阅读说明：本技术 双膦酸盐型自凝固复合骨移植物及其制备方法 (Bisphosphonate type self-coagulation composite bone graft and preparation method thereof ) 是由 严永刚 邓光进 于 2020-05-21 设计创作，主要内容包括：本发明属于骨修复材料领域,具体涉及用于骨质疏松修复和重建的双膦酸盐型自凝固复合骨移植物及其制备方法。本发明提供一种双膦酸盐型自凝固复合骨移植物,所述复合骨移植物为双膦酸钙盐/自凝固钙盐复合物,所述双膦酸钙盐/自凝固钙盐复合物由固相部分和液相部分组成,固液比为1：0.3～1.0g/ml；其中,固相部分包括双膦酸钙盐和自凝固钙盐,各原料的质量比为：双膦酸钙盐：自凝固钙盐＝1：1～6。本发明首次将双膦酸盐与自凝固复合材料复合得到一种骨移植物,该自凝固复合骨移植物具有优良的可注射性、合适的凝固时间、缓慢的双膦酸根释放和优异的生物活性,可以用于骨质疏松和创伤引起的骨缺损的修复和重建与功能恢复。(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.)

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.

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.