CN111544655A - Bisphosphonate type self-coagulation composite bone graft and preparation method thereof - Google Patents
Bisphosphonate type self-coagulation composite bone graft and preparation method thereof Download PDFInfo
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- CN111544655A CN111544655A CN202010433195.4A CN202010433195A CN111544655A CN 111544655 A CN111544655 A CN 111544655A CN 202010433195 A CN202010433195 A CN 202010433195A CN 111544655 A CN111544655 A CN 111544655A
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- Prior art keywords
- calcium
- self
- bisphosphonate
- sodium
- degradation
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- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 54
- 239000002131 composite material Substances 0.000 title claims abstract description 51
- 229940122361 Bisphosphonate Drugs 0.000 title claims abstract description 46
- 150000004663 bisphosphonates Chemical class 0.000 title claims abstract description 39
- 238000005345 coagulation Methods 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title abstract description 4
- 159000000007 calcium salts Chemical class 0.000 claims abstract description 46
- 239000002994 raw material Substances 0.000 claims abstract description 21
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 claims abstract description 17
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 235000019691 monocalcium phosphate Nutrition 0.000 claims abstract description 16
- 239000007790 solid phase Substances 0.000 claims abstract description 16
- 229910000389 calcium phosphate Inorganic materials 0.000 claims abstract description 15
- 208000001132 Osteoporosis Diseases 0.000 claims abstract description 14
- WPSNYFRQPAASHJ-UHFFFAOYSA-J [Ca+2].[Ca+2].[O-]P([O-])=O.[O-]P([O-])=O Chemical compound [Ca+2].[Ca+2].[O-]P([O-])=O.[O-]P([O-])=O WPSNYFRQPAASHJ-UHFFFAOYSA-J 0.000 claims abstract description 10
- 239000007791 liquid phase Substances 0.000 claims abstract description 7
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 53
- 238000000498 ball milling Methods 0.000 claims description 32
- -1 diphosphonic acid calcium salt Chemical class 0.000 claims description 31
- 238000000034 method Methods 0.000 claims description 26
- 238000003756 stirring Methods 0.000 claims description 26
- 239000011575 calcium Substances 0.000 claims description 22
- 239000000243 solution Substances 0.000 claims description 19
- 239000008215 water for injection Substances 0.000 claims description 16
- OGSPWJRAVKPPFI-UHFFFAOYSA-N Alendronic Acid Chemical compound NCCCC(O)(P(O)(O)=O)P(O)(O)=O OGSPWJRAVKPPFI-UHFFFAOYSA-N 0.000 claims description 15
- ZOMBKNNSYQHRCA-UHFFFAOYSA-J calcium sulfate hemihydrate Chemical compound O.[Ca+2].[Ca+2].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O ZOMBKNNSYQHRCA-UHFFFAOYSA-J 0.000 claims description 15
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 14
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 claims description 14
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- BCAARMUWIRURQS-UHFFFAOYSA-N dicalcium;oxocalcium;silicate Chemical compound [Ca+2].[Ca+2].[Ca]=O.[O-][Si]([O-])([O-])[O-] BCAARMUWIRURQS-UHFFFAOYSA-N 0.000 claims description 13
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- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 8
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- UGEPSJNLORCRBO-UHFFFAOYSA-N [3-(dimethylamino)-1-hydroxy-1-phosphonopropyl]phosphonic acid Chemical compound CN(C)CCC(O)(P(O)(O)=O)P(O)(O)=O UGEPSJNLORCRBO-UHFFFAOYSA-N 0.000 claims description 5
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims description 5
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- MPBVHIBUJCELCL-UHFFFAOYSA-N Ibandronate Chemical compound CCCCCN(C)CCC(O)(P(O)(O)=O)P(O)(O)=O MPBVHIBUJCELCL-UHFFFAOYSA-N 0.000 claims description 4
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
- A61L2300/112—Phosphorus-containing compounds, e.g. phosphates, phosphonates
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Abstract
The invention belongs to the field of bone repair materials, and particularly relates to a bisphosphonate type self-setting composite bone graft for repairing and reconstructing osteoporosis and a preparation method thereof. The invention provides a bisphosphonate type self-coagulation composite bone graft, which is a calcium bisphosphonate/self-coagulation calcium salt composite, wherein the calcium bisphosphonate/self-coagulation calcium salt composite consists of a solid phase part and a liquid phase part, and the solid-liquid ratio is 1: 0.3-1.0 g/ml; wherein the solid phase part comprises calcium biphosphate and self-solidifying calcium salt, and the mass ratio of the raw materials is as follows: calcium bisphosphate salt: self-setting calcium salt 1: 1 to 6. The invention firstly compounds the bisphosphonate and the self-solidifying composite material to obtain the bone graft, and the self-solidifying composite bone graft has excellent injectability, proper solidification time, slow diphosphonate release and excellent biological activity, and can be used for repairing and reconstructing bone defects caused by osteoporosis and trauma and restoring functions.
Description
Technical Field
The invention belongs to the field of bone repair materials, and particularly relates to a bisphosphonate type self-setting composite bone graft for repairing and reconstructing osteoporosis and a preparation method thereof.
Background
Osteoporosis (OP) is a systemic metabolic disease of bone characterized by low bone mass and microstructural destruction of bone tissue, resulting in increased bone fragility and susceptibility to fracture. The disease can be developed in all age periods and is divided into primary and secondary types. Primary osteoporosis refers to the absence of other diseases that cause the disease; secondary osteoporosis is a decrease in the amount of bone tissue due to various systemic or endocrine metabolic diseases. Vertebral compression fractures often occur unconsciously, and can also be induced by coughing, sneezing, minor trauma, and the like. Within weeks of fresh vertebral body fracture, local pain appears, and tapping pain appears in physical signs. The height of a plurality of vertebral body compressors becomes shorter due to the occurrence of humpback (rolling over); pain and deformity manifest more severely when non-vertebral body is fractured.
Bone resorption inhibitors include 4 classes of estrogens, estrogen receptor modulators, bisphosphonates, calcitonin, and the like, often used alone or in turn; however, the effect on "bone microarchitecture", "bone fragility", "incidence of fracture" is not known, although the increase in bone density is more effective when 2 or more are combined and when sufficient amounts are applied (e.g. a sufficient dose of female hormone replacement is combined with a dose of alendronate sodium (fosfomi) of 10mg per day).
For bisphosphonate drugs, both oral and injectable, the following problems exist:
(1) bisphosphonates have high water solubility, such as 10mg/mL in water of sodium alendronate, are metabolized too quickly, reach a peak in the stomach or blood for a short time, then decay rapidly, and are discharged outside the body, the use efficiency of the drug is low, and frequent administration is required;
(2) irritation and side effects caused by high water solubility, such as erosion of rabbit stomach caused by sodium aminophosphonate; alendronate increases the incidence of indomethacin-induced antral ulcers; the alendronate can also enhance the indomethacin-induced gastric injury of rats and delay the healing of gastric ulcer; alendronate (0.04-0.1 mg/kg twice weekly or 0.1mg/kg weekly) partially blocks the establishment of bone metastases of human PC-3ML cells and leads to tumor formation in the peritoneum and other soft tissues;
(3) basic elements of bone formation are free calcium ions and free phosphate ions, insoluble calcium salts such as hydroxyapatite and the like are gradually formed through the regulation of DNA so as to form bone tissues, single bisphosphonate such as alendronate directly acts on osteoclasts, and more bone forming elements are difficult to provide besides the speed limiting step in the cholesterol biosynthesis pathway;
(4) bone loss and damage due to osteoporosis or other causes, require immediate filling of the re-defect site, free from further fracture and trauma due to voids, and thus, curing the fixed and filled bone repair and reconstruction material in situ facilitates bone reconstruction and restoration.
Disclosure of Invention
In view of the above circumstances, the present invention is directed to a bisphosphonate type self-setting composite bone graft and a method for preparing the same, which combines a bisphosphonate and a self-setting composite material for the first time to obtain a bone graft, and the self-setting composite bone graft has excellent injectability, appropriate setting time, slow bisphosphonates release and excellent biological activity, and can be used for the repair and reconstruction of bone defects caused by osteoporosis and trauma and functional recovery.
The technical scheme of the invention is as follows:
the invention provides a bisphosphonate type self-coagulation composite bone graft, which is a calcium bisphosphonate/self-coagulation calcium salt composite, wherein the calcium bisphosphonate/self-coagulation calcium salt composite consists of a solid phase part and a liquid phase part, and the solid-liquid ratio is 1: 0.3-1.0 g/ml; wherein the solid phase part comprises calcium biphosphate and self-solidifying calcium salt, and the mass ratio of the raw materials is as follows: calcium bisphosphate salt: self-setting calcium salt 1: 1 to 6.
Further, the self-setting calcium salt is a composite of two or more self-setting calcium salts.
Still further, the self-setting calcium salt is selected from tricalcium silicate (Ca)3SiO5，C3S), dicalcium silicate (2 CaO. SiO)2，C2S), calcium citrate (C)12H10Ca3O14) Calcium alginate (C)18H24CaO19) Calcium hydrogen phosphate (CaHO)4P) or calcium sulfate hemihydrate (CaH)2O5S.H2O) at least two of them.
Further, the liquid phase component is at least one of water for injection, normal saline or glucose injection.
Further, the calcium biphosphate is prepared by the following method: the neutral or slightly alkaline calcium compound and the sodium diphosphonate are subjected to ion exchange reaction to obtain the slightly soluble or insoluble calcium diphosphonate.
Further, the sodium bisphosphonate is selected from: at least one of alendronate sodium, neridronate sodium salt, olpadronate sodium, risedronate sodium, ibandronate sodium or pamidronate sodium.
Further, the neutral or slightly basic calcification is selected from: anhydrous calcium chloride (CaCl)2) Tricalcium silicate (Ca)3SiO5，C3S), dicalcium silicate (2 CaO. SiO)2，C2S), calcium oxide (CaO), calcium hydroxide [ Ca (OH) ]2]Calcium citrate (C)12H10Ca3O14) Or calcium hydrogen phosphate (CaHO)4P).
Further, the solid phase portion further comprises a humectant selected from the group consisting of: at least one of sodium alginate, hyaluronic acid or gelatin.
Further, the addition amount of the humectant is 0.5-5% of the total mass of the solid phase part (solid).
Further, the pH value of the diphosphonic acid calcium salt/self-coagulation calcium salt compound entering blood, body fluid, simulated body fluid and the like is 7-8.
The second technical problem to be solved by the present invention is to provide a method for preparing the above dual phosphate type self-setting composite bone graft, wherein the method comprises: the method comprises the following steps of taking a bisphosphonate calcium salt, a self-setting calcium salt and a curing liquid as raw materials, uniformly stirring and forming the raw materials to obtain a bisphosphonate self-setting composite bone graft; wherein, the solid-to-liquid ratio is 1: 0.3-1.0 g/ml; the mass ratio of the calcium biphosphate to the self-solidifying calcium salt is as follows: calcium bisphosphate salt: self-setting calcium salt 1: 1 to 6.
Further, the self-setting calcium salt is a composite of two or more self-setting calcium salts.
Still further, the self-setting calcium salt is selected from tricalcium silicate (C)a3SiO5，C3S), dicalcium silicate (2 CaO. SiO)2，C2S), calcium citrate (C)12H10Ca3O14) Calcium alginate (C)18H24CaO19) Calcium hydrogen phosphate (CaHO)4P) or calcium sulfate hemihydrate (CaH)2O5S.H2O) at least two of them.
Further, the solidifying liquid is at least one of water for injection, normal saline or glucose injection.
Further, the raw materials also comprise a humectant which is selected from: at least one of sodium alginate, hyaluronic acid or gelatin.
Further, the addition amount of the humectant is 0.5-5% of the total amount of the solid phase part (solid).
Further, in the method, the calcium biphosphate and the self-setting calcium (added with the humectant) are uniformly mixed by a ball milling method before being stirred, mixed and molded with the curing liquid; wherein the ball milling time is 2-12 hours; preferably 4 to 8 hours, and the ball milling speed is 100 to 200 revolutions per minute, preferably 120 to 150 revolutions per minute.
Further, the self-solidifying calcium salt is dried under vacuum at 120 ℃ for 5 to 24 hours, preferably 8 to 12 hours before use, and free moisture is removed.
Further, in the above method, the calcium bisphosphonate is prepared by the following method: the neutral or slightly alkaline calcium compound and the sodium diphosphonate are subjected to ion exchange reaction to obtain the slightly soluble or insoluble calcium diphosphonate.
Further, the method for preparing the calcium biphosphate comprises the following steps: dissolving sodium diphosphonate (NaR) in water at normal temperature, then adding a neutral or slightly-alkaline calcium compound (CaX), fully stirring for 0.5-2.0 hours, sealing and placing for 12-24 hours, then centrifuging to remove supernatant, washing for at least 4 times (the water washing ratio is 1: 5), and then freeze-drying or vacuum-drying to obtain calcium diphosphonate (CaR), and grinding into fine powder with the particle size of below 300 mu m for later use; wherein the molar ratio of the sodium diphosphate salt to the calcium compound is as follows: NaR: CaX ═ 1: 0.5 to 2.0 (mol).
Further, the sodium bisphosphonate is selected from: at least one of alendronate sodium, neridronate sodium salt, olpadronate sodium, risedronate sodium, ibandronate sodium or pamidronate sodium.
Further, the neutral or slightly basic calcification is selected from: anhydrous calcium chloride (CaCl)2) Tricalcium silicate (Ca)3SiO5，C3S), dicalcium silicate (2 CaO. SiO)2，C2S), calcium oxide (CaO), calcium hydroxide [ Ca (OH) ]2]Calcium citrate (C)12H10Ca3O14) Or calcium hydrogen phosphate (CaHO)4P).
A third technical problem to be solved by the present invention is to provide the use of the above-mentioned bisphosphonate type self-setting composite bone graft, which can be used for osteoporosis repair and reconstruction.
The invention has the beneficial effects that:
the bisphosphonate and the self-setting calcium salt form the bisphosphonate self-setting composite bone graft with the treatment function for the first time, the time effect of the bisphosphonate self-setting composite bone graft for releasing diphosphonate can reach more than 30 weeks, the bone formation period is met, the release amount can be regulated and controlled, and the setting time can be randomly adjusted according to the proportion within 20-60 minutes; the compressive strength reaches 2MPa after 1 hour of curing, and reaches 20MPa after 24 hours; the combined use of various self-solidifying calcium compounds and calcium biphosphate salt makes it have good plastic solidification performance. The proper setting time, good injectability, higher mechanical strength and slow bisphosphonate release enable the calcium-negative-bisphosphonate self-setting composite bone graft to be widely applied to the aspect of bone defects caused by osteoporosis or other factors.
Detailed Description
The invention provides a bisphosphonate type self-coagulation composite bone graft, which is a calcium bisphosphonate/self-coagulation calcium salt composite, wherein the calcium bisphosphonate/self-coagulation calcium salt composite consists of a solid phase part and a liquid phase part, and the solid-liquid ratio is 1: 0.3-1.0 g/ml; wherein the solid phase part comprises diphosphonic acid calcium salt (CaR) and self-solidifying calcium salt, and the mass ratio of the raw materials is as follows: calcium bisphosphate salt: self-setting calcium salt 1: 1 to 6.
Further, the self-setting calcium salt is a composite of two or more self-setting calcium salts. The invention selects the diphosphonate calcium salt and two or more than two self-setting calcium salts to be compounded in a proper proportion, and aims to simultaneously give consideration to pH, strength, degradation and setting time. The proportion of the two or more self-coagulating calcium salts needs to consider that the pH value of the formed composite calcium salt after being dissolved in water or simulated body fluid and other solidifying liquid is in the range of 7.0-8.0 so as to meet the requirements of the human blood environment and the osteogenic environment.
The calcium biphosphate is prepared by the following method: carrying out ion exchange reaction on a neutral or slightly-alkaline calcium compound and sodium biphosphate to obtain slightly-soluble or insoluble calcium biphosphate; thereby reducing the dissolution of the sodium diphosphonate, delaying the dissolution, controlling the release speed and prolonging the effective time.
Further, the solid phase portion further comprises a humectant selected from the group consisting of: at least one of sodium alginate, hyaluronic acid or gelatin. The addition amount of the humectant is 0.5-5% of the total amount of the solid phase part (solid). The compression strength and the formability of the bisphosphonate type self-coagulation composite bone graft for repairing and reconstructing osteoporosis can be improved by adding a proper amount of sodium alginate, hyaluronic acid or gelatin.
In the invention, the diphosphonic acid calcium salt with lower solubility is obtained by modifying through ion exchange by using the diphosphonic acid sodium salt with higher solubility, wherein the calcium compound used for modification is required to be alkaline or slightly alkaline, because acidic or neutral calcium salts such as monocalcium phosphate, calcium gluconate, calcium chloride, calcium nitrate and the like have high solubility, the solubility of the diphosphonic acid calcium salt formed after the reaction with the diphosphonic acid sodium salt is increased due to the fact that the pH value is less than 7, and the content of the filtrate is higher during centrifugation or washing, so that the yield and the later ratio of the diphosphonic acid calcium salt are influenced.
In the invention, the solubility of the calcium biphosphonate (CaR) after ion exchange is far less than that of the sodium biphosphonate, the calcium biphosphonate and other solidified calcium salts form a compound together, and the compound is hydrated after meeting water to form a linked solidification form bridged by hydrated crystal water, so that a compound with toughness and strength is formed.
The second technical problem to be solved by the present invention is to provide a method for preparing the above dual phosphate type self-setting composite bone graft, wherein the method comprises: the method comprises the following steps of taking a bisphosphonate calcium salt, a self-setting calcium salt and a curing liquid as raw materials, uniformly stirring and forming the raw materials to obtain a bisphosphonate self-setting composite bone graft; wherein, the solid-to-liquid ratio is 1: 0.3 to 1.0 g/ml. In the invention, the solid-liquid ratio is a core factor influencing the strength and the setting time of the bone cement, and if the solid-liquid ratio is too low, the viscosity is too large and the bone cement is difficult to form; if the solid-to-liquid ratio is too high, the coagulation time is too long to satisfy clinical requirements, and the strength thereof is also reduced.
A third technical problem to be solved by the present invention is to provide the use of the above-mentioned bisphosphonate type self-setting composite bone graft, which can be used for osteoporosis repair and reconstruction.
The following examples are given to further illustrate the embodiments of the present invention and are not intended to limit the scope of the present invention.
Example 1
Dissolving 32.51g of alendronate sodium in 500ml of water, stirring, adding 5.55g of anhydrous calcium chloride, fully stirring for 2 hours, and standing for 12 hours; then centrifuging to remove supernatant, washing with water for four times, and freeze drying to obtain 26.5g alendronate calcium salt (CaR).
Respectively mixing 10g of alendronate calcium salt (CaR), 10g of tricalcium silicate (CaSS1) dried at 120 ℃, 20g of calcium sulfate hemihydrate (CaSS2) and 0.5g of hyaluronic acid, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 39.75g of the complex were obtained.
Weighing 10g of the obtained compound, adding 5ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding; after 10 minutes the sample was taken out,
after 2 hours and 24 hours, measuring the compressive strength and the stress application speed of 2mm/mim at 25 ℃ and 55% humidity; and testing the degradation speed and the bone cell growth rate.
And (3) testing results:
2-hour compressive strength: 2.8MPa, 24-hour compressive strength: 25 MPa.
According to the following steps of 1:30 (g/ml) (ratio of sample to in vitro simulated solution) was put into a shaker of SBF (SBF solution used for in vitro simulated solution) at 37 ℃ and shaken at a speed of 60 times/min for degradation experiments: 7.25% of degradation in the first day, 15.16% of degradation in the first week, 22.22% of degradation in the second week, 29.38% of degradation in the third week and 32.15% of degradation in the fourth week; degradation is 36.09% in the fifth week; degradation at 41.44% in the eighth week; degradation in the twelfth week is 51.66%; 60.33% degradation in the sixteenth week; 68.38% degradation in the twentieth week; 75.79% degradation in the twenty-sixth week.
Soaking in 37 deg.C water for injection according to 0.2g/ml standard for 72 hr, filtering to obtain extractive solution, culturing mouse osteoblast with the extractive solution with original concentration and diluted 5 times, and observing and analyzing cell morphology and cell growth and differentiation rate for 24 hr, 48 hr and 72 hr; at 72 hours, the results of cell proliferation rate at original concentration and 5-fold dilution are: 101% and 120%.
Example 2
Dissolving 32.51g of alendronate sodium in 500ml of water, stirring, adding 8.31g of calcium citrate after all the alendronate sodium is dissolved, fully stirring for 2 hours, and standing for 12 hours; then centrifuged to remove the supernatant, washed with water for four times and freeze-dried to obtain 27.1g of alendronate calcium salt (CaR).
Respectively mixing 15g of alendronate calcium salt (CaR), 10g of calcium citrate (CaSS1) dried at 120 ℃, 5g of calcium hydrophosphate (CaSS2), 20g of calcium sulfate hemihydrate (CaSS3) and 0.5g of hyaluronic acid, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 49.25g of the complex was obtained.
Weighing 20g of the obtained compound, adding 11ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 2.1MPa, 24-hour compressive strength: 21.5 MPa.
According to the following steps of 1:30 (g/ml) were put on a 37 ℃ SBF shaker and shaken at a speed of 60 times/min for degradation experiments: 6.12% of degradation in the first day, 13.25% of degradation in the first week, 23.37% of degradation in the second week, 30.49% of degradation in the third week and 35.65% of degradation in the fourth week; degradation in the fifth week is 41.16%; degradation is 45.32% in the eighth week; 53.52% degradation in the twelfth week; degradation at 63.12% in the sixteenth week; 70.28% degradation in the twentieth week; degradation is 78.35% in twenty-sixth week.
Soaking in 37 deg.C water for injection according to 0.2g/ml standard for 72 hr, filtering to obtain extractive solution, culturing mouse osteoblast with the extractive solution with original concentration and diluted 5 times, and observing and analyzing cell morphology and cell growth and differentiation rate for 24 hr, 48 hr and 72 hr; at 72 hours, the results of the original concentration and the 5-fold dilution cell proliferation rate are respectively: 105% and 118%.
Example 3
Dissolving 59.8g neridronic acid sodium salt in 1000ml water, stirring, adding 7.75g tricalcium silicate (C)3S), fully stirring for 2 hours, and standing for 12 hours; then the supernatant was removed by centrifugation, washed four times with water by centrifugation and freeze-dried to obtain 58.5g of calcium neridronate (CaR).
Respectively mixing 20g of neridronic acid calcium salt (CaR), 15g of dicalcium silicate (CaSS1) dried at 120 ℃, 10g of calcium citrate (CaSS2), 20g of calcium sulfate hemihydrate (CaSS3) and 0.5g of hyaluronic acid, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 63.80g of complex were obtained.
Weighing 20g of the obtained compound, adding 10ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 3.3MPa, 24-hour compressive strength: 22 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. 8.15% of degradation in the first day, 16.22% of degradation in the first week, 21.37% of total degradation in the second week, 31.50% of total degradation in the third week and 35.49% of degradation in the fourth week; 38.13% degradation in the fifth week; degradation is 42.21% in the eighth week; 49.49% degradation in the twelfth week; 59.36% degradation in the sixteenth week; degradation at twenty weeks 67.58%; 73.89% degradation in the twenty-sixth week.
Injecting and soaking for 72 hours at 37 ℃ according to the standard of 0.2g/ml, filtering to obtain extracting solution, culturing mouse osteoblasts by using the extracting solution with the original concentration and diluted by 5 times respectively, and observing and analyzing cell morphology and cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 111% and 130%.
Example 4
Dissolving 32.51g of alendronate sodium in 500ml of water, stirring, adding 3.80g of calcium hydroxide after all the alendronate sodium is dissolved, fully stirring for 2 hours, and standing for 12 hours; then centrifuging to remove supernatant, washing with water for four times, and freeze drying to obtain 26.4g alendronate calcium salt (CaR).
Respectively mixing 10g of alendronate calcium salt (CaR), 10g of tricalcium silicate (CaSS1) dried at 120 ℃, 20g of calcium sulfate hemihydrate (CaSS2), 10g of calcium hydrophosphate (CaSS3) and 0.5g of hyaluronic acid, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 49.80g of the complex was obtained.
Weighing 10g of the obtained compound, adding 5.5ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 3.1MPa, 24-hour compressive strength: 29 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. 6.63% of degradation in the first day, 13.78% of degradation in the first week, 20.55% of degradation in the second week, 25.25% of degradation in the third week and 29.60% of degradation in the fourth week; 33.87% degradation in the fifth week; 38.66% in the eighth week; 43.04% degradation in the twelfth week; 49.11% degradation in the sixteenth week; 52.58% degradation in the twentieth week; 55.82% degradation in the twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 99 percent and 116 percent.
Example 5
Weighing 26.31g of sodium olpadronate, dissolving in 500ml of water, stirring, adding 5.55g of anhydrous calcium chloride after complete dissolution, fully stirring for 2 hours, and standing for 12 hours; then centrifuging to remove supernatant, washing with water for four times, and freeze drying to obtain 26.0g calcium olpadronate (CaR).
Respectively mixing 15g of olpadronate calcium salt (CaR), 10g of calcium citrate (CaSS1) dried at 120 ℃, 10g of calcium hydrophosphate (CaSS2), 15g of calcium sulfate hemihydrate (CaSS3) and 0.6g of hyaluronic acid, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 49.60g of complex were obtained.
Weighing 10g of the obtained compound, adding 5ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding; taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 2.1 MPa; 24-hour compressive strength: 20 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. 8.11% of degradation in the first day, 15.97% of degradation in the first week, 23.62% of degradation in the second week, 30.39% of degradation in the third week and 33.85% of degradation in the fourth week; degradation in the fifth week was 39.39%; degradation is 45.33% in the eighth week; 55.36% degradation in the twelfth week; degradation at 63.19% in the sixteenth week; 71.55% degradation in the twentieth week; 78.86% degradation in the twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 98 percent and 116 percent.
Example 6
Weighing 30.51g of sodium risedronate, dissolving in 500ml of water, stirring, adding 2.81g of high-grade pure calcium oxide after complete dissolution, fully stirring for 2 hours, and standing for 12 hours; the supernatant was then removed by centrifugation, washed four times with water and freeze dried to give 30.10g of calcium risedronate (CaR).
Respectively mixing 15g of calcium risedronate (CaR), 20g of tricalcium silicate (CaSS1) dried at 120 ℃, 5g of calcium hydrophosphate (CaSS2), 20g of calcium sulfate hemihydrate (CaSS3) and 0.5g of hyaluronic acid, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 59.05g of complex were obtained.
Weighing 10g of the obtained compound, adding 6ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 2.2MPa, 24-hour compressive strength: 35 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. 6.68% of degradation in the first day, 15.39% of degradation in the first week, 21.05% of degradation in the second week, 25.36% of degradation in the third week and 29.13% of degradation in the fourth week; 33.22% degradation in the fifth week; degradation is 38.02% in the eighth week; 43.10% degradation in the twelfth week; 50.53% degradation in the sixteenth week; 55.36% degradation in the twentieth week; degradation is 61.15% in twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 98 percent and 125 percent.
Example 7
The same as example 1, wherein 0.5g of the aqueous gelatin retention agent was replaced by 0.5g of gelatin.
And (3) testing results: 2-hour compressive strength: 1.1MPa, 24-hour compressive strength: 15 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. 9.21% of degradation in the first day, 17.26% of degradation in the first week, 25.65% of degradation in the second week, 32.56% of degradation in the third week and 37.11% of degradation in the fourth week; 45.01% degradation in the fifth week; degradation is 50.23% in the eighth week; 55.86% degradation in the twelfth week; 63.25% degradation in the sixteenth week; degradation at 73.58% in the twentieth week; degradation is 79.92% in twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 101% and 120%.
Example 8
As in example 2, 0.5g of hyaluronic acid water retaining agent was replaced by 0.6g of sodium alginate.
And (3) testing results: 2-hour compressive strength: 2.8MPa, 24-hour compressive strength: 25 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. 7.25% of degradation in the first day, 15.16% of degradation in the first week, 22.22% of degradation in the second week, 29.38% of degradation in the third week and 32.15% of degradation in the fourth week; degradation is 36.09% in the fifth week; degradation at 41.44% in the eighth week; degradation in the twelfth week is 51.66%; 60.33% degradation in the sixteenth week; 68.38% degradation in the twentieth week; 75.79% degradation in the twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 101% and 120%.
Example 9
In the same manner as in example 1, the solidification solution was changed from pure water to physiological saline.
And (3) testing results: 2-hour compressive strength: 0.9MPa, 24-hour compressive strength: 13.5 MPa.
8.30% of degradation in the first day, 17.06% of degradation in the first week, 23.55% of degradation in the second week, 31.68% of degradation in the third week and 35.35% of degradation in the fourth week; degradation is 41.11% in the fifth week; degradation at 46.32% in the eighth week; degradation is 58.76% in the twelfth week; degradation at the sixteenth week is 65.32%; degradation at 72.39% in the twentieth week; degradation is 78.93% in twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 102 percent and 120 percent.
Example 10
In the same manner as in example 2, the solidifying solution was changed from pure water to glucose injection solution.
And (3) testing results: 2-hour compressive strength: 2.1MPa, 24-hour compressive strength: 21.5 MPa.
According to the following steps of 1:30 (by mass) were placed in a 37 ℃ SBF shaker and shaken at a speed of 60 times/min. Degradation experiments were performed. 6.12% of degradation in the first day, 13.25% of degradation in the first week, 23.37% of degradation in the second week, 30.49% of degradation in the third week and 35.65% of degradation in the fourth week; degradation in the fifth week is 41.16%; degradation is 45.32% in the eighth week; 53.52% degradation in the twelfth week; degradation at 63.12% in the sixteenth week; 70.28% degradation in the twentieth week; degradation is 78.35% in twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results of the original concentration and the 5-fold dilution cell proliferation rate are respectively: 105% and 118%.
Comparative example 1
A comparison of sodium alendronate. 10g of sodium alendronate is ground (less than 120 meshes), 5ml of water is added to be prepared into slurry, and the slurry is put into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Half an hour later, ejection from the mold occurred and cracking occurred. Put into a shaking table with SBF at 37 ℃ according to the mass ratio of 1:30, and shake at the speed of 60 times/min. After half an hour, the dispersion was complete and after two hours the dissolution was complete. No other tests were entered except for the measurable concentration of sodium alendronate.
Comparative example 2
Calcium salt of alendronate. 10g of calcium alendronate is ground (less than 120 meshes), 5ml of water is added to be prepared into slurry, and the slurry is put into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Half an hour later, the product was ejected from the mold without cracking. After two hours, the compressive strength was determined to be 0.9 MPa. Put into a shaking table with SBF at 37 ℃ according to the mass ratio of 1:30, and shake at the speed of 60 times/min. The solution is cracked after half an hour, and after two hours, the solution is totally dispersed but not totally dissolved, the solution is in a turbid state, the shape cannot be kept in SBF, the strength is not high, and the solution is not suitable for clinical use.
Comparative example 3
Bisphosphonate open position comparison. Respectively weighing 20g of tricalcium silicate (CaSS1), 20g of calcium sulfate hemihydrate (CaSS2) and 0.5g of hyaluronic acid, mixing, ball-milling at the ball-milling speed of 150r/m, and filtering in a 120-mesh sieve after ball-milling for 6 hours; 39.75g of the complex were obtained.
Weighing 10g of the obtained compound, adding 5ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 3.9MPa, 24-hour compressive strength: 35 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. 5.35% of degradation in the first day, 25.16% of degradation in the first week, 28.51% of degradation in the second week, 32.33% of degradation in the third week and 41.11% of degradation in the fourth week; 45.09% degradation in the fifth week; 46.63% degradation in the eighth week; 46.95% degradation in the twelfth week; degradation at 46.33% in the sixteenth week; 46.15% degradation in the twentieth week; 46.23% degradation in the twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; at 72 hours, the results for the original concentration and 5-fold dilution were: 50 percent and 85 percent.
The compound bisphosphonate is absent, has better compressive strength, poorer biological performance, unsatisfactory cell differentiation and proliferation, fast degradation in the early stage, and stable and difficult degradation in the fifth week, and is not beneficial to the continuous regeneration and reconstruction of bone tissues.
Comparative example 4
Alendronate calcium salt and humectant are compared for lack of position. 20g of calcium citrate, 15g of calcium hydrogen phosphate (CaSS2) and 15g of calcium sulfate hemihydrate are mixed and ball-milled at a ball milling rate of 150r/m, and are filtered in a 120-mesh sieve after ball milling for 6 hours. 49.00g of the complex was obtained.
Weighing 10g of the obtained compound, adding 6ml of water, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding. Taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 1.8MPa, 24-hour compressive strength: 14.5 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. SBF is put into the cured product within 5 hours, so that a good shape can be protected, and the product is gradually degraded from the surface; after 24 hours of drying, the pellets will crack and become small when exposed to SBF for half an hour.
7.55% of degradation in the first day, 15.22% of degradation in the first week, 23.97% of degradation in the second week, 31.69% of degradation in the third week and 35.65% of degradation in the fourth week; degradation is carried out by 40.15% in the fifth week; degradation is 45.55% in the eighth week; 51.51% degradation in the twelfth week; 56.02% degradation in the sixteenth week; 60.18% degradation in the twentieth week; degradation was 63.33% in the twenty-sixth week.
Soaking the raw materials in 37-degree water for injection for 72 hours according to the standard of 0.2g/ml, filtering to obtain extract, culturing mouse osteoblasts by using the extract with the original concentration and diluted by 5 times, and observing and analyzing the cell morphology and the cell growth and differentiation rate for 24 hours, 48 hours and 72 hours; 72h, the results of the original concentration and the diluted 5-fold cell proliferation rate are respectively: 80% and 95%.
The humectant lacks position, which can cause stress cracking after solidification, bisphosphonate lacks position, and causes degradation and reduction of cell proliferation rate.
Comparative example 5
Comparison of the changes in the curing fluid in the absence of calcium biphosphate.
Mixing 20g of calcium citrate, 15g of calcium hydrophosphate (CaSS2), 15g of calcium sulfate hemihydrate and 0.5g of sodium alginate, carrying out ball milling at the ball milling speed of 150r/m, and filtering in a 120-mesh sieve after ball milling for 6 hours; 49.00g of the complex was obtained.
Weighing 10g of the obtained compound, adding 6ml of glucose injection, uniformly stirring, and then placing the slurry body into a polytetrafluoroethylene mold with the diameter of 6mm and the diameter of 12mm for molding; taking out the sample after 10 minutes, and measuring the compressive strength after 2 hours and 24 hours; and testing the degradation speed and the bone cell growth rate.
And (3) testing results: 2-hour compressive strength: 0.8MPa, 24-hour compressive strength: 12.2 MPa.
According to the following steps of 1:30 (g/ml) was placed on a 37 ℃ SBF shaker at a shaking speed of 60 times/min. Degradation experiments were performed. SBF is put into the cured product within 5 hours, so that a good shape can be protected, and the product is gradually degraded from the surface; after 24 hours of drying, the pellets will crack and become small when exposed to SBF for half an hour.
8.96% of degradation in the first day, 18.25% of degradation in the first week, 25.36% of degradation in the second week, 35.44% of degradation in the third week and 39.64% of degradation in the fourth week; 45.15% degradation in the fifth week; degradation is 50.33% in the eighth week; 56.71% degradation in the twelfth week; 61.12% degradation in the sixteenth week; 66.37% degradation in the twentieth week; degradation was 72.32% in the twenty-sixth week.
Soaking in water for injection at 37 deg.C for 72 hr according to 0.2g/ml standard, filtering to obtain extractive solution, culturing mouse osteoblast with the extractive solution at original concentration and diluted 5 times, and observing and analyzing cell morphology and cell growth and differentiation rate for 24 hr, 48 hr and 72 hr; 72h, the results of the original concentration and the diluted 5-fold cell proliferation rate are respectively: 88 percent and 98 percent.
Bisphosphonate position deficiency and cell proliferation rate are reduced.
Claims (10)
1. The bisphosphonate type self-coagulation composite bone graft is characterized in that the composite bone graft is a calcium bisphosphonate/self-coagulation calcium salt composite, the calcium bisphosphonate/self-coagulation calcium salt composite consists of a solid phase part and a liquid phase part, and the solid-liquid ratio is 1: 0.3-1.0 g/ml; wherein the solid phase part comprises calcium biphosphate and self-solidifying calcium salt, and the mass ratio of the raw materials is as follows: calcium bisphosphate salt: self-setting calcium salt 1: 1 to 6.
2. The bisphosphonate self-coagulating composite bone graft of claim 1, wherein the self-coagulating calcium salt is a composite of two or more self-coagulating calcium salts;
further, the self-setting calcium salt is selected from at least two of tricalcium silicate, dicalcium silicate, calcium citrate, calcium alginate, calcium hydrogen phosphate or calcium sulfate hemihydrate; or:
the liquid phase component is at least one of water for injection, normal saline or glucose injection; or:
the pH value of the diphosphonic acid calcium salt/self-coagulation calcium salt compound entering blood, body fluid or simulated body fluid is 7-8.
3. The bisphosphonate self-coagulating composite bone graft according to claim 1 or 2, wherein the bisphosphonate calcium salt is prepared by a method comprising: carrying out ion exchange reaction on a neutral or slightly-alkaline calcium compound and sodium biphosphate to obtain slightly-soluble or insoluble calcium biphosphate;
further, the sodium bisphosphonate is selected from: at least one of alendronate sodium, neridronate sodium, olpadronate sodium, risedronate sodium, ibandronate sodium, or pamidronate sodium;
further, the neutral or slightly basic calcification is selected from: at least one of anhydrous calcium chloride, tricalcium silicate, dicalcium silicate, calcium oxide, calcium hydroxide, calcium citrate, or calcium hydrogen phosphate.
4. The bisphosphonate self-coagulating composite bone graft according to any one of claims 1 to 3, wherein the solid phase portion further comprises a humectant;
further, the humectant is selected from: at least one of sodium alginate, hyaluronic acid or gelatin;
further, the addition amount of the humectant is 0.5-5% of the mass of the solid phase part.
5. The method for preparing the bisphosphonate type self-setting composite bone graft of any one of claims 1 to 4, wherein the method comprises: the method comprises the following steps of taking a bisphosphonate calcium salt, a self-setting calcium salt and a curing liquid as raw materials, uniformly stirring and forming the raw materials to obtain a bisphosphonate self-setting composite bone graft; wherein, the solid-to-liquid ratio is 1: 0.3-1.0 g/ml; the mass ratio of the calcium biphosphate to the self-solidifying calcium salt is as follows: calcium bisphosphate salt: self-setting calcium salt 1: 1 to 6.
6. The method for preparing a bisphosphonate type self-coagulating composite bone graft according to claim 5, wherein the self-coagulating calcium salt is a composite of two or more self-coagulating calcium salts;
further, the self-setting calcium salt is selected from at least two of tricalcium silicate, dicalcium silicate, calcium citrate, calcium alginate, calcium hydrogen phosphate or calcium sulfate hemihydrate; or:
the liquid phase component is at least one of water for injection, normal saline or glucose injection.
7. The method for preparing a bisphosphonate type self-coagulating composite bone graft according to claim 5 or 6, wherein the raw materials further comprise a humectant selected from: at least one of sodium alginate, hyaluronic acid or gelatin;
further, the addition amount of the humectant is 0.5-5% of the mass of the solid phase part.
8. The method for preparing the bisphosphonate type self-setting composite bone graft according to claim 5 or 6, wherein the calcium bisphosphonate and the self-setting calcium are uniformly mixed by a ball milling method before being stirred, mixed and formed with the curing solution; wherein the ball milling time is 2-12 hours, preferably 4-8 hours; the ball milling speed is 100-200 r/min, preferably 120-150 r/min.
9. The method for preparing the bisphosphonate type self-coagulating composite bone graft according to any one of claims 5 to 7, wherein the bisphosphonate calcium salt is prepared by a method comprising: carrying out ion exchange reaction on a neutral or slightly-alkaline calcium compound and sodium biphosphate to obtain slightly-soluble or insoluble calcium biphosphate;
further, the method for preparing the calcium biphosphate comprises the following steps: dissolving sodium biphosphate in water at normal temperature, adding a neutral or slightly-alkaline calcium compound, fully stirring for 0.5-2.0 hours, sealing and placing for 12-24 hours, centrifuging to remove supernatant, washing with water for at least 4 times, and freeze-drying or vacuum-drying to obtain calcium biphosphate; wherein the molar ratio of the sodium diphosphate salt to the calcium compound is as follows: sodium diphosphate salt: calcium compound 1: 0.5 to 2.0;
further, the sodium bisphosphonate is selected from: at least one of alendronate sodium, neridronate sodium, olpadronate sodium, risedronate sodium, ibandronate sodium, or pamidronate sodium;
further, the neutral or slightly basic calcification is selected from: at least one of anhydrous calcium chloride, tricalcium silicate, dicalcium silicate, calcium oxide, calcium hydroxide, calcium citrate, or calcium hydrogen phosphate.
10. The bisphosphonate self-coagulation composite bone graft is used for repairing and reconstructing osteoporosis, and the bisphosphonate self-coagulation composite bone graft is the composite bone graft according to any one of claims 1 to 4, or the composite bone graft prepared by the method according to any one of claims 5 to 9.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112316153A (en) * | 2020-11-24 | 2021-02-05 | 中山大学附属第七医院(深圳) | Anti-tumor pharmaceutical composition and application thereof |
| CN112336915A (en) * | 2020-12-11 | 2021-02-09 | 中鼎凯瑞科技成都有限公司 | Cyclic-phospho-adenyl functional self-solidifying composite bone implant and its preparation method |
| CN112707934A (en) * | 2020-12-30 | 2021-04-27 | 华南理工大学 | Alendronate calcium complex and preparation method thereof |
| CN113577383A (en) * | 2021-07-21 | 2021-11-02 | 西南交通大学 | Metal-organic/inorganic hybrid coating for promoting bone regeneration and regulating corrosion on degradable metal surface and preparation method thereof |
| CN114349786A (en) * | 2021-12-28 | 2022-04-15 | 中国科学院上海硅酸盐研究所 | Alendronate calcium ultralong nanowire and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4753652A (en) * | 1984-05-04 | 1988-06-28 | Children's Medical Center Corporation | Biomaterial implants which resist calcification |
| WO2010024549A2 (en) * | 2008-08-29 | 2010-03-04 | 한스바이오메드 주식회사 | Bone filler containing a sustained-release therapeutic agent for osteoporosis |
| CN101687059A (en) * | 2007-02-14 | 2010-03-31 | 格拉夫蒂斯公司 | Injectable calcium phosphate cement for releasing bone resorption inhibitor |
| CN103463678A (en) * | 2013-08-30 | 2013-12-25 | 中国人民解放军第四军医大学 | Multifunctional medical biological bone cement |
| CN110755680A (en) * | 2019-11-25 | 2020-02-07 | 中鼎凯瑞科技成都有限公司 | Vitamin D-loaded calcium citrate/calcium sulfate composite bone cement and preparation method thereof |
| CN111068109A (en) * | 2019-12-20 | 2020-04-28 | 华南理工大学 | Method for constructing calcium biphosphate crystals on surface of composite support and product |
-
2020
- 2020-05-21 CN CN202010433195.4A patent/CN111544655B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4753652A (en) * | 1984-05-04 | 1988-06-28 | Children's Medical Center Corporation | Biomaterial implants which resist calcification |
| CN101687059A (en) * | 2007-02-14 | 2010-03-31 | 格拉夫蒂斯公司 | Injectable calcium phosphate cement for releasing bone resorption inhibitor |
| WO2010024549A2 (en) * | 2008-08-29 | 2010-03-04 | 한스바이오메드 주식회사 | Bone filler containing a sustained-release therapeutic agent for osteoporosis |
| CN103463678A (en) * | 2013-08-30 | 2013-12-25 | 中国人民解放军第四军医大学 | Multifunctional medical biological bone cement |
| CN110755680A (en) * | 2019-11-25 | 2020-02-07 | 中鼎凯瑞科技成都有限公司 | Vitamin D-loaded calcium citrate/calcium sulfate composite bone cement and preparation method thereof |
| CN111068109A (en) * | 2019-12-20 | 2020-04-28 | 华南理工大学 | Method for constructing calcium biphosphate crystals on surface of composite support and product |
Non-Patent Citations (1)
| Title |
|---|
| 张惠琳: "《一种新型骨修复材料—难溶性阿仑膦酸钙的研制及其成骨性能研究》", 《中国优秀博硕士学位论文全文数据库(硕士)医药卫生科技辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112316153A (en) * | 2020-11-24 | 2021-02-05 | 中山大学附属第七医院(深圳) | Anti-tumor pharmaceutical composition and application thereof |
| CN112316153B (en) * | 2020-11-24 | 2023-08-25 | 中山大学附属第七医院(深圳) | Antitumor pharmaceutical composition and application thereof |
| CN112336915A (en) * | 2020-12-11 | 2021-02-09 | 中鼎凯瑞科技成都有限公司 | Cyclic-phospho-adenyl functional self-solidifying composite bone implant and its preparation method |
| CN112707934A (en) * | 2020-12-30 | 2021-04-27 | 华南理工大学 | Alendronate calcium complex and preparation method thereof |
| CN113577383A (en) * | 2021-07-21 | 2021-11-02 | 西南交通大学 | Metal-organic/inorganic hybrid coating for promoting bone regeneration and regulating corrosion on degradable metal surface and preparation method thereof |
| CN114349786A (en) * | 2021-12-28 | 2022-04-15 | 中国科学院上海硅酸盐研究所 | Alendronate calcium ultralong nanowire and preparation method and application thereof |
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