Background technology
Owing to factors such as wound, infection, tumor make bone lose some sclerotin, form bigger gap, it is damaged to be called bone.Damaged all being difficult to of most bones heals, and forms bone does not connect at last.Because defect gap is big, osteoblast is difficult to get over the gap and normal agglutination can not take place, only by the fibrous tissue filling.And damaged treatment is a difficulty and the challenging problem in orthopaedics field for bone, clinically mainly by reaching certain therapeutic purposes from body, allosome tissue's transplanting and biological substitution material.The most frequently used autograft at present because the source is not enough, wound is big, for unfavorable factors such as district's complication, limited and used clinically; And allosome tissue transplant exist allosome rejection, source limited, have a factor such as risk of disease transmission; So, seek and not damage self, target that the person pursues that the substitute that can reach the functional effect of expection bone defect repair again becomes the clinical position.Organizational project learns a skill provides new thinking and method for solving this difficult problem.
The basic skills of organizational project is: be adsorbed in good biocompatibility after the high concentration histiocyte amplification with In vitro culture, can be by on the tissue engineering bracket material of human body degraded and absorbed, the function of this material is to provide vivosphere for cell, make cell obtain enough nutrient substance, carry out gas exchange, and cell is grown by the three-dimensional rack of prefabricated form, then with the sick position of decreasing of the complex implanting to human body of cell and biomaterial, in biological support degraded and absorbed process, the cell of plantation continues the propagation breeding, form new respective organization with original specific function and form and organ, to reach the purpose of repairing wound and rebuilding function.
It is an important branch of organizational project that bone tissue engineer is learned, and is expected at first be applied clinical.Formed at present comparatively perfect theory and technology route gradually about seed cell, cell carrier support and tissue construction.But undeniable is that these adjusted range bone tissue engineers finally still have certain distance in Clinical Application.Therefore the focus of present stage research still concentrates on suitable seed cell, material and the construct in vitro mode sought, and seeks three's best of breed, and the natural process that simulation human body bone is repaired is reappeared bone structure and function to greatest extent.At present, the focus of bone tissue engineer research concentrates on seed cell, timbering material, bioactie agent three aspects.Wherein the effect of timbering material in organizational project provides the framework of a kind of porous material as tissue regeneration, cultured cell in vitro is planted on it, and return and to implant, guide the continuous growth of required tissue, frame materials fades away simultaneously, finally in former framework region, form and bear complete lived tissue again, realize permanent reparation.
Ideal timbering material need meet following condition (1) material surface structure and character helps cell absorption, propagation and differentiation; (2) degrade by controllable speed; (3) excellent biological compatibility; (4) easily make three-dimensional porous shape, and easily be processed into irregular geometric shape; (5) have certain mechanical strength and can support physical stress; (6) can keep the differentiation of pair cell, can not make cell produce variation.The preparation method of osseous tissue engineering stephanoporate stent material and process are at home to be mature on the whole at present, the timbering material that adopts mainly contains synthetic material and natural biological derived material and composite, wherein with poly-acetic acid (polyglycolicAcid, PGA), polylactic acid (polylactic Acid, PLA), tricalcium phosphate (tricalciumphosphate, TCP), (hydroxy lapatite, HA) grade is comparatively commonly used for hydroxyapatite.By deeply it is found that of research, above-mentioned different materials its biological degradability and ossification in further research process all have weak point; Though PGA, PLA have better biocompatibility, degradability and absorbability, but there are shortcomings such as expense costliness, poor plasticity, and the acidic metabolite after the degraded can reduce polymer pH value on every side, influence the growth of cell and tissue, also can cause the immunoreation of fibrosis and generation surrounding tissue.Be difficult to after HA exist to implant absorb alternative, be stranded in and hinder the reconstruction of osseous tissue and shortcoming such as reparation fully in the body for a long time. we might as well widen thinking and seek some new material-medical grade calcium sulfate (Medical-grade Calcium Sulfate) so, it has excellent biological compatibility, degradability and ossification, makes its raw material as the preparation support will obtain good effect and using value.
Summary of the invention
Purpose of the present invention: invent complex stephanoporate bracket of a kind of calcium sulfate and apatite complex and preparation method thereof.
Through multiple discovering, calcium sulfate is a kind of bone guided material, and mainly as the implant in space, it can recover the form profile of bone, stops soft tissue to be grown into.It provides the substrate of bone guided for blood vessel and osteoblastic growing into.
The preparation method of the complex stephanoporate bracket of a kind of calcium sulfate and apatite complex, wherein, the step of preparation is as follows:
1) the compound slurry of preparation sulfate hemihydrate calcium powder and ground apatite complex;
2) the calcium sulfate slurry evenly is coated on polyurethane foam after, after super-dry, high-temperature process, obtain the ceramic porous support of calcium sulfate;
3) the ceramic porous support of above-mentioned calcium sulfate is soaked in the above-mentioned compound slurry after the hanging, places natural drying under the indoor environment, behind the heat drying, obtain the complex stephanoporate bracket of calcium sulfate of the present invention and apatite complex then.
The preparation method of above-mentioned compound slurry is as follows: with mixing of sulfate hemihydrate calcium powder and ground apatite complex, the powder body of calcium sulfate powder body, apatite complex, tobramycin powder are with (1.5-2.5): (0.8-1.2): evenly obtain compound slurry after (0.8-1.2) adding the carboxymethyl cellulose rheological agent of 0.5-2% (preferred 1%) and 25-45% (preferred 35%) deionized water after the mixed of (preferred 2: 1: 1).
The composition and the preparation method of above-mentioned apatite complex are as follows: apatite mixes with collagen protein, and ratio is 3: 2.
Above-mentioned polyurethane foam is: 45ppi polyurethane foam, aperture are between the ratio of 150-500 μ m and porosity 84.29%, and the elasticity height, pore is even, the porosity is high, has tridimensional network.
The processing method of above-mentioned polyurethane foam is: foams are cut into the cylinder fritter of ∮ 5mm * 15mm, be immersed in the sodium hydroxide solution of 10% concentration after cleaning up with distilled water, hydrolysis process 4h under 60 ℃ of temperature, film between the network in the scumming body, then, rub repeatedly and clean, dry standby with flushing with clean water.
The preparation method of above-mentioned calcium sulfate slurry is as follows: will add the silica sol binder of its weight 10-50% (preferred 15%) in the calcium sulfate powder body, and the deionized water of the 20-60% of its weight (preferred 40%), fully mix homogeneously obtains the calcium sulfate slurry.
Above-mentioned half-H 2 O calcium sulphate preparation method is as follows: the sulfate dihydrate calcium powder is mixed with deionized water, place in the airtight pressure vessel, the polyacrylamide crystal modifier that adds convention amount simultaneously, at 90-160 degree centigrade and 1.5-4.5 atmospheric pressure, make calcium sulphate dihydrate dissolving recrystallize form the crystal modification of half-H 2 O calcium sulphate, take out physical filtering then fast, put into air dry oven under 140-160 ℃ of temperature freeze-day with constant temperature 4-6 hour, sieving through grinding is prepared from.
Described polyacrylamide is coated in the polyurethane foam surface, forms a polymeric layer, strengthened the covering property between polyurethane foam.
The above-mentioned the 2nd) one step process is as follows: the ∮ 5mm that will cut out * 15mm polyurethane foam cylinder fritter is soaked in the calcium sulfate slurry, treat to take out after porose stem evenly sticks the calcium sulfate slurry on the polyurethane foam, make the calcium sulfate base substrate, then base substrate is placed natural drying 24h under the indoor environment, dry 24h under 110 ℃ in the electric drying oven with forced convection; The burning volatilization of polyurethane foam is the key point of ceramic porous rack forming, according to organic foam body GH-1GI curve determination, its plastic removal temperature is about 300 ℃, when base substrate carries out sintering 300 to 600 ℃ the time programming rate be controlled at 1 ℃/min, thereby the gas of having avoided the oxidation of organic foam body to produce damages base substrate; And then rise to 1000 ℃ of insulation 2h, and the back is with the furnace temperature cooling, and polyurethane at high temperature is completely decomposed of volatile, and the ceramic porous support of calcium sulfate prepares successfully.
The above-mentioned the 3rd) one step process is as follows: the ceramic porous support of calcium sulfate places natural drying 10-40 hour (preferred 24 hours) under the indoor environment after being soaked in the compound slurry further hanging, 80 ℃ dry 5-20 hour (preferred 12 hours) down; After this step repeats 3-5 time, just make on all hole walls of the ceramic porous support of calcium sulfate and sticked compound material uniformly, calcium sulfate prepared with the composite type biological stephanoporate bracket of apatite complex and finished this moment.Aseptic preservation is standby behind cobalt-60 radiation sterilization.
Beneficial effect
After scanning, Electronic Speculum SEM shape appearance figure can be measured, and the compound support of gained aperture is distributed between 150~300 μ m, interconnect between the hole, and the material internal micropore enriches, is evenly distributed, and presents loose coralliform structure.This similar is in the spongy bone framework of biological living, and it helps the biodegradation of growing into of osseous tissue and material self.
Calculate through test, the porous support mean porosities is 62.59%-79.36%, and comprcssive strength is 27-53MPa (the biomechanical strength spongy bone of normal human's osseous tissue is 7.09MPa, Compact bone 177-221MPa).
When medical calcium sulfate is absorbed, new bone is moulding and recover anatomic characteristic and architectural characteristic.Its most important advantage is that its natural infiltration rate is suitable with new bone formation speed.Along with the absorption of medical calcium sulfate implant, new bone recovers to dissect character and construction features gradually.
The specific embodiment
Embodiment 1
Raw-material preparation: (1) calcium sulfate is a kind of very common raw material of industry, and wide material sources and cheapness very can be divided into 3 types: common calcium sulfate (calcium sulphate dihydrate), dead plaster and half-H 2 O calcium sulphate on chemical property; The latter is formed by the calcium sulphate dihydrate deep processing, and the crystal structure of half-H 2 O calcium sulphate is even, purity is high, can be used for the research and the treatment in medical science orthopaedics field, also claims medical calcium sulfate; Concrete preparation method is: the half-H 2 O calcium sulphate preparation method is as follows: the sulfate dihydrate calcium powder is mixed with deionized water, place in the airtight pressure vessel, the polyacrylamide crystal modifier that adds convention amount simultaneously, at 160 degrees centigrade and 1.5 atmospheric pressure, make calcium sulphate dihydrate dissolving recrystallize form the crystal modification of half-H 2 O calcium sulphate, take out fast then, carry out physical filtering, put into air dry oven freeze-day with constant temperature 4 hours under 160 ℃ of temperature, sieving through grinding is prepared from.(2) the apatite complex is refrigerated to-70 ℃, puts into vacuum again and continues cooling, reduces to gross weight to residual moisture and gets final product below 5%.(3) instruments such as prepared silicon sol adhesive, carboxymethyl cellulose rheological agent, polyurethane foam, QM21SP2 ball mill, 101A-3E electric drying oven with forced convection, R23-115-12 molding box formula resistance furnace.
The manufacturing process of compound material:
A. preliminary preparation: the piece of (1) calcium sulfate and apatite complex is put into ball mill respectively, and ball milling 1h obtains its powder body; (2) add 15% silica sol binder, 40% deionized water in the calcium sulfate powder body, fully mix homogeneously obtains the MSC slurry; (3) powder body of calcium sulfate powder body, apatite complex, 2: 1: 1 ratio of tobramycin powder and 1% carboxymethyl cellulose rheological agent and 35% deionized water are put into and are obtained being uniformly dispersed compound slurry behind the ball grinder ball milling 1h; (4) the 45ppi polyurethane foam is selected in this experiment for use, and the aperture is between the ratio of 150-500 μ m and porosity 84.29%, and the elasticity height, pore is even, the porosity is high, has tridimensional network.Foams are cut into the cylinder fritter of the required φ 5mm * 15mm of experiment, be immersed in the sodium hydroxide solution of 10% concentration after cleaning up with distilled water, hydrolysis process 4h under 60 ℃ of temperature, film between the network in the scumming body, then, rub repeatedly and clean, dry standby with flushing with clean water.In order to improve the adhesiveness between organic foam and the slurry, polyacrylamide is coated in the organic foam surface, form a polymeric layer, strengthened the covering property between organic foam.
B. technological process:
The φ 5mm that (1) will cut out * 15mm polyurethane foam cylinder fritter is soaked in the calcium sulfate slurry, treat to take out after porose stem evenly sticks the calcium sulfate slurry on the polyurethane foam, make the calcium sulfate base substrate, then base substrate is placed natural drying 24h under the indoor environment, dry 24h under 110 ℃ in the electric drying oven with forced convection; The burning volatilization of polyurethane foam is the key point of ceramic porous rack forming, according to organic foam body GH-1GI curve determination, its plastic removal temperature is about 300 ℃, when base substrate carries out sintering 300 to 600 ℃ the time programming rate be controlled at 1 ℃/min, thereby the gas of having avoided the oxidation of organic foam body to produce damages base substrate; And then rise to 1000 ℃ of insulation 2h, and the back is with the furnace temperature cooling, and polyurethane at high temperature is completely decomposed of volatile, and the ceramic porous support of calcium sulfate prepares successfully.
(2) after the ceramic porous support of calcium sulfate is soaked in the compound slurry further hanging, placed under the indoor environment natural drying 10 hours, in the electric drying oven with forced convection 80 ℃ dry 5 hours down; After this step repeats 3 times, just make on all hole walls of the ceramic porous support of calcium sulfate and sticked compound material uniformly, this moment, the composite type biological stephanoporate bracket preparation of calcium sulfate/apatite complex was finished.Aseptic preservation is standby behind cobalt-60 radiation sterilization.
Embodiment 2
Raw-material preparation: (1) calcium sulfate is a kind of very common raw material of industry, and wide material sources and cheapness very can be divided into 3 types: common calcium sulfate (calcium sulphate dihydrate), dead plaster and half-H 2 O calcium sulphate on chemical property; The latter is formed by the calcium sulphate dihydrate deep processing, and the crystal structure of half-H 2 O calcium sulphate is even, purity is high, can be used for the research and the treatment in medical science orthopaedics field, also claims medical calcium sulfate; Concrete preparation method is: the half-H 2 O calcium sulphate preparation method is as follows: the sulfate dihydrate calcium powder is mixed with deionized water, place in the airtight pressure vessel, the polyacrylamide crystal modifier that adds convention amount simultaneously, at 90 degrees centigrade and 4.5 atmospheric pressure, make calcium sulphate dihydrate dissolving recrystallize form the crystal modification of half-H 2 O calcium sulphate, take out fast then, carry out physical filtering, put into air dry oven freeze-day with constant temperature 6 hours under 140 ℃ of temperature, sieving through grinding is prepared from.(2) the apatite complex is refrigerated to-100 ℃, puts into vacuum again and continues cooling, reduces to gross weight to residual moisture and gets final product below 5%.(3) instruments such as prepared silicon sol adhesive, carboxymethyl cellulose rheological agent, polyurethane foam, QM21SP2 ball mill, 101A-3E electric drying oven with forced convection, R23-115-12 molding box formula resistance furnace.
The manufacturing process of compound material:
A. preliminary preparation: the piece of (1) calcium sulfate and apatite complex is put into ball mill respectively, and ball milling 1h obtains its powder body; (2) add its weight 50% silica sol binder, 60% deionized water in the calcium sulfate powder body, fully mix homogeneously obtains the MSC slurry; (3) powder body of calcium sulfate powder body, apatite complex, 2.5: 0.8: 0.8 ratio of tobramycin powder and 2% carboxymethyl cellulose rheological agent and 45% deionized water are put into and are obtained being uniformly dispersed compound slurry behind the ball grinder ball milling 1h; (4) the 45ppi polyurethane foam is selected in this experiment for use, and the aperture is between the ratio of 150-500 μ m and porosity 84.29%, and the elasticity height, pore is even, the porosity is high, has tridimensional network.Foams are cut into the cylinder fritter of the required φ 5mm * 15mm of experiment, be immersed in the sodium hydroxide solution of 10% concentration after cleaning up with distilled water, hydrolysis process 4h under 60 ℃ of temperature, film between the network in the scumming body, then, rub repeatedly and clean, dry standby with flushing with clean water.In order to improve the adhesiveness between organic foam and the slurry, polyacrylamide is coated in the organic foam surface, form a polymeric layer, strengthened the covering property between organic foam.
B. technological process:
The φ 5mm that (1) will cut out * 15mm polyurethane foam cylinder fritter is soaked in the calcium sulfate slurry, treat to take out after porose stem evenly sticks the calcium sulfate slurry on the polyurethane foam, make the calcium sulfate base substrate, then base substrate is placed natural drying 24h under the indoor environment, dry 24h under 110 ℃ in the electric drying oven with forced convection; The burning volatilization of polyurethane foam is the key point of ceramic porous rack forming, according to organic foam body GH-1GI curve determination, its plastic removal temperature is about 300 ℃, when base substrate carries out sintering 300 to 600 ℃ the time programming rate be controlled at 1 ℃/min, thereby the gas of having avoided the oxidation of organic foam body to produce damages base substrate; And then rise to 1000 ℃ of insulation 2h, and the back is with the furnace temperature cooling, and polyurethane at high temperature is completely decomposed of volatile, and the ceramic porous support of calcium sulfate prepares successfully.
(2) after the ceramic porous support of calcium sulfate is soaked in the compound slurry further hanging, placed under the indoor environment natural drying 40 hours, in the electric drying oven with forced convection 80 ℃ dry 20 hours down; After this step repeats 5 times, just make on all hole walls of the ceramic porous support of calcium sulfate and sticked compound material uniformly, this moment, the composite type biological stephanoporate bracket preparation of calcium sulfate/apatite complex was finished.Aseptic preservation is standby behind cobalt-60 radiation sterilization.
Embodiment 3
Raw-material preparation: (1) calcium sulfate is a kind of very common raw material of industry, and wide material sources and cheapness very can be divided into 3 types: common calcium sulfate (calcium sulphate dihydrate), dead plaster and half-H 2 O calcium sulphate on chemical property; The latter is formed by the calcium sulphate dihydrate deep processing, and the crystal structure of half-H 2 O calcium sulphate is even, purity is high, can be used for the research and the treatment in medical science orthopaedics field, also claims medical calcium sulfate; Concrete preparation method is: the half-H 2 O calcium sulphate preparation method is as follows: the sulfate dihydrate calcium powder is mixed with deionized water, place in the airtight pressure vessel, the polyacrylamide crystal modifier that adds convention amount simultaneously, at 150 degrees centigrade and 2 atmospheric pressure, make calcium sulphate dihydrate dissolving recrystallize form the crystal modification of half-H 2 O calcium sulphate, take out fast then, carry out physical filtering, put into air dry oven freeze-day with constant temperature 4.5 hours under 150 ℃ of temperature, sieving through grinding is prepared from.(2) the apatite complex is refrigerated to-90 ℃, puts into vacuum again and continues cooling, reduces to gross weight to residual moisture and gets final product below 5%.(3) instruments such as prepared silicon sol adhesive, carboxymethyl cellulose rheological agent, polyurethane foam, QM21SP2 ball mill, 101A-3E electric drying oven with forced convection, R23-115-12 molding box formula resistance furnace.
The manufacturing process of compound material:
A. preliminary preparation: the piece of (1) calcium sulfate and apatite complex is put into ball mill respectively, and ball milling 1h obtains its powder body; (2) add 15% silica sol binder, 40% deionized water in the calcium sulfate powder body, fully mix homogeneously obtains the MSC slurry; (3) powder body of calcium sulfate powder body, apatite complex, 1.5: 1.2: 1.2 ratio of tobramycin powder and 0.5% carboxymethyl cellulose rheological agent and 35% deionized water are put into and are obtained being uniformly dispersed compound slurry behind the ball grinder ball milling 1h; (4) the 45ppi polyurethane foam is selected in this experiment for use, and the aperture is between the ratio of 150-500 μ m and porosity 84.29%, and the elasticity height, pore is even, the porosity is high, has tridimensional network.Foams are cut into the cylinder fritter of the required φ 5mm * 15mm of experiment, be immersed in the sodium hydroxide solution of 10% concentration after cleaning up with distilled water, hydrolysis process 4h under 60 ℃ of temperature, film between the network in the scumming body, then, rub repeatedly and clean, dry standby with flushing with clean water.In order to improve the adhesiveness between organic foam and the slurry, polyacrylamide is coated in the organic foam surface, form a polymeric layer, strengthened the covering property between organic foam.
B. technological process:
The φ 5mm that (1) will cut out * 15mm polyurethane foam cylinder fritter is soaked in the calcium sulfate slurry, treat to take out after porose stem evenly sticks the calcium sulfate slurry on the polyurethane foam, make the calcium sulfate base substrate, then base substrate is placed natural drying 24h under the indoor environment, dry 24h under 110 ℃ in the electric drying oven with forced convection; The burning volatilization of polyurethane foam is the key point of ceramic porous rack forming, according to organic foam body GH-1GI curve determination, its plastic removal temperature is about 300 ℃, when base substrate carries out sintering 300 to 600 ℃ the time programming rate be controlled at 1 ℃/min, thereby the gas of having avoided the oxidation of organic foam body to produce damages base substrate; And then rise to 1000 ℃ of insulation 2h, and the back is with the furnace temperature cooling, and polyurethane at high temperature is completely decomposed of volatile, and the ceramic porous support of calcium sulfate prepares successfully.
(2) after the ceramic porous support of calcium sulfate is soaked in the compound slurry further hanging, placed under the indoor environment natural drying 30 hours, in the electric drying oven with forced convection 80 ℃ dry 15 hours down; After this step repeats 3 times, just make on all hole walls of the ceramic porous support of calcium sulfate and sticked compound material uniformly, this moment, the composite type biological stephanoporate bracket preparation of calcium sulfate/apatite complex was finished.Aseptic preservation is standby behind cobalt-60 radiation sterilization.
After scanning, Electronic Speculum SEM shape appearance figure can be measured, and the compound support of gained aperture is distributed between 150~300 μ m, interconnect between the hole, and the material internal micropore enriches, is evenly distributed, and presents loose coralliform structure.This similar is in the spongy bone framework of biological living, and it helps the biodegradation of growing into of osseous tissue and material self.
Calculate through test, the porous support mean porosities is 62.59%-79.36%, and comprcssive strength is 27-53MPa (the biomechanical strength spongy bone of normal human's osseous tissue is 7.09MPa, Compact bone 177-221MPa).
When medical calcium sulfate is absorbed, new bone is moulding and recover anatomic characteristic and architectural characteristic.Its most important advantage is that its natural infiltration rate is suitable with new bone formation speed.Along with the absorption of medical calcium sulfate implant, new bone recovers to dissect character and construction features gradually.