observation gained compound rest bone grafting material shape
Looks, as a result see Fig. 4, material surface there are many irregular holes as we can see from the figure, is inlaid with hole with PMMA surface
The nano HA particle of needle-like.
The structure of material provides preferable support for Bone Ingrowth obtained by Fig. 3,4 results show.
It is molten that gained PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are placed in the artificial body fluid SBF prepared
Soaked in liquid, change a SBF solution week about, freeze-drying is taken out after 28d, with sigma Flied emission transmission electron microscope observings
Its pattern, with its mechanical property data of MTS810 ceramic tests system acquisition, as a result see Fig. 5 and Fig. 6.
Fig. 5 is the SEM figures in the compound rest bone grafting material cross section after immersion, by Fig. 5 it can be seen that material
After 28d is soaked in SBF, there is massive deposit that one layer is abundant, that volume is larger in PMMA and acicular nanometer HA surfaces, it was demonstrated that
Prepared compound support, which has, promotes mineralising and potential bone guided effect.
Fig. 6 is the mechanical property result of the material after immersion, the results showed that before and after the material soaks 28d in SBF,
Its modulus of elasticity and compression strength no difference of science of statistics, and in the range of the mechanical property value in human body cancellous bone, there is mould
The potential quality of anthropomorphic body cancellous bone.
Embodiment 4
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg gentamicin sulphates（GM）With 300mg hydroxyapatites（HA）In well mixed CS-GP
In solution, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/GM mixed solutions.
（5）CS-GP/Nano-HA/GM mixed solutions are placed in 37 DEG C of insulating box, CS-GP/Nano- is formed after 5min
HA/GM gels.
（6）By PMMA pulvis and liquor according to mass ratio 1:1 ratio is stirred, and it is plastic to form homogeneous dough
Thing.
（7）By this dough plastic according to 3:10 volume ratio is sufficiently mixed with above-mentioned CS-GP/Nano-HA/GM gels
Uniformly, it is subsequently injected into the cylinder mold of polytetrafluoroethylene (PTFE)（Internal diameter 6mm, high 12mm）It is interior, so as to moulding.
（8）Overnight after solidification, the demoulding, PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are dried to obtain.
It is molten that gained PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are placed in the artificial body fluid SBF prepared
Soaked in liquid, change a SBF solution week about, freeze-drying is taken out after 28d, with MTS810 ceramic tests system acquisition its
Mechanical property data, as a result see Fig. 7.As a result before and after showing that the material soaks 28d in SBF, its modulus of elasticity and compression strength
No difference of science of statistics, and in the range of the mechanical property value in human body cancellous bone, there is the potential quality of simulation human body cancellous bone.
Embodiment 5
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg gentamicin sulphates（GM）With 300mg hydroxyapatites（HA）In well mixed CS-GP
In solution, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/GM mixed solutions.
（5）CS-GP/Nano-HA/GM mixed solutions are placed in 37 DEG C of insulating box, CS-GP/Nano- is formed after 5min
HA/GM gels.
（6）By PMMA pulvis and liquor according to mass ratio 1:1 ratio is stirred, and it is plastic to form homogeneous dough
Thing.
（7）By this dough plastic according to 6:5 volume ratio is sufficiently mixed with above-mentioned CS-GP/Nano-HA/GM gels
Uniformly, it is subsequently injected into the cylinder mold of polytetrafluoroethylene (PTFE)（Internal diameter 6mm, high 12mm）It is interior, so as to moulding.
（8）Overnight after solidification, the demoulding, PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are dried to obtain.
It is molten that gained PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are placed in the artificial body fluid SBF prepared
Soaked in liquid, change a SBF solution week about, freeze-drying is taken out after 28d, with MTS810 ceramic tests system acquisition its
Mechanical property data, as a result see Fig. 8.As a result before and after showing that the material soaks 28d in SBF, its modulus of elasticity and compression strength
No difference of science of statistics, and in the range of the mechanical property value in human body cancellous bone, there is the potential quality of simulation human body cancellous bone.
Embodiment 6
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg gentamicin sulphates（GM）With 300mg hydroxyapatites（HA）In well mixed CS-GP
In solution, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/GM mixed solutions.
（5）CS-GP/Nano-HA/GM mixed solutions are placed in 37 DEG C of insulating box, CS-GP/Nano- is formed after 5min
HA/GM gels.
（6）By PMMA pulvis and liquor according to mass ratio 1:1 ratio is stirred, and it is plastic to form homogeneous dough
Thing.
（7）By this dough plastic according to 3:4 volume ratio is sufficiently mixed with above-mentioned CS-GP/Nano-HA/GM gels
Uniformly, it is subsequently injected into the cylinder mold of polytetrafluoroethylene (PTFE)（Internal diameter 6mm, high 12mm）It is interior, so as to moulding.
（8）Overnight after solidification, the demoulding, PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are dried to obtain.
（9）The PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials crossed with disinfection with high pressure steam are soaked in
It is dual anti-containing 10% hyclone and 1%（Penicillin and streptomycin sulphate）A-MEM culture mediums in, be placed in 37 DEG C of incubator cultures
Leaching liquor is made in 24h.Recovered simultaneously with this a-MEM culture medium and pass on MC3T3 cells.
（10）By 150-200 μ L MC3T3 cells according to 1 × 104The density in/hole is inoculated into 96 orifice plates, after cultivating 24h
Cell attachment is treated, culture medium is suctioned out, replaces with above-mentioned steps（9）Leaching liquor（Experimental group）, containing 10% hyclone and 1% pair
Anti- a-MEM culture mediums as a control group, continue to cultivate, and every group sets 6 secondary orifices.
（11）At the 1st and the 3rd day, 20 μ L 5mg/mL MTT solution is added per hole, suctions out, replaces with after cultivating 4h
200 μ L DMSO solutions, lucifuge mixing 15min, measures the spectrophotometric value at 490nm, and calculate relative growth ratio at room temperature
（RGR）, i.e. ODExperimental group/ODControl group, whether the compound rest bone grafting material obtained with detecting the present embodiment have cytotoxicity.
As a result Fig. 9 is seen, from the figure, it can be seen that the RGR of the MC3T3 cells of the 1st day and the 3rd day is all higher than 100, and with
Time growth, OD values can also increase, it was demonstrated that this compound rest bone grafting material has good to MC3T3 cytotoxics
Biocompatibility.
Embodiment 7
（1）At room temperature by chitosan（CS）It is dissolved in the CS hydrochloric acid for preparing that mass concentration is 2% in 0.1mol/L hydrochloric acid solutions
Solution, after dissolving completely, 15min is stirred in ice bath.
（2）At the same time, by sodium glycero-phosphate（GP）It is dissolved in distilled water and prepares the GP solution that mass concentration is 50%, together
Sample places 15min in ice bath.
（3）In the case where being stirred continuously, 0.5mL GP solution is slowly dropped into 4.5mL CS hydrochloric acid solutions, keeps solution without mixed
Turbid, stirring 20min obtains well mixed CS-GP solution.
（4）Successively add 25mg gentamicin sulphates（GM）With 300mg hydroxyapatites（HA）In well mixed CS-GP
In solution, and being ultrasonically treated makes it well mixed obtain CS-GP/Nano-HA/GM mixed solutions.
（5）CS-GP/Nano-HA/GM mixed solutions are placed in 37 DEG C of insulating box, CS-GP/Nano- is formed after 5min
HA/GM gels.
（6）By PMMA pulvis and liquor according to mass ratio 1:1 ratio is stirred, and it is plastic to form homogeneous dough
Thing.
（7）By this dough plastic according to 3:4 volume ratio is sufficiently mixed with above-mentioned CS-GP/Nano-HA/GM gels
Uniformly, it is subsequently injected into the cylinder mold of polytetrafluoroethylene (PTFE)（Internal diameter 6mm, high 12mm）It is interior, so as to moulding.
（8）Overnight after solidification, the demoulding, PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials are dried to obtain.
（9）The PMMA/CS-GP/Nano-HA/GM compound rest bone grafting materials crossed with disinfection with high pressure steam are soaked in
It is dual anti-containing 10% cow's serum and 1%（Penicillin and streptomycin sulphate）A-MEM culture mediums in, be placed in 37 DEG C of incubator culture 24h
Leaching liquor is made.Recovered simultaneously with this a-MEM culture medium and pass on mesenchymal stem cells MSCs（BMSC）Cell；
（10）By 150-200 μ L BMSC cells according to 1 × 104The density in/hole is inoculated into 96 orifice plates, is treated after cultivating 24h
Cell attachment, culture medium is suctioned out, replace with above-mentioned steps（9）Leaching liquor（Experimental group）, it is dual anti-containing 10% hyclone and 1%
A-MEM culture mediums as a control group, continue to cultivate.Every group sets 6 secondary orifices.
（11）At the 1st and the 3rd day, 20 μ L 5mg/mL MTT solution is added per hole, suctions out, replaces with after cultivating 4h
200 μ L DMSO solutions, lucifuge mixing 15min, measures the spectrophotometric value at 490nm, and calculate relative growth ratio at room temperature
（RGR）, i.e. ODExperimental group/ODControl group, whether the compound rest bone grafting material obtained with detecting the present embodiment have cytotoxicity.
As a result Figure 10 is seen, from the figure, it can be seen that the RGR of the BMSC cells of the 1st day and the 3rd day is all higher than 100, and with
Time growth, OD values can also increase, it was demonstrated that this compound rest bone grafting material has good to BMSC cytotoxics
Biocompatibility.
Claims (9)
- A kind of 1. polymethyl methacrylate base compound rest bone grafting material of injectable-porous-load medicine, it is characterised in that： The injectable being mixed to get for the chitosan-based thermo-sensitive gel of polymethyl methacrylate and load bone guiding material and medicine The bone cement compound of porous three-dimensional structure；Described chitosan-based thermo-sensitive gel gathers for the shell of the sodium glycero-phosphate containing 1-5wt% Sugared gel.
- 2. polymethyl methacrylate base compound rest bone grafting material according to claim 1, it is characterised in that：It is described Polymethyl methacrylate be by the pulvis of polymethyl methacrylate and liquor in mass ratio 1:1 ratio is stirred Arrive.
- 3. polymethyl methacrylate base compound rest bone grafting material according to claim 1, it is characterised in that：It is described Bone guided material be calcium phosphate ceramic, calcium phosphate bone cement and calcium silicon materials in one or more；Described medicine is anti- One or more in bacterium medicine, anti-inflammatory drug, antineoplastic, Peptides and proteins.
- 4. polymethyl methacrylate base compound rest bone grafting material according to claim 3, it is characterised in that：It is described The amount of chitosan-based thermo-sensitive gel load bone guiding material be 5-100mg/mL, the amount of carrying medicament is 1-150mg/mL.
- 5. polymethyl methacrylate base compound rest bone grafting material according to claim 1, it is characterised in that：Poly- first Base methyl acrylate and the volume ratio of load bone guiding material and the chitosan-based thermo-sensitive gel of medicine are 3-12:10.
- 6. the preparation method of the polymethyl methacrylate base compound rest bone grafting material described in claim any one of 1-5, It is characterized in that：Comprise the following steps：(1) the chitosan-based composite solution of the sodium glycero-phosphate containing 1-5wt% is prepared；(2) toward addition bone guided material and medicine in well mixed chitosan-based composite solution, and supersound process makes its mixing Uniformly；(3) composite solution is placed in incubator and forms it into gel；(4) pulvis of polymethyl methacrylate and liquor are stirred to form homogeneous dough plastic；(5) dough plastic obtained by (4) is well mixed with gel obtained by (3), be injected into mould, the cured, demoulding, It is dried to obtain the polymethyl methacrylate base compound rest bone grafting material of-porous-the load medicine of injectable.
- 7. the preparation method of polymethyl methacrylate base compound rest bone grafting material according to claim 6, it is special Sign is：Bone guided material, the final concentration of medicine are respectively 5- in the mixed solution of step (2) addition bone guided material and medicine 100mg/mL、1-150mg/mL。
- 8. the preparation method of polymethyl methacrylate base compound rest bone grafting material according to claim 6, it is special Sign is：The mass ratio of the pulvis of polymethyl methacrylate and liquor is 1 in step (4):1.
- 9. the preparation method of polymethyl methacrylate base compound rest bone grafting material according to claim 6, it is special Sign is：The volume ratio of dough plastic and gel is 3-12 in step (5):10.
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| US9974883B2 (en) | 2016-08-11 | 2018-05-22 | University of Central Oklahoma | Method and apparatus to control the heterogeneous flow of bone cement and improve osseointegration of cemented implant |
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| CN113087861B (en) * | 2021-03-31 | 2022-07-08 | 四川大学 | Modified hydrogel with mild photothermal effect and preparation method and application thereof |
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